Late blight resistance genes and methods of use

Abstract

Compositions and methods for enhancing plant resistance to plant diseases caused by Phytophthora species are provided. The compositions include nucleic acid molecules encoding resistance (R) gene products and variants thereof and plants, seeds and plant cells comprising such nucleic acid molecules. Methods for enhancing plant resistance to plant diseases caused by Phytophthora species include introducing nucleic acid molecules encoding R gene products into plant cells. Methods for using the plants in agriculture to limit plant diseases are also provided.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 435,451, filed December 16, 2016, which is incorporated herein by reference in its entirety.

[0003] References to sequence lists submitted as text files

[0004] An official copy of the sequence list was submitted electronically via EFS-Web as a sequence list in ASCII format, filenamed 070294-0127SEQLST.TXT, created on December 11, 2017, and 440 kilobytes in size, and was submitted along with the specification. The sequence list contained in this ASCII format file is part of the specification and is incorporated herein by reference in its entirety. Invention Field

[0005] This invention relates to the field of gene isolation and plant improvement, and particularly to enhancing plant resistance to plant diseases through the use of disease-resistant genes. Background of the Invention

[0007] Late blight, caused by the oomycete pathogen *Phytophthora infestans*, is a devastating disease of cultivated potato (*Solanum tuberosum*) and tomato (*Solanum lycopersicum*), resulting in billions of dollars in losses annually (Jones (2014) Philos. Trans. R. Soc. Lond. B Biol. Sci. 369: 20130087-20130087). It is estimated that in Europe alone, the cost of late blight in potato production exceeds €1 billion, including the costs of controlling and managing the damage caused by the pathogen (Haverkort (2008) Potato Res. 51: 47-57).

[0008] Plant breeders typically introduce one Rpi (resistance to Phytophthora) gene at a time into cultivated potatoes from wild relatives. However, this process is both laborious and slow, and so far, the Rpi gene has been overcome by new strains of Phytophthora in a shorter time than it takes to breed new potato varieties containing it (Jones et al., 2014). Transgenic approaches allow for the simultaneous introduction of several genes ('gene stacking'), thus providing more durable resistance. Several major genes conferring resistance to late blight have been reported; however, due to the rapid evolution of Phytophthora, cloning additional Rpi genes remains necessary.

[0009] Cloned Rpi genes and their functional alleles include, for example, Rpi-blb1 / RB from Solanum demissum (van der Vossen et al. (2003) Plant J. 36: 867-882; Song et al. (2003) PNAS 100: 9128-9133) and their homologs Rpi-sto1 and Rpi-pta1 from S. stoloniferum and S. papita, respectively (Vleeshouwers et al. (2008) PLOS ONE 3: e2875); Rpi-blb2 from Solanum demissum (van der Vossen EA et al. (2005) Plant J. 36: 867-882; Song et al. (2003) PNAS 100: 9128-9133) J.44:208-222); Rpi-blb3 from S. bulbocastanum and its homologs Rpi-abpt and R2-like and R2 from S. bulbocastanum (Lokossou et al. (2009) MPMI22:630-641) and R2 from S. bulbocastanum ((2010) “Functional genomics of Phytophthora infestans effectors and Solanum resistance genes,” Ph.D. Thesis, Wageningen The Univ., Wageningen describes other homologs from S. edinense, S. schenckii and S. hjertingii, namely Rpi-edn1.1, Rpi-edn1.2, Rpi-snk1.1, Rpi-snk1.2 and Rpi-hjt1.1-Rpi-hjt1.3; Rpi-bt1 from yam (Oosumi et al. (2009) Amer. J. Potato Res. 86:456-465); R1 from yam (Ballvora et al. (2002) Plant J. 30:361-71); R3a and R3b from yam (Huang et al. (2005) Plant J. 30:361-71 respectively). J.42:261-271; Li et al. (2011) MPMI24:1132-114); Rpi-vnt1.1, Rpi-vnt1.2, Rpi-vnt1.3 from S. venturii (Foster et al. (2009) MPMI 22:589-600; Pel et al. (2009) MPMI 22:601-615; WO2009013468); Rpi-mcq1 from S. mochiquense (WO2009013468); from S. mochiquense (S. mochiquense)Rpi-chc from *S. pimpinellifolium* (WO 2011 / 034433) and Ph-3 from *S. pimpinellifolium* (Zhang et al. (2014) Theor. Appl. Genet. 127:1353-1364).

[0010] Black nightshade (Solanum nigrum) and closely related species are generally considered non-hosts of Phytophthora infectivity. They are not infected under laboratory conditions and infection is rarely observed in the field (Lebecka (2009) Eur. J. Plant Pathol. 124:345-348). However, there is a report on the susceptibility of black nightshade to Phytophthora infectivity, and the Mendelian segregation of resistance when susceptible lines are crossed with resistant lines and the F1 offspring are self-crossed to produce F2 offspring (Lebecka (2008) Eur. J. Plant Pathol. 120:233-240; Lebecka (2009) Eur. J. Plant Pathol. 124:345-348). This resistance under strong pathogen pressure suggests that the resistance genes present in black nightshade may have unique efficacy and recognition specificity, making them worthy of cloning and characterization. Black nightshade is a hexaploid plant of complex polyploid origin, making classic map-based cloning both laborious and time-consuming.

[0011] Recently, a cloning of the novel Rpi gene Rpi-amr3i from the Mexican registry of *Solanum nigrum* was reported (Witek et al. (2016) Nat. Biotechnol. 34:656). *Solanum nigrum* is a worldwide herbaceous flowering plant and is reported to be the presumed diploid ancestor of *Solanum nigrum* (Poczai and Hyvonen (2010) Mol. Biol. Rep. 38:1171-1185). Due to the rapid evolution of pathogenic races of *Phytophthora* that can overcome existing Rpi genes, additional novel Rpi genes will soon be needed to combat late blight in potatoes and tomatoes. Because cloning new Rpi genes from diploid Solanaceae species such as Solanum nigrum is expected to take less time than cloning Rpi genes from Solanaceae species with complex polyploid genomes such as S. nigrum, using diploid Solanaceae species as the source of Rpi genes allows researchers to clone new Rpi genes more quickly, providing plant breeders with new sources of resistance to late blight caused by Phytophthora blight. Summary of the Invention

[0012] This invention provides a nucleic acid molecule containing a resistance (R) gene capable of conferring resistance, particularly to plants in the Solanaceae family, to at least one race of *Phytophthora* species (sp.) known to cause plant diseases. In one embodiment, the invention provides a nucleic acid molecule comprising the R gene, referred herein as Rpi-amr1e, and variants thereof, including, for example, alleles of Rpi-amr1e, homologs of Rpi-amr1e, and other naturally occurring and non-naturally occurring variants of Rpi-amr1e. In another embodiment, the invention provides a nucleic acid molecule comprising the R gene, referred herein as Rpi-amr6b, and variants thereof, including, for example, alleles of Rpi-amr6b, homologs of Rpi-amr6b, and other naturally occurring and non-naturally occurring variants of Rpi-amr6b. In another embodiment, the present invention provides a nucleic acid molecule comprising an R gene, referred herein as Rpi-amr7d, and variants thereof, including, for example, alleles and homologs of Rpi-amr7d, as well as other naturally occurring and non-naturally occurring variants of Rpi-amr7d. In yet another embodiment, the present invention provides a nucleic acid molecule comprising an R gene, referred herein as Rpi-amr8c, and variants thereof, including, for example, alleles of Rpi-amr8c, homologs of Rpi-amr8c, and other naturally occurring and non-naturally occurring variants of Rpi-amr8c.

[0013] The present invention further provides plants, plant cells, and seeds containing one or more of the heteropolynucleotides of the present invention in their genomes. The heteropolynucleotides contain nucleotide sequences encoding resistance (R) proteins of the present invention. Such R proteins are encoded by R genes of the present invention (particularly Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c, as well as alleles, homologs, and other naturally and non-naturally occurring variants of such R genes). In a preferred embodiment, the plants and seeds are transgenic Solanaceae plants and seeds transformed with one or more of the heteropolynucleotides of the present invention. Preferably, such Solanaceae plants contain enhanced resistance to at least one race of a species of Phytophthora, known to cause plant diseases in Solanaceae plants, compared to the resistance of control plants lacking the heteropolynucleotides. The Solanaceae plants of the present invention include, but are not limited to, domesticated Solanaceae plants, including, for example, domesticated potato and tomato varieties.

[0014] This invention provides a method for enhancing the resistance of plants (particularly Solanaceae plants) to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. Such a method comprises introducing a heteropolynucleotide containing the nucleotide sequence of the R gene of this invention into at least one plant cell. Preferably, the heteropolynucleotide, or a portion thereof, is stably integrated into the genome of the plant cell. The method may optionally further comprise regenerating the plant cell into a plant whose genome contains the heteropolynucleotide. Preferably, the plant contains enhanced resistance to plant diseases caused by at least one race of a species of the genus *Phytophthora*, relative to a control plant that does not contain the heteropolynucleotide. More preferably, the plant contains enhanced resistance to plant diseases caused by at least two, three, four, five, or more races of a species of the genus *Phytophthora*, relative to a control plant that does not contain the heteropolynucleotide.

[0015] The present invention further provides a method for identifying Solanaceae plants exhibiting newly conferred or enhanced resistance to plant diseases caused by at least one race of a species of the genus Phytophthora. The method comprises detecting in Solanaceae plants the presence of alleles, homologs, and other naturally occurring and non-natural variants of Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and / or Rpi-amr8c and / or such R genes.

[0016] A method for limiting plant diseases caused by at least one race of a species of the genus *Phytophthora* is also provided for using the plants of the present invention in agricultural crop production. The method includes planting the plants of the present invention (e.g., seedlings), tubers, or seeds, wherein said plants, tubers, or seeds contain at least one R gene nucleotide sequence of the present invention. The method also includes growing the plants under conditions favorable to plant growth and development, and optionally harvesting at least one fruit, tuber, leaf, or seed from the plants.

[0017] Additionally, plants, plant parts, seeds, plant cells, other host cells, expression cassettes, and vectors comprising one or more nucleic acid molecules of the present invention are provided. Brief description of the attached diagram

[0019] Figure 1This is a genetic linkage map and a map of co-segregating BAC clones. The upper horizontal bar represents a bulked segregant analysis of 94 susceptible F2 plants combined with RenSeq, RAD-seq, and whole-genome shotgun (WGS), which led to the development of flanking and co-segregating markers. Markers with only numbers in their names or beginning with 'Ren' are RenSeq markers, those with 'WGS' are WGS-derived markers, and those with 'RAD' are RAD-seq-based markers. The middle horizontal bar represents the genes of 1793 F2 plants carrying the markers RAD_3 and WGS_1, which identified 118 informative recombinants, which were further phenotypically and genotyped using additional markers. This analysis confirmed that WGS_2,56766, and 46418 co-segregated with resistance. The lower horizontal bar is a schematic diagram of the contiguous groups of two BAC clones with the co-segregating marker WGS2, obtained from screening of a BAC library. Open reading frame prediction identified 11 potential coding sequences, of which 9 were confirmed as nucleotide-binding domain-rich leucine-containing proteins (NLRs) (ai). Solid black arrows indicate expressed NLRs, while white arrows represent pseudogenes.

[0020] Figure 2 This is a photographic illustration showing the resistance to Phytophthora infestans conferred by candidate Rpi-amr1e in transient complementation assays in leaves of *N. Benzylene*. The top two leaves are the third leaves of *N. Benzylene* plants, infiltrated with the vector pICSLUS0003::35S overexpressing Rpi-amr3i (positive control), Rpi-amr1 candidate, or GFP (negative control), and inoculated with *P. Benzylene* strain 88069 24 hours later. No growth of *P. Benzylene* was observed for Rpi-amr3i and Rpi-amr1e (illustrated), while growth of *P. Benzylene* was unchanged at all other infiltration sites for Rpi-amr1e candidates and GFP controls. Rpi-amr1c is shown as an example. The photograph was taken 6 days post-inoculation. The bottom two leaves are from a transient complementation assay in which the Rpi-amr1e genome construct (natural promoter and terminator) and *P. Benzylene* were used at the same level as under the 35S promoter. The vector overexpressing GFP was used as a negative control. Experiments were performed as previously described (Witek et al. (2016) Nat. Biotechnol. 34:656). Photographs were taken 6 days post-inoculation.

[0021] Figure 3This is a photographic illustration of leaves from a stable transgenic potato plant (Maris Piper cultivar) carrying the Rpi-amr1e gene, expressed under the control of a natural regulatory element, exhibiting resistance to the pathogenic Phytophthora 88069. The transgenic tetraploid potato “Maris Piper,” expressing Rpi-amr1e under the control of a natural regulatory element, is resistant to Phytophthora 88069. The transgenic line shows a high hazard rate (HR) at the inoculation site. In contrast, the control wild-type Maris Piper plants show large necrotic lesions and bud formation. Each leaflet was inoculated with a droplet containing 100–200 zoospores. Photographs were taken 6 days post-inoculation.

[0022] Figure 4 This is a photographic illustration of leaves from a stable transgenic potato plant (Marispabo cultivar) carrying the Rpi-amr1e gene, under the control of a natural regulatory element, exhibiting resistance to the pathogenic Phytophthora 88069. The transgenic tetraploid potato 'Marispabo' expressing Rpi-amr1e under the control of a natural regulatory element is resistant to Phytophthora 88069. Transgenic lines 1, 2, 3, 4A, 6, 10, 12, 14A, and 14B showed no symptoms or exhibited HR at the inoculation site. In contrast, the control wild-type 'Marispabo' potato plant and transgenic line 15 showed typical symptoms of late blight with bud formation. Each leaflet was inoculated with 3-4 droplets containing 1000-2000 zoospores; photographs were taken 10 days post-inoculation.

[0023] Figure 5 This is a photographic illustration showing the conferral of resistance to Phytophthora in leaves of *N. Benzyl* var. *bensemmeliensis* in a transient complementation assay using the Rpi-amr1e allele from another resistant *Solanum nigrum* registry number. The third leaf of a *N. Benzyl* plant was inoculated with the vector pICSLUS0001, which overexpresses the Rpi-amr1e allele from resistant lines SP1032, SP1123, SP2272, SP2307, and SP3408 (from top to bottom, the two leaves on the left) or GFP (negative control, the leaf on the right). Twenty-four hours later, the leaf was inoculated with *P. Benzyl* strain 88069. No *P. Benzyl* growth was observed for any of the tested Rpi-amr1e alleles, while *P. Benzyl* growth was unchanged at the inoculation site in the GFP control. The photograph was taken 10 days post-inoculation.

[0024] Figure 6This is a photographic illustration of leaves of stable transgenic *Nicotiana bensemere* plants expressing resistance to *Phytophthora* race 88069, under the control of natural regulatory elements. Transgenic *Nicotiana bensemere* plants expressing Rpi-amr1e_1032 (top) or Rpi-amr1e_2307 (bottom) under the control of natural regulatory elements are resistant to *Phytophthora* isolate 88069. The transgenic line (left) is asymptomatic at the inoculation site. In contrast, the control wild-type (WT, right) *Nicotiana bensemere* shows typical symptoms of late blight with bud formation. Each leaflet was inoculated with 3-4 droplets containing 1000-2000 zoospores; photographs were taken 14 days post-inoculation.

[0025] Figure 7 This is a photographic illustration of the transient expression of Rpi-amr6b, Rpi-amr1e, and Rpi-amr6b-s in *N. Benzema* after infection with *Phytophthora* isolates. Four-week-old leaves of *N. Benzema* were inoculated with the binary vector pICSLUS000435S, which overexpresses the late blight resistance gene Rpi-amr3 (positive control), and the binary vector pICSLUS0002, which expresses candidate Rpi-amr6b or negative control Rpi-amr6b-s from a susceptible parent with its own natural promoter and terminator. Twenty-four hours after inoculation, the leaves were inoculated with *Phytophthora* isolate 88069. *Phytophthora* grew well on the leaves for the negative control susceptibility allele of Rpi-amr6b; however, it could not grow or showed limited growth on leaves inoculated with Rpi-amr3 (positive control) and candidate Rpi-amr6b.

[0026] Figure 8 This is a photographic illustration of leaves of a stable transgenic potato plant (Marispibo cultivar) carrying Rpi-amr6b under the control of a natural regulatory element, exhibiting resistance to the pathogenic Phytophthora 88069. The transgenic tetraploid potato 'Marispibo' expressing Rpi-amr6b under the control of a natural regulatory element is resistant to Phytophthora 88069 (left). The transgenic line showed no symptoms or showed HR at the inoculation site. In contrast, the control wild-type Marispibo plant showed typical symptoms of late blight with bud formation (right leaf). Each leaflet was inoculated with 3-4 droplets containing 500-1000 zoospores; photographs were taken 10 days post-inoculation.

[0027] Figure 9This is a photographic illustration of the transient expression of Rpi-amr7d, Rpi-amr1e, and Rpi-amr6b-s in *N. Benzema* after infection with *Phytophthora* isolates. Four-week-old *N. Benzema* leaves were inoculated with the binary vector pICSLUS000435S overexpressing the late blight resistance gene Rpi-amr3 (positive control) and the binary vector pICSLUS0002 expressing candidate Rpi-amr67d or the negative control Rpi-amr6b-s from a susceptible parent with its own natural promoter and terminator. Twenty-four hours after inoculation, the leaves were inoculated with *Phytophthora* isolate 88069. *Phytophthora* grew well on the leaves for the negative control susceptibility allele of Rpi-amr6b; however, it could not grow or showed limited growth on leaves inoculated with Rpi-amr3 (positive control) and candidate Rpi-amr7d.

[0028] Figure 10 This is a photographic illustration of a leaf of the stable transgenic potato cultivar 'Marispibo' carrying the Rpi-amr7d gene, under the control of a natural regulatory element, exhibiting resistance to the pathogenic Phytophthora 88069. The transgenic tetraploid potato 'Marispibo' expressing Rpi-amr7d under the control of a natural regulatory element is resistant to the pathogenic Phytophthora 88069. The transgenic line (left) shows no symptoms or HR at the inoculation site. In contrast, the control wild-type (WT, right) 'Marispibo' plant shows typical symptoms of late blight with bud formation. Each leaflet was inoculated with 3-4 droplets containing 1000-2000 zoospores; the photograph was taken 10 days post-inoculation.

[0029] Figure 11 This is a photographic illustration of the transient expression of Rpi-amr8c, Rpi-amr1e, and Rpi-amr6b-s in *N. Benzema* leaves after inoculation with *Phytophthora* isolates. Four-week-old *N. Benzema* leaves were inoculated with the binary vector pICSLUS000435S overexpressing the late blight resistance gene Rpi-amr3 (positive control) and the binary vector pICSLUS0002 expressing either candidate Rpi-amr8c or the negative control Rpi-amr6b-s from a susceptible parent with its own natural promoter and terminator. Twenty-four hours after inoculation, the leaves were inoculated with *Phytophthora* isolate 88069. *Phytophthora* grew well on the leaves for the negative control susceptibility allele of Rpi-amr6b, but could not grow or grew only limitedly on leaves inoculated with Rpi-amr3 (positive control) and candidate Rpi-amr8c.

[0030] Figure 12This is a photographic illustration of a leaf of the stable transgenic potato cultivar 'Marispibo' carrying Rpi-amr8c under the control of a natural regulatory element, exhibiting resistance to Phytophthora 88069. The transgenic tetraploid potato 'Marispibo' expressing Rpi-amr8c under the control of a natural regulatory element is resistant to Phytophthora 88069. The transgenic line (left) shows no symptoms or shows HR at the inoculation site. In contrast, the control wild-type (WT, right) 'Marispibo' shows typical symptoms of late blight with bud formation. Each leaflet was inoculated with 3-4 droplets containing 1000-2000 zoospores; photographs were taken 10 days post-inoculation.

[0031] Figure 13 This is a photographic illustration of the transient expression of Rpi-amr9d, Rpi-amr1e, and Rpi-amr6b-s in *N. Benzema* after infection with pathogenic *Phytophthora* isolates. Four-week-old *N. Benzema* leaves were inoculated with the binary vector pICSLUS000435S overexpressing the late blight resistance gene Rpi-amr3 (positive control) and the binary vector pICSLUS000294 expressing candidate Rpi-amr9d or the negative control Rpi-amr6b-s from a susceptible parent with its own natural promoter and terminator. Twenty-four hours after inoculation, the leaves were inoculated with *Phytophthora* strain 88069. *Phytophthora* grew well on the leaves for the negative control susceptibility allele of Rpi-amr6b, but could not grow or grew only limitedly on leaves inoculated with Rpi-amr3 (positive control) and candidate Rpi-amr9d.

[0032] Figure 14 This is a photographic illustration of leaves from a stable transgenic *Nicotiana benthamiana* plant carrying the Rpi-amr9d gene, under the control of a natural regulatory element, exhibiting resistance to *Phytophthora* race 88069. The transgenic *Nicotiana benthamiana* plant expressing Rpi-amr9d under the control of a natural regulatory element is resistant to *Phytophthora* isolate 88069. The transgenic line (left) did not show symptoms at the inoculation site. In contrast, the control wild-type (WT, right) *Nicotiana benthamiana* showed typical symptoms of late blight with spore formation. Each leaflet was inoculated with 3-4 droplets containing 1000-2000 zoospores; photographs were taken 14 days post-inoculation.

[0033] Figure 15 This is a phylogenetic tree of all functional Rpi-amr1e alleles and homologs (i.e., Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c), which shows that Rpi-amr1e and Rpi-amr1e_SP2272 are the genes most distant from other functional alleles and homologs, while the remaining genes form two distinct categories.

[0034] sequence list

[0035] The nucleotide and amino acid sequences listed in the attached sequence listing are displayed using standard letter abbreviations for nucleotide bases and three-letter codes for amino acids. Nucleotide sequences follow the standard convention of starting from the 5' end of the sequence and moving forward (i.e., from left to right in each row) to the 3' end. Only one strand of each nucleotide sequence is shown, but it should be understood that any mention of a strand shown includes the complementary strand. Amino acid sequences follow the convention of starting from the amino terminus of the sequence and moving forward (i.e., from left to right in each row) to the carboxyl terminus.

[0036] SEQ ID NO:1 shows the nucleotide sequence of the R gene Rpi-amr1e of Solanum nigrum.

[0037] SEQ ID NO:2 shows the amino acid sequence of Rpi-amr1e (the R protein encoded by Rpi-amr1e).

[0038] SEQ ID NO:3 shows the nucleotide sequence of the coding region of the Rpi-amr1e cDNA. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:3. The natural stop codon for this cDNA is TGA.

[0039] SEQ ID NO:4 shows the nucleotide sequence of the Rpi-amr1e allele from the Solanum nigrum registry number A14750130.

[0040] SEQ ID NO:5 shows the amino acid sequence of the R protein encoded by the Rpi-amr1e allele from the Solanum nigrum registry number A14750130.

[0041] SEQ ID NO:6 shows the nucleotide sequence of the cDNA coding region of the Rpi-amr1e allele from the Solanum nigrum registry number A14750130. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:6. The natural stop codon for this cDNA is TGA.

[0042] SEQ ID NO:7 shows the nucleotide sequence of the Rpi-amr1e allele from the Registry No. Veg422 of Solanum lyratum.

[0043] SEQ ID NO:8 shows the amino acid sequence of the R protein encoded by the Rpi-amr1e allele from the Solanum nigrum registry number Veg422.

[0044] SEQ ID NO:9 shows the nucleotide sequence of the cDNA coding region of the Rpi-amr1e allele from the genus *Solanum nigrum* (accession number Veg422). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:9. The natural stop codon for this cDNA is TGA.

[0045] SEQ ID NO:10 shows the nucleotide sequence of the Rpi-amr1e allele from the Solanum nigrum registry number Wang2058.

[0046] SEQ ID NO:11 lists the amino acid sequence of the R protein encoded by the Rpi-amr1e allele from the Solanum nigrum registry number Wang2058.

[0047] SEQ ID NO:12 shows the nucleotide sequence of the cDNA coding region of the Rpi-amr1e allele from the Solanum nigrum registry number Wang2058. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:12. The natural stop codon for this cDNA is TGA.

[0048] SEQ ID NO:13 shows the nucleotide sequence of the Rpi-amr1e allele from the genus *Solanum nigrum* accession number sn27.

[0049] SEQ ID NO:14 lists the amino acid sequence of the R protein encoded by the Rpi-amr1e allele from the Solanum nigrum registry number sn27.

[0050] SEQ ID NO:15 shows the nucleotide sequence of the cDNA coding region of the Rpi-amr1e allele from the genus *Solanum nigrum* registry number sn27. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:15. The natural stop codon for this cDNA is TGA.

[0051] SEQ ID NO:16 shows the nucleotide sequence of the Rpi-amr1e allele from the Solanum nigrum registry number SOLA425.

[0052] SEQ ID NO:17 shows the amino acid sequence of the R protein encoded by the Rpi-amr1e allele from the Solanum nigrum registry number SOLA425.

[0053] SEQ ID NO:18 shows the nucleotide sequence of the cDNA coding region of the Rpi-amr1e allele from the Solanum nigrum registry number SOLA425. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:18. The natural stop codon for this cDNA is TAA.

[0054] SEQ ID NO:19 shows the nucleotide sequence of the Rpi-amr1e allele from the Solanum nigrum registry number A14750006.

[0055] SEQ ID NO:20 shows the amino acid sequence of the R protein encoded by the Rpi-amr1e allele from the Solanum nigrum registry number A14750006.

[0056] SEQ ID NO:21 shows the nucleotide sequence of the cDNA coding region of the Rpi-amr1e allele from the Solanum nigrum registry number A14750006. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:21. The natural stop codon for this cDNA is TGA.

[0057] SEQ ID NO:22 shows the nucleotide sequence of the R gene Rpi-amr1e. The promoter region spans nucleotides 1-1633, and the terminator region spans nucleotides 6443-7349.

[0058] SEQ ID NO:23 shows the nucleotide sequence of the cDNA coding region of splice variant 1 of Rpi-amr1e (SEQ ID NO:22). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:23. The natural stop codon for this cDNA is TAA.

[0059] SEQ ID NO:24 shows the amino acid sequence of the R protein encoded by the splice variant 1 cDNA shown in SEQ ID NO:23.

[0060] SEQ ID NO:25 shows the nucleotide sequence of the cDNA coding region of splice variant 2 of Rpi-amr1e (SEQ ID NO:22). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:25. The natural stop codon for this cDNA is TAA.

[0061] SEQ ID NO:26 shows the nucleotide sequence of the cDNA coding region of splice variant 3 of Rpi-amr1e (SEQ ID NO:22). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:26. The natural stop codon for this cDNA is TAA.

[0062] SEQ ID NO:27 shows the nucleotide sequence of the cDNA coding region of splice variant 4 of Rpi-amr1e (SEQ ID NO:22). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:27. The natural stop codon for this cDNA is TGA.

[0063] SEQ ID NO:28 shows the amino acid sequence of the R protein encoded by the splice variant 2 cDNA shown in SEQ ID NO:25.

[0064] SEQ ID NO:29 shows the amino acid sequence of the R protein encoded by the splice variant 3 cDNA shown in SEQ ID NO:26.

[0065] SEQ ID NO:30 shows the amino acid sequence of the R protein encoded by the splice variant 4 cDNA shown in SEQ ID NO:27.

[0066] SEQ ID NO:31 shows the nucleotide sequence of the SP1032 allele of the R gene Rpi-amr1e. The promoter region spans nucleotides 1-1823, and the terminator region spans nucleotides 6944-7913.

[0067] SEQ ID NO:32 shows the nucleotide sequence of the SP1123 allele of the R gene Rpi-amr1e. The promoter region spans nucleotides 49-1577, and the terminator region spans nucleotides 6705-7662.

[0068] SEQ ID NO:33 shows the nucleotide sequence of the SP2272 allele of the R gene Rpi-amr1e. The promoter region spans nucleotides 641-1745, and the terminator region spans nucleotides 6802-7770.

[0069] SEQ ID NO:34 shows the nucleotide sequence of the SP2307 allele of the R gene Rpi-amr1e. The promoter region spans nucleotides 1-1991, and the terminator region spans nucleotides 9253-9596.

[0070] SEQ ID NO:35 shows the nucleotide sequence of the SP3408 allele of the R gene Rpi-amr1e. The promoter region spans nucleotides 1-1405, and the terminator region spans nucleotides 7567-8398.

[0071] SEQ ID NO:36 shows the nucleotide sequence of the cDNA coding region of the SP1032 allele of Rpi-amr1e. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:36. The natural stop codon for this cDNA is TAA.

[0072] SEQ ID NO:37 shows the nucleotide sequence of the cDNA coding region of the SP1123 allele of Rpi-amr1e. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:37. The natural stop codon for this cDNA is TAA.

[0073] SEQ ID NO:38 shows the nucleotide sequence of the cDNA coding region of the SP2272 allele of Rpi-amr1e. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:38. The natural stop codon for this cDNA is TAA.

[0074] SEQ ID NO:39 shows the nucleotide sequence of the cDNA coding region of the SP2307 allele of Rpi-amr1e. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:39. The natural stop codon for this cDNA is TAA.

[0075] SEQ ID NO:40 shows the nucleotide sequence of the cDNA coding region of the SP3408 allele of Rpi-amr1e. If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:40. The natural stop codon for this cDNA is TAA.

[0076] SEQ ID NO:41 shows the amino acid sequence of the R protein encoded by the SP1032 cDNA sequence shown in SEQ ID NO:36.

[0077] SEQ ID NO:42 shows the amino acid sequence of the R protein encoded by the SP1123 cDNA sequence shown in SEQ ID NO:37.

[0078] SEQ ID NO:43 shows the amino acid sequence of the R protein encoded by the SP2272 cDNA sequence shown in SEQ ID NO:38.

[0079] SEQ ID NO:44 shows the amino acid sequence of the R protein encoded by the SP2307 cDNA sequence shown in SEQ ID NO:39.

[0080] SEQ ID NO:45 shows the amino acid sequence of the R protein encoded by the SP3408 cDNA sequence shown in SEQ ID NO:40.

[0081] SEQ ID NO:46 shows the nucleotide sequence of the R gene Rpi-amr6b from Solanum nigrescens accession number A14750423. The promoter region spans nucleotides 1-2030, and the terminator region spans nucleotides 7162-8005.

[0082] SEQ ID NO:47 shows the amino acid sequence of the R protein encoded by splice variant 1 of Rpi-amr6b (SEQ ID NO:46). The cDNA of splice variant 1 is shown in SEQ ID NO:49.

[0083] SEQ ID NO:48 shows the amino acid sequence of the R protein encoded by splice variant 2 of Rpi-amr6b (SEQ ID NO:46). The cDNA of splice variant 2 is shown in SEQ ID NO:50.

[0084] SEQ ID NO:49 shows the nucleotide sequence of the cDNA coding region of splice variant 1 of Rpi-amr6b (SEQ ID NO:46). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:49. The natural stop codon for this cDNA is TAA.

[0085] SEQ ID NO:50 shows the nucleotide sequence of the cDNA coding region of splice variant 1 of Rpi-amr6b (SEQ ID NO:46). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:50. The natural stop codon for this cDNA is TAA.

[0086] SEQ ID NO:51 shows the nucleotide sequence of the R gene Rpi-amr7d from the accession number A54750014 of *Solanum nigrum*. The promoter region spans nucleotides 1-1960, and the terminator region spans nucleotides 7032-7842.

[0087] SEQ ID NO:52 shows the amino acid sequence of the R protein encoded by splice variant 1 (SEQ ID NO:51) of Rpi-amr7d (SEQ ID NO:51). The cDNA of splice variant 1 is shown in SEQ ID NO:54.

[0088] SEQ ID NO:53 shows the amino acid sequence of the R protein encoded by splice variant 2 of Rpi-amr7d (SEQ ID NO:51). The cDNA of splice variant 2 is shown in SEQ ID NO:55.

[0089] SEQ ID NO:54 shows the nucleotide sequence of the cDNA coding region of splice variant 1 of Rpi-amr7d (SEQ ID NO:51). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:54. The natural stop codon for this cDNA is TAA.

[0090] SEQ ID NO:55 shows the nucleotide sequence of the cDNA coding region of splice variant 2 of Rpi-amr7d (SEQ ID NO:51). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:55. The natural stop codon for this cDNA is TAA.

[0091] SEQ ID NO:56 shows the nucleotide sequence of the R gene Rpi-amr8c from the Solanum nigrum registry number SOLA 226. The promoter region spans nucleotides 1-1953, and the terminator region spans nucleotides 7078-7456.

[0092] SEQ ID NO:57 shows the amino acid sequence of the R protein encoded by splice variant 1 of Rpi-amr8c (SEQ ID NO:56). The cDNA of splice variant 1 is shown in SEQ ID NO:60.

[0093] SEQ ID NO:58 shows the amino acid sequence of the R protein encoded by splice variant 2 of Rpi-amr8c (SEQ ID NO:56). The cDNA of splice variant 2 is shown in SEQ ID NO:59.

[0094] SEQ ID NO:59 shows the nucleotide sequence of the cDNA coding region of Rpi-amr8c splice variant 2 (SEQ ID NO:56). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:59. The natural stop codon for this cDNA is TAA.

[0095] SEQ ID NO:60 shows the nucleotide sequence of the cDNA coding region of splice variant 1 of Rpi-amr8c (SEQ ID NO:56). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:60. The natural stop codon for this cDNA is TAA.

[0096] SEQ ID NO:61 shows the nucleotide sequence of the R gene Rpi-amr9d from the Solanum nigrum registry number SOLA425. The promoter region spans nucleotides 1-1991, and the terminator region spans nucleotides 9269-9596.

[0097] SEQ ID NO:62 shows the amino acid sequence of the R protein encoded by splice variant 1 of Rpi-amr9d (SEQ ID NO:61). The cDNA of splice variant 1 is shown in SEQ ID NO:60.

[0098] SEQ ID NO:63 shows the amino acid sequence of the R protein encoded by splice variant 2 of Rpi-amr9d (SEQ ID NO:61). The cDNA of splice variant 2 is shown in SEQ ID NO:60.

[0099] SEQ ID NO:64 shows the nucleotide sequence of the cDNA coding region of splice variant 2 of Rpi-amr9d (SEQ ID NO:61). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:64. The natural stop codon for this cDNA is TAA.

[0100] SEQ ID NO:65 shows the nucleotide sequence of the cDNA coding region of splice variant 1 of Rpi-amr9d (SEQ ID NO:61). If desired, a stop codon (e.g., TAA, TAG, TGA) can be operatively linked to the 3' end of the nucleic acid molecule containing SEQ ID NO:65. The natural stop codon for this cDNA is TAA. Invention Details

[0102] The invention will now be described more fully below with reference to the accompanying drawings, which show some, but not all, embodiments of the invention. In fact, these inventions may be embodied in many different forms and should not be construed as limited to the embodiments shown herein; rather, these embodiments are provided to enable this disclosure to meet applicable legal requirements. The same numerals always refer to the same elements.

[0103] Benefiting from the teachings presented in the foregoing description and the accompanying drawings, those skilled in the art will conceive of many modifications and other embodiments of the invention illustrated herein. Therefore, it should be understood that the invention is not limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terminology is used herein, it is used only in a general and descriptive sense and not for purposes of limitation.

[0104] This invention relates to the isolation of plant resistance (R) genes (particularly R genes conferring resistance to late blight caused by one or more races of Phytophthora in the Solanaceae family). As disclosed below, using a map-based cloning method, the R gene, referred to herein as Rpi-amr1e, was isolated from *Solanum nigrum* Registry No. 954750184 (a diploid, non-tuber relative of potato) using fine mapping of 1793 F2 plants and sequencing of co-segregated BAC clones. Using previously described long-read sequencing (Eid et al. (2008) Science 323:133-138; Sharon et al. (2013) Nat. Biotechnol. 31:1009-14; both are incorporated herein by reference), additional Rpi-amr1e alleles from Veg422, A14750130, Wang 2058, sn27, A14750006, and SOLA425, the accession number for *Solanum nigrum*, were isolated using a method involving R gene sequence capture (RenSeq). The isolation of additional Rpi-amr1e alleles from accession numbers 954750174, A14750130, and 954750172 is disclosed in Example 8 below. In Examples 9-16 below, the isolation of three additional R genes, which are homologs of Rpi-amr1e, is also disclosed: Rpi-amr6b from Solanumnigrescens accession number A14750423; Rpi-amr7d from Solanum nigrum accession number A54750014; and Rpi-amr8c from Solanum nigrum accession number SOLA 226.

[0105] This invention provides nucleic acid molecules comprising the nucleotide sequences of R genes (particularly Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c, and alleles, homologs, orthologs, and other naturally occurring variants of such R genes, as well as synthetic or artificial (i.e., non-naturally occurring) variants thereof). As used herein, unless otherwise stated or obvious from the context of use, such nucleic acid molecules are referred to herein as “Rpi-amr nucleic acid molecules” or “Rpi-amr genes.” Similarly, unless otherwise stated or obvious from the context of use, the nucleotide sequences of Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c, and the alleles, homologs, orthologs, and other naturally occurring variants of such R genes, as well as synthetic or artificial (i.e., non-naturally occurring) variants thereof, are referred to herein as “Rpi-amr nucleotide sequences.”

[0106] The Rpi-amr nucleotide sequence of the present invention is the nucleotide sequence of the R gene, which is also referred to herein as the R gene nucleotide sequence. Preferably, such nucleotide sequences of the R gene encode the R protein. The Rpi-amr nucleotide sequences of the present invention include, but are not limited to, the nucleotide sequences, cDNA sequences, and nucleotide sequences containing only the coding region of wild-type Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c genes containing a natural promoter and a 3' adjacent region containing the coding region. Examples of such Rpi-amr nucleotide sequences include the nucleotide sequences shown in SEQ ID NOS:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, and 65, and variations thereof. In embodiments where the native Rpi-amr gene promoter is not used to drive the expression of the nucleotide sequence encoding the R protein, a heterologous promoter can be operatively linked to the nucleotide sequence encoding the R protein of the present invention to drive the expression of the nucleotide sequence encoding the R protein in plants.

[0107] Preferably, the R protein encoded by the Rpi-amr nucleotide sequence of the present invention is a functional R protein or one or more portions or one or more domains thereof, which is capable of conferring enhanced resistance to plant diseases caused by at least one race of at least one species of the genus Phytophthora in plants containing said R protein (particularly Solanaceae plants). In some preferred embodiments, the R protein of the present invention is capable of conferring broad-spectrum resistance to at least one race of Phytophthora pathogens, but preferably multiple races, and includes, for example, Rpi-amr1e (SEQ ID NO:2) (the R protein encoded by Rpi-amr1e (SEQ ID NO:1)) and R proteins (SEQ ID NO:5, 8, 11, 14, 17, 20, 41, 42, 43, 44 and 45) encoded by alleles of Rpi-amr1e (SEQ ID NO:4, 7, 10, 13, 16, 19, 31, 32, 33, 34 and 35, respectively). Such R proteins of the present invention include, but are not limited to, R proteins comprising the amino acid sequences shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, and 63, and / or encoded by the Rpi-amr nucleotide sequences shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, and 65.

[0108] Similarly, the preferred Rpi-amr gene, Rpi-amr nucleic acid molecule, and Rpi-amr1e allele of the present invention can confer enhanced resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora* in plants (especially Solanaceae plants) containing the Rpi-amr gene, Rpi-amr nucleic acid molecule, or Rpi-amr1e allele. In some preferred embodiments, the Rpi-amr gene, Rpi-amr nucleic acid molecule, and Rpi-amr1e allele of the present invention can confer broad-spectrum resistance to at least one race of pathogenic *Phytophthora*, but broad-spectrum resistance to multiple races is preferred. Such Rpi-amr genes, Rpi-amr nucleic acid molecules, and Rpi-amr1e alleles include, but are not limited to, Rpi-amr genes, Rpi-amr nucleic acid molecules, and Rpi-amr1e alleles comprising nucleotide sequences selected from the group consisting of: SEQ ID NO: 1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, or 65; and nucleotide sequences encoding ...8, 9, 10, 11, 64, or 65. The nucleotide sequence of the amino acid sequence shown in NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63.

[0109] The present invention also provides plants comprising heteropolynucleotides, said heteropolynucleotides comprising the R gene nucleotide sequence of the present invention. Preferably, such R gene nucleotide sequence encodes the full-length R protein of the present invention, or at least one or more functional portions or one or more domains thereof. In some embodiments, such heteropolynucleotides of the present invention are stably integrated into the plant genome, and in other embodiments, the plant is transformed by a transient transformation method, and the heteropolynucleotides are not stably integrated into the plant genome.

[0110] In other embodiments, the method of the present invention is used to produce plants comprising heterologous polynucleotides containing the R gene nucleotide sequence of the present invention. The method involves genome editing to modify the nucleotide sequence of a natural or non-natural gene in the genome of the plant. The natural or non-natural gene contains a nucleotide sequence different (i.e., not identical) from the R gene nucleotide sequence of the present invention, and after modification by a method further detailed below, the modified natural or non-natural gene contains the R gene nucleotide sequence of the present invention. Typically, such methods involve using a plant whose genome contains a natural or non-natural gene, wherein the natural or non-natural gene contains a nucleotide sequence homologous to the R gene nucleotide sequence of the present invention, and further involve introducing a nucleic acid molecule containing at least a portion of the R gene nucleotide sequence of the present invention into the plant. Preferably, the nucleotide sequence of the natural or non-natural gene has about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher nucleotide sequence identity with at least one R gene nucleotide sequence of the present invention. Such natural or non-natural genes can be, for example, R genes, particularly Rpi-amr genes, or non-functional homologs of such R genes that cannot or are unknown whether they can confer resistance to plant diseases. It has been recognized that plants produced through the genome engineering disclosed herein are stably transformed plants when the modified natural or non-natural gene is stably integrated into the plant genome.

[0111] Methods for stable and transient transformation of plants and genome editing are disclosed elsewhere herein or are otherwise known in the art. In a preferred embodiment of the invention, the plant is a stably transformed potato or tomato plant containing heteropolynucleotides of the invention stably integrated into its respective genome and further containing enhanced resistance to late blight caused by at least one race of Phytophthora. In a more preferred embodiment of the invention, the plant is a stably transformed potato or tomato plant containing heteropolynucleotides of the invention stably integrated into its respective genome and further containing enhanced resistance to late blight caused by at least two, three, four, five, six, or more races of Phytophthora.

[0112] In some embodiments, the plant of the present invention comprises a heteropolynucleotide containing a nucleotide sequence encoding the R protein of the present invention and a heteropromoter operatively linked to express the nucleotide sequence encoding the R protein. The selection of the heteropromoter can depend on many factors, such as, for example, desired timing, localization and expression patterns, and responsiveness to specific biotic or abiotic stimuli. Target promoters include, but are not limited to, pathogen-inducible promoters, constitutive promoters, tissue-preferred promoters, wound-inducible promoters, and chemically regulated promoters.

[0113] In some embodiments of the invention, the plants of the invention, particularly Solanaceae plants, may contain one, two, three, four, five, six, or more nucleotide sequences encoding R proteins. Typically, but not necessarily, the two or more R proteins are different from each other. For the purposes of this invention, an R protein is different from another R protein when two R proteins have different amino acid sequences. In some embodiments of the invention, each of the different R proteins for resistance to plant diseases caused by a single species of the genus *Phytophthora* has one or more differences in resistance characteristics, such as, for example, resistance to different races and / or groups of races of the same species of *Phytophthora* or even resistance to different species of the genus *Phytophthora*. It is recognized that by combining two, three, four, five, six, or more nucleotide sequences with each nucleotide sequence encoding different races or certain species (spp.) of the different R proteins for resistance to a single species of *Phytophthora*, Solanaceae plants containing broad-spectrum resistance to multiple races or even multiple species of the genus *Phytophthora* can be produced. This nightshade plant, especially potato or tomato plants, can be used in agriculture in areas where multiple races of a species of Phytophthora are prevalent, such as Phytophthora pathogenica.

[0114] Examples of R genes that can combine with one or more Rpi-amr nucleotide sequences of the present invention in a single potato plant include, but are not limited to, the following cloned Rpi gene: Rpi-amr3i (accession number KT373889; SEQ ID NO. 2016 / 182881). NO:1), Rpi-blb1 (also known as "RB"; accession numbers FB764493.1 and AY336128.1), Rpi-sto1 (accession number EU884421), Rpi-pta1 (accession number EU884422), Rpi-blb2 (accession number DQ122125), Rpi-blb3 (accession number FJ536326), Rpi-abpt (accession number FJ536324), R2-like (accession number FJ536323), R2 (accession number FJ536325), Rpi-edn1.1 (accession number GU563963), Rpi-edn1.2, Rpi-snk1.1, Rpi-sn k1.2, Rpi-hjt1.1-Rpi-hjt1.3 (login number GU563971-3), Rpi-bt1 (login number FJ188415), R1 (login number AF447489), R3a (login number AY849382), R3b (login number JF900492), Rpi-vnt1.1 (login number FJ423044), Rpi-vnt1.2 (login number FJ423045), Rpi-vnt1.3 (login number FJ423046), Rpi-mcq1 (login number GN043561), Rpi-chc, Ph-3 (login number KJ563933), and R8 (login number KU530153). Nucleotide sequences corresponding to the accession numbers of the genes listed above or any genes or proteins disclosed elsewhere in this document are available from publicly available online nucleotide and amino acid sequence databases, such as GenBank and EMBL databases (available on the World Wide Web at ncbi.nlm.nih.gov / genbank and ebi.ac.uk, respectively).

[0115] The plants of the present invention containing multiple R genes can be produced, for example, by transforming plants already containing one or more other R gene nucleotide sequences with a heteropolynucleotide containing at least one of the Rpi-amr nucleotide sequences of the present invention, said Rpi-amr nucleotide sequences including, for example, one or more of the Rpi-amr1e nucleotide sequence, Rpi-amr6b nucleotide sequence, Rpi-amr7d nucleotide sequence, and Rpi-amr8c nucleotide sequence. Such plants already containing one or more other R gene nucleotide sequences may contain the R genes naturally occurring in the genome or the plant, introduced through sexual reproduction, or introduced through transformation of the plant or its ancestors with R gene nucleotide sequences. Alternatively, said one or more other R gene nucleotide sequences can be introduced into the plants of the present invention, which already contain the heteropolynucleotides of the present invention, by, for example, transformation or sexual reproduction.

[0116] In other embodiments, two or more distinct R gene sequences can be introduced into plants by stably transforming them with a heteropolynucleotide or vector containing two or more R gene nucleotide sequences. This approach has been recognized as preferred for plant breeding because the two or more R gene nucleotide sequences are expected to be closely linked and thus segregate at a single locus. Alternatively, the heteropolynucleotides of the present invention can be integrated into the plant genome adjacent to another R gene nucleotide sequence using genome modification methods based on homologous recombination described elsewhere herein or known in the art.

[0117] The present invention also provides a method for enhancing plant resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. The method includes modifying at least one plant cell to contain a heteropolynucleotide, and optionally regenerating a plant from the modified plant containing the heteropolynucleotide. In a first aspect, the method for enhancing plant resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora* includes introducing the heteropolynucleotide of the present invention into at least one plant cell (particularly plant cells from Solanaceae plants). In some embodiments, the heteropolynucleotide is stably integrated into the genome of the plant cell.

[0118] In a second aspect, a method for enhancing plant resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora* involves modifying the nucleotide sequence of a natural or non-natural gene in the genome of a plant cell using genome editing methods to include the heteropolynucleotide of the present invention. The method includes introducing a nucleic acid molecule into a plant cell, wherein the nucleic acid molecule contains a nucleotide sequence comprising at least a portion of the Rpi-amr nucleotide sequence of the present invention, and wherein at least a portion of the nucleotide sequence of the natural or non-natural gene is replaced by at least a portion of the nucleotide sequence of the nucleic acid molecule. Therefore, the method of the present invention relates to gene substitution to produce the heteropolynucleotide of the present invention in the genome of a plant cell.

[0119] If desired, the methods of the first and / or second aspects may further include regenerating plant cells into plants whose genomes contain heterologous polynucleotides. Preferably, such regenerated plants contain enhanced resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*, relative to the resistance of control plants to the plant diseases.

[0120] The method of the present invention for enhancing plant resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora* may further include producing a plant comprising two, three, four, five, six or more nucleotide sequences encoding R proteins (preferably each nucleotide sequence encoding a different R protein). Such a plant comprising multiple R gene nucleotide sequences comprises one or more additional R gene nucleotide sequences of the present invention and / or any other nucleotide sequence encoding an R protein known in the art. It should be recognized that the methods of the first and / or second aspects can be used to produce such a plant comprising multiple nucleotide sequences encoding R proteins. Furthermore, it is recognized that the heteropolynucleotides of the present invention may comprise, for example, one or more Rpi-amr nucleotide sequences of the present invention or at least one Rpi-amr nucleotide sequence of the present invention and one or more nucleotide sequences encoding an R protein known in the art.

[0121] The plants disclosed herein can be used in agricultural crop production, particularly in areas where such plant diseases are prevalent and known to negatively impact or at least potentially negatively impact agricultural yields, to restrict plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. The method of the invention includes planting the plant of the invention (e.g., seedlings), tubers, or seeds, wherein the plant, tuber, or seed contains at least one R gene nucleotide sequence of the invention. The method also includes growing the plant derived from the seedling, tuber, or seed under conditions favorable to plant growth and development, and optionally harvesting at least one fruit, tuber, leaf, or seed from the plant.

[0122] The present invention further provides a method for identifying Solanaceae plants exhibiting newly conferred or enhanced resistance to plant diseases caused by at least one race of a species of the genus *Phytophthora*. This method can be used to cultivate Solanaceae plants resistant to plant diseases (e.g., late blight) caused by certain species of *Phytophthora*. Such resistant plants can be used for agricultural production of fruits, tubers, leaves, and / or seeds for human or livestock consumption or other uses. The method includes detecting the presence of the Rpi-amr nucleotide sequence of the present invention in a Solanaceae plant or at least a portion thereof or in cells. In some embodiments of the invention, detecting the presence of the Rpi-amr nucleotide sequence includes detecting the entire Rpi-amr nucleotide sequence in genomic DNA isolated from a Solanaceae plant. However, in a preferred embodiment, detecting the presence of the Rpi-amr nucleotide sequence includes detecting the presence of at least one marker within the Rpi-amr nucleotide sequence. In other embodiments of the invention, detecting the presence of the Rpi-amr nucleotide sequence includes detecting the presence of the R protein encoded by the Rpi-amr nucleotide sequence using an immunological detection method, for example, involving an antibody specific to the R protein.

[0123] In methods for identifying Solanaceae plants exhibiting newly conferred or enhanced resistance to plant diseases caused by at least one race of a species of the genus *Phytophthora*, the detection of the presence of the Rpi-amr nucleotide sequence in the Solanaceae plant may involve one or more of the following molecular biological techniques disclosed elsewhere herein or known in the art, including but not limited to isolating genomic DNA and / or RNA from the plant, amplifying nucleic acid molecules containing the Rpi-amr nucleotide sequence and / or label thereof by PCR amplification, sequencing nucleic acid molecules containing the Rpi-amr nucleotide sequence and / or label thereof, identifying the Rpi-amr nucleotide sequence, label, or transcript of the Rpi-amr nucleotide sequence by nucleic acid hybridization, and performing an immunological assay for detecting the R protein encoded by the Rpi-amr nucleotide sequence. It should be recognized that oligonucleotide probes and PCR primers can be designed to identify the Rpi-amr nucleotide sequence of the present invention, and such probes and PCR primers can be used in methods disclosed elsewhere herein or known in the art to rapidly identify one or more plants containing the Rpi-amr nucleotide sequence of the present invention in a plant population.

[0124] Depending on the desired outcome, the heteropolynucleotides of the present invention can be stably integrated into the genome of plant cells or unstablely integrated into the genome of plant cells. For example, if the desired outcome is to produce stably transformed plants with enhanced resistance to plant diseases caused by at least one race of a species of Phytophthora, then the heteropolynucleotides can be fused, for example, into plant transformation vectors suitable for stably integrating the heteropolynucleotides into the genome of plant cells. Typically, stably transformed plant cells are regenerated into transformed plants containing the heteropolynucleotides in their genome. Such stably transformed plants are able to transfer the heteropolynucleotides to subsequent generations of offspring plants through sexual and / or asexual reproduction. Plant transformation vectors, methods for stably transforming plants with introduced heteropolynucleotides, and methods for regenerating plants from transformed plant cells and tissues are well known in the fields of monocotyledonous and dicotyledonous plants or described elsewhere herein.

[0125] In other embodiments of the invention where it is not desired to stably integrate heteropolynucleotides into the plant genome, the heteropolynucleotides can be introduced into one or more plant cells of the plant using a transient transformation method. Such transient transformation methods include, for example, virus-based methods involving the use of viral particles or at least viral nucleic acids. Typically, such virus-based methods involve constructing a modified viral nucleic acid (containing the heteropolynucleotides of the invention operably linked to the viral nucleic acid), and then contacting the plant with a modified virus containing the modified viral nucleic acid, or with the viral nucleic acid itself. The modified virus and / or modified viral nucleic acid can be applied to the plant or parts thereof, for example, according to conventional methods used in agriculture, such as by spraying, irrigation, dusting, etc. The modified virus and / or modified viral nucleic acid can be applied by spraying, atomizing, dusting, sowing, or watering in the form of a sprayable solution, powder, suspension or dispersion, emulsion, oil dispersion, paste, dusting product, sowing material, or granules. It has been recognized that, prior to application to a plant or one or more parts thereof, it may be necessary to prepare a formulation containing the modified virus and / or modified viral nucleic acid. Methods for preparing insecticide formulations are generally known in the art or described elsewhere herein.

[0126] This invention provides nucleic acid molecules comprising the Rpi-amr nucleotide sequence. Preferably, such nucleic acid molecules are capable of conferring enhanced resistance to plant diseases caused by at least one race of a species of the genus *Phytophthora* on host plants, particularly Solanaceae host plants. Therefore, such nucleic acid molecules can be used in agricultural production to restrict plant diseases caused by at least one race of a species of the genus *Phytophthora*. The nucleic acid molecules of this invention include, but are not limited to, nucleic acid molecules comprising at least one Rpi-amr nucleotide sequence disclosed herein, and also include additional orthologs and other variants of the Rpi-amr nucleotide sequence capable of conferring resistance to plant diseases caused by at least one race of a species of the genus *Phytophthora*. Methods for determining plant resistance to plant diseases caused by at least one race of a species of the genus *Phytophthora* are known in the art or disclosed herein, including, for example, the decomposition assay (DLA) using decomposition leaves of *Tobacco Bensem's leaves* as described elsewhere herein.

[0127] The present invention also provides plants and their cells, particularly Solanaceae plants and their cells, said plants and their cells comprising Rpi-amr1e, Rpi-amr6b, Rpi-amr7d and / or Rpi-amr8c and / or alleles, homologs and other naturally and non-naturally occurring variants of such R genes, and produced by methods not involving the introduction of recombinant DNA into the plant or its cells. Such methods may include, for example, interspecific hybridization involving two or more different plant species. In a preferred embodiment, the plant is a Solanaceae plant.

[0128] In some embodiments, the Solanaceae plant is any Solanaceae plant other than *Solanum nigrescens* or *Solanum nigrescens*. In some other embodiments, the Solanaceae plant is any Solanaceae plant other than *Solanum nigrescens* and *S. nigrescens*. In other embodiments, the Solanaceae plant is any Solanaceae plant other than *Solanum nigrescens* containing alleles of *Rpi-amr1e* having the nucleotide sequences shown in SEQ ID NO:1 and / or 22 and / or alleles of *Rpi-amr1e* having the nucleotide sequences shown in SEQ ID NO:4, 7, 10, 13, 16, 19, 31, 32, 33, 34 and 35, wherein *Rpi-amr1e* and / or one or more alleles of *Rpi-amr1e* are endogenous or natural genes present at one or more natural locations in its genome.

[0129] While the Rpi-amr nucleotide sequences shown in SEQ ID NO:4, 7, 10, 13, 16, 19, 31, 32, 33, 34, and 35 are considered to be nucleotide sequences of the allele of Rpi-amr1e (SEQ ID NO:1) of *Solanum nigrum*, it should be understood that the present invention does not rely on such Rpi-amr nucleotide sequences corresponding to alleles present at the Rpi-amr1e locus in *Solanum nigrum* and / or other Solanaceae plants. Such Rpi-amr nucleotide sequences and Rpi-amr nucleic acid molecules and Rpi-amr genes containing such Rpi-amr nucleotide sequences can be used in the methods and compositions of the present invention disclosed herein, regardless of whether any such Rpi-amr nucleotide sequence corresponds to the allele of Rpi-amr1e in *Solanum nigrum* and / or other Solanaceae plants.

[0130] In other embodiments, the Solanaceae plant is any Solanaceae plant other than the *Solanum nigrum* plant containing the nucleotide sequence Rpi-amr7d shown in SEQ ID NO:51, wherein Rpi-amr7d is an endogenous or natural gene present at one or more natural locations in its genome. In other embodiments, the Solanaceae plant is any Solanaceae plant other than the *Solanum nigrum* plant containing the nucleotide sequence Rpi-amr8c shown in SEQ ID NO:56, wherein Rpi-amr8c is an endogenous or natural gene present at one or more natural locations in its genome. In yet another embodiment, the Solanaceae plant is any Solanaceae plant other than *S. nigrescens* plant containing the nucleotide sequence Rpi-amr6b shown in SEQ ID NO:46.

[0131] Additionally, a method is provided for introducing at least one Rpi-amr gene of the present invention into plants, particularly Solanaceae plants, which lack the at least one Rpi-amr gene in their genome. The Rpi-amr genes of the present invention include, for example, Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c, as well as alleles, homologs, and other naturally and non-naturally occurring variants of such R genes, and / or R genes containing the nucleotide sequences shown in SEQ ID NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, or 65, and / or encoding R genes containing SEQ ID NO:1, 3, 4, 6, 6, 65, and / or encoding SEQ ID NO:1, 3, 4, 6 ... The method involves crossing (i.e., cross-pollinating) a first plant containing at least one copy of the Rpi-amr gene of the present invention with a second Solanaceae plant lacking the Rpi-amr gene in its genome. The first and second plants can be the same species or different Solanaceae species, although in a preferred embodiment, the first and second plants are Solanaceae plants. For example, the first plant can be *Solanum nigrum*, and the Solanaceae plant can be potato or tomato (*Solanum lycopersicum*). This hybridization of the first species of plant with the second species of plant is called interspecific hybridization and can be used to infiltrate one or more target genes (e.g., Rpi-amr1e) from one species into a related species lacking the one or more target genes, and typically involves multiple generations of backcrossing of offspring with related species and selection of offspring containing one or more target genes in each generation. Such methods of interspecific hybridization, gene introgression, and backcrossing are well known in the art and can be used in the methods of this invention. See “Principals of Cultivar Development,” Fehr, 1993, Macmillan Publishing Company, New York; and “Fundamentals of Plant Genetics and Breeding,” Welsh, 1981, John Wiley & Sons, Inc., New York.

[0132] In the method of the present invention for introducing at least one Rpi-amr gene of the present invention into a plant lacking said at least one Rpi-amr gene in its genome, the first plant or the second plant may be a pollen donor plant. For example, if the first plant is a pollen donor plant, the second plant is a pollen recipient plant. Similarly, if the second plant is a pollen donor plant, the first plant is a pollen recipient plant. After hybridization, the pollen recipient plant is grown under conditions favorable to plant growth and development, and grown for a sufficient time to allow the seeds to mature or to achieve other desired growth stages for subsequent in vitro germination processes (e.g., embryo rescue described below). The seeds can then be harvested, and those containing said Rpi-amr genes are identified by any method known in the art, including, for example, methods described elsewhere herein for identifying Solanaceae plants exhibiting newly conferred or enhanced resistance to plant diseases caused by at least one race of a species of the genus Phytophthora. In some embodiments, the first plant is a Solanum nigrum plant containing one or more Rpi-amr genes, and the second plant is a Solanum nigrum plant lacking said one or more Rpi-amr genes. In a preferred embodiment, the first plant is a *Solanum nigrum* species containing one or more Rpi-amr genes, or another Solanaceae species containing one or more Rpi-amr genes in its genome, and the second plant is a Solanaceae species other than *Solanum nigrum*. Preferred Solanaceae plants include potato, tomato, eggplant, pepper, tobacco, and petunia.

[0133] However, it should be recognized that in certain embodiments of the invention involving interspecific hybridization, it may be advantageous to harvest seeds produced by such interspecific hybridization at an immature growth stage and then germinate the immature seeds in culture (i.e., in vitro), wherein the seeds are germinated in culture using methods known in the art as “embryo rescue” methods. See Reed (2005) “Embryo Rescue,” in Plant Development and Biotechnology, Trigiano and Gray, eds. (PDF). CRC Press, Boca Raton, pp. 235-239; and Sharma et al. (1996) Euphaitica 89:325-337. It should also be recognized that “embryo rescue methods are typically used when mature seeds produced by interspecific hybridization show little or no germination (thus producing few or no interspecific hybrids).

[0134] The method of the present invention can be used to produce plants with enhanced resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. Typically, the method of the present invention will enhance or increase the resistance of the test plant to the plant disease by at least 25%, 50%, 75%, 100%, 150%, 200%, 250%, 500%, or more compared to the resistance of a control plant to the same race of a species of *Phytophthora*. Unless otherwise stated or apparent from the context of use, the control plant of the present invention is a plant that does not contain the heteropolynucleotide and / or Rpi-amr1e nucleotide sequence of the present invention. Preferably, the control plant is substantially identical (e.g., the same species, subspecies, and variety) to the plant containing the heteropolynucleotide, except that the control does not contain the heteropolynucleotide or Rpi-amr nucleotide sequence of the present invention. In some embodiments, the control will contain the heteropolynucleotide but not one or more Rpi-amr nucleotide sequences present in the heteropolynucleotide of the present invention.

[0135] In addition, the present invention provides plants, seeds, and plant cells transformed by the methods of the present invention and / or containing heterologous polynucleotides of the present invention. Descendant plants and their seeds containing heterologous polynucleotides of the present invention are also provided. The present invention also provides fruits, seeds, tubers, leaves, stems, roots, and other plant parts produced by the transformed plants and / or descendant plants of the present invention, as well as food and other agricultural products containing said plants or any one or more of them, or food and other agricultural products produced or derived from said plants or any one or more of them, including but not limited to fruits, tubers, leaves, stems, roots, and seeds. Other agricultural products include, for example, tobacco products (e.g., cigarettes, cigars, pipes, and chewing tobacco) produced from tobacco leaves, and food and industrial starch products produced from potato tubers. It should be recognized that such foods can be consumed or used by humans and other animals, including but not limited to pets (e.g., dogs and cats), livestock (e.g., pigs, cattle, chickens, turkeys, and ducks), and animals produced in freshwater and marine aquaculture systems (e.g., fish, shrimp, prawns, crayfish, and lobsters).

[0136] Non-limiting examples of the compositions and methods of the present invention are as follows:

[0137] 1. A nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of:

[0138] (a) The nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 31, 32, 33, 34, 35, 46, 51, 56 or 61;

[0139] (b) A nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63, and optionally, wherein said nucleotide sequence is not naturally occurring;

[0140] (c) The nucleotide sequence shown in SEQ ID NO: 3, 6, 9, 12, 15, 18, 21, 23, 25, 26, 27, 36, 37, 38, 39, 40, 49, 50, 54, 55, 59, 60, 64 or 65;

[0141] (d) A nucleotide sequence having at least 90% sequence identity with at least one of the nucleotide sequences shown in SEQ ID NOS:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, and 65, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus *Phytophthora*, and optionally, wherein the nucleotide sequence is not naturally occurring; and

[0142] (e) A nucleic acid molecule comprising a nucleotide sequence having at least 90% sequence identity with at least one amino acid sequence of the amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 and 63, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus Phytophthora, and optionally, wherein the nucleotide sequence is not naturally occurring.

[0143] 2. The nucleic acid molecule of implementation scheme 1, wherein the nucleic acid molecule is an isolated nucleic acid molecule.

[0144] 3. An expression cassette comprising a nucleic acid molecule according to embodiment 1 or 2 and an operatively linked heterologous promoter.

[0145] 4. A vector comprising a nucleic acid molecule of embodiment 1 or 2 or an expression cassette of embodiment 3.

[0146] 5. The vector of implementation scheme 4, which also contains an additional R gene.

[0147] 6. A host cell that has been transformed with a nucleic acid molecule of embodiment 1 or 2, an expression cassette of embodiment 3, or a vector of embodiment 4 or 5.

[0148] 7. The host cell of embodiment 6, wherein the host cell is a plant cell, bacterial cell, fungal cell or animal cell.

[0149] 8. The host cell of implementation scheme 6 or 7, wherein the host cell is a Solanaceae plant cell.

[0150] 9. A plant or plant cell comprising a nucleic acid molecule according to embodiment 1 or 2, an expression cassette according to embodiment 3, or a vector according to embodiment 4 or 5.

[0151] 10. The plant or plant cell of embodiment 9, wherein the plant is a Solanaceae plant and the plant cell is a Solanaceae plant cell.

[0152] 11. The plant of embodiment 10, wherein the solanaceous plant is not Solanum americanum and / or Solanum nigrescens, or wherein the solanaceous plant is selected from potato, tomato, eggplant, pepper, tobacco and petunia.

[0153] 12. A plant comprising a heteropolynucleotide stably integrated into its genome, said heteropolynucleotide comprising a nucleotide sequence selected from the group consisting of:

[0154] (a) The nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 31, 32, 33, 34, 35, 46, 51, 56 or 61;

[0155] (b) A nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63;

[0156] (c) The nucleotide sequence shown in SEQ ID NO: 3, 6, 9, 12, 15, 18, 21, 23, 25, 26, 27, 36, 37, 38, 39, 40, 49, 50, 54, 55, 59, 60, 64 or 65;

[0157] (d) A nucleotide sequence having at least 90% sequence identity with at least one nucleotide sequence shown in SEQ ID NOS:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, and 65, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus *Phytophthora*; and

[0158] (e) A nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity with at least one amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 and 63, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus Phytophthora.

[0159] 13. The plant of embodiment 12, wherein the heteropolynucleotide comprises a nucleotide sequence of any one of (b)-(e) and further comprises a promoter operatively linked to express the nucleotide sequence in the plant.

[0160] 14. The plant of embodiment 13, wherein the promoter is selected from pathogen-inducible promoters, constitutive promoters, tissue-preferred promoters, wound-inducible promoters, and chemically regulated promoters.

[0161] 15. A plant of any of the embodiments of 12-14, wherein the plant is a plant of the Solanaceae family.

[0162] 16. The plant of any one of embodiments 12-15, wherein the solanaceous plant is selected from potato, tomato, eggplant, pepper, tobacco and petunia.

[0163] 17. A plant according to any one of embodiments 12-16, wherein the plant comprises enhanced resistance to plant diseases caused by at least one race of at least one species of the genus Phytophthora, relative to a control plant.

[0164] 18. The plant of embodiment 17, wherein the plant comprises enhanced resistance to late blight caused by at least one race of Phytophthora infestans relative to the control plant.

[0165] 19. The plant of any one of embodiments 12-18, wherein the plant is a potato or tomato plant.

[0166] 20. A method for enhancing plant resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*, the method comprising modifying at least one plant cell to contain a heteropolynucleotide, said heteropolynucleotide comprising a nucleotide sequence selected from the group consisting of:

[0167] (a) The nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 31, 32, 33, 34, 35, 46, 51, 56 or 61;

[0168] (b) A nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63;

[0169] (c) The nucleotide sequence shown in SEQ ID NO: 3, 6, 9, 12, 15, 18, 21, 23, 25, 26, 27, 36, 37, 38, 39, 40, 49, 50, 54, 55, 59, 60, 64 or 65;

[0170] (d) A nucleotide sequence having at least 90% sequence identity with at least one nucleotide sequence shown in SEQ ID NOS:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, and 65, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus *Phytophthora*; and

[0171] (e) A nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity with at least one amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 and 63, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus Phytophthora.

[0172] 21. The method of embodiment 20, wherein the heteropolynucleotide is stably integrated into the genome of a plant cell.

[0173] 22. The method of embodiment 20 or 21, wherein the plant cells are regenerated into a plant whose genome contains the heterologous polynucleotide.

[0174] 23. The method of any one of embodiments 20-22, wherein modifying at least one plant cell to contain a heteropolynucleotide comprises introducing the heteropolynucleotide into at least one plant cell.

[0175] 24. The method of any one of embodiments 20-23, wherein the heteropolynucleotide comprises a nucleotide sequence of any one of (b)-(e) and further comprises a promoter operatively linked to express the nucleotide sequence in a plant.

[0176] 25. The method of implementation scheme 24, wherein the promoter is selected from pathogen-inducible promoters, constitutive promoters, tissue-optimized promoters, wound-inducible promoters, and chemically regulated promoters.

[0177] 26. The method of any one of embodiments 20-22, wherein modifying at least one plant cell to include a heterologous polynucleotide comprises using genome editing to modify the nucleotide sequence of a natural or non-natural gene in the genome of the plant cell to include a nucleotide sequence of any one of (a)-(e).

[0178] 27. The method of embodiment 26, wherein the modification further comprises introducing a nucleic acid molecule into the plant cell, wherein the nucleic acid molecule comprises a nucleotide sequence containing at least a portion of a nucleotide sequence of any one of (a)-(e).

[0179] 28. The method of embodiment 27, wherein at least a portion of the nucleotide sequence of the natural or non-natural gene is replaced by at least a portion of the nucleotide sequence of the nucleic acid molecule.

[0180] 29. The method of any one of embodiments 22-28, wherein, relative to a control plant, the plant containing the heteropolynucleotide comprises enhanced resistance to plant diseases caused by at least one race of at least one species of the genus Phytophthora.

[0181] 30. The method of any one of embodiments 22-29, wherein the plant containing the heteropolynucleotide comprises enhanced resistance to late blight caused by at least two races of Phytophthora blight, relative to the control plant.

[0182] 31. The method of any one of embodiments 20-30, wherein the plant is a potato or tomato plant.

[0183] 32. A plant produced by any one of the methods described in embodiments 20-31.

[0184] 33. The fruit, tuber, leaf, or seed of a plant according to any one of embodiments 9-19 and 32, wherein the fruit, tuber, leaf, or seed contains the heteropolynucleotide.

[0185] 34. A method for restricting plant diseases caused by at least one race of at least one species of the genus Phytophthora in crop production, the method comprising planting seedlings, tubers or seeds of any one of embodiments 9-19 and 32, and cultivating the seedlings, tubers or seeds under conditions favorable to the growth and development of the plant from which they are produced, wherein the seedlings, tubers or seeds contain the nucleic acid molecule, expression cassette, vector or heteropolynucleotide.

[0186] 35. The method of embodiment 34, further comprising harvesting at least one fruit, tuber, leaf and / or seed from said plant.

[0187] 36. A method for identifying a Solanaceae plant exhibiting newly conferred or enhanced resistance to a plant disease caused by at least one race of at least one species of the Phytophthora genus, the method comprising detecting the presence of an Rpi-amr nucleotide sequence in the plant or in at least one part or cell thereof.

[0188] 37. The method of implementation scheme 36, wherein the plant disease is late blight caused by at least one race of Phytophthora.

[0189] 38. The method of implementation scheme 36 or 37, wherein the nightshade plant is a potato or tomato plant.

[0190] 39. The method of any one of embodiments 36-38, wherein the presence of the Rpi-amr nucleotide sequence is detected by detecting at least one marker within the Rpi-amr nucleotide sequence.

[0191] 40. The method of any one of embodiments 36-39, wherein the Rpi-amr nucleotide sequence comprises or is composed of the nucleotide sequences shown in SEQ IDNOS:11, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64 and 65.

[0192] 41. The method of any one of embodiments 36-39, wherein detecting the presence of the Rpi-amr nucleotide sequence comprises a method selected from PCR amplification, nucleic acid sequencing, nucleic acid hybridization, and an immunoassay for detecting the R protein encoded by the Rpi-amr nucleotide sequence.

[0193] 42. A plant of the Solanaceae family, which is identified by means of any one of embodiments 36-41.

[0194] 43. The Solanaceae plant of embodiment 42, wherein the Solanaceae plant is not Solanum nigrum and / or Solanumnigrescens.

[0195] 44. Fruits, tubers, leaves or seeds of Solanaceae plants according to implementation plan 42 or 43.

[0196] 45. A plant or plant cell comprising: (i) at least one of Rpi-amr1e, an allele of Rpi-amr1e, Rpi-amr7d, and Rpi-amr8c, wherein the plant is not a Solanum nigrescens plant and the plant cell is not a Solanum nigrescens plant cell; or (ii) Rpi-amr6b, wherein the plant is not a Solanum nigrescens plant and the plant cell is not a Solanum nigrescens plant cell.

[0197] 46. ​​The plant or plant cell of embodiment 45, wherein the plant is a Solanaceae plant and the plant cell is a Solanaceae plant cell.

[0198] 47. A method for introducing at least one Rpi-amr gene into a plant, the method comprising:

[0199] (a) Crossing a first plant containing at least one copy of at least one Rpi-amr gene in its genome with a second plant lacking said at least one Rpi-amr gene in its genome, thereby producing at least one offspring plant; and

[0200] (b) Select at least one progeny plant that contains at least one Rpi-amr gene in its genome.

[0201] 48. The method of embodiment 47, wherein the first plant is a Solanum nigrescens plant and the second plant is not a Solanum nigrescens plant, or wherein the first plant is a Solanum nigrescens plant and the second plant is not a Solanum nigrescens plant.

[0202] 49. The method of implementation scheme 47 or 48, wherein the second plant is a potato (Solanum tuberosum) plant or a tomato (Solanum lycopersicum) plant.

[0203] 50. The method of any one of embodiments 47-49, wherein at least one Rpi-amr gene comprises a nucleotide sequence selected from the group consisting of:

[0204] (a) The nucleotide sequence shown in SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 31, 32, 33, 34, 35, 46, 51, 56 or 61;

[0205] (b) A nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63;

[0206] (c) The nucleotide sequence shown in SEQ ID NO: 3, 6, 9, 12, 15, 18, 21, 23, 25, 26, 27, 36, 37, 38, 39, 40, 49, 50, 54, 55, 59, 60, 64 or 65;

[0207] (d) A nucleotide sequence having at least 90% sequence identity with at least one nucleotide sequence shown in SEQ ID NOS:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, and 65, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus *Phytophthora*, and

[0208] (e) A nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity with at least one amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 and 63, wherein the nucleic acid molecule is capable of conferring resistance in a plant containing the nucleic acid molecule to a plant disease caused by at least one race of at least one species of the genus Phytophthora.

[0209] 51. The method of any one of embodiments 47-50, wherein selecting at least one progeny plant includes detecting the presence of the Rpi-amr nucleotide sequence in the progeny plant or at least a portion thereof or in cells using the method of any one of embodiments 36-41.

[0210] 52. The method of any one of embodiments 47-51, further comprising (i) backcrossing at least one selected progeny plant of (b) with a Solanaceae plant that is of the same species and genotype as the second Solanaceae plant, or that is of the same species as the second Solanaceae plant and lacks the at least one Rpi-amr gene in its genome, wherein at least one progeny plant is produced by the backcross; and (ii) selecting at least one progeny plant produced by the backcross of (i) that contains the at least one Rpi-amr gene in its genome.

[0211] 53. Offspring plants according to any one of implementation schemes 47-52.

[0212] 54. The offspring plants of embodiment 53, wherein the Solanaceae plant is not Solanum nigrum and / or Solanumnigrescens.

[0213] 55. Fruits, tubers, leaves or seeds of Solanaceae plants according to implementation plan 53 or 54.

[0214] 56. Use in agriculture of any of the plants, fruits, tubers, leaves or seeds described in any of Implementation Schemes 9-19, 32, 33, 42-46 and 53-55.

[0215] 57. A food for humans or animals comprising or produced using any one of embodiments 9-19, 32, 33, 42-46 and 53-54, of a plant, fruit, tuber, leaf and / or seed, or produced using said plant, fruit, tuber, leaf and / or seed.

[0216] 58. A polypeptide comprising an amino acid sequence selected from the group consisting of:

[0217] (a) The amino acid sequence shown in SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63;

[0218] (b) An amino acid sequence encoded by the nucleotide sequence shown in SEQ ID NO: 1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64 or 65; and

[0219] (c) An amino acid sequence having at least 90% sequence identity with at least one of the amino acid sequences shown in SEQ ID NOS:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 and 63, wherein a polypeptide containing said amino acid sequence is capable of conferring resistance in a plant containing said polypeptide to a plant disease caused by at least one race of at least one species of the genus Phytophthora.

[0220] Further embodiments of the methods and compositions of the present invention are described elsewhere in this document.

[0221] Unless otherwise expressly stated or obvious from the context of use, the methods and compositions of the present invention can be used with any plant species, including, for example, monocots, dicots and conifers. Examples of target plant species include, but are not limited to, maize (corn), species of the genus *Brassica* (e.g., Brassica napus, Brassica oleracea, Brassica rapa), especially those *Brassica* species used as a source of seed oil, alfalfa (alfalfa), rice, black wheat (× rye or *Wheat* × rye), sorghum (bicolor sorghum, *Sorghum vulgare*), sorghum bran (*Eriocheir sinensis*), millet (e.g., pearl millet), millet (*Panicum miliaceum*), foxtail millet (*Setaria italica*), dragon claw millet (*Eleusine coracana*), switchgrass (*Panicum virgatum*), sunflower (*Helianthus annuus*), safflower (*Carthamus tinctorius*), wheat (*Triticum*). Soybeans (Glycinemax), Tobacco (Nicotiana tabacum), Potatoes (Solanum tuberosum), Peanuts (Arachis hypogaea), Cotton (Gossypium huberutum, Gossypium hirsutum), Strawberries (e.g., Fragaria × ananassa, Fragaria vesca, Fragaria moschata, Fragaria virginiana, Fragaria chiloensis), Sweet potatoes (Ipomoea batatus), Yams (certain species of Dioscorea). spp.), round potato (D. rotundata), yellow potato (D. cayenensis), ginseng (D. alata), long yam (D. polystachya), yellow herb (D. bulbifera), D. esculenta, D. dumetorum, D. trifida), cassava (Manihotesculenta)), coffee (certain species of the genus Coffea (Coffea spp.)).), coconut (Coconut (Cocos nucifera)), oil palm (e.g., oil palm (Elaeis guineensis, American oil palm (Elaeis oleifera)), pineapple (Ananascomosus)), citrus tree (some species of Citrus spp.)), cocoa tree (Theobroma cacao)), tea (Camellia sinensis)), banana (some species of Musa spp.)), avocado (Perseaamericana)), fig (Ficus casica)), guava (Psidium guajava)), mango (Manaifera indica)), olive (Olea europaea)), papaya (Caricapapaya)), cashew (Anacardium occidentale)), macadamia nut (Macadamia integrifolia)), almond (Prunus alba) (amygdalus), date palm (Phoenix dactylifera), cultivated forms of beets (beets, garden beets, chard or leaf beets, forage beets or fodder beets), sugarcane (some species of the genus Saccharum spp.), oats (Avena sativa), barley (Hordeum vulgare), hemp (canna, Indian hemp, bulrush hemp), poplar (some species of the genus Populus spp.), eucalyptus (some species of the genus Eucalyptus spp.), Arabidopsis thaliana, rooted Arabidopsis thaliana (Arabidopsis rhizogenes), tobacco of Bensem, two-spike short-stalked grass vegetables, ornamental plants, conifers and other trees. In specific implementation schemes, the plants used in this invention are crop plants (e.g., potatoes, tobacco, tomatoes, corn, sorghum, wheat, millet, rice, barley, oats, sugarcane, alfalfa, soybeans, peanuts, sunflowers, cotton, safflower, certain species of Brassica, lettuce, strawberries, apples, citrus, etc.).

[0222] Vegetables include tomatoes, eggplants (also known as "aubergine" or "brinjal") (Solanum melongena), peppers (Capsicum annuum), lettuce (e.g., romaine lettuce), green beans (kidney beans), lima beans (beans), peas (some species of Lathyrus spp.), chickpeas (Cicerarietinum), and members of the Cucumber genus such as cucumber (C. sativus), cantaloupe (C. cantalupensis), and honeydew melon (C. melon). Ornamental plants include rhododendrons (some species of the genus *Rhododendron*), hydrangeas (*Macrophylla hydrangea*), hibiscus (*Hibiscus rosa-anensis*), roses (some species of the genus *Rosa*), tulips (some species of the genus *Tulipa*), daffodils (some species of the genus *Narcissus*), petunias (*Petunia hybrida*), carnations (*Aianthus caryophyllus*), poinsettias (*Euphorbia pulcherrima*), and chrysanthemums. Fruit trees and related plants include, for example, apples, pears, peaches, plums, oranges, grapefruits, limes, pomelos, palms, and bananas. Nut trees and related plants include, for example, almonds, cashews, walnuts, pistachios, macadamia nuts, nuts, hazelnuts, and pecans.

[0223] In specific implementations, the plants of the present invention are crop plants, such as, for example, corn, soybean, wheat, rice, cotton, alfalfa, sunflower, brassica (some species of the genus Brassica, especially Brassica napus, Brassica oleracea, and Brassica oleracea), rapeseed, sorghum, millet, barley, black wheat, safflower, peanut, sugarcane, tobacco, potato, tomato, and pepper.

[0224] The preferred plants of this invention are plants of the Solanaceae family. As used herein, the term "Solanaceae plant" refers to plants that are members of the Solanaceae family. Such Solanaceae plants include, for example, domesticated and non-domesticated members of the Solanaceae family. The Solanaceae plants of this invention include, but are not limited to, potato, eggplant (Solanum melongena), petunia (certain species of the genus Petunia, such as Petunia x hybrid or Petunia hybrida), groundcherry (Physalis philadelphica), peruviana (Physalis peruviana), a species of the genus Physalis (Physalis sp.), white sage (Solanum dulcamara), garden blueberry (Solanum scabrum), gboma eggplant (Solanum macrocarpon), pepper (certain species of the genus Capsicum; such as chili pepper (Capsicum annuum), bell pepper (C. baccatum), Chinese pepper (C. chinense), small red pepper (C. frutescens), C. pubescens, etc.), tomato (Solanum The plant species include *Lycopersicum* or *Lycopersicon esculentum*, tobacco (some species of the genus *Nicotiana*, such as *N. tabacum*, *N. benthamiana*), *Solanum nigrum*, *Solanum nigrescens*, *Solanum stoloniferum*, *Solanum papita*, *Solanum bulbocastanum*, *Solanum schenckii*, *Solanum hjertingii*, *Solanum venture*, *Solanum mochiquense*, *Solanum chacoense*, and *Solanum impinellifolium*. In a preferred embodiment of the method and composition of the present invention, the solanaceae plants are agriculturally grown solanaceae plants, including but not limited to potatoes, tomatoes, physalis, physalis, eggplants, peppers, tobacco, and petunias. In a more preferred embodiment, the solanaceae plants are potatoes and tomatoes. In an even more preferred embodiment, the preferred plant is potato.In some other embodiments of the methods and compositions disclosed herein, the preferred Solanaceae plants are all Solanaceae plants except for *Solanum nigrescens* and / or *Solanum nigrescens*.

[0225] The term "Solanaceae plant" is intended to include any Solanaceae plant at any stage of maturity or development, and, unless the context clearly indicates otherwise, any cell, tissue, or organ (plant part) obtained or derived from any such plant. Solanaceae plant parts include, but are not limited to, fruits, stems, tubers, roots, flowers, ovules, stamens, leaves, embryos, meristematic zones, callus tissue, anther cultures, gametophytes, sporophytes, pollen, microspores, protoplasts, etc. As used herein, the term "tuber" is intended to refer to the whole tuber or any part thereof, such as, for example, a slice or portion of a potato tuber containing one or more buds (i.e., "eyes") suitable for field cultivation to produce potato plants. The invention also includes seeds produced from the Solanaceae plants of the invention.

[0226] The compositions and methods of the present invention enable us to produce plants with enhanced resistance to at least one species of at least one species of the genus *Phytophthora*. In a preferred embodiment of the invention, the species of *Phytophthora* is *Phytophthora infestans*. In other embodiments, the species of *Phytophthora* is a species of the genus capable of causing plant disease on at least one plant. For the purposes of this invention, certain species of *Phytophthora* include, but are not limited to, *Phytophthora infestans*, *Phytophthora parasitica*, *Phytophthora ramorum*, *Phytophthora ipomoeae*, *Phytophthora mirabilis*, *Phytophthora capsici*, *Phytophthora porri*, *Phytophthora sojae*, *Phytophthora palmivora*, and *Phytophthora phaseoli*.

[0227] In one embodiment of the invention, the nucleotide sequence encoding the R protein has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence identity with the complete nucleotide sequence or fragment thereof shown in at least one of SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 31, 32, 33, 34, 35, 46, 51, 56 and 61. In another embodiment of the invention, the nucleotide sequence encoding the R protein has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence identity with the complete nucleotide sequence or fragment thereof shown in at least one of SEQ IDNOS:3, 6, 9, 12, 15, 18, 21, 23, 25, 26, 27, 36, 37, 38, 39, 40, 49, 50, 54, 55, 59, 60, 64 and 65.

[0228] This invention comprises isolated or substantially purified compositions of polynucleotides (also referred to herein as “nucleic acid molecules,” “nucleic acids,” etc.) or proteins (also referred to herein as “polypeptides”). The “isolated” or “purified” polynucleotides or proteins, or their biologically active portions, are substantially or substantially free of components that typically accompany or interact with polynucleotides or proteins found in their natural environment. Therefore, when produced by recombinant technology, the isolated or purified polynucleotides or proteins are substantially free of other cellular material or culture medium, or when chemically synthesized, are substantially free of chemical precursors or other chemicals. Preferably, the “isolated” polynucleotides are free of polynucleotides (preferably protein-coding sequences) naturally located flanking the polynucleotide in the genomic DNA of the organism from which the polynucleotide is derived (i.e., sequences located at the 5' and 3' ends of the polynucleotide). For example, in various embodiments, the isolated polynucleotides may contain nucleotide sequences of less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb naturally located flanking the polynucleotide in the genomic DNA of the cell from which the polynucleotide is derived. Proteins that are essentially free of cellular material include protein formulations containing less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of contaminating proteins. When recombinantly producing the proteins of the present invention or their bioactive portions, the optimal culture medium represents less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of chemical precursors or non-protein target chemicals.

[0229] Fragments and variants of the disclosed polynucleotides and proteins thus encoded are also included in this invention. "Fragment" is intended to refer to a portion of a polynucleotide or amino acid sequence and thus a portion of the protein encoded therefrom. A polynucleotide fragment containing the coding sequence may encode a protein fragment that retains the biological activity of the full-length or native protein. Alternatively, a polynucleotide fragment that can be used as a hybridization probe typically does not encode a protein that retains biological activity or promoter activity. Therefore, fragments of nucleotide sequences can be at least about 20 nucleotides, about 50 nucleotides, about 100 nucleotides, and up to the full-length polynucleotide of this invention.

[0230] In some embodiments of the invention, the disclosed polynucleotides and fragments and variants of the proteins encoded therein are those fragments and variants capable of conferring resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. Preferably, polynucleotides containing fragments of the natural R polynucleotide of the invention are capable of conferring resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. Similarly, proteins or polypeptides containing the natural R protein of the invention are preferably capable of conferring resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*.

[0231] The polynucleotide fragments of the natural R polynucleotide contain at least 16, 20, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1500, 2000, 3000, 4000, 5000, 6000, 7000, 8000, or 9000 consecutive nucleotides, or the number of nucleotides present in the full-length R polynucleotide disclosed herein.

[0232] The term "variant" is intended to denote substantially similar sequences. For polynucleotides, variants include polynucleotides with deletions (i.e., truncation) at the 5' and / or 3' ends; and / or deletions and / or additions of one or more nucleotides at one or more internal sites in the native polynucleotide; and / or substitutions of one or more nucleotides at one or more sites in the native polynucleotide. As used herein, a "native" polynucleotide or polypeptide comprises either a naturally occurring nucleotide sequence or an amino acid sequence, respectively. For polynucleotides, conserved variants include those sequences that encode the amino acid sequence of one of the R proteins of the present invention due to the degeneracy of the genetic code. Naturally occurring allelic variants can be identified using known molecular biology techniques, such as polymerase chain reaction (PCR) and hybridization techniques described below. Variant polynucleotides also include synthetically derived polynucleotides, such as those produced, for example, by site-directed mutagenesis but still encoding the R protein of the present invention. Typically, as determined by the sequence alignment procedures and parameters described elsewhere herein, a variant of a particular polynucleotide of the present invention will have at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with that particular polynucleotide. In certain embodiments of the invention, specific polynucleotide variants of the invention will have at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity with at least one nucleotide sequence selected from SEQ ID NO: 1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64 and 65, and optionally include non-naturally occurring nucleotide sequences that are identical to ...8, 95, 96, 97%, 98%, 99% or greater, and optionally include non-naturally occurring nucleotide sequences that are identical to SEQ ID NO: The nucleotide sequences shown in NO:1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64 and / or 65 differ from each other in that at least one nucleotide modification is selected from the substitution of at least one nucleotide, the addition of at least one nucleotide, and the deletion of at least one nucleotide.It should be understood that the addition of at least one nucleotide may be the addition of one or more nucleotides within the nucleotide sequence of the present invention (e.g., SEQ ID NO: 1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64, or 65), the addition of one or more nucleotides to the 5' end of the nucleotide sequence of the present invention, and / or the addition of one or more nucleotides to the 3' end of the nucleotide sequence of the present invention.

[0233] Variants of a particular polynucleotide (i.e., a reference polynucleotide) of the present invention can also be assessed by comparing the percentage of sequence identity between a polypeptide encoded by a variant polynucleotide and a polypeptide encoded by a reference polynucleotide. Thus, for example, polynucleotides encoding polypeptides having a given percentage of sequence identity with at least one polypeptide having an amino acid sequence selected from SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, and 63 are disclosed. The percentage of sequence identity between any two polypeptides can be calculated using the sequence alignment procedures and parameters described elsewhere herein. When any given polynucleotide pair of the present invention is evaluated by comparing the percentage of sequence identity shared by the two polypeptides they encode, the percentage of sequence identity between the two encoded polypeptides is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more of sequence identity. In certain embodiments of the invention, variants of specific polypeptides of the invention will have at least one of the amino acid sequences shown in SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, or 63 having at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher sequence identity, and optionally include non-naturally occurring amino acid sequences that are selected from SEQ ID NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, or 63 ... have at least about 60%, 65%, 75%, 80%, 85%, 90%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher, and optionally include non-naturally occurring amino acid sequences that are selected from At least one amino acid sequence of SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, and 63 differs from at least one amino acid modification selected from substitution of at least one amino acid, addition of at least one amino acid, and deletion of at least one amino acid. It should be understood that the addition of at least one amino acid can be the addition of one or more amino acids within the amino acid sequence of the present invention (e.g., SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, or 63), the addition of one or more amino acids to the N-terminus of the amino acid sequence of the present invention, and / or the addition of one or more amino acids to the C-terminus of the amino acid sequence of the present invention.

[0234] "Variant" protein is intended to refer to a protein derived from a natural protein by the deletion (so-called truncation) of one or more amino acids at the N-terminus and / or C-terminus of the natural protein, the deletion and / or addition of one or more amino acids at one or more internal sites in the natural protein, or the substitution of one or more amino acids at one or more sites in the natural protein. Such variants can be generated, for example, by genetic polymorphism or artificial manipulation. As determined by the sequence alignment procedures and parameters described elsewhere herein, a biologically active variant of the R protein will have at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity with the amino acid sequences of the present invention (e.g., the amino acid sequences shown in SEQ ID NO: 2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62 or 63). The bioactive variants of the protein of the present invention may differ from the protein in having as few as 1-15 amino acid residues, as few as 1-10, such as 6-10, as few as 5, as few as 4, 3, 2 or even 1 amino acid residue.

[0235] The proteins of the present invention can be modified in various ways, including amino acid substitution, deletion, truncation, and insertion. Methods for such operations are generally known in the art. Methods for mutagenesis and polynucleotide alteration are well known in the art. See, for example, Kunkel (1985) Proc. Natl. Acad. Sci. USA 82:488-492; Kunkel et al. (1987) Methods in Enzymollette. 154:367-382; U.S. Patent No. 4,873,192; Walker and Gaastra, editors (1983) Techniques in Molecular Biology (MacMillan Publishing Company, New York) and references cited therein. Guidance on appropriate amino acid substitutions that do not affect the biological activity of the target protein can be found in the model of Dayhoff et al. (1978) Atlas of Protein Sequence and Structure (Natl. Biomed. Res. Found., Washington, DC) (incorporated herein by reference). Conservative substitution, such as replacing one amino acid with another amino acid that has similar properties, may be the best approach.

[0236] Therefore, the genes and polynucleotides of the present invention include naturally occurring sequences as well as mutants and other variant forms. Similarly, the proteins of the present invention include naturally occurring proteins as well as their variants and modified forms. More preferably, such variants confer enhanced resistance to plants containing said variants or portions thereof to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. In some embodiments, the mutations produced in the DNA encoding the variant will not place the sequence outside the reading frame. Most ideally, the mutations will not produce complementary regions that could generate secondary mRNA structures. See EP patent application publication number 75,444.

[0237] Deletions, insertions, and substitutions of the protein sequences included herein are not expected to produce fundamental changes in the protein's characteristics. However, when the exact effect of a substitution, deletion, or insertion is difficult to predict before doing so, those skilled in the art will understand that the effect will be assessed by conventional screening assays. That is, activity can be assessed by the assays disclosed below.

[0238] Variant polynucleotides and proteins also include sequences and proteins derived from mutagenesis and recombination processes, such as DNA shuffling. Such DNA shuffling strategies are known in the art. See, for example, Stemmer (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751; Stemmer (1994) Nature 370:389-391; Cramer et al. (1997) Nature Biotech. 15:436-438; Moore et al. (1997) J. Mol. Biol. 272:336-347; Zhang et al. (1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Cramer et al. (1998) Nature 391:288-291; and U.S. Patent Nos. 5,605,793 and 5,837,458.

[0239] The polynucleotides of this invention can be used to isolate corresponding sequences from other organisms, particularly other plants. Thus, methods such as PCR and hybridization can be used to identify such sequences based on their sequence homology with the sequences shown herein. Sequences isolated based on their sequence identity with the complete sequences shown herein or their variants and fragments are included in this invention. Such sequences include sequences that are orthologs of the disclosed sequences. "Ortholog" is intended to represent genes that originate from a common ancestral gene and are found in different species due to speciation. Genes found in different species are considered orthologs when their nucleotide sequences and / or the protein sequences they encode share at least 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater sequence identity. The function of orthologs is generally highly conserved across species. Therefore, isolated polynucleotides encoding R proteins having at least 60% amino acid sequence identity with at least one of the full-length amino acid sequences of R proteins disclosed herein or known in the art are included in this invention.

[0240] In one embodiment, the ortholog of the present invention has a nucleotide sequence comprising at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater of at least one nucleotide sequence comprising a nucleotide sequence selected from those shown in SEQ ID NO: 1, 3, 4, 6, 7, 9, 10, 12, 13, 15, 16, 18, 19, 21, 22, 23, 25, 26, 27, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 46, 49, 50, 51, 54, 55, 56, 59, 60, 61, 64 and 65, and / or encoding a ... Proteins in which at least one amino acid sequence of the amino acid sequences shown in NO:2, 5, 8, 11, 14, 17, 20, 24, 28, 29, 30, 41, 42, 43, 44, 45, 47, 48, 52, 53, 57, 58, 62, and 63 has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater amino acid sequence identity.

[0241] In PCR methods, oligonucleotide primers can be designed for PCR reactions to amplify corresponding DNA sequences from cDNA or genomic DNA extracted from any target plant. Methods for designing PCR primers and PCR cloning are generally known in the art and disclosed in Sambrook et al. (1989) *Molecular Cloning: A Laboratory Manual* (2nd ed., Cold Spring Harbor Laboratory Press, Plainview, New York). See also Innis et al., ed. (1990) *PCR Protocols: A Guide to Methods and Applications* (Academic Press, New York); Innis and Gelfand, ed. (1995) *PCR Strategies* (Academic Press, New York); and Innis and Gelfand, ed. (1999) *PCR Methods Manual* (Academic Press, New York). Known PCR methods include, but are not limited to, methods using paired primers, nested primers, single-specific primers, degenerate primers, gene-specific primers, vector-specific primers, partially mismatched primers, etc.

[0242] In hybridization techniques, all or part of a known polynucleotide is used as a probe, which selectively hybridizes with other corresponding polynucleotides present in a group of clonal genomic DNA fragments or cDNA fragments (i.e., a genome or cDNA library) from a selected organism. Hybridization probes can be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and can utilize detectable groups such as… 32 P or any other detectable marker. Therefore, for example, probes for hybridization can be prepared by synthesizing oligonucleotides based on the polynucleotide markers of this invention. Methods for preparing probes for hybridization and for constructing cDNA and genomic libraries are generally known in the art and are disclosed in Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2nd edition, Cold Spring Harbor Laboratory Press, Plainview, New York).

[0243] For example, the complete polynucleotides or one or more portions thereof disclosed herein can be used as probes capable of specifically hybridizing with the corresponding polynucleotides and messenger RNA. To achieve specific hybridization under various conditions, such probes comprise a unique sequence within the target gene or cDNA sequence, and preferably are at least about 10 nucleotides in length, most preferably at least about 20 nucleotides in length. Such probes can be used to amplify the corresponding polynucleotide of a specific target gene from selected plants via PCR. This technique can be used to isolate additional coding sequences from desired plants or as a diagnostic assay to determine the presence of coding sequences in plants. Hybridization techniques include screening of plated DNA libraries (plasters or colonies; see, for example, Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Plainview, New York). Detailed instructions on nucleic acid hybridization can be found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, New York); and Ausubel et al., eds., Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience, New York).

[0244] It should be understood that the R protein-coding sequences of the present invention comprise polynucleotide molecules containing nucleotide sequences substantially identical to any one or more of the nucleotide sequences in SEQ ID NO: 1 and 3. The term “substantially identical” is used herein to mean that the first amino acid or nucleotide sequence contains a sufficient or minimum number of identical or equivalent (having similar side chains) amino acid residues or residues relative to the second amino acid or nucleotide sequence, such that the first and second amino acid or nucleotide sequences have common structural domains and / or common functional activities. For example, amino acid or nucleotide sequences containing at least about 45%, 55%, or 65% identical, preferably 75% identical, more preferably 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical common structural domains are defined herein as substantially identical.

[0245] To determine the percentage of identity between two amino acid sequences or two nucleic acids, the sequences are aligned for optimal comparison. The percentage of identity between two sequences is a function of the number of common positions shared by the sequences (i.e., percentage of identity = number of common positions / total number of positions (e.g., overlapping positions) × 100). In one embodiment, the two sequences are of the same length. The percentage of identity between two sequences can be determined using techniques similar to those described below, with or without gaps. When calculating percentage identity, exact matches are typically counted.

[0246] Mathematical algorithms can be used to determine the percentage of identity between two sequences. A preferred, non-limiting example of a mathematical algorithm for comparing two sequences is the improved version of the algorithm in Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 90:5873-5877, as described in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 87:2264. This algorithm is incorporated into the NBLAST and XBLAST programs in Altschul et al. (1990) J. Mol. Biol. 215:403. A BLAST nucleotide search can be performed using the NBLAST program with a score of 100 and a word length of 12 to obtain nucleotide sequences homologous to the polynucleotide molecule of the present invention. A BLAST protein search can be performed using the XBLAST program with a score of 50 and a word length of 3 to obtain amino acid sequences homologous to the protein molecule of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used as described by Altschul et al. (1997) Nucleic Acids Res. 25:3389. Alternatively, PSI-Blast can be used to perform iterative searches to detect long-distance relationships between molecules. See Altschul et al. (1997) (ibid.). When using the BLAST, Gapped BLAST, and PSI-Blast procedures, the default parameters of the respective procedures (e.g., XBLAST and NBLAST) can be used. BLAST, Gapped BLAST, and PSI-Blast, XBLAST, and NBLAST are available at ncbi.nlm.nih.gov on the World Wide Web. Another preferred, non-limiting example of a mathematical algorithm for comparing sequences is the algorithm of Myers and Miller (1988) CABIOS 4:11-17. This algorithm was incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When comparing amino acid sequences using the ALIGN program, the PAM120 weighted residue table can be used, with a gap length penalty of 12 and a gap penalty of 4. Manual alignment can also be performed via inspection.

[0247] Unless otherwise stated, the sequence identity / similarity values ​​provided herein refer to the full-length sequences of this invention and to the multiple alignments performed using the algorithm Clustal W (Nucleic Acid Research, 22(22):4673-4680, 1994) with the program AlignX included in the software package Vector NTI Suite version 7.0 (InforMax, Inc., Bethesda, MD, USA) using default parameters; or any equivalent program thereof. “Equivalent program” means any sequence comparison program that, for any two sequences, produces an alignment with the same nucleotide or amino acid residue match and the same percentage of sequence identity when compared to the corresponding alignment generated by CLUSTALW (version 1.83) using default parameters (available at the European Institute of Bioinformatics website ebi.ac.uk / Tools / clustalw / index on the World Wide Web).

[0248] The use of the term "polynucleotide" is not intended to limit the invention to polynucleotides containing DNA. Those skilled in the art will recognize that polynucleotides can include ribonucleotides and combinations of ribonucleotides and deoxyribonucleotides. Such deoxyribonucleotides and ribonucleotides include naturally occurring molecules and synthetic analogs. The polynucleotides of the present invention also include all forms of sequences, including but not limited to single-stranded, double-stranded, hairpin, stem-loop, and other similar structures.

[0249] Heteronucleotides or polynucleotide constructs containing the R protein-coding region can be provided in expression cassettes for expression in target plants or other organisms or non-human host cells. The cassette will include 5' and 3' regulatory sequences operatively linked to the R protein-coding region. "Operably linked" is intended to indicate a functional connection between two or more elements. For example, an operative link between a target polynucleotide or gene and a regulatory sequence (i.e., a promoter) is a functional link that allows expression of the target polynucleotide. The operatively linked elements can be adjacent or non-adjacent. When used to refer to the connection of two protein-coding regions, operative linking is intended to mean that the coding regions are in the same reading frame. The cassette may additionally contain at least one additional gene to be co-transformed into an organism. Alternatively, one or more additional genes may be provided on multiple expression cassettes. Such expression cassettes have multiple restriction sites and / or recombination sites for insertion into the R protein-coding region, placing it under the transcriptional regulation of the regulatory region. The expression cassette may additionally contain a selectable marker gene.

[0250] The expression cassette will include, in a 5'-3' transcriptional orientation, transcription and translation initiation regions (i.e., promoters), the R protein-coding region of this invention, and transcription and translation termination regions (i.e., termination regions) that are functional in plants or other organisms or non-human host cells. The regulatory regions (i.e., promoters, transcriptional regulatory regions, and translation termination regions) and / or the R protein-coding regions of this invention may be native to the host cell or similar to each other. Alternatively, the regulatory regions and / or the R protein-coding regions of this invention may be heterologous to the host cell or to each other.

[0251] As used herein, “heterologous” for a nucleic acid molecule, polynucleotide, nucleotide sequence, or polynucleotide construct means a nucleic acid molecule, polynucleotide, nucleotide sequence, or polynucleotide construct derived from a foreign species, or, if from the same species, modified from its natural form in terms of composition and / or genomic loci through intentional human intervention. For example, a promoter operatively linked to a heterologous polynucleotide is from a species different from the species from which the polynucleotide originates, or, if from the same / similar species, one or both are substantially modified from their original form and / or genomic loci, or the promoter is not a natural promoter for the operatively linked polynucleotide. As used herein, a chimeric gene contains a coding sequence operatively linked to a transcription start region, which is heterologous to the coding sequence.

[0252] As used herein, “natural gene” is intended to mean a gene that is naturally present at its natural or native location in the plant genome. Such a natural gene has not been genetically engineered or otherwise modified at its nucleotide sequence and / or location in the plant genome by human intervention, nor has it been introduced into the plant genome by artificial methods such as, for example, plant transformation.

[0253] As used herein, "non-natural gene" is intended to refer to a gene that has been introduced into a plant by artificial means and / or contains a nucleotide sequence that is not naturally present in a plant. Non-natural genes include, for example, genes introduced into a plant through plant transformation (e.g., R genes). Additionally, a modified gene is a non-natural gene when, for example, a natural gene in a plant genome is modified by genome editing methods to contain a nucleotide sequence that differs from (i.e., is not identical to) the nucleotide sequence of the natural gene.

[0254] This invention provides a host cell comprising at least the nucleic acid molecule, expression cassette, and vector of this invention. In a preferred embodiment of the invention, the host cell is a plant cell. In other embodiments, the host cell is selected from bacterial, fungal, and animal cells. In some embodiments, the host cell is a non-human animal cell. However, in some other embodiments, the host cell is a human cell cultured in vitro.

[0255] Although using a heterologous promoter to express the R protein may be optimal, the natural promoter of the corresponding R gene can be used.

[0256] The termination region can be natural for the transcription initiation region, natural for the target R protein coding region that can be operatively linked, natural for the plant host, or derived from another source (i.e., foreign or heterologous for the promoter, target R protein, and / or plant host) or any combination thereof. Convenient termination regions can be obtained from Ti plasmids of Agrobacterium tumefaciens, such as octopaline synthase and carmine synthase termination regions. See also Guerineau et al. (1991) Mol. Gen. Genet. 262: 141-144; Proudfoot (1991) Cell 64: 671-674; Sanfacon et al. (1991) Genes Dev. 5: 141-149; Mogen et al. (1990) Plant Cell 2: 1261-1272; Munroe et al. (1990) Gene 91: 151-158; Ballas et al. (1989) Nucleic Acids Res. 17: 7891-7903; and Joshi et al. (1987) Nucleic Acids Res. 15: 9627-9639.

[0257] When appropriate, polynucleotides can be optimized to increase expression in transformed plants. That is, polynucleotides can be synthesized using plant-preferred codons to improve expression. For a discussion of host-preferred codon selection, see, for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11. Methods for synthesizing plant-preferred genes are available in the art. See, for example, U.S. Patents 5,380,831 and 5,436,391, and Murray et al. (1989) Nucleic Acids Res. 17:477-498, which are incorporated herein by reference.

[0258] Other known sequence modifications can enhance gene expression in the host cell. These include the removal of sequences encoding pseudopolyadenylation signals, exon-intron splicing site signals, transposon-like repeat sequences, and other well-characterized sequences that may be detrimental to gene expression. The GC content of the sequence can be adjusted to the average level of a given host cell, calculated by referencing known genes expressed in the host cell. Where possible, sequences are modified to avoid predicted hairpin secondary mRNA structures.

[0259] The expression box may additionally contain a 5' leader sequence. Such leader sequences can enhance the translation. Translational leader sequences are known in the art and include: parvovirus leader sequences, such as the EMCV leader sequence (5' untranslated region of encephalomyocarditis) (Elroy Stein et al. (1989) Proc. Natl. Acad. Sci. USA 86:6126-6130); potato γ-virus leader sequences, such as the TEV leader sequence (tobacco etch virus) (Gallie et al. (1995) Gene 165(2):233-238), the MDMV leader sequence (maize dwarf mosaic virus) (Virology 154:9-20), and human immunoglobulin heavy chain binding protein (BiP) (Macejak et al. (1991) Nature 353:90-94); and untranslated leader sequences of the capsid protein mRNA from alfalfa mosaic virus (AMV RNA 4) (Jobling et al. (1987) Nature). 325:622-625); Tobacco mosaic virus leader sequence (TMV) (Gallie et al. (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York), pp. 237-256); and maize chlorotic mottle virus leader sequence (MCMV) (Lommel et al. (1991) Virology 81:382-385). See also Della Cioppa et al. (1987) Plant Physiol. 84:965-968.

[0260] In preparing expression cassettes, various DNA fragments can be manipulated to provide a DNA sequence in the appropriate orientation and, where appropriate, a suitable reading frame. For this purpose, adaptors or linkers can be used to ligate the DNA fragments, or other manipulations can be involved to provide convenient restriction sites, remove redundant DNA, remove restriction sites, etc. For this purpose, in vitro mutagenesis, primer repair, restriction, annealing, and re-substitution (e.g., conversion and transversion) can be employed.

[0261] Many promoters are available for carrying out this invention. Promoters can be selected based on the desired results. Nucleic acids can be combined with constitutive, tissue-biased, or other promoters for expression in plants. Such constitutive promoters include, for example, the core CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroy et al. (1990) Plant Cell 2:163-171); ubiquitin (Christensen et al. (1989) Plant Mol. Biol. 12:619-632 and Christensen et al. (1992) Plant Mol. Biol. 18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet. 81:581-588); MAS (Velten et al. (1984) EMBO J. 3:2723-2730); ALS promoter (US Patent No. 5,659,026), etc. Other constitutive promoters include, for example, U.S. Patents 5,608,149, 5,608,144, 5,604,121, 5,569,597, 5,466,785, 5,399,680, 5,268,463, 5,608,142, and 6,177,611.

[0262] Tissue-biased promoters can be used to target and enhance the expression of R protein coding sequences in specific plant tissues. These tissue-biased promoters include, but are not limited to, leaf-biased promoters, root-biased promoters, seed-biased promoters, and stem-biased promoters. Tissue-biased promoters include Yamamoto et al. (1997) Plant J. 12(2): 255-265; Kawamata et al. (1997) Plant Cell Physiol. 38(7): 792-803; Hansen et al. (1997) Mol. GenGenet. 254(3): 337-343; Russell et al. (1997) Transgenic Res. 6(2): 157-168; Rinehart et al. (1996) Plant Physiol. 112(3): 1331-1341; Van Camp et al. (1996) Plant Physiol. 112(2): 525-535; Canevascini et al. (1996) Plant Physiol. 112(2): 513-524; Yamamoto et al. (1994) Plant Cell Physiol. 35(5):773-778; Lam (1994) Results Probl. Cell Differ. 20:181-196; Orozco et al. (1993) Plant Mol Biol. 23(6):1129-1138; Matsuoka et al. (1993) Proc Natl. Acad. Sci. USA 90(20):9586-9590; and Guevara-Garcia et al. (1993) Plant J. 4(3):495-505. If needed, such promoters can be modified for weak expression.

[0263] Generally, gene expression from inducible promoters, particularly pathogen-inducible promoters, is beneficial. Such promoters include those derived from pathogenesis-associated proteins (PR proteins) that are induced upon pathogen infection, such as PR proteins, SAR proteins, β-1,3-glucanase, chitosanase, etc. See, for example, Redolfi et al. (1983) Neth. J. Plant Pathol. 89:245-254; Uknes et al. (1992) Plant Cell 4:645-656; and Van Loon (1985) Plant Mol. Virol. 4:111-116. See also WO 99 / 43819, incorporated herein by reference.

[0264] Of interest are promoters expressed locally at or near the site of pathogen infection. See, for example, Marineau et al. (1987) Plant Mol. Biol. 9:335-342; Matton et al. (1989) Molecular Plant-Microbe Interactions 2:325-331; Somsisch et al. (1986) Proc. Natl. Acad. Sci. USA 83:2427-2430; Somsisch et al. (1988) Mol. Gen. Genet. 2:93-98; and Yang (1996) Proc. Natl. Acad. Sci. USA 93:14972-14977. See also Chen et al. (1996) Plant J. 10: 955-966; Zhang et al. (1994) Proc. Natl. Acad. Sci. USA 91: 2507-2511; Warner et al. (1993) Plant J. 3: 191-201; Siebertz et al. (1989) Plant Cell 1: 961-968; U.S. Patent No. 5,750,386 (nematode-induced promoter); and the references cited therein. Of particular interest is the inducible promoter of the maize PRms gene, whose expression is induced by the pathogen Fusarium moniliforme (see, for example, Cordero et al. (1992) Physiol. Mol. Plant Path. 41: 189-200).

[0265] In addition, wound-inducible promoters can be used in the heteropolynucleotides of this invention when pathogens enter plants through wounds or insect damage. Such wound-inducing promoters include the potato glycoprotein inhibitor (pin II) gene (Ryan (1990) Ann. Rev. Phytopath. 28:425-449; Duan et al. (1996) Nature Biotechnology 14:494-498); wun1 and wun2, US Patent No. 5,428,148; win1 and win2 (Stanford et al. (1989) Mol. Gen. Genet. 215:200-208); systemin (McGurl et al. (1992) Science 225:1570-1573); WIP1 (Rohmeier et al. (1993) Plant Mol. Biol. 22:783-792; Eckelkamp et al. (1993) FEBS Letters 323:73-76); MPI gene (Corderok et al. (1994) Plant J.6(2):141-150, etc., are incorporated into this paper by reference.

[0266] Chemically regulated promoters can be used to regulate gene expression in plants through the application of exogenous chemical regulators. Depending on the purpose, the promoter can be a chemically induced promoter, in which the application of the chemical induces gene expression, or a chemically repressive promoter, in which the application of the chemical represses gene expression. Chemically induced promoters are known in the art and include, but are not limited to, the maize In2-2 promoter (which is activated by a benzenesulfonamide herbicide safener), the maize GST promoter (which is activated by a hydrophobic electrophilic compound used as a pre-germination herbicide), and the tobacco PR-1a promoter (which is activated by salicylic acid). Other promoters of interest that regulate chemical substances include steroid-responsive promoters (see, for example, Schena et al. (1991) Proc. Natl. Acad. Sci. USA 88:10421-10425 and McNellis et al. (1998) Plant J. 14(2):247-257) including glucocorticoid-inducible promoters and tetracycline-inducible and tetracycline-repressive promoters (see, for example, Gatz et al. (1991) Mol. Gen. Genet. 227:229-237 and U.S. Patent Nos. 5,814,618 and 5,789,156), which are incorporated herein by reference.

[0267] The expression cassette may also contain selection marker genes for selecting transformed cells. Selection marker genes are used to select transformed cells or tissues. Marker genes include genes encoding antibiotic resistance, such as genes encoding neomycin phosphotransferase II (NEO) and hygromycin phosphotransferase (HPT), and genes conferring resistance to herbicides such as glufosinate, bromobenzonitrile, imidazolinone, and 2,4-dichlorophenoxyacetate (2,4-d)). Other selection markers include phenotypic markers such as β-galactosidase and fluorescent proteins, such as green fluorescent protein (GFP) (Su et al. (2004) Biotechnol Bioeng 85:610-9 and Fetter et al. (2004) Plant Cell 16:215-28), cyan fluorescent protein (CYP) (Bolte et al. (2004) J. Cell Science 117:943-54 and Kato et al. (2002) Plant Physiol 129:913-42), and yellow fluorescent protein (PhiYFP). TMFrom Evrogen, see Bolte et al. (2004) J. Cell Science 117:943-54. For other selection markers, see Yarranton (1992) Curr. Opin. Biotech. 3:506-511; Christopherson et al. (1992) Proc. Natl. Acad. Sci. USA 89:6314-6318; Yao et al. (1992) Cell 71:63-72; Reznikoff (1992) Mol. Microbiol. 6:2419-2422; Barkley et al. (1980) in The Operon, pp. 177-220; Hu et al. (1987) Cell 48:555-566; Brown et al. (1987) Cell 49:603-612; Figge et al. (1988) Cell 52:713-722; Deuschle et al. (1989) Proc. Natl. Acad. Aci. USA 86:5400-5404; Fuerst et al. (1989) Proc. Natl. Acad. Sci. USA 86: 2549-2553; Deuschle et al. (1990) Science 248: 480-483; Gossen (1993) Ph. D. Thesis, University of Heidelberg; Reines et al. (1993) Proc.Natl.Acad.Sci.USA90:1917-1921; Labow et al. (1990) Mol.Cell.Biol.10:3343-3356; Zambretti et al. (1992) Proc.Natl.Acad.Sci.USA 89:3952-3956; Baim et al. (1991) Proc.Natl.Acad.Sci.USA 88:5072-5076; Wyborski et al. (1991) Nucleic Acids Res. 19: 4647-4653; Hillenand-Wissman (1989) Topics Mol. Struc. Biol. 10: 143-162; Degenkolb et al. (1991) Antimicrob. Agents Chemother.35:1591-1595; Kleinschnidt et al. (1988) Biochemistry 27:1094-1104; Bonin (1993) Ph.D.Thesis, University of Heidelberg; Gossen et al. (1992) Proc.Natl.Acad.Sci.USA 89:5547-5551; Oliva et al. (1992) Antimicrob. Agents Chemother. 36:913-919; Hlavka et al. (1985) Handbook of Experimental Pharmacology, Vol. 78 (Springer-Verlag, Berlin); Gill et al. (1988) Nature 334:721-724. Such publications are incorporated herein by reference.

[0268] The above list of selectable marker genes is not intended to be limiting. Any selectable marker gene may be used in this invention.

[0269] Many plant transformation vectors and methods for transforming plants are available. See, for example, An, G. et al. (1986) Plant Pysiol., 81:301-305; Fry, J. et al. (1987) Plant Cell Rep. 6:321-325; Block, M. (1988) Theor. Appl Genet. 76:767-774; Hinchee, et al. (1990) Stadler. Genet. Symp. 203-212. 203-212; Cousins, et al. (1991) Aust. J. Plant Physiol. 18:481-494; Chee, PP and Slightom, JL (1992) Gene. 118:255-260; Christou, et al. (1992) Trends. Biotechnol. 10:239-246; D'Halluin, et al. (1992) Bio / Technol. 10:309-314; Dhir, et al. (1992) Plant Physiol. 99:81-88; Casas, et al. (1993) Proc. Nat. Acad Sci. USA 90:11212-11216; Christou, P. (1993) In Vitro Cell. Dev. Biol. - Plant; 29P:119-124; Davies, et al. (1993) Plant Cell Rep. 12:180-183; Dong, J.A. and Mchughen, A. (1993) Plant Sci. 91:139-148; Franklin, CI. and Trieu, TN (1993) Plant. Physiol. 102:167; Golovkin, et al. (1993) Plant Sci. 90:41-52; Guo Chin, Sci. Bull. 38:2072-2078; Asano, et al. (1994) Plant Cell Rep. 13; Ayeres NM and Park, WD (1994) Crit. Rev. Plant. Sci. 13: 219-239; Barcelo, et al. (1994) Plant. J. 5: 583-592; Becker, et al. (1994) Plant. J. 5: 299-307; Borkowska, et al. (1994) Acta. Physiol Plant. 16: 225-230; Christou, P. (1994) Agro. Food. Ind. Hi Tech. 5: 17-27; Eapen, et al. (1994) Plant Cell Rep.13:582-586; Hartman, et al. (1994) Bio-Technology 12:919923; Ritala, et al. (1994) Plant. Mol. Biol. 24:317-325; and Wan, YC and Lemaux, PG (1994) Plant Physiol. 104:3748.

[0270] The method of this invention relates to introducing heteropolynucleotides or polynucleotide constructs into plants. "Introduction" is defined as presenting the heteropolynucleotide or polynucleotide construct to a plant in a manner that allows the construct to enter the interior of plant cells. The method of this invention does not depend on a specific method for introducing the heteropolynucleotide or polynucleotide construct into the plant, as long as the heteropolynucleotide or polynucleotide construct can enter the interior of at least one cell of the plant. Methods for introducing heteropolynucleotides or polynucleotide constructs into plants are known in the art, including but not limited to stable transformation methods, transient transformation methods, and virus-mediated methods.

[0271] "Stable transformation" refers to the integration of a heterologous polynucleotide or polynucleotide construct introduced into a plant into the plant's genome and its ability to be inherited by its offspring. "Transient transformation" refers to the introduction of a heterologous polynucleotide or polynucleotide construct into a plant that does not integrate into the plant's genome. It should be recognized that both stable and transient transformation methods involve the introduction of one or more nucleic acid molecules (e.g., DNA), particularly one or more recombinant nucleic acid molecules (e.g., recombinant DNA), into a plant, plant cell, or other host cell or organism.

[0272] To transform plants and plant cells, the nucleotide sequences of the present invention are inserted into any vector known in the art suitable for expressing nucleotide sequences in plants or plant cells using standard techniques. The choice of vector depends on the preferred transformation technique and the target plant species to be transformed.

[0273] Methods for constructing plant expression cassettes and introducing foreign nucleic acids into plants are generally known in the art and have been previously described. For example, tumor-inducing (Ti) plasmid vectors can be used to introduce exogenous DNA into plants. Other methods for exogenous DNA delivery involve PEG-mediated protoplast transformation, electroporation, microinjection of whiskers, and bioprojectiles or particle bombardment for direct DNA uptake. Such methods are known in the art (as per U.S. Patent No. 5,405,765 to Vasil et al.; Bilang et al. (1991) Gene 100:247-250; Scheid et al. (1991) Mol. Gen. Genet., 228:104-112; Guerche et al. (1987) Plant Science 52:111-116; Neuhause et al. (1987) Theor. Appl Genet. 75:30-36; Klein et al. (1987) Nature 327:70-73; Howell et al. (1980) Science 208:1265; Horsch et al. (1985) Science 227:1229-1231; DeBlock et al. (1989) Plant Physiology 91:694-701; Methods for Plant) Molecular Biology (Weissbach and Weissbach, eds.) Academic Press, Inc. (1988) and Methods in Plant Molecular Biology (Schuler and Zielinski, eds.) Academic Press, Inc. (1989). The transformation method depends on the plant cells to be transformed, the stability of the vector used, the expression level of the gene product, and other parameters.

[0274] Other suitable methods for introducing nucleotide sequences into plant cells and subsequently inserting them into the plant genome include microinjection as described by Crossway et al. (1986) Biotechniques 4:320-334; electroporation as described by Riggs et al. (1986) Proc. Natl. Acad. Sci. USA 83:5602-5606; Agrobacterium-mediated transformation as described by Townsend et al., U.S. Patent No. 5,563,055, and Zhao et al., U.S. Patent No. 5,981,840; and EMBO as described by Paszkowski et al. (1984). Direct gene transfer as described in J.3:2717-2722, and ballistic particle acceleration as described in, for example, Sanford et al., U.S. Patent No. 4,945,050; Tomes et al., U.S. Patent No. 5,879,918; Tomes et al., U.S. Patent No. 5,886,244; Bidney et al., U.S. Patent No. 5,932,782; Tomes et al. (1995) “Direct DNA Transfer into Intact Plant Cells via Microprojectile Bombardment,” in Plant Cell, Tissue, and Organ Culture: Fundamental Methods, ed. Gamborg and Phillips (Springer-Verlag, Berlin); McCabe et al. (1988) Biotechnology 6:923-926; and Lec1 transformation (WO 00 / 28058). See also Weissinger et al. (1988) Ann. Rev. Genet. 22:421-477; Sanford et al. (1987) Particulate Science and Technology 5:27-37 (onion); Christou et al. (1988) Plant Physiol. 87:671-674 (soybean); McCabe et al. (1988) Bio / Technology 6:923-926 (soybean); Finer and McMullen (1991) In Vitro Cell Dev. Biol. 27P:175-182 (soybean); Singh et al. (1998) Theor. Appl. Genet. 96:319-324 (soybean); Datta et al. (1990) Biotechnology 8:736-740 (rice); Klein et al. (1988) Proc. Natl.Acad. Sci. USA 85:4305-4309 (maize); Klein et al. (1988) Biotechnology 6:559-563 (maize); Tomes, U.S. Patent No. 5,240,855; Buising et al., U.S. Patent Nos. 5,322,783 and 5,324,646; Tomes et al. (1995) “Direct DNA Transfer into Intact Plant Cells via Microprojectile Bombardment,” in Plant Cell, Tissue, and Organ Culture: Fundamental Methods, ed. Gamborg (Springer-Verlag, Berlin) (maize); Klein et al. (1988) Plant Physiol. 91:440-444 (maize); Fromm et al. (1990) Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren et al. (1984) Nature (London) 311:763-764; Bowen et al., US Patent No. 5,736,369 (Cereals); Bytebier et al. (1987) Proc. Natl. Acad. Sci. USA 84:5345-5349 (Liliaceae); De Wet et al. (1985) in The Experimental Manipulation of Ovule Tissues, ed. Chapman et al. (Longman, New York), pp. 197-209 (Pollen); Kaeppler et al. (1990) Plant Cell Reports 9:415-418 and Kaeppler et al. (1992) Theor. Appl. Genet. 84:560-566 (Whisker-mediated transformation); D'Halluin et al. (1992) Plant Cell 4:1495-1505 (electroporation); Li et al. (1993) Plant Cell Reports 12:250-255 and Christou and Ford (1995) Annals of Botany 75:407-413 (rice); Osjoda et al. (1996) Nature Biotechnology 14:745-750 (maize, via Agrobacterium tumefaciens)); all of these are incorporated herein by reference.

[0275] The polynucleotides of the present invention can be introduced into plants by contacting the plants with viruses or viral nucleic acids. Typically, such methods involve incorporating the heterologous polynucleotides or polynucleotide constructs of the present invention into viral DNA or RNA molecules. Additionally, it should be recognized that the promoters of the present invention also include promoters for transcription by viral RNA polymerase. Methods for introducing polynucleotide constructs into plants and expressing proteins encoded therein (including viral DNA or RNA molecules) are known in the art. See, for example, U.S. Patents 5,889,191, 5,889,190, 5,866,785, 5,589,367, and 5,316,931; which are incorporated herein by reference.

[0276] If necessary, modified viruses or modified viral nucleic acids can be prepared in the formulation. Such formulations are prepared in known ways (see, for example, US 3,060,084, EP-A 707 445 (for liquid concentrates), Browning, “Agglomeration”, Chemical Engineering, Dec. 4, 1967, 147-48, Perry's Chemical Engineer's Handbook, 4th edition, McGraw-Hill, New York, 1963, pp. 8-57, etc.; WO 91 / 13546, US 4,172,714, US 4,144,050, US 3,920,442, US 5,180,587, US 5,232,701, US 5,208,030, GB 2,095,558, US 3,299,566, Klingman, Weed Control as a Science, John Wiley and Sons, Inc., New York). York, 1961, Hance et al., Weed Control Handbook, 8th edition, Blackwell Scientific Publications, Oxford, 1989; and Mollet, H., Grubemann, A., Formulation technology, Wiley VCH Verlag GmbH, Weinheim (Germany), 2001, 2. D.A. Knowles, Chemistry and Technology of Agrochemical Formulations, Kluwer Academic Publishers, Dordrecht, 1998 (ISBN 0-7514-0443-8), for example, by extending active compounds with adjuvants suitable for formulating agrochemicals (such as solvents and / or carriers, emulsifiers, surfactants and dispersants, preservatives, defoamers, antifreeze agents, and optionally, colorants and / or binders and / or gelling agents for seed treatment formulations).

[0277] In specific embodiments, the polynucleotides, polynucleotide constructs, and expression cassettes of the present invention can be provided to plants using various transient transformation methods known in the art. Such methods include, for example, microinjection or particle bombardment. See, for example, Crossway et al. (1986) Mol Gen. Genet. 202:179-185; Nomura et al. (1986) Plant Sci. 44:53-58; Hepler et al. (1994) PNAS Sci. 91:2176-2180 and Hush et al. (1994) J. Cell Science 107:775-784, all of which are incorporated herein by reference. Alternatively, polynucleotides can be transiently transformed into plants using techniques known in the art. Such techniques include viral vector systems and transient expression mediated by *Agrobacterium tumefaciens* as described elsewhere herein.

[0278] The transformed cells can be grown into plants using conventional methods. See, for example, McCormick et al. (1986) Plant Cell Reports 5:81-84. These plants can then be grown and pollinated with the same or different transformed lines, and the resulting hybrids with constitutive expression of the desired phenotypic trait can be identified. Two or more generations can be grown to ensure stable maintenance and inheritance of the expression of the desired phenotypic trait, and then the seeds can be harvested to ensure the achievement of the desired phenotypic trait expression. Thus, the present invention provides transformed seeds (also known as “transgenic seeds”) having the heteropolynucleotides or polynucleotide constructs of the present invention (e.g., expression cassettes of the present invention stably integrated into their genome).

[0279] Any method known in the art for modifying DNA in a plant genome can be used to modify the nucleotide sequence of a plant genome in situ, for example, to generate or insert resistance genes or even replace or modify its endogenous resistance genes or alleles. Such methods include, but are not limited to, genome editing (or gene editing) techniques, such as those involving directed mutagenesis, homologous recombination, and mutation breeding. Targeted mutagenesis or similar techniques are disclosed in U.S. Patents Nos. 5,565,350, 5,731,181, 5,756,325, 5,760,012, 5,795,972, 5,871,984, and 8,106,259; all of which are incorporated herein by reference in their entirety. Methods involving gene modification or substitution, including homologous recombination, may involve inducing double breaks in DNA using zinc finger nucleases (ZFNs), TAL (transcription activator-like) effector nucleases (TALENs), regularly clustered short palindromic repeats / CRISPR-associated nucleases (CRISPR / Cas nucleases), or homing endonucleases (which have been engineered to produce double-strand breaks at specific recognition sequences in the genomes of plants, other organisms, or host cells). See, for example, Durai et al., (2005) Nucleic Acids Res 33:5978-90; Mani et al., (2005) Biochem Biophys Res Comm 335:447-57; U.S. Patents 7,163,824, 7,001,768 and 6,453,242; Arnould et al., (2006) J Mol Biol 355:443-58; Ashworth et al., (2006) Nature 441:656-9; Doyon et al., (2006) J Am Chem Soc 128:2477-84; Rosen et al., (2006) Nucleic Acids Res 34:4791-800; and Smith et al., (2006) Nucleic Acids Res 34:e149; U.S. Patent Application Publication No. 2009 / 0133152 and U.S. Patent Application Publication No. 2007 / 0117128; all of which are incorporated herein by reference in their entirety.

[0280] Unless otherwise stated or obvious from the context of use, the term "gene substitution" is intended to mean the substitution of any part of a first polynucleotide or nucleic acid molecule (e.g., a chromosome) involving homologous recombination with a second polynucleotide or nucleic acid molecule using gene editing techniques as described elsewhere herein, wherein at least a portion of the nucleotide sequence of the first polynucleotide or nucleic acid molecule is replaced by the second polynucleotide or nucleic acid molecule. It should be recognized that such gene substitution can result in additions, deletions, and / or modifications in the nucleotide sequence of the first polynucleotide or nucleic acid molecule, and can involve the substitution of one or more complete genes, the substitution of one or more portions of a gene, or the substitution of non-genetic sequences in the first polynucleotide or nucleic acid molecule.

[0281] TAL effector nucleases (TALENs) can be used to generate double-strand breaks at specific recognition sequences in the plant genome for gene modification or substitution via homologous recombination. TAL effector nucleases are a class of sequence-specific nucleases that can be used to generate double-strand breaks at specific target sequences in the genomes of plants or other organisms. TAL effector nucleases are generated by fusing natural or engineered transcription activator-like (TAL) effectors or their functional portions with the catalytic domain of an endonuclease, such as FokI. The unique modular DNA-binding domain of TAL effectors allows for the design of proteins with the potential specificity for recognizing any given DNA. Therefore, the DNA-binding domain of TAL effector nucleases can be engineered to recognize specific DNA target sites, thereby generating double-strand breaks at desired target sequences. See also WO 2010 / 079430; Morbitzer et al. (2010) PNAS 10.1073 / pnas.1013133107; Scholze & Boch (2010) Virulence 1:428-432; Christian et al. Genetics (2010) 186:757-761; Li et al. (2010) Nuc. Acids Res. (2010) doi:10.1093 / nar / gkq704 and Miller et al. (2011) Nature Biotechnology 29:143-148; all of which are incorporated herein by reference.

[0282] The CRISPR / Cas nuclease system can also be used to generate double-strand breaks at specific recognition sequences in plant genomes for gene modification or substitution via homologous recombination. CRISPR / Cas nucleases are RNA-guided (simple guide RNA, sgRNA) DNA endonuclease systems that perform sequence-specific double-strand breaks in DNA segments homologous to the designed RNA. Sequence specificity can be designed (Cho SW et al., Nat. Biotechnol. 31:230-232, 2013; Cong L. et al., Science 339:819-823, 2013; Mali P. et al., Science 339:823-826, 2013; Feng Z. et al., Cell Research:1-4, 2013).

[0283] In addition, ZFNs can be used to generate double-strand breaks at specific recognition sequences in plant genomes for gene modification or substitution via homologous recombination. Zinc finger nucleases (ZFNs) are fusion proteins comprising a portion of the FokI restriction endonuclease protein responsible for DNA cleavage and zinc finger proteins that recognize specific, designed genomic sequences and cleave double-stranded DNA at these sequences, thereby generating free DNA ends (Urnov FD et al., Nat RevGenet. 11:636-46, 2010; Carroll D., Genetics. 188:773-82, 2011).

[0284] Disrupting DNA using site-specific nucleases (such as those described above, for example) can increase the rate of homologous recombination in the disrupted regions. Therefore, the coupling of such effectors with nucleases, as described above, enables the generation of targeted changes in the genome, including additions, deletions, and other modifications.

[0285] The nucleic acid molecules, expression cassettes, vectors, and heteropolynucleotides of the present invention can be used for transformation and / or genome editing of any plant species (including but not limited to monocots and dicots).

[0286] As used herein, the term "plant" includes seeds, plant cells, plant protoplasts, plant cell tissue cultures from which regenerate plants, plant callus, plant masses, and whole plant cells in plants or plant parts such as embryos, pollen, ovules, seeds, tubers, propagules, leaves, flowers, branches, fruits, roots, root tips, anthers, etc. Progeny, variants, and mutants of regenerated plants are also included within the scope of this invention, provided that these parts contain introduced polynucleotides. As used herein, unless otherwise expressly stated or apparent from the context of use, "progeny" and "progeny plant" include any subsequent generations of a plant, whether produced by sexual and / or asexual reproduction.

[0287] As used herein, the terms “transgenic plant” and “transformed plant” are equivalent terms to “plant” as described above, wherein a plant comprises a heterologous nucleic acid molecule, heterologous polynucleotide, or heterologous polynucleotide construct introduced into the plant by any stable and transient transformation method disclosed elsewhere herein or otherwise known in the art. Such transgenic and transformed plants also refer to, for example, the plant in which the heterologous nucleic acid molecule, heterologous polynucleotide, or heterologous polynucleotide construct was first introduced, and any of its progeny plants comprising said heterologous nucleic acid molecule, heterologous polynucleotide, or heterologous polynucleotide construct.

[0288] In some embodiments of the invention, the method involves planting seedlings and / or tubers, then growing such seedlings and tubers to produce plants derived therefrom, and optionally harvesting one or more portions of the plant. As used herein, “seedling” refers to an incompletely mature plant that is typically grown in a greenhouse or other controlled or semi-controlled (e.g., cold frame) environment and then planted or replanted outdoors or in a greenhouse to produce harvestable plant parts (such as, for example, tomato fruit, potato tubers, or tobacco leaves). As used herein, unless otherwise stated or apparent from the context, “tuber” refers to the whole tuber or one or more portions thereof. The preferred tuber of the invention is a potato tuber.

[0289] In the present invention's method involving the cultivation of tubers, a portion of the tuber preferably comprises a sufficient portion of the tuber, thereby enabling that portion to grow into a plant under conditions favorable to the growth and development of the plant derived from the tuber. It should be recognized that such favorable conditions for the growth and development of crop plants, particularly Solanaceae crops, are generally known in the art.

[0290] In some embodiments of the invention, plant cells are transformed with a heteropolynucleotide encoding the R protein of the invention. As used herein, the term “expression” refers to the biosynthesis of a gene product, including transcription and / or translation of said gene product. “Expressing” or “producing” a protein or polypeptide from a DNA molecule means transcribing and translating the coding sequence to produce a protein or polypeptide, while “expressing” or “producing” a protein or polypeptide from an RNA molecule means translating the coding sequence of RNA to produce a protein or polypeptide. Examples of heteropolynucleotides and nucleic acid molecules encoding the R protein are described elsewhere herein.

[0291] The use of the terms “DNA” or “RNA” in this document is not intended to limit the invention to polynucleotide molecules comprising DNA or RNA. Those skilled in the art will recognize that the methods and compositions of the present invention comprise polynucleotide molecules composed of deoxyribonucleotides (i.e., DNA), ribonucleotides (i.e., RNA), or combinations of ribonucleotides and deoxyribonucleotides. Such deoxyribonucleotides and ribonucleotides include naturally occurring molecules and synthetic analogs, including but not limited to nucleotide analogs or modified backbone residues or linkages that are synthetic, naturally occurring, or non-natural, have binding properties similar to a reference nucleic acid, and are metabolized in a manner similar to a reference nucleotide. Examples of such analogs include, but are not limited to, phosphate thioesters, aminophosphate esters, methylphosphonates, chiral methylphosphonates, 2-O-methylribonucleotides, and peptide-nucleic acids (PNAs). The polynucleotide molecules of the present invention also include all forms of polynucleotide molecules, including but not limited to single-stranded, double-stranded, hairpin, stem-loop, and so on. Furthermore, those skilled in the art will understand that the nucleotide sequences disclosed herein also include complementary sequences to the exemplary nucleotide sequences.

[0292] This invention relates to compositions and methods for enhancing plant resistance to plant diseases, particularly compositions and methods for enhancing plant resistance to plant diseases caused by at least one race of at least one species of the genus *Phytophthora*. "Disease resistance" is defined as the avoidance by which a plant avoids disease symptoms as a result of plant-pathogen interactions. That is, prevention of pathogen-induced plant diseases and related disease symptoms, or alternatively, minimization or mitigation of pathogen-induced disease symptoms.

[0293] The following examples are provided for illustrative purposes and not for limitation. Example

[0294] Example 1: Population development for testing the genetic basis of resistance of Solanum nigrum to Phytophthora infestans.

[0295] Recently, a clone of Rpi-amr3i from the Mexican Registry of Solanum spp. has been reported (Witek et al. (2016) Nat. Biotechnol. 34:656; see also WO2016 / 182881; both of which are incorporated herein by reference). To identify resistance to Phytophthora in other Solanum spp. genes, we investigated the immune response to Phytophthora in a group of Solanum spp. (2n) Registries obtained from seed collections (Table 1).

[0296] Table 1. Registry numbers for pathogenic Phytophthora resistance linked to the Rpi-amr1 locus

[0297]

[0298] *RU - Radboud University Radboud, Radboud, Netherlands; IPK - Leibniz Institute for Plant Genetics and Crop Science (IPK-Gatersleben), Germany; NHM - Natural History Museum, London, UK; SBG - Shanghai Botanical Garden, Shanghai, China; NN - Nickys Nursery Ltd, UK.

[0299] Central America is a region encompassing southern North America and northern South America, including Mexico, Belize, Costa Rica, El Salvador, Guatemala, Honduras, Nicaragua, Panama, Colombia, and Venezuela.

[0300] Susceptibility to the pathogen was assessed in detached leaf assays (DLA) using three highly virulent strains of *Phytophthora* (06_3928A, 88069, and EC1). Accession number 954750186 was sensitive to all tested isolates (supporting hyphal growth and sporulation (Witek et al. ((2016) Nat Biotechnol 34:656)). All other accessions retained complete resistance, with no obvious signs of infection or only small hypersensitive (HR) sites present as localized cell death at the *Phytophthora* inoculation site.

[0301] To determine the genetic basis of resistance in *Solanum nigrum* with few flowers, we crossed seven resistance accession numbers (i.e., 954750184, sn27, Veg422, A14750006, SOLA 425, Wang 2058, and A14750130) as male parents with the susceptible line 954750186. The resistant F1 plants were self-pollinated, and we tested the responses of 60 to 100 plants in each F2 generation to 06_3928A and 88069. We found that the progeny of six hybrids segregated in a single ratio, indicating the presence of a single (semi-)dominant resistance gene (fit 3:1 or 2:1). One hybrid showed a 15:1 segregation ratio, indicating the presence of two or more unlinked R genes (Table 1).

[0302] Example 2: Mapping of potential resistance genes in a population derived from resistance accession number 954750184

[0303] We initially focused on F2 and F3 populations derived from a hybrid of the resistant parent 954750184. Bulked susceptible (BS) genomic DNA samples were generated from 94 of the most susceptible F2 and F3 plants. We then performed Illumina-based RenSeq (76 bp PE reads) and additional RAD-seq experiments on the BS gDNA and DNA from the resistant (R) and susceptible (S) parents. Additionally, we performed whole-genome shotgun sequencing (WGS) on the R and S samples using Illumina HiSeq 90 bp PE reads. We used our previously published computational trait mapping pipeline (Jupe et al. ((2013) Plant J. 76:530) and Witek et al. ((2016) Nat Biotechnol 34:656)) to perform single nucleotide polymorphism (SNP) recall and detection of polymorphisms associated with resistance. DNA screening of 94 susceptible F2 and F3 plants yielded a set of markers derived from these analyses, identifying 12 markers associated with resistance responses. These markers were located flanking the R locus, 7.5 cM to one side and 4.3 cM to the other. Although four markers were found to co-segregate with resistance, two markers were found to be located approximately 1 cM on either side; the CAPS marker RAD_3 (BslI) was distal, and the PCR-marker CLC_1 (WGS_1) was proximal. Figure 1 (Upper horizontal bar). Both markers were subsequently used to genotype 1793 F2 plants and identify 259 recombinants (118 homozygous susceptible-heterozygous; 110 homozygous resistant-heterozygous).

[0304] Further genotyping was performed on 118 informative recombinants (homozygous susceptible to one side and heterozygous to the other) using eight linkage markers. Figure 1 The middle horizontal bar was used to test their response to pathogenic Phytophthora isolates EC1 and 06_3928A in DLA. This identified single recombination events for each marker flanking CLC_3(WGS_3) and RAD1, and CLC_2(WGS_2), 56766, and 46418 co-segregated with the resistance locus. Figure 1 (middle horizontal bar).

[0305] Example 3: BAC Clonal Selection

[0306] Linkage map comparison with the potato reference genome identified the homologous CNL-3 NLR gene subfamily within a cosegregating locus. This cluster contains 18 members on potato reference chromosome 11. The marker WGS_2, derived from NLR sequences derived from *Solanum nigrum* WGS data, is orthologous to the CNL-3 cluster. WGS_2 was then used to probe two BAC clones (outsourced to BioS&T; Quebec, Canada; see biost.com). While the cosegregating marker WGS_2 was present on both 5G and 12H of the two derived BAC clones, another cosegregating marker, WGS_3, was present only on 12H. Differences between the two BAC clones were further identified using HindIII digestion. Both were subsequently sequenced on the PacBio RS platform and assembled into single contigs of 125,327 bp (5G) and 144,006 bp (12H), and further assembled into a single contig of 192,456 bp.

[0307] The open reading framework identified 11 potential coding sequences, 9 of which were NLRs, such as those identified by mapping of R parent RenSeq reads and NLR-parser analysis (Steuernagel et al. (2015) Bioinformatics 31:1665). Figure 1 (Lower horizontal bar). All nine sequences had over 80% identity and belonged to the CNL-3 subgroup. Mapping of the cDNA RenSeq readings of the R parent identified seven NLRs as expressed, and they were further considered as candidate NLRs (Rpi-amr1a, b, c, d, e, g, and h).

[0308] Example 4: Transient expression of seven NLR genes in *Nicotiana benthamiana* revealed resistance genes conferring resistance to *Phytophthora infestans*.

[0309] We cloned the open reading frames of seven candidate NLRs into binary expression vectors controlled by the 35S promoter and transformed them into Agrobacterium. These constructs were transiently expressed in detached leaves of *Nicotiana benzinum* and subsequently inoculated with *Phytophthora infestans* isolate 88069 as described by Witek et al. ((2016) Nat Biotechnol 34:656). *Phytophthora infestans* growth was observed 6 days after inoculation on GFP-infiltrated control leaves and on all other constructs except the Rpi-amr3i control and the candidate gene Rpi-amr1e, as well as on the candidate gene Rpi-amr1e. Leaves infiltrated with 35S:Rpi-amr1e showed no to small HR at 6 days post-inoculation (dpi). Figure 2(Left upper leaf). Transient delivery of candidate Rpi-am1e under its natural promoter and terminator elements (1.7kb 5' and 1.3kb 3', nucleotides 1 to 1665 and 4429 to 5732 of SEQ ID NO:1, respectively), followed by pathogenic Phytophthora infection ( Figure 2 (lower left leaf), showing the same level of resistance as the 35S:Rpi-amr1e construct ( Figure 2 (Left upper leaf). This transient expression system identified the candidate Rpi-amr1e as a functional Rpi-amr1 gene.

[0310] Example 5: Potato lines stably transformed with 35S::Rpi-amr1 exhibit resistance to multiple pathogenic Phytophthora strains.

[0311] We used the transformation method described by Kumar et al. ((1996) Plant J. 9:147) to generate stable transgenic plants in the tetraploid cultivar Marispabo using the Rpi-amr1e construct with a natural regulatory element. The transgenic plants showed resistance to the pathogenic Phytophthora race 88069. Figure 3 This result confirms that the cloned gene is a functional Rpi-amr1 gene, which confers resistance to pathogenic Phytophthora in situ in plants.

[0312] Example 6: Resistance was linked to the Rpi-amr1 locus in 6 other populations.

[0313] Genotyping of 10–20 susceptible F2 plants from populations of resistance accessions sn27, Veg422, A14750006, SOLA 425, Wang 2058, and A14750130 showed resistance linked to the Rpi-amr1 locus. To test whether Rpi-amr1e orthologs conferred resistance, we performed SMRT RenSeq on resistance accessions and assembled NLRs as described by Witek et al. ((2016) Nat Biotechnol 34:656). We then mapped all assembled contigs to the coding sequence of Rpi-amr1e, allowing 10% mismatches and gaps, and selected the closest transcribed orthologs (see Table 2 for percentage of amino acid sequence identity), as identified by mapping cDNA RenSeq reads. In the three resistant parents (Veg422, A14750130, and Wang 2058), the identified genes showed 100% amino acid identity with Rpi-amr1e, while the remaining accessions showed over 94% identity with functional Rpi-amr1e (Table 2). We cloned polymorphic genes controlled by the 35S promoters (sn27 and A14750006) or the natural regulatory element (SOLA425) into binary expression vectors. These constructs were transiently expressed in detached leaves of *Nicotiana bensemergère* and inoculated with *Phytophthora infestans* isolate 88069 (24 hours post-inoculation), with resistance assessed at 6 dpi. Compared to the GFP control inoculation, all tested genes conferred enhanced resistance to *Phytophthora infestans* similar to Rpi-amr1e.

[0314] Table 2. Percentage of amino acid sequence identity of proteins encoded by cloned Rpi-amr1e orthologs.

[0315]

[0316] Example 7: Full-length Rpi-amr1e confers strong resistance to multiple pathogenic Phytophthora isolates in stable transgenic potato plants.

[0317] We used the TopHat splice site mapper (Trapnell et al. (2009) Bioinformatics 25:1105-1111) for RNA-Seq readings to map cDNA RenSeq data to BAC contigs and detect two dominant splice variants of the Rpi-amr1e gene (SEQ ID NO:22). The most abundant form, supported by over 80% of the cDNA readings, consists of 4 exons (SEQ ID NO:23) and encodes a protein of 1013 amino acids (SEQ ID NO:24). The remaining cDNA readings showed that several other splice variants corresponding to various 3' truncated forms of SEQ ID NO:23 are possible. We confirmed this by rapid 3' amplification (RACE) PCR of the cDNA ends and observed the following CDS sequences: SEQ ID NO:25, SEQ ID NO:26 and SEQ ID NO:27, which encode a protein of 1004 amino acids (SEQ ID NO:28), a protein of 925 amino acids (SEQ ID NO:29) and a protein of 868 amino acids (SEQ ID NO:30), respectively.

[0318] We constructed stable transgenic potato plants (cultivar Marispabo) carrying the Rpi-amr1e gene (SEQ ID NO:23) using the transformation method described above. We recovered 10 transgenic lines, the presence of which was confirmed by PCR using gene-specific primers. In the DLA, 9 lines showed resistance to the pathogenic Phytophthora 88069 isolate. Figure 4 The selected resistant strain (strain 6) was further phenotyped using other highly virulent pathogenic Phytophthora isolates US23, EC3626, NL14307, NL14538, NL14518 and NL14327, and showed strong resistance (data not shown).

[0319] Example 8: Allele mining identified five additional functional alleles of the resistance-conferring Rpi-amr1e in transient complementation assays of *Nicotiana sambac*.

[0320] In addition to the Rpi-amr1e alleles disclosed in Example 6 above (accession number sn27, also referred to herein as SP1032; accession number SOLA425, also referred to herein as SP2307; and accession number A14750006, also referred to herein as SP1123), we also found three other populations derived from the resistant parents 954750174 (also referred to herein as SP2272), A14750130 (SP3400), and 954750172 (SP3408), where resistance co-segregates with the Rpi-amr1e locus. To test whether the Rpi-amr1e allele is involved in this resistance, we performed SMRT RenSeq and searched for the closest Rpi-amr1e transcriptional homologs as described in Example 6. The gene from SP3408 showed less than 92.5% identity with Rpi-amr1e. The remaining two candidate sequences were more divergent and showed 89.3% identity with Rpi-amr1e at the amino acid level; however, they were 100% identical to each other. We cloned two new Rpi-amr1e alleles, along with three previously reported alleles (SP1032, SOLA425, and SP1123), into the previously described binary vector under their natural regulatory elements: SP1032 (SEQ ID NO:31), SP1123 (SEQ ID NO:32), SP2272 (SEQ ID NO:33), SP2307 (SEQ ID NO:34), and SP3408 (SEQ ID NO:35). In transient complementation assays, all genes conferred resistance to the pathogenic *Phytophthora* isolate 88069. Figure 5 Additionally, we generated stable transgenic *Nicotiana bensemeria* carrying the Rpi-amr1e allele from SP1032 and SP2272 (SEQ ID NO:31 and SEQ ID NO:33, respectively). We recovered 12 independent transgenic lines for each construct and performed phenotypic analysis in DLA using *Phytophthora infestans* isolate 88069. For both constructs, 10 of the 12 lines showed strong resistance (…). Figure 6 ).

[0321] We used the AUGUSTUS gene prediction software (Stanke et al. (2008) Bioinformatics 24:637-644) and also annotated the coding sequences of the functional Rpi-amr1e alleles by alignment with the coding sequence of Rpi-amr1e (SEQ ID NO:23). The predicted CDS sequences of accession numbers SP1032 (SEQ ID NO:36), SP1123 (SEQ ID NO:37), SP2272 (SEQ ID NO:38), SP2307 (SEQ ID NO:39), and SP3408 (SEQ ID NO:40) encode proteins of 986 amino acids (SEQ ID NO:41), 987 amino acids (SEQ ID NO:42), 976 amino acids (SEQ ID NO:43), 986 amino acids (SEQ ID NO:44), and SP3408 (SEQ ID NO:45), respectively.

[0322] Example 9: Development of a segregating population derived from S. nigrescens accession number A14750423

[0323] We investigated the immune response of *S. nigrescens* (2n) accession number A14750423 (also referred to herein as SP3409; country of origin, Mauritius) to *Phytophthora virulence*. In the descaling assay (DLA) using highly virulent *Phytophthora virulence* isolates (06_3928A, 88069, EC1, and NL07434), plants with accession number SP3409 maintained complete resistance (R parent), with no obvious signs of infection or only small sites of hypersensitive response (HR) in the form of localized cell death at the *Phytophthora virulence* inoculation site.

[0324] To determine the genetic basis of resistance, we crossed the resistant strain SP3409 as the male parent with the susceptible strain SP2271 (the S parent reported by Witek et al. (2016) Nat. Biotechnol. 34:656-660) as the female parent. The heterozygous F1 offspring did not show segregation of resistance to Phytophthora 06_3928A and EC1 (6-8 plants were tested for each F1), and these heterozygous F1 offspring were self-pollinated to produce F2 offspring. We tested the resistance of 90 F2 offspring to Phytophthora 88069 and found that the F2 offspring segregated at a ratio of 3:1, indicating the presence of a single dominant resistance gene, which we named Rpi-amr6. Therefore, this F2 population was selected for R gene identification.

[0325] Example 10: Identification of candidate genes by combining RenSeq mapping with PacBio and MiSeq sequencing

[0326] We successfully cloned the R gene without constructing a BAC library using a Solanum NLR decoy library (Witek et al. (2016) Nat. Biotechnol. 34:656-660) by applying the previously described method. To define the complementary sequence of the NLR from the resistant SP3409 parental line, we captured 3-4 kb gDNA fragments and sequenced them in two SMRT cells. This resulted in over 32 kb of insert reads (ROIs). De novo assembly of the ROIs was performed using Geneious, and analysis using an NLR-parser (Steuernagel et al. (2015) Bioinformatics, 31:1665-1667) identified 287 full-length and 555 partial NLRs. To identify candidate NLRs for ligation, we performed Illumina RenSeq on gDNA from 42 susceptible individuals from F2 plants (mixed susceptible, BS), as described (Jupe et al. (2013) Plant J. 76:530-544; Witek et al. (2016) Nat. Biotechnol. 34:656-660). Illumina MiSeq runs generated 744,943, 2,824,501, 678,099, and 1,597,558 paired-end reads from resistant (R) parents, susceptible (S) parents, mixed susceptible, and resistant parents, respectively. After performing initial QC, we mapped the MiSeq data (R, S, and BS parents) to the splicing of PacBio data from the R parents. We used our previously published computer feature mapping pipeline (Jupe et al. (2013) Plant J. 76:530-544; Witek et al. (2016) Nat. Biotechnol. 34:656-660). SNPs were called and detected to detect polymorphisms associated with disease resistance. Briefly, we called homozygous SNPs between the S and R parents and looked for contigs showing a lack of R-specific alleles (less than 5% of R alleles in BS). Transcriptionally active NLRs and their intron / exon structures were annotated with cDNA RenSeq reads as described above (Andolfo et al. (2014) BMCPlantBiol. 14:120; Witek et al. (2016) Nat. Biotechnol. 34:656-660). These identified five candidate NLRs for Rpi-amr6. We further confirmed the cosegregation of these sequences using gene-specific markers (data not shown).

[0327] Example 11: Transient expression of the NLR gene isolated from *Tobacco Bensemium* reveals a pathogenic *Phytophthora* resistance gene.

[0328] We cloned the open reading frame of the candidate NLR for Rpi-amr6 into a binary expression vector under natural regulatory elements, and then introduced the vector into *Agrobacterium tumefaciens* strain AGL-1. These constructs were transiently expressed in *Tobacco Bensemium* leaves that were detached after 24 hours and inoculated with *Phytophthora infestans* isolates 88069 and US-23 as described by Witek et al. ((2016) Nat. Biotechnol. 34:656-660). At 6 dpi, *Phytophthora infestans* growth was restricted in the candidate construct Rpi-amr6b (SEQ ID NO:46) and the positive control construct Rpi-amr3, while symptoms of *Phytophthora infestans* infection were observed in GFP-infiltrated control leaves and the remaining candidate genes. Figure 7 This transient expression system identified candidate Rpi-amr6b as a functional gene.

[0329] Example 12: Stable transformant potato lines carrying Rpi-amr6b and tests of their resistance to multiple pathogenic Phytophthora races.

[0330] Using a potato transformation method described by Kumar et al. ((1996) Plant J, 9:147-158), stable transgenic potato plants were produced in the Marispai background using constructs carrying the Rpi-amr6b gene at its natural regulatory element (SEQ ID NO:1) in the pICSLUS0001 binary vector (Witek et al. (2016) Nat Biotechnol 34:656-660). We recovered five stable transgenic lines showing transgenic activity in PCR tests using Rpi-amr6b-specific primers. Compared to wild-type potato plants, one line showed enhanced resistance to Phytophthora race 88069 (…). Figure 8 Resistance to US23, NL14538 and 14327 (data not shown).

[0331] Example 13: Rpi-amr6b and Rpi-amr1e are homologous

[0332] Rpi-amr6b contains a 5,131 bp open reading frame (ORF). Mapping patterns of the cDNA data indicate that Rpi-amr6b underwent additional splicing, and the two splicing forms can be distinguished. The dominant (i.e., most abundant) transcript variant consists of five exons (SEQ ID NO:49) encoding a 961-amino acid protein (SEQ ID NO:47). A longer transcript (SEQ ID NO:50) was also detected, encoding a 978-amino acid protein (SEQ ID NO:48). Both proteins contain typical features of CC-NB-LRR class resistance proteins, including a coiled-coil domain (CC; amino acids 4-114), a nucleotide-binding domain (NB-ARC; amino acids 153-437), and two potentially leucine-rich repeat sequences (LRR; amino acids 683-800).

[0333] Example 14: GenSeq mapping reveals the mapping location of Rpi-amr6b on the potato double haploid (DM) genome.

[0334] To determine the chromosomal location of Rpi-amr6b, we used enrichment-based genotyping (called GenSeq (Chen et al., “Identification and rapid mapping of a gene conferring broad-spectrumlate blight resistance in the diploid potato species Solanum verrucosum through DNA capture technologies,” Theor. Appl. Genet., 2017)), which utilizes targeting 1,143 conserved orthologs (COS) (Lindqvist-Kreuze et al. (2013) BMC Genetics, 14:51-51) and 837 single-copy genes identified in the potato reference genome (DM) (PotatoGenome Sequencing). 19,716 biotinylated RNA baits were obtained from the Consortium (2011) Nature 475:189-195. Target enrichment and Illumina sequencing were performed as described above, yielding 536,335, 1,200,699, and 658,651 paired-end reads for the gDNA of the R parent (SP3409), S parent (SP2271), and BS, respectively. After performing initial QC, we mapped the MiSeq data to the potato DM sequence. To find linkage regions, we annotated the presence / absence of R alleles in BS (less than 5%; Jupe et al. (2013) Plant Homozygous SNPs between the S and R parents (J.76:530-544). Based on these SNPs, we developed genetic markers and confirmed that Rpi-amr6b maps to a 78Mb region on chromosome 1 in the DM sequence, a region different from that of Rpi-amr1e, which maps to the top of chromosome 11 (approximately 6Mb in the DNA reference genome).

[0335] Example 15: Cloning Rpi-amr7d from Solanum nigrum accession number A54750014 using the RenSeq method

[0336] In our screening, *Solanum nigrum* Registry No. A54750014 (also referred to herein as SP1101) showed strong resistance to *Phytophthora* isolate 88069 in DLA assays. All 60 plants from the F2 progeny derived from crosses with susceptible SP2271 were resistant, indicating the presence of two or more resistance genes. To isolate these genes, we backcrossed F1 plants with susceptible SP2271 (BC1) and then performed another backcross with SP2271 (BC2). The resistant BC2F1 plants were self-pollinated to produce BC2F2, and the segregation rate was tested on 90 plants in the population. Another 600 plants from the selected population were sown and phenotypically typed in DLA for their response to *Phytophthora*. gDNA from 133 susceptible plants was isolated and RenSeq pipelined as described above to identify linked candidate genes. Additionally, using the standard SNP recall pipeline described above, we performed RenSeq and SNP recall on mixed resistance (BR) samples (BR, 20 R plants from the segregating population) to identify fixed susceptibility loci. In short, for homozygous SNPs between S and R parents that were absent in BS (as described above), we counted allele ratios from the BR data. SNPs showing less than 5% R alleles were annotated as fixed and excluded from further analysis. This revealed eight candidate genes encoding NLR, which were cloned and tested in the transient assay described above. Candidate Rpi-amr7d (SEQ ID NO: 51) conferred resistance in the transient assay ( Figure 9 This was used to produce stable transgenic potato plants in the Marispibo background. We recovered six lines showing the presence of Rpi-amr7d through PCR screening using gene-specific markers. Five-week-old plants were evaluated using DLA assays with Phytophthora 8869 and US23; five lines showed strong resistance. Figure 10 ).

[0337] Rpi-amr7d contains a 5131 bp ORF. Mapping of cDNA reads indicated that Rpi-amr7d undergoes additional splicing, and the two splicing forms could be distinguished. The dominant transcript variant consists of five exons (SEQ ID NO:54) encoding a protein of 961 amino acids (SEQ ID NO:52). An additional transcript (SEQ ID NO:55) was detected, encoding a protein of 978 amino acids (SEQ ID NO:53). Both proteins contain typical features of CC-NB-LRR class resistance proteins, including a coiled-coil domain (CC; amino acids 2-120), a nucleotide-binding domain (NB-ARC; amino acids 154-432), and two potentially leucine-rich repeat sequences (LRR; amino acids 683-800).

[0338] Example 16: Cloning Rpi-amr8c from SOLA 226, a genus of Solanum lyratum, using the RenSeq method

[0339] *Solanum nigrum* Registry No. SOLA 226 (also referred to herein as SP2300) showed strong resistance to *Phytophthora* isolate 88069 in DLA assays. In the F2 population derived from a cross between resistant SP2300 and susceptible SP2271 plants, we observed a segregation ratio of 15:1 (resistance to susceptibility), indicating the presence of two unlinked dominant resistance genes. We showed co-segregation of the resistance with a previously cloned Rpi-amr3 gene (Witek et al. (2016) Nat. Biotechnol. 34:656-660; see also WO2016 / 182881 patent application). We screened the F2 population using an Rpi-amr3 gene-specific marker and selected resistant plants lacking Rpi-amr3. The plants were self-pollinated and screened with *Phytophthora* isolate 88069 to detect families segregating at a 3:1 ratio (resistance to susceptibility). From one of the populations separated at a 3:1 ratio, 600 plants were phenotypically typed using DLA for response to Phytophthora pathogenica. gDNA from 114 susceptible (BS) and 20 resistant (BR) plants was isolated, and a RenSeq pipeline was performed as described above to identify linked candidate genes. Additionally, RenSeq and SNP calling were performed on BR resistant samples for the standard SNP calling pipeline described above to identify fixed susceptible loci. Briefly, for homozygous SNPs between S and R parents that were absent in BS (as described above), allele ratios were counted from BR data. SNPs showing less than 5% R alleles were marked as fixed and excluded from further analysis. This resulted in 10 candidate genes encoding NLR, which were cloned and tested in a transient assay as described above. Candidate Rpi-amr8c (SEQ ID NO: 56) conferred resistance in the transient assay. Figure 11 ), and was used to produce stable transgenic potato plants in the Marispaibo cultivar background as described above. Twelve lines were evaluated in a DLA assay using the pathogenic Phytophthora isolate US23, and three lines showed strong resistance ( Figure 12 Five strains showed partial resistance to Phytophthora infestans, while the remaining four strains were susceptible.

[0340] Rpi-amr8c contains a 5125 bp ORF. Mapping of cDNA data indicates that Rpi-amr8c undergoes alternative splicing, and two splicing forms can be distinguished. The dominant transcript variant consists of five exons (SEQ ID NO:59) encoding a protein of 960 amino acids (SEQ ID NO:58). Another transcript (SEQ ID NO:60) was detected, encoding a protein of 986 amino acids (SEQ ID NO:57). Both proteins contain typical features of CC-NB-LRR class resistance proteins, including a coiled-coil domain (CC; amino acids 2-120), a nucleotide-binding domain (NB-ARC; amino acids 153-431), and two potentially leucine-rich repeat sequences (LRR; amino acids 683-800).

[0341] Example 17: Cloning Rpi-amr9d from SOLA 425, a genus of Solanum lyratum, using the RenSeq method.

[0342] The genus *Solanum nigrum*, accession number SOLA 425 (also referred to herein as SP2307), showed strong resistance to *Phytophthora* isolate 88069 in DLA assays. In the F2 population derived from a cross between resistant SP2307 and susceptible SP2271 plants, we observed a segregation ratio of 9:1 (resistance to susceptibility), indicating the presence of more than one resistance gene. The resistant F2 plants were self-pollinated, and the resulting F3 population, screened with *Phytophthora* isolate 88069, was used to detect families segregating at a 3:1 ratio (resistance to susceptibility). From one population showing a 3:1 segregation ratio, 600 plants were phenotyped in DLA for *Phytophthora* response, and gDNA from 117 susceptible (BS) and 20 resistant (BR) plants was isolated. RenSeq pipelines were performed as described above to identify linked candidate genes. In addition to standard SNP calls as described above, we also performed RenSeq and SNP calls on BR samples to identify fixed susceptibility loci. In short, for homozygous SNPs between S and R parents that were not present in BS (as defined above), we calculated the allele ratios from BR data. SNPs showing less than 5% R alleles were noted as fixed and excluded from further analysis. This resulted in 10 genes encoding NLR, which were cloned as described above and tested in a transient assay. Candidate Rpi-amr9d (SEQ ID NO: 61) conferred resistance in the transient assay. Figure 13 ), and used to produce stable transgenic tobacco plants of the same species. Phytophthora spp. 8869 was used to phenotypically analyze 12 independent transgenic events using DLA assays; 11 of these transgenic events showed strong resistance (…). Figure 14 ).

[0343] Rpi-amr9d contains a 7357 bp ORF. Mapping of cDNA data indicated that the Rpi-amr9d gene undergoes alternative splicing, and two splicing forms could be distinguished. The dominant transcript variant consists of five exons (SEQ ID NO: 64) encoding a protein of 986 amino acids (SEQ ID NO: 63). An additional transcript (SEQ ID NO: 65) was detected, encoding a protein of 1011 amino acids (SEQ ID NO: 62). Both proteins contain typical features of CC-NB-LRR class resistance proteins, including a coiled-coil domain (SEQ ID NO: 62, amino acids 2-145; SEQ ID NO: 63, amino acids 2-120), a nucleotide-binding domain (SEQ ID NO: 62, amino acids 179-457; SEQ ID NO: 63, amino acids 154-432), and two potential leucine-rich repeat sequences (SEQ ID NO: 62, amino acids 683-800; SEQ ID NO: 63, amino acids 683-986). Alignment of the nucleotide sequence of Rpi-amr9d with that of its homolog Rpi-amr1e (i.e., Rpi-amr1_2307 (see Examples 6 and 8 above)) cloned from the resistance line SP2307 showed that the two genes are 100% identical.

[0344] Example 18: Rpi-amr1e, Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c belong to the same evolutionary branch of CC-NLR.

[0345] We aligned the full-length amino acid sequences of all Rpi-amr1e functional homologs as well as Rpi-amr6b, Rpi-amr7d, and Rpi-amr8c (Table 3) and generated a phylogenetic tree. Figure 15 All genes are closely related, sharing 88.7% to 97.4% identity. Alignment of the NB-ARC domains of these genes with the NB-ARC domains of cloned functional NLRs (as described in Witek et al. (2016) Nat. Biotechnol. 34:656-660) indicates that these genes belong to the previously uncharacterized clade CNL-3 (not shown).

[0346] Our data show that there is extensive allelic variation in the functional Rpi-amr1e of *Solanum nigrum*, with differences between different alleles reaching up to 12%. Therefore, it is likely that various alleles can recognize unrelated pathogenic *Phytophthora* effectors. This was demonstrated for the wheat mycotoxin resistance locus (MLA), where different alleles of the Mla immune receptor recognize sequence-independent, non-toxic genes of homologous pathogens (Lu et al. (2016) PNAS18:E6486-E6495).

[0347] Table 3. Percentage of amino acid identity of the full-length protein encoded by the cloned Rpi-amr1e homolog.

[0348]

[0349] The articles “a” and “an” used in this article refer to one or more (i.e., at least one) grammatical objects of the article. For example, “an element” means one or more elements.

[0350] Throughout this specification, the word “comprising” or variations thereof, such as “comprises” or “comprising”, shall be understood to imply inclusion of the said element, integer or step, or group of elements, integers or steps, but not to exclude any other element, integer or step, or group of elements, integers or steps.

[0351] All publications and patent applications mentioned in this specification represent the level of skill of a person skilled in the art to which this invention pertains. All publications and patent applications are incorporated herein by reference to the extent that each individual publication or patent application is specifically and individually indicated by reference.

[0352] Although the foregoing invention has been described in considerable detail by way of description and examples for purposes of clarity, it will be apparent that certain changes and modifications may be made within the scope of the appended claims. sequence list <110> Two Blades Foundation <120> Late blight resistance genes and their usage <130> 070294.0127 <150> US 62 / 435,451 <151> 2016-12-16 <160> 65 <170> PatentIn version 3.5 <210> 1 <211> 5732 <212> DNA <213> Solanum americanum <400> 1 ctacatctat ttgcctacca taaaattacc ggtcaagcaa cgtaatttgt aagtcccata 60 gaatttttaa cagatgcatt ggttcatggt tgaggtgaag aacagatgta gcagtttttt 120 gttgaagccg ttagtttctt gacagactat aacagaggaa tgtaagttgc aacttttgag 180 gatagtttag ggatgatttt tgttcattat gtagtataag gatgtagttt aagtttgagg 240 tataattgag ggatgttttt gctatttact ctacaaaatt ccacacagta ttattaattt 300 tctaaggacc gtttggccat gtgatatgaa atcatgatat gaaattatga aatgaagtta 360 aagttttgtt tggacatgta atttggactt tttatgttgt atgttttctt ataaacataa 420 aaaacccaca agttgtaaaa ttattaaact tgtcccattt ttttattcaa ttttaccaaa 480 taaacaaaaa tttacaaaat cacataatat gctaacacaa aactattctt taaaaaatac 540 aatatttatt gatcaaactt taattcaaca aaaaataaaa ttcaacataa gttgtagtgt 600 actagtcttt aatataattc tcccacatag tacggagcaa ttatatgatc gagtatacaa 660 accaacattg ctactttgag ctataaatgg tttaaaaagt aatgatatga atcataaatt 720 ttattacata tgaaacaaat ggtgagtaga tattttttaa taaaatataa acttatgggt 780 caatttttgt atttgaaaaa tcccaaatca tgatttgaaa ttttcaaatc atgatttttg 840 aagaatttgg gattttatct catgatatga aatcatgaga tgaaatcagc ctgaaatcgc 900 atgtccaaat gctgatttca tctcatgatt tcatatcgtg atatgaaatc gcatgtccaa 960 acgcctacta agtaacttag acacgtcttg acctttata attgctccat ccatcctaat 1020 ttacttgtca aatattttct aatttgattc cccttttact tgtcatttt tacaaatcaa 1080 gaaacgacaa tttttttct ttctattata ccctcaattt attaacattg aattaatgtc 1140 cttgaaaaat atagtaagta aatatgttta aaactctatc aaattaatag gggtaaaatg 1200 gtaaactcat tataccaatt atttttttt taatagatgc gtcaaatcaa aaattgacaa 1260 gtaaaaaagag acggaaagag tatataaatt tcaaagaaaa tttgtattga aagttaattg 1320 tattcgtcca cttgacacat catgtcttaa ttactacaag ttgatttgaa taagaaatgt 1380 gagacatctc tactatcgtg gtgggaccaa tgcttgaaaa gtcgacgaaa actacaagag 1440 tgtctttatt agaaatcagg acaacttgac tctttatttt tcatttcttt tttcttactt 1500 tttcttggtc aatccatagt tttgcatcca taaaccacaa gttctgttta gatattaaat 1560 agaaaattgt ccaaattcat ttagaaaaat gtggacataa atcatttaga caaaacctct 1620 tatagcctaa gcagagacat ttctcttcca gcaaacaaaa gagaaatggc tttgctgct 1680 ctttcttcac ttatctatac attgcaacaa ctcttcaaac ctaatcaatc ttcgtttgt 1740 caaagctgct gtacgcaaca acattgtcaa tctctctgtc aaaatctttc tgctctgcaa 1800 cttttccttg acgatactac aacaaaggat attgaaactc ttaaggtatg taattatcta 1860 atctcttact catcttatat ttattcaata ataattaatt tatcgagttt caattttaag 1920 gttatagaaa agaggatcag agatgtagta tacaaagcag aagataaagt tgattcaagc 1980 ctaagaagca tcattctagc tgattgcaca gagaataaag aaggggcttg taaattcttt 2040 gaggaagaat tgctaaaagt ggaaaaagat gttgattctc tcagcaaaga ggtgatgcag 2100 atcaatgtta acaagcatgg aagcagatct gcagaactaa caacaattcc tccctctcca 2160 gaaaaaagta caaatgagga acatactatt gttgggatgg aggatgagta caacaccata 2220 cttgatcgcc tcactgccca aacagacgag ttgactgtca taccaatttt tggtatgggc 2280 ggtataggta agacaactct tgccagaaag gtttatgatg attcatctat tcgttctcga 2340 tttgatagac atgtatgggt cactacctct gaagaattca atgagagacg aatgcttctc 2400 gaagttgttt cttcaattac tactggaagc aatcaagaaa agagcgatga tcaactaatg 2460 gagattgtgt atagaggtct taagggtagg agatttctaa ttgtcataga tgatatttgg 2520 agtactgagg cttgggacca aatgcaaaga atattccaa atgatgacaa taaaagccga 2580 attctactaa ctacacggct caagtatgtt gctgattatg tcagcagtcc tgattttcca 2640 cctcatagta agtcttttct aagtcttgat gatagttgga atctattcac cgaaaaagta 2700 ttcaacaaag atacctgtcc tcctcaccta gaagaaacag ggaagcatat tgtacaacaa 2760 tgtcgaggat tacctctctc ggttgttgta gttgctggac ttgttggaaa aatggaccca 2820 acgcatgaca attgggagaa cgttgaggaa aatctgaact cattcattgg tactgtatcc 2880 gaacggtgcc aatcaattct ttctttaagc tacaattact tgccccaata tttgagggct 2940 tgttttctct atgttggatg ttttcctgaa gataaagaga ttgatgtttc caagttgatt 3000 aggctatgga ttgctgagca attcgtaaag gcgagaagca ataaaaggtt agaagtggta 3060 gcagaggagt atctagaaga gttaattgat agaagcctaa ttttgagtgg tagacaaagg 3120 gctaatggaa ggatgaaaac ttgcaaaatt catgatcttc ttcgccaact atgcctaagt 3180 gaagctcata ctgaaaatat tagtcatatc atgaatagaa atgtccccgt gtcctcagaa 3240 gccatagatg atcaacggcg agtgattgtt ccattggaac ttgaagagga accagtttat 3300 cctacaagga atagcagtgg tattacaagt acaacccgca cctttatttc aatggaaata 3360 tgcctaagag aagctcagac cgaagccata tatgatcaac ggcgagtgat ccttctgtct 3420 aaacgacata ggattgatac aatccgcacc attattccat tcaaagatac ttttccgaaa 3480 gagattgtt ccattgtttc acagttgaag ttgcttaagg tgttggatgt attatcagtc 3540 tggtacgatg tctcttgtat aatacctcag cttgtacatt tgagatatgt tggtgcagta 3600 atttgaaag ctctttcact acccaaattg agaaatctac agaccataat tcttacaagt 3660 gttgaaacca cagagttgaa gcactcacta gatatctgga gaatgtcaga gataagacat 3720 ttggatattg taccgccact atatatatca aatcctcttg aagcagaaca acctttgttt 3780 ctcaataact 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ttttgatcga aaatttagac gaaataagca tgttgccccc gaatgaagta 4680 gccgaaaaaa tcaaaggttt gtctatttaa ttcctttttt atttcattga aagaggttca 4740 gatatattaa acaacttttt aacctttcta tttgattttt tatcgcttag atagatacaa 4800 catttatttt aacatcaatt gttattttga ttcaattatg ttcttttaat atatagtttc 4860 cactcggttc ttcaattctt ttttcaatta tagttttat tttttaattg cacatggttg 4920 atcgatttaa tttgtgagat tcacttttct ttcggtttta cgagttcttc gcgattaatg 4980 ttattcagat ttaggagtat tatatttcaa ctttcgatta gatttaggat tcaaattgtc 5040 ttctttcatt tatgatcatc atcctttcga tcttctttat ttcttttagt aaatagcttc 5100 aaaaactcta tagaaattac atataattaa caattgagaa ttgattctct cgtaatttct 5160 catgttctct ttcagttaaa atttttgatc aaaaactaaa agagattccg gagagcactg 5220 gacaataat gacacgaaaa atcatcaaaa cagaattttg cggctctggt gtagagacta 5280 gtgcaaagaa gattcaagaa gagcaagaaaa gcttgggaaa ttgtgagctt catcaagttc 5340 aaattactcc taaggtatat taaaactcca tctttgatta ttagccaatt ctttaatttc 5400 atattatttc tagactaaat taaaatatta aaaagatataa agaattttaat taatttaatt 5460 attcctttaa aatccaaaca tgatgaattt gagtaaatta aatgaatata gttcagattc 5520 atttaactaa taggccattt agttttaata tggcaacgtt ttaggctttt aacattctct 5580 ttgaatgtag gtataatgca ttaagtgcag aaattttaatc tttatttaaa aaatgaaaac 5640 atctcataat aactttggtc tttttactcc aaacatcttg taataaagtg agtgtcttct 5700 acttctccta acatctcata ataaatttct ac 5732 <210> 2 <211> 920 <212> PRT <213> Solanum photeinocarpum <400> 2 Met Ala Tyr Ala Ala Leu Ser Ser Leu Ile Tyr Thr Leu Gln Gln Leu 1 5 10 15 Phe Lys Pro Asn Gln Ser Phe Val Cys Gln Ser Cys Cys Thr Gln Gln 20 25 30 His Val Gln Ser Leu Cys Gln Asn Leu Ser Ala Leu Gln Leu Phe Leu 35 40 45 Asp Asp Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr 50 55 60 Leu Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser 65 70 75 80 Ser Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr 85 90 95 Lys Ala Glu Asp Lys Val Asp Ser Ser Leu Arg Ser Ile Ile Leu Ala 100 105 110 Asp Cys Thr Glu Asn Lys Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu 115 120 125 Leu Leu Lys Val Glu Lys Asp Val Asp Ser Leu Ser Lys Glu Val Met 130 135 140 Gln Ile Asn Val Asn Lys His Gly Ser Arg Ser Ala Glu Leu Thr Thr 145 150 155 160 Ile Pro Pro Ser Pro Glu Lys Ser Thr Asn Glu Glu His Thr Ile Val 165 170 175 Gly Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln 180 185 190 Thr Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly 195 200 205 Lys Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser 210 215 220 Arg Phe Asp Arg His Val Trp Val Thr Thr Ser Glu Glu Phe Asn Glu 225 230 235 240 Arg Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn 245 250 255 Gln Glu Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Gly Leu 260 265 270 Lys Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Glu 275 280 285 Ala Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Asn Lys Ser 290 295 300 Arg Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Ser 305 310 315 320 Ser Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp 325 330 335 Ser Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Lys Asp Thr Cys Pro 340 345 350 Pro His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Arg Gly 355 360 365 Leu Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp 370 375 380 Pro Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe 385 390 395 400 Ile Gly Thr Val Ser Glu Arg Cys Gln Ser Ile Leu Ser Leu Ser Tyr 405 410 415 Asn Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Cys 420 425 430 Phe Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp 435 440 445 Ile Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Arg Leu Glu Val 450 455 460 Val Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu 465 470 475 480 Ser Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His 485 490 495 Asp Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile 500 505 510 Ser His Ile Met Asn Arg Asn Val Pro Val Ser Ser Glu Ala Ile Asp 515 520 525 Asp Gln Arg Arg Val Ile Val Pro Leu Glu Leu Glu Glu Glu Pro Val 530 535 540 Tyr Pro Thr Arg Asn Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe 545 550 555 560 Ile Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Ala Ile Tyr 565 570 575 Asp Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr 580 585 590 Ile Arg Thr Ile Ile Pro Phe Lys Asp Thr Phe Pro Lys Glu Ile Cys 595 600 605 Ser Ile Val Ser Gln Leu Lys Leu Leu Lys Val Leu Asp Val Leu Ser 610 615 620 Val Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg 625 630 635 640 Tyr Val Gly Ala Val Ile Leu Lys Ala Leu Ser Leu Pro Lys Leu Arg 645 650 655 Asn Leu Gln Thr Ile Ile Leu Thr Ser Val Glu Thr Thr Glu Leu Lys 660 665 670 His Ser Leu Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile 675 680 685 Val Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu 690 695 700 Phe Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val 705 710 715 720 Ala Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu 725 730 735 Asp Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn 740 745 750 Leu Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile 755 760 765 Asp Pro Met Ile Phe Ser Gly Asp Ile Phe Pro Arg Asn Leu Lys Gln 770 775 780 Leu Lys Leu Ser Tyr Thr Cys Ile Pro Trp Glu Asp Met Lys Leu Leu 785 790 795 800 Ala Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asn 805 810 815 Gly Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys 820 825 830 Ser Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala 835 840 845 Gly Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Leu Leu Tyr Gly Phe 850 855 860 Lys Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu 865 870 875 880 Lys Phe Ile Lys Thr Glu Phe Cys Gly Ser Gly Val Lys Thr Ser Ala 885 890 895 Lys Lys Ile Gln Glu Glu Gln Glu Ser Leu Gly Asn Tyr Glu Leu Gln 900 905 910 Val Gln Ile Thr Pro Lys Val Cys 915 920 <210> 3 <211> 2760 <212> DNA <213> Solanum photeinocarpum <400> 3 atggcttatg ctgctctttc ttcacttatc tatacattgc aacaactctt caaacctaat 60 caatctttcg tttgtcaaag ctgctgtacg caacaacatg ttcaatctct ctgtcaaaat 120 ctttctgctc tgcaactttt ccttgacgat actacaacaa aggatattga aactcttaag 180 gtatgtaatt atctaatctc ttactcatct tatatttatt caataataat taatttatcg 240 agtttcaatt ttaaggttat agaaaagagg atcagagatg tagtatacaa agcagaagat 300 aaagttgatt caagcctaag aagcatcatt ctagctgatt gcacagagaa taaagaaggg 360 gcttgtaaat tctttgagga agaattgcta aaagtggaaa aagatgttga ttctctcagc 420 aaagaggtga tgcagatcaa tgttaacaag catggaagca gatctgcaga actaacaaca 480 attcctccct ctccagaaaa aagtacaaat gaggaacata ctattgttgg gatggaggat 540 gagtacaaca ccatacttga tcgcctcact gcccaaacag acgagttgac tgtcatacca 600 atttttggta tgggcggtat aggtaagaca actcttgcca gaaaggttta tgatgattca 660 tctattcgtt ctcgatttga tagacatgta tgggtcacta cctctgaaga attcaatgag 720 agacgaatgc ttctcgaagt tgtttcttca attactactg gaagcaatca agaaaagagc 780 gatgatcaac taatggagat tgtgtataga ggtcttaagg gtaggagatt tctaattgtc 840 atagatgata tttggagtac tgaggcttgg gaccaaatgc aaagaatatt tccaaatgat 900 gacaataaaa gccgaattct actaactaca cggctcaagt atgttgctga tttgtcagc 960 agtcctgatt ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta 1020 ttcaccgaaa aagtattcaa caaagatacc tgtcctcctc acctagaaga aacagggaag 1080 catattgtac aacaatgtcg aggattacct ctctcggttg ttgtagttgc tggacttgtt 1140 ggaaaaatgg acccaacgca tgacaattgg gagaacgttg aggaaaatct gaactcattc 1200 attggtactg tatccgaacg gtgccaatca attctttctt taagctacaa ttacttgcc 1260 1320 gtttccaagt tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa 1380 1440 agtggtagac aaagggctaa tggaaggtg aaaacttgca aaattcatga tcttcttcgc 1500 1560 cccgtgtcct cagaagccat agatgatcaa cggcgagtga ttgttccatt ggaacttgaa 1620 gaggaaccag tttatcctac aaggaatagc agtggtatta caagtacaac ccgcaccttt 1680 atttcaatgg aaatatgcct aagagaagct cagaccgaag ccatatatga tcaacggcga 1740 gtgatccttt tgtctaaacg acataggatt gatacaatcc gcaccattat tccattcaaa 1800 gatacttttc cgaaagagat ttgttccatt gtttcacagt tgaagttgct taaggtgttg 1860 gatgtattat cagtctggta cgatgtctct tgtataatac ctcagcttgt acatttgaga 1920 tatgttggtg cagtaatttt gaaagctctt tcactaccca aattgagaaa tctacagacc 1980 father caagtgttga aaccacagag ttgaagcact cactagatat ctggagaatg tcagagata gacatttgga tattgtaccg ccactatata tatcaaatcc tcttgaagca 2160. 2160. 2160. 2160. 2160. 2160. 2160. 2160. 2160 gcgaaaatca taagaagaac tcccaatcta aaaaagctaa agattttaga taaatctaag catcctgact ggcctgatat tcttgattct ctcaatcttc tagaggagct ggagacacta 2340. snow snow snow snow snow snow snow snow aatctcaagc aactgaaatt atcatatact tgtataccat gggaagatat gaaattgctg gctaatttac ccaatcttga ggtgttcaag ggtcattatg cattcaatgg aacagattgg aaactagatg aagatgttgt gttttgcaaa ttaaaatctc tacgactgta tgagcgtgga gatttgcaaa ggtgggaagc tgctggtagt gataattttc caatgcttga gcaactatta ttgtatgggt tcaaaaaact ggagagatt ccggagagta ttggagaat aatgacacta aaatttatca aaacagaatt ttgcggctct ggtgtaaaga caagtgcaaa gaaaattcaa gaagagcaag aaagcctggg aaattatgag cttcaagttc aaattactcc taaggtatgt 2760 <210> 4 <211> 5957 <212> DNA <213> Solanum photeinocarpum <400> 4 attggtgatt cgacgtttga atttttaaga taattatatc tttttcaatg atacccgcaa 60 gggtgatcga gttggttgag cagataattt ctaaatgggg tcataggttc aaaatcctct 120 acatgtttgc tcggtagctc gtcatatgga acttgcttag tatagtttac cttttcagtg 180 tgatttgcgc gctattacgt tgaagcagtg ttatcgtata cacacctaaa ccctgaagaa 240 tcatgactgc gagcttcctt gacaacaaat tttttttatc tatttcaatg acattactat 300 tagatcaatt tttttatatc acatgatttg atccatccta catctatttg cctaccataa 360 aattaccggt caagcaacgt aatttgtaag tcccatagaa tttttaacag atgcattggt 420 tcatggttga ggtgaagaac agatgtagca gttttttgtt gaagccgtta gtttcttgac 480 agactataac agaggaatgt aagttgcaac ttttgaggat agtttaggga tgatttttgt 540 tcattatgta gtataaggat gtagtttaag tttgaggtat aattgaggga tgtttttgct 600 attackcta caaattcca cacagtatta ttaatttct aaggaccgtt tggccatgtg 660 attgaaatc atgatatgaa attatgaat gaagttaag ttttgttttgg acatgtaatt 720 tggacttttt atgttgtatg ttttcttata aacataaaaa acccacaagt tgtaaaatta 780 ttaacttgt cccatttttt tattcaattt taccaaata acaaaattt acaaatcac 840 aatatatgct aacacaaac tattcttta aaatacaat atttattgat caacttta 900 ttcacaaaa aaaaatttc aacataagtt gtagtgtact agtctttaat atattctcc 960 cacatagtac ggagcaatta tatgatcgag tatacaacc aacattgcta ctttgagcta 1020 taaatggttt aaaagtaat gattgaatc aataatttta tcatatga aacaatggt 1080 gagtagatat ttttaata ataataact tattgggtca ttttgtatt tgaaaaatcc 1140 caaatcatga tttgaattt tcaatcatg atttttgag aatttgggat ttatctcat 1200 gattgaat catgagatga aatcagcctg aaatcgcatg tccaatgct gatttcatct 1260 catgatttca tatcgtgata tgaaatcgca tgtccaacg cctactaagt aacttagaca 1320 cgtcttgacc ttttatatt gctccatcca tcctaattta cttgtcaaat attttctaat 1380 ttgattcccc tttactgt cattttttac aaatcaagaa acgacaattt ttttctttc 1440 tattataccc tcaatttatt aacattgaat taatgtcctt gaaaaata gtaagtaat 1500 atgtttaaaa ctctatcaa ttaatagggg taaaatgta aactcattat accaattatt 1560 tttttttta tagatgcgtc aaatcaaaa ttgacaagta aaagagacg gaagagtat 1620 aaatttca aagaaaattt gtattgaaag ttaattgtat tcgtccactt gandacatcat 1680 gtcttaatta ctacaagttg atttgaataa gaatgtgag acacctctac tatcgtggtg 1740 ggaccaatgc tgaaaagtc gacgaaaact acagagtgt ctttattaga atcaggaca 1800 acttgactct ttattttca ttctttttt cttactttt cttggtcaat ccatagtttt 1860 gcatccataa accacaagtt ctgtttagat attaataga aaattgtcca aattcattta 1920 gaaaaatgtg gacaataatc atttagacaaacccttt agcctaagca gagacattc 1980 tcttccagca aaaaagag aaatggctta tgctgctt tctcactta tctatacatt 2040 gcaacaactc ttcaaaccta atcaatcttt cgtttgtcaa agctgctgta cgcaacaaca 2100 tgttcaatct ctctgtcaaa atctttctgc tctgcaactt ttccttgacg atactacaac 2160 aaaggatatt gaaactctta aggtatgtaa ttatctaatc tcttactcat cttatattta 2220 ttcaataata attaatttat cgagtttcaa ttttaaggtt atagaaaaga ggatcagaga 2280 tgtagtatac aaagcagaag ataaagttga ttcaagccta agaagcatca ttctagctga 2340 ttgcacagag aataaagaag gggcttgtaa attctttgag gaagaattgc taaaagtgga 2400 aaaagatgtt gattctctca gcaaagaggt gatgcagatc aatgttaaca agcatggaag 2460 cagatctgca gaactaacaa caattcctcc ctctccagaa aaaagtacaa atgaggaaca 2520 tactattgtt gggatggagg atgagtacaa caccatactt gatcgcctca ctgcccaaac 2580 agacgagttg actgtcatac caatttttgg tatgggcggt ataggtaaga caactcttgc 2640 cagaaaggtt tatgatgatt catctattcg ttctcgattt gatagacatg tatgggtcac 2700 tacctctgaa gaattcaatg agagacgaat gcttctcgaa gttgtttctt caattactac 2760 tggaagcaat caagaaaaga gcgatgatca actaatggag attgtgtata gaggtcttaa 2820 gggtaggaga tttctaattg tcatagatga tatttggagt actgaggctt gggaccaaat 2880 gcaaagaata tttccaaatg atgacaataa aagccgaatt ctactacta cacggctcaa 2940 gtatgttgct gattatgtca gcagtcctga ttttccacct catagtaagt cttttctaag 3000 tcttgatgat agttggaatc tattcaccga aaaagtattc aacaaagata cctgtcctcc 3060 tcacctagaa gaaacaggga agcatattgt acaacaatgt cgaggattac ctctctcggt 3120 tgttgtagtt gctggacttg ttggaaaaat ggacccaacg catgacaatt gggagaacgt 3180 tgaggaaaat ctgaactcat tcattggtac tgtatccgaa cggtgccaat caattctttc 3240 tttaagctac aattacttgc cccaatattt gagggcttgt tttctctatg ttggatgttt 3300 tcctgaagat aaagagattg atgtttccaa gttgattagg ctatggattg ctgagcaatt 3360 cgtaaggcg agaagcaata aaaggttaga agtggtagca gaggattac tagagagtt 3420 aattgataga agcctaattt tgagtggtag acaaagggct aatggagga tgaaaacttg 3480 caaaattcat gatcttcttc gccaactatg cctaagtgaa gctcatactg aaaattag 3540 tcatatcatg aatagaaatg tccccgtgtc ctcagaagcc agatatgatc aacggcgagt 3600 gattgttcca ttggaacttg aagaggaacc agtttatcct acaaggaata gcagtggtat 3660 tacaagtaca acccgcacct ttatttcaat ggaaatatgc ctaagagaag ctcagaccga 3720 agccatatat gatcaacggc gagtgatcct tctgtctaaa cgacatagga ttgatacaat 3780 ccgcaccatt attccattca aagatacttt tccgaaagag atttgttcca ttgtttcaca 3840 gttgaagttg cttaaggtgt tggatgtatt atcagtctgg tacgatgtct cttgtataat 3900 acctcagctt gtacatttga gatatgttgg tgcagtaatt ttgaaagctc tttcactacc 3960 caaattgaga aatctacaga ccataattct tacaagtgtt gaaaccacag agttgaagca 4020 ctcactagat atctggagaa tgtcagagat aagacatttg gatatgtac cgccactata 4080 tatatcaaat cctcttgaag cagaacaacc tttgtttctc aataacttgc acacactttt 4140 tctccgttgc tctcctttg ttgcgaaaat cataagaaga actcccaatc taaaaaagct 4200 aaagatttta gataaatcta agcatcctga ctggcctgat attcttgatt ctctcaatct 4260 tctagaggag ctggagacac tacaaatatc aacagaagaa aacattgacc cgatgattttt 4320 ctctggggat atttccctc gtaatctcaa gcaactgaaa tttcatata cttgtatacc 4380 atgggaagat atgaaattgc tggctaattt acccaatctt gaggtgttca agggtcatta 4440 tgcattcaat ggaacagatt ggaaactaga tgaagatgtt gtgttttgca aattaaaatc 4500 tctacgactg tatgagcgtg gagatttgca aaggtgggaa gctgctggta gtgataattt 4560 tccaatgctt gagcaactat tattgtatgg gttcaaaaaa ctggaagaga ttccggagag 4620 tattggagaa ataatgacac taaatttat caaaacagaa ttttgcggct ctggtgtaaa 4680 gacaagtgca aagaaaattc aagaagagca agaaagcctg ggaaattatg agcttcaagt 4740 tcaaattact cctaaggtat gttgaaactc aatctttgat tatactctc cactcggtta 4800 gatttaggat tcaaattgtc tacttacatt tatgatcatc acccttgcgg tcttcttatt 4860 tcttttagta atagcttca aaactctac agaaattaca cataattaac aattgacaat 4920 tgattctc ttattgataa tttgtaattt tctctttcag ttaagatttt tgatcgaaaa 4980 tttagacgaa ataagcatgt tgcccccgaa tgaagtagcc gaaaaaatca aaggtttgtc 5040 tatttaattc cttttttatt tcattgaaag aggttcagat atattaaaca actttttaac 5100 ctttctattt gatttttat cgcttagata gatacaacat ttattttaac atcaattgtt 5160 atttgattc aattatgttc ttttaatata tagttccac tcggttcttc aattcttttt 5220 tcaattatag ttttattttt ttaattgcac atggttgatc gatttaattt gtgagattca 5280 cttttctttc ggttttacga gttcttcgcg attaatgtta ttcagattta ggagtattat 5340 atttcaactt tcgattagat ttaggattca aattgtcttc tttcatttat gatcatcatc 5400 ctttcgatct tctttattc ttttagtaaa tagcttcaaa aactctatag aaattacata 5460 taattaacaa ttgagaattg attctctcgt aatttctcat gttctctttc agttaaaatt 5520 tttgatcaaa aactaaaaga gattccggag agcactggac aataatgac acgaaaaatc 5580 atcaaaacag aatttgcgg ctctggtgta gagactagtg caaagaagat tcaagaagag 5640 caagaaagct tgggaaattg tgagcttcat caagttcaaa ttactcctaa ggtatattaa 5700 aactccatct ttgattatta gccaattctt taatttcata ttatttctag actaaattaa 5760 aatattaaaa gatataaaga atttaattaa tttaattatt cctttaaaat ccaaacatga 5820 tgaatttgag taaattaaat gaatatagtt cagattcatt taactaatag gccatttagt 5880 tttaatatgg caacgtttta ggcttttaac attctctttg aatgtaggta taatgcatta 5940 agtgcagaaa tttaatc 5957 <210> 5 <211> 920 <212> PRT <213> Solanum photeinocarpum <400> ⑤ Met Ala Tyr Ala Ala Leu Ser Ser Leu Ile Tyr Thr Leu Gln Gln Leu 1 5 10 15 Phe Lys Pro Asn Gln Ser Phe Val Cys Gln Ser Cys Cys Thr Gln Gln 20 25 30 His Val Gln Ser Leu Cys Gln Asn Leu Ser Ala Leu Gln Leu Phe Leu 35 40 45 Asp Asp Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr 50 55 60 Leu Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser 65 70 75 80 Ser Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr 85 90 95 Lys Ala Glu Asp Lys Val Asp Ser Ser Leu Arg Ser Ile Ile Leu Ala 100 105 110 Asp Cys Thr Glu Asn Lys Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu 115 120 125 Leu Leu Lys Val Glu Lys Asp Val Asp Ser Leu Ser Lys Glu Val Met 130 135 140 Gln Ile Asn Val Asn Lys His Gly Ser Arg Ser Ala Glu Leu Thr Thr 145 150 155 160 Ile Pro Pro Ser Pro Glu Lys Ser Thr Asn Glu Glu His Thr Ile Val 165 170 175 Gly Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln 180 185 190 Thr Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly 195 200 205 Lys Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser 210 215 220 Arg Phe Asp Arg His Val Trp Val Thr Thr Ser Glu Glu Phe Asn Glu 225 230 235 240 Arg Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn 245 250 255 Gln Glu Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Gly Leu 260 265 270 Lys Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Glu 275 280 285 Ala Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Asn Lys Ser 290 295 300 Arg Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Ser 305 310 315 320 Ser Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp 325 330 335 Ser Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Lys Asp Thr Cys Pro 340 345 350 Pro His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Arg Gly 355 360 365 Leu Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp 370 375 380 Pro Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe 385 390 395 400 Ile Gly Thr Val Ser Glu Arg Cys Gln Ser Ile Leu Ser Leu Ser Tyr 405 410 415 Asn Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Cys 420 425 430 Phe Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp 435 440 445 Ile Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Arg Leu Glu Val 450 455 460 Val Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu 465 470 475 480 Ser Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His 485 490 495 Asp Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile 500 505 510 Ser His Ile Met Asn Arg Asn Val Pro Val Ser Ser Glu Ala Ile Asp 515 520 525 Asp Gln Arg Arg Val Ile Val Pro Leu Glu Leu Glu Glu Glu Pro Val 530 535 540 Tyr Pro Thr Arg Asn Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe 545 550 555 560 Ile Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Ala Ile Tyr 565 570 575 Asp Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr 580 585 590 Ile Arg Thr Ile Ile Pro Phe Lys Asp Thr Phe Pro Lys Glu Ile Cys 595 600 605 Ser Ile Val Ser Gln Leu Lys Leu Leu Lys Val Leu Asp Val Leu Ser 610 615 620 Val Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg 625 630 635 640 Tyr Val Gly Ala Val Ile Leu Lys Ala Leu Ser Leu Pro Lys Leu Arg 645 650 655 Asn Leu Gln Thr Ile Ile Leu Thr Ser Val Glu Thr Thr Glu Leu Lys 660 665 670 His Ser Leu Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile 675 680 685 Val Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu 690 695 700 Phe Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val 705 710 715 720 Ala Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu 725 730 735 Asp Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn 740 745 750 Leu Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile 755 760 765 Asp Pro Met Ile Phe Ser Gly Asp Ile Phe Pro Arg Asn Leu Lys Gln 770 775 780 Leu Lys Leu Ser Tyr Thr Cys Ile Pro Trp Glu Asp Met Lys Leu Leu 785 790 795 800 Ala Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asn 805 810 815 Gly Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys 820 825 830 Ser Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala 835 840 845 Gly Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Leu Leu Tyr Gly Phe 850 855 860 Lys Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu 865 870 875 880 Lys Phe Ile Lys Thr Glu Phe Cys Gly Ser Gly Val Lys Thr Ser Ala 885 890 895 Lys Lys Ile Gln Glu Glu Gln Glu Ser Leu Gly Asn Tyr Glu Leu Gln 900 905 910 Val Gln Ile Thr Pro Lys Val Cys 915 920 <210> 6 <211> 2760 <212> DNA <213> Solanum photeinocarpum <400> 6 atggcttatg ctgctctttc ttcacttatc tatacattgc aacaactctt caaacctaat 60 caatctttcg tttgtcaaag ctgctgtacg caacaacatg ttcaatctct ctgtcaaaat 120 ctttctgctc tgcaactttt ccttgacgat actacaacaa aggatattga aactcttaag 180 gtatgtaatt atctaatctc ttactcatct tatatttatt caataataat taatttatcg 240 agtttcaatt ttaaggttat agaaaagagg atcagagatg tagtatacaa agcagaagat 300 aaagttgatt caagcctaag aagcatcatt ctagctgatt gcacagagaa taaagaaggg 360 gcttgtaaat tctttgagga agaattgcta aaagtggaaa aagatgttga ttctctcagc 420 aaagaggtga tgcagatcaa tgttaacaag catggaagca gatctgcaga actaacaaca 480 attcctccct ctccagaaaa aagtacaaat gaggaacata ctattgttgg gatggaggat 540 gagtacaaca ccatacttga tcgcctcact gcccaaacag acgagttgac tgtcatacca 600 atttttggta tgggcggtat aggtaagaca actcttgcca gaaaggttta tgatgattca 660 tctattcgtt ctcgatttga tagacatgta tgggtcacta cctctgaaga attcaatgag 720 agacgaatgc ttctcgaagt tgtttcttca attactactg gaagcaatca agaaaagagc 780 gatgatcaac taatggagat tgtgtataga ggtcttaagg gtaggagatt tctaattgtc 840 atagatgata tttggagtac tgaggcttgg gaccaaatgc aaagaatatt tccaaatgat 900 gacaataaaa gccgaattct actaactaca cggctcaagt atgttgctga tttgtcagc 960 agtcctgatt ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta 1020 ttcaccgaaa aagtattcaa caaagatacc tgtcctcctc acctagaa aacagggaag 1080 catattgtac aacaatgtcg aggattacct ctctcggttg ttgtagttgc tggacttgtt 1140 ggaaaaatgg acccaacgca tgacaattgg gagaacgttg aggaaaatct gaactcattc 1200 attggtactg tatccgaacg gtgccaatca attctttctt taagctacaa ttacttgccc 1260 1320 gtttccaagt tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa 1380 1440 agtggtagac aaagggctaa tggaaggtg aaaacttgca aaattcatga tcttcttcgc 1500 1560 cccgtgtcct cagaagccat agatgatcaa cggcgagtga ttgttccatt ggaacttgaa 1620 gaggaaccag tttatcctac aaggaatagc agtggtatta caagtacaac ccgcaccttt 1680 atttcaatgg aaatatgcct aagagaagct cagaccgaag ccatatatga tcaacggcga 1740 gtgatccttt tgtctaaacg acataggatt gatacaatcc gcaccattat tccattcaaa 1800 gatacttttc cgaaagagat ttgttccatt gtttcacagt tgaagttgct taaggtgttg 1860 gatgtattat cagtctggta cgatgtctct tgtataatac ctcagcttgt acatttgaga 1920 tatgttggtg cagtaatttt gaaagctctt tcactaccca aattgagaaa tctacagacc 1980 2040 2100 gaacaacctt tgtttctcaa taacttgcac acactttttc tccgttgctc tccttttgtt 2160 gcgaaaatca taagagaac tcccaatcta aaaaagctaa agattttaga taaatctaag 2220 catcctgact ggcctgatat tcttgattct ctcaatcttc tagaggagct ggagacacta 2280 caaatatcaa cagaagaaaa cattgacccg atgattttct ctggggatat tttccctcgt 2340 aatctcaagc aactgaaatt atcatatact tgtataccat gggaagatat gaaattgctg 2400 gctaatttac ccaatcttga ggtgttcaag ggtcattatg cattcaatgg aacagattgg 2460 aaactagatg aagatgttgt gttttgcaaa ttaaaatctc tacgactgta tgagcgtgga 2520 gatttgcaaa ggtgggaagc tgctggtagt gataattttc caatgcttga gcaactatta 2580 ttgtatgggt tcaaaaaact ggaagagatt ccggagagta ttggagaaat aatgacacta 2640 aaatttatca aaacagaatt ttgcggctct ggtgtaaaga caagtgcaaa gaaaattcaa 2700 gaagagcaag aaagcctggg aaattatgag cttcaagttc aaattactcc taaggtatgt 2760 <210> 7 <211> 5948 <212> DNA <213> Solanum photeinocarpum <400> 7 [[ID=2))0]]ttgcttagta tagtttacct tttcagtgtg atttgcgcgc tattacgttg aagcagtgtt 60 atcgtataca cacctaaacc ctgaagaatc atgactgcga gcttccttga caacaaattt 120 tttttatcta tttcaatgac attactatta gatcaatttt tttatatcac atgatttgat 180 ccatcctaca tctatttgcc taccataaaa ttaccggtca agcaacgtaa tttgtaagtc 240 ccatagaatt tttaacagat gcattggttc atggttgagg tgaagaacag atgtagcagt 300 tttttgttga agccgttagt ttcttgacag actataacag aggaatgtaa gttgcaactt 360 ttgaggatag tttagggatg atttttgttc attatgtagt ataaggatgt agtttaagtt 420 tgaggtataa ttgagggatg tttttgctat ttactctaca aaattccaca cagtattatt 480 aattttctaa ggaccgtttg gccatgtgat atgaaatcat gatatgaaat tatgaaatga 540 agttaaagtt ttgtttggac atgtaatttg gactttttat gttgtatgtt ttcttataaa 600 cataaaaaac ccacaagttg taaaattatt aaacttgtcc cattttttta ttcaatttta 660 ccaaataaac aaaaatttac aaaatcacat aatatgctaa cacaaaacta ttctttaaaa 720 aatacaatat ttattgatca aactttaatt caacaaaaaa taaaattcaa cataagttgt 780 agtgtactag tctttaatat aattctccca catagtacgg agcaattata tgatcgagta 840 tacaaaccaa cattgctact ttgagctata aatggtttaa aaagtaatga tatgaatcat 900 aaattttatt acatatgaaa caaatggtga gtagatattt tttaataaaa tataaactta 960 tgggtcaatt tttgtatttg aaaaatccca aatcatgatt tgaaattttc aaatcatgat 1020 ttttgaagaa tttgggattt tatctcatga tatgaaatca tgagatgaaa tcagcctgaa 1080 atcgcatgtc caaatgctga tttcatctca tgatttcata tcgtgatatg aaatcgcatg 1140 tccaaacgcc tactaagtaa cttagacacg tcttgacctt ttataattgc tccatccatc 1200 ctaatttact tgtcaaatat tttctaattt gattcccctt ttacttgtca ttttttacaa 1260 atcaagaaac gacaattttt tttctttcta ttataccctc aatttattaa cattgaatta 1320 atgtccttga aaaatatagt aagtaaaatat gtttaaaact ctatcaaatt aataggggta 1380 aaatggtaaa ctcatattac caattatttt ttttttaata gatgcgtcaa atcaaaaatt 1440 gacaagtaaa aagagacgga aagagtatat aaatttcaaa gaaaatttgt attgaaagtt 1500 aattgtattc gtccacttga cacatcatgt cttaattact acaagttgat ttgaataaga 1560 aatgtgagac atctctacta tcgtggtggg accaatgctt gaaaagtcga cgaaaactac 1620 aagagtgtct ttattagaaa tcaggacaac ttgactcttt attttcatt tcttttttct 1680 tactttttct tggtcaatcc atagttttgc atccataaac cacaagttct gtttagatat 1740 taaatagaa attgtccaaa ttcatttaga aaaatgtgga cataaatcat ttagacaaaa 1800 cctcttatag cctaagcaga gatattctc ttccagcaa CAAAGAGAA atggcttatg 1860 ctgctctttc ttcacttc tatacattgc aaaactctt caactaat caatctttcg 1920 ttgtcaaag ctgctgtacg siacaatg ttcaatctct ctgtcaaat ctttctgctc 1980 tgcaactttt ccttgacgat actacaaca aggatattga aactcttaag gtatgtaatt 2040 atctaatctc ttactcatct tatattatt caataat taatttatcg agtttchcatt 2100 tttaggttat agaaagagg atcagagatg tagtataca agcagagat aaagttgatt 2160 caagcctaag aagcatcatt ctagctgatt gcacagagaa taagaaggg gcttgtaaat 2220 tctttgagga agaattgcta aaagtggaa aagatgttga ttctcagc aaagaggtga 2280 tgcagatcaa tgttaacaag catggaagca gatctgcaga actacaca attcctccct 2340 ctccagaaaa agtacaat gaggacata ctattgttgg gatggagat gaggacaca 2400 ccatacttga tcgcctcact gcccaacag acgagttgac tgtcatacca atttttggta 2460 tgggcggtat aggtaagaca actctgcca gaaggttta tgatgattca tctattcgtt 2520 ctcgatttga tagacatgta tgggtcacta cctctgaaga attcaatgag agacgaatgc 2580 ttctcgaagt tgttcttca attactactg gaagcaatca agaaaagagc gatgatcaac 2640 taatggagat tgtgtataga ggtcttaagg gtaggagatt tctaattgtc atagatgata 2700 tttggagtac tgaggcttgg gaccaaatgc aaagaatatt tccaaatgat gacaataaaa 2760 gccgaattct actaactaca cggctcaagt atgttgctga ttatgtcagc agtcctgatt 2820 ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta ttcaccgaaa 2880 aagtattcaa caaagatacc tgtcctcctc acctagaaga aacagggaag catattgtac 2940 aacaatgtcg aggattacct cctcggttg ttgtagttgc tggacttgtt ggaaaaatgg 3000 acccaacgca tgacaattgg gagaacgttg aggaaaatct gaactcattc attggtactg 3060 tatccgaacg gtgccaatca attctttctt taagctacaa ttacttgccc caatatttga 3120 gggcttgttt tctctatgtt ggatgttttc ctgaagataa agagattgat gtttccaagt 3180 tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa aggttagaag 3240 3300 aaagggctaa tggaaggtg aaaacttgca aaattcatga tcttcttcgc caactatgcc 3360 3420 3480 tttatcctac aaggaatagc agtggtatta caagtacaac ccgcaccttt atttcaatgg 3540 aaatatgcct aagagaagct cagaccgaag ccatatatga tcaacggcga gtgatccttc 3600 tgtctaaacg acataggatt gatacaatcc gcaccattat tccattcaaa gatacttttc 3660 cgaaagagat ttgttccatt gtttcacagt tgaagttgct taaggtgttg gatgtattat 3720 cagtctggta cgatgtctct tgtataatac ctcagcttgt acatttgaga tatgttggtg 3780 footattttt gaaagctctt tcactaccca aattgagaaa tctacagacc ataattctta 3840 3900 3960 tgtttctcaa taacttgcac acactttttc tccgttgctc tcctttgtt gcgaaaatca 4020 taagaaac tcccaatcta aaaaagctaa agattttaga taaatctaag catcctgact 4080 ggcctgatat tcttgattct ctcaatcttc tagaggagct ggagacacta caaatatcaa 4140 cagaagaaaa cattgacccg atgattttct ctggggatat tttccctcgt aatctcaagc 4200 aactgaaatt atcatatact tgtataccat gggaagatat gaaattgctg gctaatttac 4260 ccaatcttga ggtgttcaag ggtcattatg cattcaatgg aacagattgg aaactagatg 4320 aagatgttgt gttttgcaaa ttaaaatctc tacgactgta tgagcgtgga gatttgcaaa 4380 ggtgggaagc tgctggtagt gataattttc caatgcttga gcaactatta ttgtatgggt 4440 tcaaaaaact ggaagagatt ccggagagta ttggagaaat aatgacacta aaatttatca 4500 aaacagaatt ttgcggctct ggtgtaaaga caagtgcaaa gaaaattcaa gaagcaag 4560 aaagcctggg aaattatgag cttcaagttc aaattactcc taaggtatgt tgaaactcaa 4620 tctttgatta atactctcca ctcggttaga tttaggattc aaattgtcta cttacattta 4680 tgatcatcac ccttgcggtc ttcttatttc ttttagtaaa tagcttcaaa aactctacag 4740 aaattacaca taattaacaa ttgacaattg attctctctt attgataatt tgtaattttc 4800 tctttcagtt aagatttttg atcgaaaatt tagacgaaat aagcatgttg cccccgaatg 4860 aagtagccga aaaaatcaaa ggtttgtcta tttaattcct tttttatttc attgaaagag 4920 gttcagatat attaaacaac tttttaacct ttctatttga ttttttatcg cttagataga 4980 tacaacattt attttaacat caattgttat tttgattcaa ttatgttctt ttaatatata 5040 gtttccactc ggttcttcaa ttcttttttc aattatagtt ttatttttt aattgcacat 5100 ggttgatcga tttaatttgt gagattcact tttctttcgg ttttacgagt tcttcgcgat 5160 taatgttat cagatttagg agtattatat ttcaactttc gattagattt aggattcaaa 5220 ttgtcttctt tcatttatga tcatcatcct ttcgatcttc tttatttctt ttagtaaata 5280 gcttcaaaaa ctctatagaa attacatata attaacaatt gagaattgat tctctcgtaa 5340 tttctcatgt tctctttcag ttaaaatttt tgatcaaaaa ctaaaagaga ttccggagag 5400 cactggacaa ataatgacac gaaaaatcat caaaacagaa ttttgcggct ctggtgtaga 5460 gactagtgca aagaagattc aagaagagca agaaagcttg ggaaattgtg agcttcatca 5520 agttcaaatt actcctaagg tatattaaaa ctccatcttt gattattagc caattcttta 5580 atttcatatt atttctagac taaattaaaa tattaaaaga tataaagaat ttaattaatt 5640 taattattcc tttaaaatcc aaacatgatg aatttgagta aattaaatga atatagttca 5700 gattcattta actaataggc catttagttt taatatggca acgttttagg cttttaacat 5760 tctctttgaa tgtaggtata atgcattaag tgcagaaatt taatctttat ttaaaaaatg 5820 aaaacatctc ataataactt tggtcttttt actccaaaca tcttgtaata aagtgagtgt 5880 cttctacttc tcctaacatc tcataataaa tttctactat tcctaacatc tcattataaa 5940 taagtctt 5948 <210> 8 <211> 920 <212> PRT <213> Solanum photeinocarpum <400> 8 Met Ala Tyr Ala Ala Leu Ser Ser Leu Ile Tyr Thr Leu Gln Gln Leu 1 5 10 15 Phe Lys Pro Asn Gln Ser Phe Val Cys Gln Ser Cys Cys Thr Gln Gln 20 25 30 His Val Gln Ser Leu Cys Gln Asn Leu Ser Ala Leu Gln Leu Phe Leu 35 40 45 Asp Asp Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr 50 55 60 Leu Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser 65 70 75 80 Ser Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr 85 90 95 Lys Ala Glu Asp Lys Val Asp Ser Ser Leu Arg Ser Ile Ile Leu Ala 100 105 110 Asp Cys Thr Glu Asn Lys Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu 115 120 125 Leu Leu Lys Val Glu Lys Asp Val Asp Ser Leu Ser Lys Glu Val Met 130 135 140 Gln Ile Asn Val Asn Lys His Gly Ser Arg Ser Ala Glu Leu Thr Thr 145 150 155 160 Ile Pro Pro Ser Pro Glu Lys Ser Thr Asn Glu Glu His Thr Ile Val 165 170 175 Gly Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln 180 185 190 Thr Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly 195 200 205 Lys Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser 210 215 220 Arg Phe Asp Arg His Val Trp Val Thr Thr Ser Glu Glu Phe Asn Glu 225 230 235 240 Arg Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn 245 250 255 Gln Glu Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Gly Leu 260 265 270 Lys Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Glu 275 280 285 Ala Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Asn Lys Ser 290 295 300 Arg Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Ser 305 310 315 320 Ser Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp 325 330 335 Ser Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Lys Asp Thr Cys Pro 340 345 350 Pro His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Arg Gly 355 360 365 Leu Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp 370 375 380 Pro Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe 385 390 395 400 Ile Gly Thr Val Ser Glu Arg Cys Gln Ser Ile Leu Ser Leu Ser Tyr 405 410 415 Asn Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Cys 420 425 430 Phe Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp 435 440 445 Ile Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Arg Leu Glu Val 450 455 460 Val Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu 465 470 475 480 Ser Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His 485 490 495 Asp Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile 500 505 510 Ser His Ile Met Asn Arg Asn Val Pro Val Ser Ser Glu Ala Ile Asp 515 520 525 Asp Gln Arg Arg Val Ile Val Pro Leu Glu Leu Glu Glu Glu Pro Val 530 535 540 Tyr Pro Thr Arg Asn Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe 545 550 555 560 Ile Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Ala Ile Tyr 565 570 575 Asp Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr 580 585 590 Ile Arg Thr Ile Ile Pro Phe Lys Asp Thr Phe Pro Lys Glu Ile Cys 595 600 605 Ser Ile Val Ser Gln Leu Lys Leu Leu Lys Val Leu Asp Val Leu Ser 610 615 620 Val Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg 625 630 635 640 Tyr Val Gly Ala Val Ile Leu Lys Ala Leu Ser Leu Pro Lys Leu Arg 645 650 655 Asn Leu Gln Thr Ile Ile Leu Thr Ser Val Glu Thr Thr Glu Leu Lys 660 665 670 His Ser Leu Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile 675 680 685 Val Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu 690 695 700 Phe Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val 705 710 715 720 Ala Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu 725 730 735 Asp Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn 740 745 750 Leu Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile 755 760 765 Asp Pro Met Ile Phe Ser Gly Asp Ile Phe Pro Arg Asn Leu Lys Gln 770 775 780 Leu Lys Leu Ser Tyr Thr Cys Ile Pro Trp Glu Asp Met Lys Leu Leu 785 790 795 800 Ala Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asn 805 810 815 Gly Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys 820 825 830 Ser Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala 835 840 845 Gly Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Leu Leu Tyr Gly Phe 850 855 860 Lys Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu 865 870 875 880 Lys Phe Ile Lys Thr Glu Phe Cys Gly Ser Gly Val Lys Thr Ser Ala 885 890 895 Lys Lys Ile Gln Glu Glu Gln Glu Ser Leu Gly Asn Tyr Glu Leu Gln 900 905 910 Val Gln Ile Thr Pro Lys Val Cys 915 920 <210> 9 <211> 2760 <212> DNA <213> Solanum photeinocarpum <400> 9 atggcttatg ctgctctttc ttcacttatc tatacattgc aacaactctt caaacctaat 60 caatctttcg ttgtcaaag ctgctgtacg caacaatg ttcaatctct ctgtcaaat 120 ctttctgctc tgcaactttt ccttgacgat actacaaca aggatattga acctcttaag 180 gtatgtaatt atctaatctc ttactcatct tatattatt caataataat taatttatcg 240 agttcaatt taggttt agaaaagagg atcagagg tagtataca agahaagat 300 aaagttgatt caagcctaag aagcatcatt ctagctgatt gcacagagaa taaagaaggg 360 gcttgtaaat tctttgagga agaattgcta aaagtggaa aagatgttga ttctctcagc 420 aaagaggtga tgcagatcaa tgttacaag catggagca gatctgcaga actacaca 480 attcctccct ctccagaaaa aagtacaaat gaggaacata ctattgttgg gatggagat 540 gagtacaaca ccatacttga tcgcctcact gcccaacag acgagttgac tgtcatacca 600 atttttggta tgggcggtat aggtaagaca actctgcca gaaaggttta tgatgattca 660 tctattcgtt ctcgatttga tagacatgta tggtcacta cctctgaaga attcaatgag 720 agacgaatgc ttctcgaagt tgttttctca attackacg gaagcaatca agaaagagc 780 gatgatcaac taatggagat tgtgtataga ggtcttaagg gtaggagatt tctaattgtc 840 atagatgata tttggagtac tgaggcttgg gaccaaatgc aaagaatatt tccaaatgat 900 gacaataaaa gccgaattct actaactaca cggctcaagt atgttgctga tttgtcagc 960 agtcctgatt ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta 1020 ttcaccgaaa aagtattcaa caaagatacc tgtcctcctc acctagaaga aacagggaag 1080 catattgtac aacaatgtcg aggattacct ctctcggttg ttgtagttgc tggacttgtt 1140 ggaaaaatgg acccaacgca tgacaattgg gagaacgttg aggaaaatct gaactcattc 1200 attggtactg tatccgaacg gtgccaatca attctttctt taagctacaa ttacttgcc 1260 caatatttga gggcttgttt tctctatgtt ggatgttttc ctgaagataa agagattgat 1320 gttccaagt tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa 1380 aggttagaag tggtagcaga ggagtatcta gaagagttaa ttgatagaag cctaattttg 1440 agtggtagac aaagggctaa tggaaggatg aaaacttgca aaattcatga tcttcttcgc 1500 1560 cccgtgtcct cagaagccat agatgatcaa cggcgagtga ttgttccatt ggaacttgaa 1620 gaggaaccag tttatcctac aaggaatagc agtggtatta caagtacaac ccgcaccttt 1680 atttcaatgg aaatatgcct aagagaagct cagaccgaag ccatatatga tcaacggcga 1740 gtgatccttt tgtctaaacg acataggatt gatacaatcc gcaccattat tccattcaaa 1800 gatacttttc cgaaagagat ttgttccatt gtttcacagt tgaagttgct taaggtgttg 1860 gatgtattat cagtctggta cgatgtctct tgtataatac ctcagcttgt acatttgaga 1920 tatgttggtg cagtaatttt gaaagctctt tcactaccca aattgagaaa tctacagacc 1980 2040 2100 gaacaacctt tgtttctcaa taacttgcac acactttttc tccgttgctc tccttttgtt 2160 gcgaaaatca taagagaac tcccaatcta aaaaagctaa agattttaga taaatctaag 2220 catcctgact ggcctgatat tcttgattct ctcaatcttc tagaggagct ggagacacta 2280 caaatatcaa cagaagaaaa cattgacccg atgattttct ctggggatat tttccctcgt 2340 aatctcaagc aactgaaatt atcatatact tgtataccat gggaagatat gaaattgctg 2400 gctaatttac ccaatcttga ggtgttcaag ggtcattatg cattcaatgg aacagattgg 2460 aaactagatg aagatgttgt gttttgcaaa ttaaaatctc tacgactgta tgagcgtgga 2520 gatttgcaaa ggtgggaagc tgctggtagt gataattttc caatgcttga gcaactatta 2580 ttgtatgggt tcaaaaaact ggaagagatt ccggagagta ttggagaaat aatgacacta 2640 aaatttatca aaacagaatt ttgcggctct ggtgtaaaga caagtgcaaa gaaaattcaa 2700 gaagagcaag aaagcctggg aaattatgag cttcaagttc aaattactcc taaggtatgt 2760 <210> 10 <211> 6049 <212> DNA <213> Solanum photeinocarpum <400> 10 ggtaattttt ttcttatcat gtataacatt tcttttatat taatagaggg gtcacatgtt 60 attcaaaaac aagtatggtg attggtgatt cgacgtttga atttttaaga taattatatc 120 tttttcaatg atacccgcaa gggtgatcga gttggttgag cagataattt ctaaatgggg 180 tcataggttc aaaatcctct acatgtttgc tcggtagctc gtcatatgga acttgcttag 240 tatagtttac cttttcagtg tgatttgcgc gctattacgt tgaagcagtg ttatcgtata 300 cacacctaaa ccctgaagaa tcatgactgc gagcttcctt gacaacaaat tttttttatc 360 tatttcaatg acattactat tagatcaatt ttttatatc acatgatttg atccatccta 420 catctatttg cctaccataa aattaccggt caagcaacgt aatttgtaag tcccatagaa 480 ttttaacag atgcattggt tcatggttga ggtgaagaac agatgtagca gttttttgtt 540 gaagccgtta gttcttgac agactataac agaggaatgt aagttgcaac ttttgaggat 600 agtttaggga tgatttttgt tcattatgta gtataaggat gtagtttaag tttgaggtat 660 aattgaggga tgtttttgct atttactcta caaaattcca cacagtatta ttaattttct 720 aaggaccgtt tggccatgtg atatgaaatc atgatatgaa attatgaaat gaagttaaag 780 ttttgttttgg acatgtaatt tggacttttt atgttgtatg ttttcttata aacataaaaa 840 accacaagt tgtaaaatta ttaaacttgt cccatttttt tattcaattt taccaaataa 900 aaaaattt acaaatcac atatatgct aacacaaac tattcttta aaatacaat 960 atttattgat caactttaa ttcaaaaa aaaaattc aacataagtt gtagtgtact 1020 agtctttaat atattctcc cacatagtac ggagcaatta tatgatcgag tatacaacc 1080 aacattgcta ctttgagcta taaatggttt aaaagtaat gatatgaatc aataatttta 1140 ttacatatga aaaaatggt gagtagatat ttttaata atattaact tatgggtca 1200 ttttgtatt tgaaaaatcc caatcatga tttgaattt tcaatcatg atttttgaag 1260 aatttgggat ttatctcat gatatgaat catgagatga atcagcctg aaatcgcatg 1320 tccaaatgct gatttcatct catgatttca tatcgtgata tgaatcgca tgtccaacg 1380 cctactaagt aacttagaca cgtcttgacc ttttatatt gctccatcca tcctaattta 1440 cttgtcaaat attttctaat ttgattcccc ttttactgt cattttttac aaatcaagaa 1500 acgacaattt ttttctttc tattataccc tcaatttatt aacattgaat taatgtcctt 1560 gaaaaata gtaagtaaat atgtttaaaa ctctatcaaa ttaatagggg taaatggta 1620 aactcattat accaattatt ttttttta tagatgcgtc aaatcaaaa ttgacaagta 1680 aaagagacg gaagagtat atatttca aaaaaattt gtattgaag tattgtat 1740 tcgtccactt gandacatcat gtctttatta ctacaagttg atttgaataa gaatgtgag 1800 acatctctac tatcgtgtg ggaccaatgc ttgaaagtc gacgaaaact acagagtgt 1860 ctttattaga atcaggaca acttgactct ttttttca tttctttt cttacttttt 1920 cttggtcaat ccatagtttt gcatccataa accacaagtt ctgtttagat attaataga 1980 aaattgtcca attcattta gaaaatgtg gataatc atttagacaa acctcttat 2040 agcctaagca gagacatttc tctccagca aaaaagag aaatggctta tgctgctt 2100 tcttcactta tctatacatt gcaacaactc ttcaaccta atcaatctt cgtttgtcaa 2160 agctgctgta cgcaacaa tgttcaatct ctctgtcaa atctttctgc tctgcaactt 2220 ttccttgacg atactacaac aaaggatatt gaaactctta aggtatgtaa ttatctaatc 2280 tcttactcat cttatattta ttcaataata attaatttat cgagtttcaa ttttaaggtt 2340 atagaaaaga ggatcagaga tgtagtatac aaagcagaag ataaagttga ttcaagccta 2400 agaagcatca ttctagctga ttgcacagag aataaagaag gggcttgtaa attctttgag 2460 gaagaattgc taaaagtgga aaaagatgtt gattctctca gcaaagaggt gatgcagatc 2520 aatgttaaca agcatggaag cagatctgca gaactaacaa caattcctcc ctctccagaa 2580 aaaagtacaa atgaggaaca tactattgtt gggatggagg atgagtacaa caccatactt 2640 gatcgcctca ctgcccaaac agacgagttg actgtcatac caatttttgg tatgggcggt 2700 ataggtaaga caactcttgc cagaaaggtt tatgatgatt catctattcg ttctcgattt 2760 gatagacatg tatgggtcac tacctctgaa gaattcaatg agagacgaat gcttctcgaa 2820 gttgtttct caattactac tggaagcaat caagaaaaga gcgatgatca actaatggag 2880 attgtgtata gaggtcttaa gggtaggaga tttctaattg tcatagatga tatttggagt 2940 actgaggctt gggaccaaat gcaaagaata tttccaaatg atgacaataa aagccgaatt 3000 ctactacta cacggctcaa gtatgttgct gattatgtca gcagtcctga ttttccacct 3060 catagtaagt cttttctaag tcttgatgat agttggaatc tattcaccga aaaagtattc 3120 aacaaagata cctgtcctcc tcacctagaa gaaacaggga agcatattgt acaacaatgt 3180 cgaggattac ctctctcggt tgttgtagtt gctggacttg ttggaaaaat ggacccaacg 3240 catgacaatt gggagaacgt tgaggaaaat ctgaactcat tcattggtac tgtatccgaa 3300 cggtgccaat caattctttc tttaagctac aattacttgc cccaatattt gagggcttgt 3360 tttctctatg ttggatgttt tcctgaagat aaagagattg atgtttccaa gttgattagg 3420 ctatggattg ctgagcaatt cgtaaaggcg agaagcaata aaagggtaga agtggtagca 3480 gaggattac tagagaagtt aattgataga agcctaattt tgagtggtag acaaagggct 3540 aatggagga tgaaaacttg caaaattcat gatcttcttc gccaactatg cctaagtgaa 3600 gctcatactg aaaattag tcatatcatg aatagaaatg tccccgtgtc ctcagaagcc 3660 atagatgatc aacggcgagt gattgttcca ttggaacttg aagaggaacc agtttatcct 3720 acaaggaata gcagtggtat tacaagtaca acccgcacct ttatttcaat ggaaatatgc 3780 ctaagagaag ctcagaccga agccatatat gatcaacggc gagtgatcct tctgtctaaa 3840 cgacatagga ttgatacaat ccgcaccatt attccattca aagatacttt tccgaaagag 3900 atttgttcca ttgtttcaca gttgaagttg cttaaggtgt tggatgtatt atcagtctgg 3960 tacgatgtct cttgtataat acctcagctt gtacatttga gatatgttgg tgcagtaatt 4020 ttgaaagctc tttcactacc caaattgaga aatctacaga ccataattct tacaagtgtt 4080 gaaaccacag agttgaagca ctcactagat atctggagaa tgtcagagat aagacatttg 4140 gatatgtac cgccactata tatatcaaat cctcttgaag cagaacaacc tttgtttctc 4200 aataacttgc acacactttt tctccgttgc tctcctttg ttgcgaaaat cataagaaga 4260 actcccaatc taaaaaagct aaaagatttta gataaatcta agcatcctga ctggcctgat 4320 attcttgatt ctctcaatct tctagaggag ctggagacac tacaaatatc aacagaagaa 4380 aacattgacc cgatgatttt ctctggggat atttccctc gtaatctcaa gcaactgaaa 4440 tttcatata cttgtatacc atgggaagat atgaaattgc tggctaattt acccaatctt 4500 gaggtgttca agggtcatta tgcattcaat ggaacagatt ggaaactaga tgaagatgtt 4560 gtgttttgca aattaaaatc tctacgactg tatgagcgtg gagatttgca aaggtgggaa 4620 gctgctggta gtgataattt tccaatgctt gagcaactat tattgtatgg gttcaaaaaa 4680 ctggaagaga ttccggagag tattggagaa ataatgacac taaatttat caaaacagaa 4740 ttttgcggct ctggtgtaaa gacaagtgca aagaaaattc aagaagagca agaaagcctg 4800 ggaaattatg agcttcaagt tcaaattact cctaaggtat gttgaaactc aatctttgat 4860 tatactctc cactcggtta gatttaggat tcaaattgtc tacttacatt tatgatcatc 4920 acccttgcgg tcttcttatt tcttttagta aatagcttca aaaactctac agaaattaca 4980 cataattaac aattgacaat tgattctctc ttattgataa tttgtaattt tctctttcag 5040 ttaagatttt tgatcgaaaa tttagacgaa ataagcatgt tgcccccgaa tgaagtagcc 5100 gaaaaaatca aaggtttgtc tatttaattc cttttttatt tcattgaaag aggttcagat 5160 atattaaaca actttttaac ctttctattt gatttttat cgcttagata gatacaacat 5220 ttattttaac atcaattgtt atttgattc aattatgttc ttttaatata tagttccac 5280 tcggttcttc aattctttt tcaattatag ttttattttt ttaattgcac atggttgatc 5340 gatttaattt gtgagattca cttttctttc ggttttacga gttcttcgcg attaatgtta 5400 ttcagattta ggagtattat atttcaactt tcgattagat ttaggattca aattgtcttc 5460 tttcattat gatcatcatc ctttcgatct tctttatttc ttttagtaaa tagcttcaaa 5520 aactctatag aaattacata taattaacaa ttgagaattg attctctcgt aatttctcat 5580 gttctctttc agttaaaatt tttgatcaaa aactaaaaga gattccggag agcactggac 5640 aaatatgac acgaaaaatc atcaaaacag aatttgcgg ctctggtgta gagactagtg 5700 caaagaagat tcaagaagag caagaaagct tgggaaattg tgagcttcat caagttcaaa 5760 ttactcctaa ggtatattaa aactccatct ttgattatta gccaattctt taatttcata 5820 ttatttctag actaaattaa aatattaaaa gatataaaga atttaattaa tttaattatt 5880 cctttaaaat ccaaacatga tgaatttgag taaattaaat gaatatagtt cagattcatt 5940 taactaatag gccatttagt tttaatatgg caacgtttta ggcttttaac attctctttg 6000 aatgtaggta taatgcatta agtgcagaaa tttaatcttt atttaaaaa 6049 <210> 11 <211> 920 <212> PRT <213> Solanum photeinocarpum <400> 11 Met Ala Tyr Ala Ala Leu Ser Ser Leu Ile Tyr Thr Leu Gln Gln Leu 1 5 10 15 Phe Lys Pro Asn Gln Ser Phe Val Cys Gln Ser Cys Cys Thr Gln Gln 20 25 30 His Val Gln Ser Leu Cys Gln Asn Leu Ser Ala Leu Gln Leu Phe Leu 35 40 45 Asp Asp Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr 50 55 60 Leu Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser 65 70 75 80 Ser Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr 85 90 95 Lys Ala Glu Asp Lys Val Asp Ser Ser Leu Arg Ser Ile Ile Leu Ala 100 105 110 Asp Cys Thr Glu Asn Lys Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu 115 120 125 Leu Leu Lys Val Glu Lys Asp Val Asp Ser Leu Ser Lys Glu Val Met 130 135 140 Gln Ile Asn Val Asn Lys His Gly Ser Arg Ser Ala Glu Leu Thr Thr 145 150 155 160 Ile Pro Pro Ser Pro Glu Lys Ser Thr Asn Glu Glu His Thr Ile Val 165 170 175 Gly Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln 180 185 190 Thr Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly 195 200 205 Lys Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser 210 215 220 Arg Phe Asp Arg His Val Trp Val Thr Thr Ser Glu Glu Phe Asn Glu 225 230 235 240 Arg Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn 245 250 255 Gln Glu Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Gly Leu 260 265 270 Lys Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Glu 275 280 285 Ala Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Asn Lys Ser 290 295 300 Arg Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Ser 305 310 315 320 Ser Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp 325 330 335 Ser Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Lys Asp Thr Cys Pro 340 345 350 Pro His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Arg Gly 355 360 365 Leu Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp 370 375 380 Pro Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe 385 390 395 400 Ile Gly Thr Val Ser Glu Arg Cys Gln Ser Ile Leu Ser Leu Ser Tyr 405 410 415 Asn Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Cys 420 425 430 Phe Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp 435 440 445 Ile Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Arg Leu Glu Val 450 455 460 Val Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu 465 470 475 480 Ser Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His 485 490 495 Asp Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile 500 505 510 Ser His Ile Met Asn Arg Asn Val Pro Val Ser Ser Glu Ala Ile Asp 515 520 525 Asp Gln Arg Arg Val Ile Val Pro Leu Glu Leu Glu Glu Glu Pro Val 530 535 540 Tyr Pro Thr Arg Asn Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe 545 550 555 560 Ile Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Ala Ile Tyr 565 570 575 Asp Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr 580 585 590 Ile Arg Thr Ile Ile Pro Phe Lys Asp Thr Phe Pro Lys Glu Ile Cys 595 600 605 Ser Ile Val Ser Gln Leu Lys Leu Leu Lys Val Leu Asp Val Leu Ser 610 615 620 Val Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg 625 630 635 640 Tyr Val Gly Ala Val Ile Leu Lys Ala Leu Ser Leu Pro Lys Leu Arg 645 650 655 Asn Leu Gln Thr Ile Ile Leu Thr Ser Val Glu Thr Thr Glu Leu Lys 660 665 670 His Ser Leu Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile 675 680 685 Val Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu 690 695 700 Phe Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val 705 710 715 720 Ala Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu 725 730 735 Asp Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn 740 745 750 Leu Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile 755 760 765 Asp Pro Met Ile Phe Ser Gly Asp Ile Phe Pro Arg Asn Leu Lys Gln 770 775 780 Leu Lys Leu Ser Tyr Thr Cys Ile Pro Trp Glu Asp Met Lys Leu Leu 785 790 795 800 Ala Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asn 805 810 815 Gly Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys 820 825 830 Ser Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala 835 840 845 Gly Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Leu Leu Tyr Gly Phe 850 855 860 Lys Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu 865 870 875 880 Lys Phe Ile Lys Thr Glu Phe Cys Gly Ser Gly Val Lys Thr Ser Ala 885 890 895 Lys Lys Ile Gln Glu Glu Gln Glu Ser Leu Gly Asn Tyr Glu Leu Gln 900 905 910 Val Gln Ile Thr Pro Lys Val Cys 915 920 <210> 12 <211> 2760 <212> DNA <213> Solanum photeinocarpum <400> 12 atggcttatg ctgctctttc ttcacttatc tatacattgc aacaactctt caaacctaat 60 caatctttcg tttgtcaaag ctgctgtacg caacaacatg ttcaatctct ctgtcaaaat 120 ctttctgctc tgcaactttt ccttgacgat actacaacaa aggatattga aactcttaag 180 gtatgtaatt atctaatctc ttactcatct tatatttatt caataataat taatttatcg 240 agtttcaatt ttaaggttat agaaaagagg atcagagatg tagtatacaa agcagaagat 300 aaagttgatt caagcctaag aagcatcatt ctagctgatt gcacagagaa taaagaaggg 360 gcttgtaaat tctttgagga agaattgcta aaagtggaaa aagatgttga ttctctcagc 420 aaagaggtga tgcagatcaa tgttaacaag catggaagca gatctgcaga actaacaaca 480 attcctccct ctccagaaaa aagtacaaat gaggaacata ctattgttgg gatggaggat 540 gagtacaaca ccatacttga tcgcctcact gcccaaacag acgagttgac tgtcatacca 600 atttttggta tgggcggtat aggtaagaca actcttgcca gaaaggttta tgatgattca 660 tctattcgtt ctcgatttga tagacatgta tgggtcacta cctctgaaga attcaatgag 720 agacgaatgc ttctcgaagt tgtttcttca attactactg gaagcaatca agaaaagagc 780 gatgatcaac taatggagat tgtgtataga ggtcttaagg gtaggagatt tctaattgtc 840 atagatgata tttggagtac tgaggcttgg gaccaaatgc aaagaatatt tccaaatgat 900 gacaataaaa gccgaattct actaactaca cggctcaagt atgttgctga tttgtcagc 960 agtcctgatt ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta 1020 ttcaccgaaa aagtattcaa caaagatacc tgtcctcctc acctagaaga aacagggaag 1080 catattgtac aacaatgtcg aggattacct ctctcggttg ttgtagttgc tggacttgtt 1140 ggaaaaatgg acccaacgca tgacaattgg gagaacgttg aggaaaatct gaactcattc 1200 attggtactg tatccgaacg gtgccaatca attctttctt taagctacaa ttacttgccc 1260 1320 gtttccaagt tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa 1380 1440 agtggtagac aaagggctaa tggaaggtg aaaacttgca aaattcatga tcttcttcgc 1500 1560 cccgtgtcct cagaagccat agatgatcaa cggcgagtga ttgttccatt ggaacttgaa 1620 gaggaaccag tttatcctac aaggaatagc agtggtatta caagtacaac ccgcaccttt 1680 atttcaatgg aaatatgcct aagagaagct cagaccgaag ccatatatga tcaacggcga 1740 gtgatccttt tgtctaaacg acataggatt gatacaatcc gcaccattat tccattcaaa 1800 gatacttttc cgaaagagat ttgttccatt gtttcacagt tgaagttgct taaggtgttg 1860 gatgtattat cagtctggta cgatgtctct tgtataatac ctcagcttgt acatttgaga 1920 tatgttggtg cagtaatttt gaaagctctt tcactaccca aattgagaaa tctacagacc 1980 2040 2100 gaacaacctt tgtttctcaa taacttgcac acactttttc tccgttgctc tccttttgtt 2160 gcgaaaatca taagagaac tcccaatcta aaaaagctaa agattttaga taaatctaag 2220 catcctgact ggcctgatat tcttgattct ctcaatcttc tagaggagct ggagacacta 2280 caaatatcaa cagaagaaaa cattgacccg atgattttct ctggggatat tttccctcgt 2340 aatctcaagc aactgaaatt atcatatact tgtataccat gggaagatat gaaattgctg 2400 gctaatttac ccaatcttga ggtgttcaag ggtcattatg cattcaatgg aacagattgg 2460 aaactagatg aagatgttgt gttttgcaaa ttaaaatctc tacgactgta tgagcgtgga 2520 gatttgcaaa ggtgggaagc tgctggtagt gataattttc caatgcttga gcaactatta 2580 ttgtatgggt tcaaaaaact ggaagagatt ccggagagta ttggagaaat aatgacacta 2640 aaatttatca aaacagaatt ttgcggctct ggtgtaaaga caagtgcaaa gaaaattcaa 2700 gaagagcaag aaagcctggg aaattatgag cttcaagttc aaattactcc taaggtatgt 2760 <210> 13 <211> 5149 <212> DNA <213> Solanum photeinocarpum <400> 13 gaacgctgtg taacatcatc tccaaccaac ctgggtcata tgagtcccaa atgcatttac 60 agcaacaaag agcaaagcaa agcacaactt tgaaatttga aaaatgatgc aagacaaata 120 tcaaaattag acagtaaaat tcacaattcc ccttaaaata aatgataaat taaagaagac 180 taaacaatag aaaaccttac aatctcaaag gcagtatgga tcggagtcgt atttggaaga 240 aattttgata caacttgaag aagattgaag attgttgttc ataaagtata ctgtgggcgg 300 gttaatctac aacctgtgtg ttttttatca tacaaaattc cacagagtat tattaatttt 360 ttaaggatcg tttggccatg tgatatgaaa ttatgagatg aagttgacgt tttgtttgga 420 catgcaattt ggacttttta tgttgtatgt tttcttataa acataagttg taaaattatt 480 aaagttgtcc cattttttta ttcaatttta ccaaataaac aaaaatttac aaaatcacat 540 aatatgctaa cacaaaacta ttctttaaaa aatacaatat ttattgatca aactttaatt 600 caacaaaaaa taaaattcaa cataagttgt agtgtactag tctttaatat aattctccca 660 cataatacgg acaatctcct cacgttgaac ttgcatttct cgatcatatg attgagtaga 720 caaaccaaca ttgctacttt gagctataaa tggtttaaaa agtaatgata tgaattataa 780 attttattta catatgaaac aaatggtaag taaatataaa tgtggggttg tttttataaa 840 atataaactt atgggtcaat ttttgtattt gaaaaatccc aaattatgat ttgaaatttt 900 caaatcatga tttttgaaga attcagtctg aaatcgcatg tccaaatgct gatttcatct 960 catgatttca tatcgtgata tgaaatcgca tgtccaaacg cctactaagt aacttagaca 1020 cgtcttgacc ttttataatt actccctcca tcctaattta cttgtcaaat attttctaat 1080 ttgattcccc ttttacttgt cattttttac aaatcaagaa acgacaattt tttttcttcc 1140 tattataccc tcaatttatt aacattgaat taatgtcctt gaaaaatata ggaagtaaat 1200 atgtttaaaa ctctatcaaa ttaatatgga taaaatggta aattcattat accaattatt 1260 atttcttaa tagatgcgtc aaatcaaaaa ttgacaagta atataaattt caaagaaaat 1320 ttgtattgaa agttaattgt attcgtccac ttgacacatc atgtcttaat tattcaacaa 1380 gtttgaatcc aatgtatatg aatgtttgaa aagtcttgac gaaaactaca agagtgtctt 1440 tattagaaat caggacaact tgactcttta tttttcattt cttttttctc actttgactt 1500 ggtcaatcca tagttttgca tccataaacc acaagttctg tttagatatt aatagaaaa 1560 ttgtccaaat tcatttagaa aaatgtggac ataaatcatt tagacaaaac ctcttagcct 1620 aagcagagac atttctcttc cagcaaacaa aagagaaatg gcatatgctg ctctttcttc 1680 acttatctat acattgcaac aactcttgaa acctaatcaa tctttggttt gtcgaagctg 1740 tacacaacaa catcttcaat ctatctatca caatctttct gctctgcaac ttttccttga 1800 cgatactacg acaaaggata ttgaaactct taaggtatgt aattatctaa tctcttactc 1860 atcttatatt tattcaataa taattaattt atcgagtttc aattttaagg ttatagaaaa 1920 gaggatcaga gatgtagtat acaaagcaga agataaagtt gattcaagcc taagaaacat 1980 catactagca gattgcacag agatagaga aggggcttgt aaattctttg aggaagaatt 2040 gctaaaagtg gaaaaagatg ttgattctct caggaaagag gtgatgcaga tcgagtttaa 2100 caagcatggt agcagatctg cagaactaac aacaattctt ccctctccag aaaaaagtac 2160 aattgaggaa catactattg ttgggatgga ggatgagtac aacaccatac ttgatcgcct 2220 cactgcccaa acagacgagt tgactgtcat accaattttt ggtatgggcg gtataggtaa 2280 gacaactcta gccagaaagg tttatgatga ttcatctatt cgttctcgat ttgatagaca 2340 tgtatgggtc actacctctg aagaattcaa tgagagacga atgcttctcg aagttgtttc 2400 ttcaattact actggaagca atcaagaaaa gagcgatgat caactaatgg agattgtgta 2460 tagaggtctt aagggtagga gatttctaat tgtcatagat gatatttgga gtactcaggc 2520 ttgggaccaa atgcaaagaa tatttccaaa tgatgacaat aaaagcagaa ttctactaac 2580 tacacggctc aagtatgttg ctgattatgt caacagtcct gattttccac ctcatagtaa 2640 gtcttttcta agtcttgatg atagttggaa tctattcacc gaaaaagtat tcaacaaaga 2700 tacctgtcct cctcacctag aagaaacagg gaagcatatt gtacaacaat gtcgaggatt 2760 acctctctcg gttgttgtag ttgctggact tgttggaaaa atggacccaa cgcatgacaa 2820 ttgggagaac gttgaggaaa atctgaactc attcattggt actgtatctg aacggtgcca 2880 atcaattctt tctttaagct acaattactt gccccagtat ttgagggctt gttttctcta 2940 tgttggatgt tttcctgaag ataaagagat tgatgtttcc aagttgatta ggctatggat 3000 tgctgagcaa ttcgtaaagg cgagaagcaa taaaattta gaagtggtgg cagaggagta 3060 tctggagaag ttaattgata gaagtctaat tttgagtggt agacaaaggg ctaatggaag 3120 gatgaaaact tgtaaaattc atgatcttct tcgccaacta tgcctaagtg aagctcatac 3180 tgaaaatatt agtcatatca tgaatagaaa tgtcctcgtg tcctcagaag ccatagatga 3240 tcaatggcga gtgattgttc cattggaact cgaagagaaa caagtttatc cgacaaggca 3300 tagcagtggt attacaagta caacccgcac ctttatttca atggaaatat gcctaagaga 3360 agctcagacc gaaaccatat atgatcaacg gcgagtgatc cttctctcta aacgacatag 3420 gattgataca atccgcacca ttattccatt cggagagatact tttccaaaag tgatttgttc 3480 cattttttcg cagttgaagt tgcttaaggt gttggatgta ttatcagtct ggtacgatgt 3540 ctcttgtata atacctcagc ttgtacattt gagatatgtt ggtgcagtaa ttttggaagc 3600 tgtttcacta tccaaattga gaaatctaca gaccataatt cttgcaagtg ttgaaaccac 3660 agagttgaag cacccagtag atatctggag aatgtcagag atcagacatt tggatattgt 3720 accgccacta tatatatcaa atcctcttga agcagaacaa cctttgtttc tcaataactt 3780 gcacacgctt tttctccgtt gctctccttt tgttgcgaaa atcataagaa gaactcccaa 3840 tctaaaaaag ctaaagattt tagataaatc taagcatcct gactggcctg atattcttga 3900 ttctctcaat cttctagagg agctggagac actacaaata tcaacagaag aaaacattga 3960 ccggatgatt ttctctgggg atattttcc tcgtaatctc aagcaactga aattatcata 4020 tacttgtata ccatgggaag atatgaaatt gctggctaat ttacccaatc ttgaggtgtt 4080 caagggtcat tatgcattcg atggaacaga ttggaacta gatgaagatg ttgtgttttg 4140 caaattaaaa tgtctacgac tgtatgagcg cggagatctg caaaggtggg aagctgctgg 4200 tagtgataat tttccaatgc ttgagcaact attactgtat ggattcaaaa agctggaaga 4260 gattccggag agtattggag aaataatgac actaaaattc attaatacag aattttgcgg 4320 ctctggtgta gagactagtg caaagaaaat tcaagaagag caagaaagct tgggaaatta 4380 tgagcttcaa cttcaaatta ctcctaaggt atgttgaaac tcaatctttg attaatactc 4440 tcccccagct agacttttca aagtaatatt tcacacgtac aatctccact cggtaagatt 4500 taggattgaa attgtcttct tacatttatg atcatcaccc ttcggttttt tttatttttt 4560 tttagtaaat agcttcatat actctacaaa agttacacat aattaacaat tgacaattga 4620 ttctctctta ttgataattt gtcattttct ctttcagtta agatttttga tcgaaaattt 4680 agacgaaaca agcatgttgc catcgaatga agtaatcgaa aaaatcaaag gtttgtctat 4740 ttaattcctt ttattacttc attgaaagag gttcagatat tttaaacaac tttttaacct 4800 ttctatttga attttttatc gctcagatag atacaacatt tattttgacg tcaaattgtt 4860 attttgattt aatcttattc tttttgtata tagtttccac tcggttcttc attttttttt 4920 caattatagt ttaatttttt ttaattgcac atggttgatt gatttaattt gtgagattca 4980 cttttctttc gattttagga gttcttcggg attaatgtta ttcagattta ggagtattat 5040 atttcaactt tcggttagat ttaggattca aattgtcttc ttacatttat gatcatcatc 5100 cttttgatgt tatttatttc ttttagtaaa tagcttcaaa aactctata 5149 <210> 14 <211> 919 <212> PRT <213> Solanum photeinocarpum <400> 14 Met Ala Tyr Ala Ala Leu Ser Ser Leu Ile Tyr Thr Leu Gln Gln Leu 1 5 10 15 Leu Lys Pro Asn Gln Ser Leu Val Cys Arg Ser Cys Thr Gln Gln His 20 25 30 Leu Gln Ser Ile Tyr His Asn Leu Ser Ala Leu Gln Leu Phe Leu Asp 35 40 45 Asp Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr Leu 50 55 60 Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser Ser 65 70 75 80 Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr Lys 85 90 95 Ala Glu Asp Lys Val Asp Ser Ser Leu Arg Asn Ile Ile Leu Ala Asp 100 105 110 Cys Thr Glu Asn Arg Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu Leu 115 120 125 Leu Lys Val Glu Lys Asp Val Asp Ser Leu Arg Lys Glu Val Met Gln 130 135 140 Ile Glu Phe Asn Lys His Gly Ser Arg Ser Ala Glu Leu Thr Thr Ile 145 150 155 160 Leu Pro Ser Pro Glu Lys Ser Thr Ile Glu Glu His Thr Ile Val Gly 165 170 175 Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln Thr 180 185 190 Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly Lys 195 200 205 Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser Arg 210 215 220 Phe Asp Arg His Val Trp Val Thr Thr Ser Glu Glu Phe Asn Glu Arg 225 230 235 240 Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn Gln 245 250 255 Glu Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Gly Leu Lys 260 265 270 Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Gln Ala 275 280 285 Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Asn Lys Ser Arg 290 295 300 Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Asn Ser 305 310 315 320 Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp Ser 325 330 335 Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Lys Asp Thr Cys Pro Pro 340 345 350 His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Arg Gly Leu 355 360 365 Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp Pro 370 375 380 Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe Ile 385 390 395 400 Gly Thr Val Ser Glu Arg Cys Gln Ser Ile Leu Ser Leu Ser Tyr Asn 405 410 415 Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Cys Phe 420 425 430 Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp Ile 435 440 445 Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Asn Leu Glu Val Val 450 455 460 Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu Ser 465 470 475 480 Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His Asp 485 490 495 Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile Ser 500 505 510 His Ile Met Asn Arg Asn Val Leu Val Ser Ser Glu Ala Ile Asp Asp 515 520 525 Gln Trp Arg Val Ile Val Pro Leu Glu Leu Glu Glu Lys Gln Val Tyr 530 535 540 Pro Thr Arg His Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe Ile 545 550 555 560 Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Thr Ile Tyr Asp 565 570 575 Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr Ile 580 585 590 Arg Thr Ile Ile Pro Phe Gly Asp Thr Phe Pro Lys Val Ile Cys Ser 595 600 605 Ile Phe Ser Gln Leu Lys Leu Leu Lys Val Leu Asp Val Leu Ser Val 610 615 620 Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg Tyr 625 630 635 640 Val Gly Ala Val Ile Leu Glu Ala Val Ser Leu Ser Lys Leu Arg Asn 645 650 655 Leu Gln Thr Ile Ile Leu Ala Ser Val Glu Thr Thr Glu Leu Lys His 660 665 670 Pro Val Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile Val 675 680 685 Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu Phe 690 695 700 Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val Ala 705 710 715 720 Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu Asp 725 730 735 Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn Leu 740 745 750 Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile Asp 755 760 765 Arg Met Ile Phe Ser Gly Asp Ile Phe Pro Arg Asn Leu Lys Gln Leu 770 775 780 Lys Leu Ser Tyr Thr Cys Ile Pro Trp Glu Asp Met Lys Leu Leu Ala 785 790 795 800 Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asp Gly 805 810 815 Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys Cys 820 825 830 Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala Gly 835 840 845 Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Leu Leu Tyr Gly Phe Lys 850 855 860 Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu Lys 865 870 875 880 Phe Ile Asn Thr Glu Phe Cys Gly Ser Gly Val Glu Thr Ser Ala Lys 885 890 895 Lys Ile Gln Glu Glu Gln Glu Ser Leu Gly Asn Tyr Glu Leu Gln Leu 900 905 910 Gln Ile Thr Pro Lys Val Cys 915 <210> 15 <211> 2757 <212> DNA <213> Solanum photeinocarpum <400> 15 atggcatatg ctgctctttc ttcacttatc tatacattgc aacaactctt gaaacctaat 60 caatctttgg tttgtcgaag ctgtacacaa caacatcttc aatctatcta tcacaatctt 120 tctgctctgc aacttttcct tgacgatact acgacaaagg atattgaaac tcttaaggta 180 tgtaattatc taatctctta ctcatcttat atttattcaa taataattaa tttatcgagt 240 ttcaatttta aggttataga aaagaggatc agagatgtag tatacaaagc agaagataaa 300 gttgattcaa gcctaagaaa catcatacta gcagattgca cagagaatag agaaggggct 360 tgtaaattct ttgaggaaga attgctaaaa gtggaaaag atgttgattc tctcaggaaa 420 gaggtgatgc agatcgagtt glasscat ggtagcagat ctgcagact aacaacatt 480 cttccctctc cagaaaaaag tacaattgag gaacatacta tgttgggat ggaggatgag 540 tacaacca tacttgatcg cctcactgcc caacagacg agttgactgt cataccaatt 600 ttggtatgg gcggtatagg tagacaact ctagccagaa aggtttag tgatcatct 660 attcgttctc gatttgatag acatgtatgg gtcactacct ctgaagaatt caatgagaga 720 cgaatgcttc tcgaagttgt ttctcaatt actactggaa gcaatcaaga aaagagcgat 780 gatcaactaa tggattgt gtatagaggt cttaagggta ggagatttct aattgtcata 840 gatgatattt ggagtactca ggcttgggac caatgcaa gatatttcc aaatgatgac 900 aaaaagca gattctact aactacacgg ctcaagtag ttgctgatta tgtcacagt 960 cctgattttc cacctcatag taagtctttt ctaagtcttg atgatagttg gatctattc 1020 accgaaaaag tattcaacaa agatacctgt cctcctcacc tagaagaaac agggaagcat 1080 attgtacaac aatgtcgagg attacctctc tcggttgttg tagttgctgg acttgttgga 1140 aaaatggacc caacgcatga caattgggag aacgttgagg aaaatctgaa ctcattcatt 1200 ggtactgtat ctgaacggtg ccaatcaatt ctttctttaa gctacaatta cttgccccag 1260 tatttgaggg cttgttttct ctatgttgga tgttttcctg aagataaaga gattgatgtt 1320 tccaagttga ttaggctatg gattgctgag caattcgtaa aggcgagaag caataaaaat 1380 ttagaagtgg tggcagagga gtatctggaa gagttaattg atagaagtct aattttgagt 1440 ggtagacaaa gggctaatgg aggatgaaa acttgtaaaa ttcatgatct tcttcgccaa 1500 ctatgcctaa gtgaagctca tactgaaaat attagtcata tcatgaatag aaatgtcctc 1560 gtgtcctcag aagccataga tgatcaatgg cgagtgattg ttccattgga actcgaagag 1620 aaacaagttt atccgacaag gcatagcagt ggtattacaa gtacaacccg cacctttat 1680 tcaatggaaa tatgcctaag agaagctcag accgaaacca tatatgatca acggcgagtg 1740 atccttctct ctaaacgaca taggattgat acaatccgca ccattattcc attcggagat 1800 acttttccaa aagtgatttg ttccattttt tcgcagttga agttgcttaa ggtgttggat 1860 gtattacag tctggtacga tgtctcttgt ataatacctc agcttgtaca tttgagatat 1920 gttggtgcag taattttgga agctgtttca ctatccaaat tgagaaatct acagaccata 1980 attcttgcaa gtgttgaaac cacagagttg aagcacccag tagatatctg gagaatgtca 2040 gagatcagac atttggatat tgtaccgcca ctatatatat caaatcctct tgaagcagaa 2100 caacctttgt ttctcaataa cttgcacacg cttttctcc gttgctctcc ttttgttgcg 2160 aaaatcataa gaagaactcc caatctaaaa aagctaaaga ttttagataa atctaagcat 2220 cctgactggc ctgatattct tgattctctc aatcttctag aggagctgga gacactacaa 2280 atatcaacag aagaaaacat tgaccggatg atttctctg gggatatttt ccctcgtaat 2340 ctcaagcaac tgaaattatc atatacttgt ataccatggg aagatatgaa attgctggct 2400 aatttaccca atcttgaggt gttcaagggt cattatgcat tcgatggaac agattggaaa 2460 ctagatgaag atgttgtgtt ttgcaaatta aaatgtctac gactgtatga gcgcggagat 2520 ctgcaaaggt gggaagctgc tggtagtgat aattttccaa tgcttgagca actattactg 2580 tatggattca aaaagctgga agagattccg gagagtattg gagaaataat gacactaaaa 2640 ttcattaata cagaattttg cggctctggt gtagagacta gtgcaaagaa aattcaagaa 2700 gagcaagaaa gcttgggaaa ttatgagctt caacttcaaa ttactcctaa ggtatgt 2757 <210> 16 <211> 5608 <212> DNA <213> Solanum photeinocarpum <400> 16 aagttggaag cttaactcat ttggctacct catatagttt acccatattt ttacccatga 60 atactcatat ttctatgggt tgaatatggg tataaaccca tattttaccc atctaaaaaa 120 tcactcacca acccattaaa atatgaacag attgggcggg ttaccaaaat atgggctctt 180 tttgccacca ctaacaacaa caaagagcaa agcaaagcac aactttgaaa tttgaaaaat 240 gatgcaagac aaatatcaaa attagacagt aaaattcaca attcccctta aaataaatga 300 taaattaaag aagactaaac aatagaaaac cttacaatct caaaggcact atggatcgga 360 gtcgtacctg tgtgtttttt atcgtacaat attccacaca gtattattaa ttttttaagg 420 accatttggc catgtgatat gaaattatga gatgaagttg aagttttatt tggacatgca 480 atttgaattt tttatgttgt atgttttctt ataaacataa aaaacccata agttgtaaaa 540 ttattaaact tgtctcattt ttttattcaa ttttaccaaa taaataaaaa tttacaaaat 600 cacataatat gctaacacaa aactattctt taaaaaatac aatatttatt gatcaaactt 660 taattcaaca aaaaataaaa ttcaacataa gttgtagtgt actagtcttt aatataattc 720 tcccacatag tacggacaat ctcctcacat tgaacttgca tttctcgatc atatgatcga 780 gtagacaaac caacattgct actttgagct ataaatggtt taaaaagtaa tgatatgaat 840 tataaatttt atttacatat gaaacaaatg gtaagtagat ataaatgtgg ggttgttttt 900 ataaaatata aacttatggg tcaatttttg tatttgaaaa atctcaaatc atgatttgaa 960 atttttaaat catgattttt gaagaatttg ggatttcatc tcatgatatg aaatcatgag 1020 atgaaatcag tctgaaattg catgtccaaa tgctgatttc atctcatgat ttcatatcgt 1080 gatatgaat aaatcgtgat atgaatcgc atgtccaac gtctactaaa taacttagac 1140 atgtcttgac ctttatat tactccctcc gtcctaattt acttgtcaa tattttctaa 1200 tttgattccc cttttacttg tcattttt caatcaaga acgacaatttttttctt 1260 cctattatac cctcaattta ttaacattga attaatgtcc ttgaaaaata tagaagtaa 1320 atagtttaa aactctatca aattaatagg gataaaatgg taattcatt ataccaatta 1380 ttattttctt atagatgcg tcaatcaa attgacaa taaaaaaaga cgaaagaaaa 1440 tttgtattga aagttaattg tattcgtcca cttgacacat catgtcttaa ttattcaca 1500 agtttgaatc caatgtatat gatcataaa tgtgagacat cttactatc gtggtggggc 1560 caatgtttga aaagtgttga cgaaaactac aagagtgtct ttattagaa tcaggacaac 1620 ttgactttttttcatt tctttttct tacttgact tggtcaatcc atgttttgc 1680 atccataaac cacaagttct gtttagatat taaatagaaa attgtccaaa ttcatttaga 1740 aaaatgtgga cataaatcat ttagacaaa cctcttagcc taagcagaga cattctctt 1800 ccaggcaaca aaagagaat ggcatatgct gctctttctt cacttact tacattgca 1860 caaccttga aacctaatca atctttggtt tgtcgaagct gtacacaca acatcttchca 1920 tctatctatc acatctttc tgctctgcaa cttttccttg acgatactac gandaaggat 1980 attgaaactc ttaggtag taattatcta atctcttact catcttatat ttattcaata 2040 atattaattt tatcgagttt cattttag gttatagaaa agagatcag agatgtagta 2100 tacaagcag agatacagt tgattcaagc ctaagaaca tcattctagc agattgcaca 2160 gagaatagag aaggggttg taaatttttt gaggagaat tgctaaagt ggaaaaagat 2220 gttgattctc tcagcaaga ggtgatgcag atcgacttta acagcatgg aagcagatct 2280 gcagaattag ggcaactga tccctcctca tcaggaaaaa gtacaattga ggaacatact 2340 attgttggga tggatga gtacaacacc atacttgatc gcctcactgc ccaacagac 2400 gagttgactg tcataccaat ttttgtatg ggcggtatag gtaagacaac tcttgccaga 2460 aaggtttatg atgattcatc tattcgttct cgatttgata gatagcatg ggtcactacc 2520 tctgaagaat tcaatgagag acgaatgctt ctcgaagttg tttcttcaat tactactgga 2580 agcaatcaag gaaagagcga tgatcaacta atggagattg tgtatagaag tctgaagggt 2640 aggagatttc taattgtcat agatgatatt tggagtactc aggcttggga ccaaatgcaa 2700 agaatttc caaatgatga caataaaagc cgaattctac taactacacg gctcaagtat 2760 gttgctgatt atgtcagcag tcctgatttt ccacctcata gtaagtcttt tctaagtctt 2820 gatgatagtt ggaatctatt caccgaaaaa gtattcaacg aagatacctg tcctcctcac 2880 ctagaagaaa cagggaagca tattgtacaa caatgtcaag gattacctct ctcggttgtt 2940 gtcgttgctg gacttgttgg aaaaatggac ccaacgcatg acaattggga gaatgttgag 3000 gaaaatctga actcattctt tggtactgta tccgaacggt gccactcaat tctttctttg 3060 agctacaatt acttgcccca atattgagg gcttgttttc tctatgttgg aggttttcct 3120 gaagataaag agattgatgt ttccaagttg attaggctat ggattgctga gcaattcgta 3180 aaggcgagaa gcaataaaag gctagaagtg gtggcagagg agtatctgga agagttaatt 3240 gatagaagtc taattttgag tggtagacaa agggctaatg gaaggatgaa aacttgcaaa 3300 attcatgatc ttcttcgcca actatgccta agtgaagctc atactgaaaa tattagtcat 3360 atcatgaata gaaatgtccc cgtgtcctca gaagccatag atgatcaacg gcgagtgatt 3420 gttccattgg aactcgaaga gaaacaagtt tatcctacaa ggcatagcag tggtattaca 3480 agtacaaccc gcacctttat ttcaatggaa atatgcctaa gagaagctca aaccgaagcc 3540 atatatgatc aacggcgagt gatccttctg tctaaacgac ataggattga tacaatccgc 3600 accattattc cattcggaga tacttttcca aaagagattt gttccatttt ttcagagttc 3660 aagttgctta aggtgttgga tgtattatca gtctggtacg atgtctcttg tataatacct 3720 cagcttgtac atttgagata tgttggtgca gtaatttttgg aagctctttc actacccaaa 3780 ttgagaaatc tacagaccat aatgcttaca agtgttgaaa ccacagagtt gaagcactca 3840 ctagatatct ggagaatgtc agagataaga catttggata ttgtaccgcc actatatata 3900 tcaaatcctc ttgaagcaga acaacctttg tttctcaata acttgcacac gctttttctc 3960 cgttgctctc cttttgttgc gaaaatcata agaagaactc cgaatctaaa aaagctaaag 4020 attttagata aatctaagca tcctgactgg cctgatattc ttgattctct caatcttcta 4080 gagagctgg agacactaca aatatcaaca gaaaaaca ttgaccggat gattttctct 4140 ggggataatt tccctcgtaa tctcaagcaa ctgaaattat caggtactaa aataccatgg 4200 gaagatatga aattgctggc taatttaccc aatcttgagg tgttcaaggg tcattatgca 4260 4320 4380 atgcttgagc aactagtact gtatgggttc gaaaaactgg aagagattcc ggagagtatt 4440 ggagaataa tgaactaaa attcatcaaa agaatttt gcggctctgg tgtagagaca 4500 agtgcaaaga aaattcaaga agagcaagaa agctggggaa attatgagct tcaacttcta 4560 attactccta tggtatgtta aaactccatc tttgataatt agactcaact atcaactttt 4620 ttcacagaga gtcaattctt tcacaagaag tcactcaact atcaattttt ttcacaaaaa 4680 gtcactcaac tttgattctt tcatagaaag ttactcaact atgggctttt tgcatagaaa 4740 accactcaac ctatttaatt atattttca tatgaaattt ttatttctaa tcaaaatttt 4800 aaaatccaaa tatatatcca tttaaaccat ttaatgaatc acccacacta aatccgatcc 4860 gctaaaaata tatatttac cttttgtttt atccttattc ccctaaatat tctctcctgt 4920 ttcctaaata ttctctcctg tttcctgatt cttctcttc tgcgttttcc ccctcaattt 4980 cagtctcttc ttttacgtat atatcaacct ctctttctcc ataagcatca tattattgtt 5040 ttctcaactt caaaaacaat ataataca acaatctctt gatttacaat ttcaataaaa 5100 aattctatgc acaccatgct tttatcaatt attatgaaaa ctgataaaat taacaaacaa 5160 ggctctcaat ctgcataact acgaatctac gcctatagtt gctctttaat tgcaattatt 5220 cattcaataa aatcaagtga tcaggatagt cctaaatcta gatatagttt gagaaatcga 5280 tggaaaattc ctagttaata gagattttaa aattgagaaa ttaaaaaaaa tgcagtgaat 5340 caggagacgg gagagaatat ttagggagat aaggataaaa caaaagtcaa tattatattt 5400 ttagcggatc gaatttagtg gggtgattca ttaaatggtt taaatggaaa tatatttaga 5460 ttttaaaatt ttgattaaaa ataaaaattt catatgaaaa atataattaa ataggttgag 5520 tgactttcta tgcaaaagcc ccatagttga gtgactttct gtgaaaaaaa ttgatagttg 5580 agtgactttc tgtgaaagta tcaaagtt 5608 <210> 17 <211> 920 <212> PRT <213> Solanum photeinocarpum <400> 17 Met Ala Tyr Ala Ala Leu Ser Ser Leu Ile Tyr Thr Leu Gln Gln Leu 1 5 10 15 Leu Lys Pro Asn Gln Ser Leu Val Cys Arg Ser Cys Thr Gln Gln His 20 25 30 Leu Gln Ser Ile Tyr His Asn Leu Ser Ala Leu Gln Leu Phe Leu Asp 35 40 45 Asp Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr Leu 50 55 60 Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser Ser 65 70 75 80 Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr Lys 85 90 95 Ala Glu Asp Thr Val Asp Ser Ser Leu Arg Asn Ile Ile Leu Ala Asp 100 105 110 Cys Thr Glu Asn Arg Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu Leu 115 120 125 Leu Lys Val Glu Lys Asp Val Asp Ser Leu Ser Lys Glu Val Met Gln 130 135 140 Ile Asp Phe Asn Lys His Gly Ser Arg Ser Ala Glu Leu Ala Thr Thr 145 150 155 160 Asp Pro Ser Ser Ser Gly Lys Ser Thr Ile Glu Glu His Thr Ile Val 165 170 175 Gly Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln 180 185 190 Thr Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly 195 200 205 Lys Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser 210 215 220 Arg Phe Asp Arg His Ala Trp Val Thr Thr Ser Glu Glu Phe Asn Glu 225 230 235 240 Arg Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn 245 250 255 Gln Gly Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Ser Leu 260 265 270 Lys Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Gln 275 280 285 Ala Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Asn Lys Ser 290 295 300 Arg Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Ser 305 310 315 320 Ser Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp 325 330 335 Ser Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Glu Asp Thr Cys Pro 340 345 350 Pro His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Gln Gly 355 360 365 Leu Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp 370 375 380 Pro Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe 385 390 395 400 Phe Gly Thr Val Ser Glu Arg Cys His Ser Ile Leu Ser Leu Ser Tyr 405 410 415 Asn Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Gly 420 425 430 Phe Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp 435 440 445 Ile Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Arg Leu Glu Val 450 455 460 Val Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu 465 470 475 480 Ser Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His 485 490 495 Asp Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile 500 505 510 Ser His Ile Met Asn Arg Asn Val Pro Val Ser Ser Glu Ala Ile Asp 515 520 525 Asp Gln Arg Arg Val Ile Val Pro Leu Glu Leu Glu Glu Lys Gln Val 530 535 540 Tyr Pro Thr Arg His Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe 545 550 555 560 Ile Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Ala Ile Tyr 565 570 575 Asp Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr 580 585 590 Ile Arg Thr Ile Ile Pro Phe Gly Asp Thr Phe Pro Lys Glu Ile Cys 595 600 605 Ser Ile Phe Ser Glu Phe Lys Leu Leu Lys Val Leu Asp Val Leu Ser 610 615 620 Val Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg 625 630 635 640 Tyr Val Gly Ala Val Ile Leu Glu Ala Leu Ser Leu Pro Lys Leu Arg 645 650 655 Asn Leu Gln Thr Ile Met Leu Thr Ser Val Glu Thr Thr Glu Leu Lys 660 665 670 His Ser Leu Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile 675 680 685 Val Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu 690 695 700 Phe Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val 705 710 715 720 Ala Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu 725 730 735 Asp Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn 740 745 750 Leu Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile 755 760 765 Asp Arg Met Ile Phe Ser Gly Asp Asn Phe Pro Arg Asn Leu Lys Gln 770 775 780 Leu Lys Leu Ser Gly Thr Lys Ile Pro Trp Glu Asp Met Lys Leu Leu 785 790 795 800 Ala Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asp 805 810 815 Gly Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys 820 825 830 Cys Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala 835 840 845 Gly Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Val Leu Tyr Gly Phe 850 855 860 Glu Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu 865 870 875 880 Lys Phe Ile Lys Thr Glu Phe Cys Gly Ser Gly Val Glu Thr Ser Ala 885 890 895 Lys Lys Ile Gln Glu Glu Gln Glu Ser Trp Gly Asn Tyr Glu Leu Gln 900 905 910 Leu Leu Ile Thr Pro Met Val Cys 915 920 <210> 18 <211> 2760 <212> DNA <213> Solanum photeinocarpum <400> 18 atggcatatg ctgctctttc ttcacttatc tatacattgc aacaactctt gaaacctaat 60 caatctttgg tttgtcgaag ctgtacacaa caacatcttc aatctatcta tcacaatctt 120 tctgctctgc aacttttcct tgacgatact acgacaaagg atattgaaac tcttaaggta 180 tgtaattatc taatctctta ctcatcttat atttattcaa taataattaa tttatcgagt 240 ttcaatttta aggttataga aaagaggatc agagatgtag tatacaaagc agaagataca 300 gttgattcaa gcctaagaaa catcattcta gcagattgca cagagaatag agaaggggct 360 tgtaaattct ttgaggaaga attgctaaaa gtggaaaaag atgttgattc tctcagcaaa 420 gaggtgatgc agatcgactt taacaagcat ggaagcagat ctgcagaatt agcaacaact 480 gatccctcct catcaggaaa aagtacaatt gaggaacata ctattgttgg gatggaggat 540 gagtacaaca ccatacttga tcgcctcact gcccaaacag acgagttgac tgtcatacca 600 atttttggta tgggcggtat aggtaagaca actcttgcca gaaaggttta tgatgattca 660 tctattcgtt ctcgatttga tagacatgca tgggtcacta cctctgaaga attcaatgag 720 agacgaatgc ttctcgaagt tgtttcttca attactactg gaagcaatca aggaaagagc 780 gatgatcaac taatggagat tgtgtataga agtctgaagg gtaggagatt tctaattgtc 840 atagatgata tttggagtac tcaggcttgg gaccaaatgc aaagaatatt tccaaatgat 900 gacaataaaa gccgaattct actaactaca cggctcaagt atgttgctga tttgtcagc 960 agtcctgatt ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta 1020 ttcaccgaaa aagtattcaa cgaagatacc tgtcctcctc acctagaaga aacagggaag 1080 catattgtac aacaatgtca aggattacct ctctcggttg ttgtcgttgc tggacttgtt 1140 ggaaaaatgg acccaacgca tgacaattgg gagaatgttg aggaaaatct gaactcattc 1200 tttggtactg tatccgaacg gtgccactca attctttctt tgagctacaa ttacttgccc 1260 1320 gtttccaagt tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa 1380 1440 agtggtagac aaagggctaa tggaaggtg aaaacttgca aaattcatga tcttcttcgc 1500 1560 cccgtgtcct cagaagccat agatgatcaa cggcgagtga ttgttccatt ggaactcgaa 1620 1680 atttcaatgg aaatatgcct aagagaagct caaaccgaag ccatatatga tcaacggcga 1740 gtgatccttt tgtctaaacg acataggatt gatacaatcc gcaccattat tccattcgga 1800 gatacttttc caaagagat ttgttccatt ttttcagagt tcaagttgct taaggtgttg 1860 gatgtattat cagtctggta cgatgtctct tgtataatac ctcagcttgt acatttgaga 1920 tatgttggtg cagtaatttt ggaagctctt tcactaccca aattgagaaa tctacagacc 1980 2040 2100 gaacaacctt tgtttctcaa taacttgcac acgctttttc tccgttgctc tccttttgtt 2160 gcgaaaatca taagagaac tccgaatcta aaaaagctaa agattttaga taaatctaag 2220 catcctgact ggcctgatat tcttgattct ctcaatcttc tagaggagct ggagacacta 2280 caaatatcaa cagaagaaaa cattgaccgg atgattttct ctggggataa tttccctcgt 2340 aatctcaagc aactgaaatt atcaggtact aaaataccat gggaagatat gaaattgctg 2400 gctaatttac ccaatcttga ggtgttcaag ggtcattatg cattcgatgg aacagattgg 2460 aaactagatg aagatgttgt gttttgcaaa ttaaaatgtc tacgactgta tgagcgcgga 2520 2580 ctgtatgggt tcgaaaaact ggaagagatt ccggagagta ttggagaaat aatgacacta 2640 aaattcatca aaacagaatt ttgcggctct ggtgtagaga caagtgcaaa gaaaattcaa 2700 gaagagcaag aaagctgggg aaattatgag cttcaacttc taattactcc tatggtatgt 2760 <210> 19 <211> 6006 <212> DNA <213> Solanum photeinocarpum <400> 19 attcacattc taaaggctat aaatatatcc ttaagtttcc tttgtaaaac ataacacaca 60 aatacaattc aggagatcgt gagaagaaga aagtatttag tgaggttttt gtagtgagaa 120 attcttggtg taaattctct ataattctat tcttgtgaaa tagagttgtt tttcctccca 180 aatatttttc atagtgatca gagaataaca acaaatgtgt ctacggactg ttaaaaaata 240 atggcataag tgggccctct ttgtaaacga taatggcatg ggtgagctca atatttagcg 300 gatgatataa atgaaatctt tttcaaagtt tgattactta tttaagtttt tttcttttta 360 tttaattttt aattcgatgt ccaatatgca caaccaaatt tccactacaa ccaaaaaaaa 420 aaaaaagaac gctgtgtaac atcatctcca accaacctgg gtcatatgag tcccaaatgc 480 atttacagca acaaagagca aagcaaagca cgactttgaa atttgaaaaa tgatgcaaga caaatcat tcacaattcc ccttaaata aatgataaat taaagaagac taaacaatag aaaaccttac aatctcaaag gcagtatgga tcggagtcgt atttggaaga aattttgata 660 caacttgaag aagattgaag attgttgttc father ctgtgggcgg gttaatctac aacctgtgtg ttttttatca tacaaaattc cacacagtat tattaattc atgcaatttg gactttttat gttgtatgtt ttcttataaa cataagttgt aaaattatta aagttgtccc 840 atttttttat tcaattttac 900. 900. atttttttat tcaattttac acaaaactat tctttaaaaa attacking tattgatcaa actttaattc aacaaaaaat 1020. aaaattcaac ataagttgta gtagttgtag tgtactagtc tttaatata ttctcccaca fathercggac aatctcctca cgttgaactt gcatttctcg atcatatgat tgagtagaca aaccaacatt gctactttga gctaaatg gtttaaaaag taatgatatg aattataat tttatttaca tatgaaacaa atgataagta aatataaatg tggggttgtt tttataaaat ataaacttat gggtcaattt ttgtatttga aaaatcccaa attatgattt gaaattttca 1260 aatcatgatt tttgaagaat tcagtctgaa atcgcatgtc caaatgctga tttcatctca 1320 tgatttcata tcgtgatatg aaatcgcatg tccaaacgcc tactaagtaa cttagacacg 1380 tcttgacctt ttataattat ccctccatcc taatttactt gtcaaatatt ttctaatttg 1440 attccctttt tacttgtcat tttttacaaa tcaagaaacg acaatttttt tttcttccta 1500 ttataccctc aatttattaa cattgaatta atgtccttga aaaatatagt aagtaaatat 1560 gtttaaaact ctatcaaatt aataggggta aaataataaa ctcatattat taattattat 1620 tttcttaata gatgcgtcaa atcaaaaatc gacaaagtaa tataaatttc aaagaaaatt 1680 tgtatcgaaa gttaattgta ttcgtcaagt ttgaatccaa tgtatatgaa tgtttgaaaa 1740 gtcttgacga aaactacaag agtgtcttta ttagaaatca ggacaacttg actctttatt 1800 tttcatttct tttttctcac tttgacttgg tcaatccata gttttgcatc cataaaccac 1860 aagttctgtt tagattata atagaaaatt gtccaaattt atttagaaaa atgtggacat 1920 aaatcattta vakaaaacct cttatagcct aagcagagac atttctctc cagcaaaca 1980 aagagaaatg gcttatgctg ctctttctctc acttatgtat acatgcaac aactcttgaa 2040 acctaatcaa tctttcgttt gtcgatactc tacacaaa cttgttcaat ctctctatca 2100 aaatcttact gctctgcaac ttttccttga ccatactacg aaaggata ttgaacact 2160 taaggtagt aattatctaa tctcttactc atcttatatt tattcataa taattaattt 2220 atcgagtttc aattttaagg tttagaaaa gaggatcaga gatgtagtat aaagcaga 2280 agataagtt gattcaagcc windows assistctagca gattgcag agatagaga 2340 aggggcttgt aaatttcttg aggagaatt gctaaagtg gaaaagatg ttgattctt 2400 cagcaagg gtgatgcaga tcgagtttta cagcatgga tgcagatctg cagaattagc 2460 aacaactgat cccctcctcat caggaaaag tacaattgag gaacatacta ttgttgggat 2520 ggaggatgag tacacacca tacttgatcg cctcactgcc caacagacg agttgactgt 2580 cataccaattt ttggtatgg gcggtatagg taagacaact cttgccagaa aggtttatga 2640 2700 caatgagaga cgaatgcttc tcgaagttgt ttctcaatt actactggaa gcaatcaagg 2760 aaagagcgat gatcaactaa tggattgt gtatagaagt ctgaagggta ggagatttct 2820 aattgtcata gatgatattt ggagtactca ggcttgggac caatgcaa gatatttcc 2880 aaatgatgac agtaaagcc gattctact aactacacgg ctcaagtag ttgctgatta 2940 tgtcagcagt cctgattttc cacctacatag taagtctttt ctaagtcttg atgatagttg 3000 gatctattc accgaaaag tattcacga agatacctgt cctcctcacc tagagaac 3060 agggaagcat attgtacaac atgtcagg attackcctc tcggttgttg tcgttgctgg 3120 acttgttgga aaatggacc caacgcatga caatgggag atgttgagg aaatctgaa 3180 ctcattcttt gctactgtat ccgaacggtg ccactcatt ctttctttga gctacaatta 3240 cttgccccaa tatttgaggg cttgttttct ctagttgga ggttttcctg aagataaga 3300 gattgatgtt tccaagttga ttaggctg gattgctgag cattcgtaa aggcgagaag 3360 caataaaagg ctagaagtgg tggcagagga gtatctggaa gagttaattg atagaagtct 3420 aattttgagt ggtagacaaa gggctaatgg aaggatgaaa acttgcaaaa ttcatgatct 3480 tcttcgccaa ctatgcctaa gtgaagctca tactgaaaat attagtcata tcatgaatag 3540 aaatgtcccc gtgtcctcag aagccataga tgatcaacgg cgagtgattg ttccattgga 3600 actcgaagag aaacaatttt atcctacaag gcatagcagt ggtattacaa gtacaacccg 3660 caccttatt tcaatggaaa tatgcctaag agaagctcaa accgaagcca tatatgatca 3720 acggcgagtg atccttctgt ctaaacgaca taggattgat acaatccgca ccattattcc 3780 attcggagat acttttccaa aagagatttg ttccattttt tcagagttca agttgcttaa 3840 ggtgttggat gtattatcag tctggtacga tgtctcttgt ataatacctc agcttgtaca 3900 tttgagatat gttggtgcag taattttgga agctctttca ctacccaaat tgagaaatct 3960 acagaccata atgcttacaa gtgttgaaac cacagagttg aagcactcac tagatatctg 4020 gagaatgtca gagataagac atttggatat tgtaccgcca ctatatatat caaatcctct 4080 tgaagcagaa caacctttgt ttctcaataa cttgcacacg ctttttctcc gttgctctcc 4140 ttttgttgcg aaaatcataa gaagaactcc gaatctaaaa aagctaaaga ttttagataa 4200 atctaagcat cctgactggc ctgatattct tgattctctc aatcttctag aggagctgga 4260 gacactacaa atatcaacag aagaaaacat tgaccggatg attttctctg gggatatttt 4320 ccctcgtaat ctcaagcaac tgaaattatc atatacttgt ataccatggg aagatatgaa 4380 attgctggct aatttaccca atcttgaggt gttcaagggt cattatgcat tcgatggaac 4440 agattggaaa ctagatgaag atgttgtgtt ttgcaaatta aaatgtctac gactgtatga 4500 gcgcggagat ctgcaaaggt gggaagcagc aggtagtgat aattttccta tgcttgagca 4560 actagtactg tatgggttcg aaaaactgga agagattccg gagagtattg gagaataat 4620 gacactaaaa ttcattaaaa cagaattttg cggctctggt gtagagacta gtgcaaagaa 4680 aattcaagaa gagcaagaaa gcttgggaaa ttatgagctt caacttcaaa ttactcctaa 4740 ggtatgttga aactcaatct ttgattaata ctctccccca gctagacttt tcaaagtaat 4800 atttcacacg tacaatctcc actcggtaag atttaggatt gaaattgtct tcttacattt 4860 atgatcatca cccttcggtt ttttttatttt ttttttagta aatagtttca tatactctac 4920 aaaagttaca cataattaac aattgacaat tgattctctc ttattgataa tttgtcattt 4980 tctctttcag ttaagatttt tgatcgaaaa tttagacgaa acaagcatgt tgccatcgaa 5040 tgaagtaatc gaaaaaatca aaggtttgtc tatttaattc cttttattac ttcattgaaa 5100 gaggttcaga tattttaaac aactttttaa cctttctatt tgaatttttt atcgctcaga 5160 tagatacaac atttattttg acgtcaaatt gttattttga tttaatctta ttctttttgt 5220 atatagttc cactcggttc ttcatttttt tttcaattat agttaattt ttttaattg 5280 cacatggttg attgatttaa tttgtgagat tcactttct ttcgatttta ggagttcttc 5340 gggattaatg ttattcagat ttaggagtat tatatttcaa ctttcggtta gatttaggat 5400 tcaaattgtc ttcttacatt tatgatcatc atccttttga tgttatttat ttcttttagt 5460 aatagcttc aaaaactcta tagaaattac atataattaa caattgacaa ttgattctct 5520 cttattgata atttctcatg tttctctttc aattaaaatt tttgatcgaa aactaaaaga 5580 gattccggag agtactggac aaataatgac acgaaaaatc atcaaaacag aattttgcag 5640 ctctggtgta gagactagtg caaagaaaat tcaagaagag caagaaagct tgggaaatta 5700 tgagcttcaa gttcaaatta ctcctagggt atgttaaact ccttctttga ttattagcca 5760 attctttaat ttcataatat ttctagacta agagcctgtc tggatggact taaaaaaaat 5820 aacttataag ttgaaaactg tttataagtc aaaaaaaata agtaggtcta ccctaactta 5880 tttttttttg acttataagt tgttttcaac ttttaagctg ttttaaataa gctaagtcaa 5940 atagatccaa ttattttttg ggcttatttt aagcacaaaa tgactttaag ttggccagcc 6000 aaatac 6006 <210> 20 <211> 920 <212> PRT <213> Solanum photeinocarpum <400> 20 Met Ala Tyr Ala Ala Leu Ser Ser Leu Met Tyr Thr Leu Gln Gln Leu 1 5 10 15 Leu Lys Pro Asn Gln Ser Phe Val Cys Arg Tyr Ser Thr Gln Gln Leu 20 25 30 Val Gln Ser Leu Tyr Gln Asn Leu Thr Ala Leu Gln Leu Phe Leu Asp 35 40 45 His Thr Thr Thr Lys Asp Ile Glu Thr Leu Lys Val Cys Asn Tyr Leu 50 55 60 Ile Ser Tyr Ser Ser Tyr Ile Tyr Ser Ile Ile Ile Asn Leu Ser Ser 65 70 75 80 Phe Asn Phe Lys Val Ile Glu Lys Arg Ile Arg Asp Val Val Tyr Lys 85 90 95 Ala Glu Asp Lys Val Asp Ser Ser Leu Arg Asn Ile Ile Leu Ala Asp 100 105 110 Cys Thr Glu Asn Arg Glu Gly Ala Cys Lys Phe Phe Glu Glu Glu Leu 115 120 125 Leu Lys Val Glu Lys Asp Val Asp Ser Leu Ser Lys Glu Val Met Gln 130 135 140 Ile Glu Phe Asn Lys His Gly Cys Arg Ser Ala Glu Leu Ala Thr Thr 145 150 155 160 Asp Pro Ser Ser Ser Gly Lys Ser Thr Ile Glu Glu His Thr Ile Val 165 170 175 Gly Met Glu Asp Glu Tyr Asn Thr Ile Leu Asp Arg Leu Thr Ala Gln 180 185 190 Thr Asp Glu Leu Thr Val Ile Pro Ile Phe Gly Met Gly Gly Ile Gly 195 200 205 Lys Thr Thr Leu Ala Arg Lys Val Tyr Asp Asp Ser Ser Ile Arg Ser 210 215 220 Arg Phe Asp Arg His Ala Trp Val Thr Thr Ser Glu Glu Phe Asn Glu 225 230 235 240 Arg Arg Met Leu Leu Glu Val Val Ser Ser Ile Thr Thr Gly Ser Asn 245 250 255 Gln Gly Lys Ser Asp Asp Gln Leu Met Glu Ile Val Tyr Arg Ser Leu 260 265 270 Lys Gly Arg Arg Phe Leu Ile Val Ile Asp Asp Ile Trp Ser Thr Gln 275 280 285 Ala Trp Asp Gln Met Gln Arg Ile Phe Pro Asn Asp Asp Ser Lys Ser 290 295 300 Arg Ile Leu Leu Thr Thr Arg Leu Lys Tyr Val Ala Asp Tyr Val Ser 305 310 315 320 Ser Pro Asp Phe Pro Pro His Ser Lys Ser Phe Leu Ser Leu Asp Asp 325 330 335 Ser Trp Asn Leu Phe Thr Glu Lys Val Phe Asn Glu Asp Thr Cys Pro 340 345 350 Pro His Leu Glu Glu Thr Gly Lys His Ile Val Gln Gln Cys Gln Gly 355 360 365 Leu Pro Leu Ser Val Val Val Val Ala Gly Leu Val Gly Lys Met Asp 370 375 380 Pro Thr His Asp Asn Trp Glu Asn Val Glu Glu Asn Leu Asn Ser Phe 385 390 395 400 Phe Gly Thr Val Ser Glu Arg Cys His Ser Ile Leu Ser Leu Ser Tyr 405 410 415 Asn Tyr Leu Pro Gln Tyr Leu Arg Ala Cys Phe Leu Tyr Val Gly Gly 420 425 430 Phe Pro Glu Asp Lys Glu Ile Asp Val Ser Lys Leu Ile Arg Leu Trp 435 440 445 Ile Ala Glu Gln Phe Val Lys Ala Arg Ser Asn Lys Arg Leu Glu Val 450 455 460 Val Ala Glu Glu Tyr Leu Glu Glu Leu Ile Asp Arg Ser Leu Ile Leu 465 470 475 480 Ser Gly Arg Gln Arg Ala Asn Gly Arg Met Lys Thr Cys Lys Ile His 485 490 495 Asp Leu Leu Arg Gln Leu Cys Leu Ser Glu Ala His Thr Glu Asn Ile 500 505 510 Ser His Ile Met Asn Arg Asn Val Pro Val Ser Ser Glu Ala Ile Asp 515 520 525 Asp Gln Arg Arg Val Ile Val Pro Leu Glu Leu Glu Glu Lys Gln Phe 530 535 540 Tyr Pro Thr Arg His Ser Ser Gly Ile Thr Ser Thr Thr Arg Thr Phe 545 550 555 560 Ile Ser Met Glu Ile Cys Leu Arg Glu Ala Gln Thr Glu Ala Ile Tyr 565 570 575 Asp Gln Arg Arg Val Ile Leu Leu Ser Lys Arg His Arg Ile Asp Thr 580 585 590 Ile Arg Thr Ile Ile Pro Phe Gly Asp Thr Phe Pro Lys Glu Ile Cys 595 600 605 Ser Ile Phe Ser Glu Phe Lys Leu Leu Lys Val Leu Asp Val Leu Ser 610 615 620 Val Trp Tyr Asp Val Ser Cys Ile Ile Pro Gln Leu Val His Leu Arg 625 630 635 640 Tyr Val Gly Ala Val Ile Leu Glu Ala Leu Ser Leu Pro Lys Leu Arg 645 650 655 Asn Leu Gln Thr Ile Met Leu Thr Ser Val Glu Thr Thr Glu Leu Lys 660 665 670 His Ser Leu Asp Ile Trp Arg Met Ser Glu Ile Arg His Leu Asp Ile 675 680 685 Val Pro Pro Leu Tyr Ile Ser Asn Pro Leu Glu Ala Glu Gln Pro Leu 690 695 700 Phe Leu Asn Asn Leu His Thr Leu Phe Leu Arg Cys Ser Pro Phe Val 705 710 715 720 Ala Lys Ile Ile Arg Arg Thr Pro Asn Leu Lys Lys Leu Lys Ile Leu 725 730 735 Asp Lys Ser Lys His Pro Asp Trp Pro Asp Ile Leu Asp Ser Leu Asn 740 745 750 Leu Leu Glu Glu Leu Glu Thr Leu Gln Ile Ser Thr Glu Glu Asn Ile 755 760 765 Asp Arg Met Ile Phe Ser Gly Asp Ile Phe Pro Arg Asn Leu Lys Gln 770 775 780 Leu Lys Leu Ser Tyr Thr Cys Ile Pro Trp Glu Asp Met Lys Leu Leu 785 790 795 800 Ala Asn Leu Pro Asn Leu Glu Val Phe Lys Gly His Tyr Ala Phe Asp 805 810 815 Gly Thr Asp Trp Lys Leu Asp Glu Asp Val Val Phe Cys Lys Leu Lys 820 825 830 Cys Leu Arg Leu Tyr Glu Arg Gly Asp Leu Gln Arg Trp Glu Ala Ala 835 840 845 Gly Ser Asp Asn Phe Pro Met Leu Glu Gln Leu Val Leu Tyr Gly Phe 850 855 860 Glu Lys Leu Glu Glu Ile Pro Glu Ser Ile Gly Glu Ile Met Thr Leu 865 870 875 880 Lys Phe Ile Lys Thr Glu Phe Cys Gly Ser Gly Val Glu Thr Ser Ala 885 890 895 Lys Lys Ile Gln Glu Glu Gln Glu Ser Leu Gly Asn Tyr Glu Leu Gln 900 905 910 Leu Gln Ile Thr Pro Lys Val Cys 915 920 <210> 21 <211> 2760 <212> DNA <213> Solanum photeinocarpum <400> 21 atggcttatg ctgctctttc ttcacttatg tatacattgc aacaactctt gaaacctaat 60 caatctttcg tttgtcgata ctctacacaa caacttgttc aatctctcta tcaaaatctt 120 actgctctgc aacttttcct tgaccatact acgacaagg atttgaac acttaaggta 180 tgtaattatc taatctctta ctcatcttat atttattca taatatta tattacgagt 240 ttcaattta aggttataga aagaggatc agagatgtag tatacaagc agagataaa 300 gttgattcaa gcctaagaaa catcattcta gcagattgca cagagaatag agaaggggct 360 tgtaaattct ttgaggaaga attgctaaaa gtggaaaag atgttgattc tctcagcaaa 420 gaggtgatgc agatcgagtt window ggatgcagat ctgcagaatt window 480 gatccctcct catcaggaaa agtacaatt gaggaacata ctattgttgg gatgaggat 540 gagtacaaca ccatacttga tcgcctcact gcccaacag acgagttgac tgtcatacca 600 atttttggta tgggcggtat aggtaagaca actctgcca gaaaggttta tgatgattca 660 tctattcgtt ctcgatttga tagacatgca tggtcacta cctctgaaga attcaatgag 720 agacgaatgc ttctcgaagt tgttctctca attackacg gaagcaatca aggaagagc 780 gatgatcaac taatggagat tgtgtataga agtctgaagg gtaggatt tctaattgtc 840 atagatgata tttggagtac tcaggcttgg gaccaaatgc aaagaatatt tccaaatgat 900 gacagtaaaa gccgaattct actaactaca cggctcaagt atgttgctga tttgtcagc 960 agtcctgatt ttccacctca tagtaagtct tttctaagtc ttgatgatag ttggaatcta 1020 ttcaccgaaa aagtattcaa cgaagatacc tgtcctcctc acctagaaga aacagggaag 1080 catattgtac aacaatgtca aggattacct ctctcggttg ttgtcgttgc tggacttgtt 1140 ggaaaaatgg acccaacgca tgacaattgg gagaatgttg aggaaaatct gaactcattc 1200 tttggtactg tatccgaacg gtgccactca attctttctt tgagctacaa ttacttgccc 1260 caatatttga gggcttgttt tctctatgtt ggaggttttc ctgaagataa agagattgat 1320 gttccaagt tgattaggct atggattgct gagcaattcg taaaggcgag aagcaataaa 1380 aggctagaag tggtggcaga ggagtatctg gaagagttaa ttgatagaag tctaattttg 1440 agtggtagac aaagggctaa tggaaggatg aaaacttgca aaattcatga tcttcttcgc 1500 caactatgcc taagtgaagc tcatactgaa aatattagtc atatcatgaa tagaaatgtc 1560 ...

Claims

1. A polynucleotide, the nucleotide sequence of which is selected from the group consisting of: (a) the nucleotide sequence shown in SEQ ID NO: 1, 13, 16, 19, 31, 32, 33, 34 or 35; (b) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 14, 17, 20, 41, 42, 43, 44 or 45; and (c) the nucleotide sequence shown in SEQ ID NO: 3, 15, 18, 21, 36, 37, 38, 39 or 40.

2. The polynucleotide of claim 1, wherein the polynucleotide is an isolated nucleic acid molecule.

3. An expression cassette comprising the polynucleotide of claim 1 or 2 and an operably linked heterologous promoter.

4. A vector comprising the polynucleotide of claim 1 or 2 or the expression cassette of claim 3.

5. The vector of claim 4, further comprising an additional R gene.

6. A method for enhancing resistance of a potato, tomato or tobacco plant to plant diseases caused by Phytophthora infestans isolate 88069, the method comprising modifying at least one potato, tomato or tobacco plant cell to contain a heterologous polynucleotide having a nucleotide sequence selected from the group consisting of: (a) the nucleotide sequence shown in SEQ ID NO: 1, 13, 16, 19, 31, 32, 33, 34 or 35; (b) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 14, 17, 20, 41, 42, 43, 44 or 45; and (c) the nucleotide sequence shown in SEQ ID NO: 3, 15, 18, 21, 36, 37, 38, 39 or 40.

7. The method of claim 6, wherein the heterologous polynucleotide is stably integrated into the genome of the plant cell.

8. The method of claim 6, wherein the plant cell is regenerated into a plant comprising the heterologous polynucleotide in its genome.

9. The method of claim 6, wherein modifying at least one plant cell to contain a heterologous polynucleotide comprises introducing the heterologous polynucleotide into at least one plant cell.

10. The method of any one of claims 6 to 9, wherein the heterologous polynucleotide consists of the nucleotide sequence of any one of (b) to (c), and further comprises a promoter operably linked thereto for expression in a plant.

11. The method of claim 10, wherein the promoter is selected from the group consisting of a pathogen-inducible promoter, a constitutive promoter, a tissue-preferred promoter, a wound-inducible promoter, and a chemically-regulated promoter.

12. The method of any one of claims 6-8, wherein modifying at least one plant cell to comprise a heterologous polynucleotide comprises using genome editing to modify the nucleotide sequence of a native or non-native gene in the genome of the plant cell to comprise the nucleotide sequence of any one of (a)-(c).

13. The method of any one of claims 6 to 9, wherein the plant is a potato or tomato plant.

14. A method for limiting plant diseases caused by Phytophthora infestans isolate 88069 in crop production, the method comprising planting seedlings, tubers or seeds of potato, tomato or tobacco plants produced according to the method of any one of claims 6 to 13, and cultivating the seedlings, tubers or seeds under conditions conducive to the growth and development of plants produced therefrom, wherein the seedlings, tubers or seeds contain the polynucleotide, expression cassette or vector.

15. The method of claim 14, further comprising harvesting at least one fruit, tuber, leaf and / or seed from the plant.

16. A method for identifying a potato, tomato or tobacco plant showing newly conferred or enhanced resistance to plant diseases caused by Phytophthora infestans isolate 88069, the method comprising detecting in the plant or in at least one part or cell thereof the presence of an Rpi-amr nucleotide sequence, wherein the Rpi-amr nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence shown in SEQ ID NO: 1, 13, 16, 19, 31, 32, 33, 34 or 35; (b) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 14, 17, 20, 41, 42, 43, 44 or 45; and (c) the nucleotide sequence shown in SEQ ID NO: 3, 15, 18, 21, 36, 37, 38, 39 or 40.

17. The method of claim 16, wherein the plant is a potato or tomato plant.

18. The method of claim 16, wherein the presence of the Rpi-amr nucleotide sequence is detected by detecting at least one marker within the Rpi-amr nucleotide sequence.

19. The method of claim 16, wherein the Rpi-amr nucleotide sequence consists of the nucleotide sequence shown in SEQ ID NO: 1, 3, 13, 15, 16, 18, 19, 21, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40.

20. The method of any one of claims 16-19, wherein detecting the presence of the Rpi-amr nucleotide sequence comprises a member selected from PCR amplification, nucleic acid sequencing, nucleic acid hybridization, and an immunological assay for detecting the R protein encoded by the Rpi-amr nucleotide sequence.

21. A method for introducing at least one Rpi-amr gene into a potato, tomato or tobacco plant, Wherein the at least one Rpi-amr gene consists of a nucleotide sequence selected from the group consisting of: (a) the nucleotide sequence shown in SEQ ID NO: 1, 13, 16, 19, 31, 32, 33, 34 or 35; (b) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 2, 14, 17, 20, 41, 42, 43, 44 or 45; and (c) the nucleotide sequence shown in SEQ ID NO: 3, 15, 18, 21, 36, 37, 38, 39 or 40; The method comprises: (i) crossing a first plant comprising at least one copy of at least one Rpi-amr gene in its genome with a second plant lacking the at least one Rpi-amr gene in its genome, thereby producing at least one progeny plant; and (ii) selecting at least one progeny plant comprising in its genome the at least one Rpi-amr gene.

22. The method of claim 21, wherein the first plant is a Solanum americanum plant and the second plant is not a Solanum americanum plant, or wherein the first plant is a Solanum nigrescens plant and the second plant is not a Solanum nigrescens plant.

23. The method of claim 21, wherein the second plant is a potato (Solanum tuberosum) plant or a tomato (Solanum lycopersicum) plant.

24. The method of claim 21, wherein selecting at least one progeny plant comprises detecting the presence of the Rpi-amr nucleotide sequence in a progeny plant or at least a part or cell thereof using a method according to any one of claims 16-20.

25. The method of any one of claims 21-24, further comprising (i) backcrossing at least one selected progeny plant of (b) with a Solanaceae plant that is of the same species and genotype as the second Solanaceae plant, or that is of the same species as the second Solanaceae plant and lacks the at least one Rpi-amr gene in its genome, wherein at least one progeny plant results from the backcross; and (ii) selecting at least one progeny plant resulting from the backcross of (i) that comprises the at least one Rpi-amr gene in its genome.

26. Use of a plant produced by the method of any one of claims 6 to 13 and 21 to 25, or a plant identified by the method of any one of claims 16 to 20 showing newly conferred or enhanced resistance to plant diseases caused by Phytophthora infestans isolate 88069, or its fruits, tubers, leaves or seeds in agriculture.

27. A polypeptide, whose amino acid sequence is selected from the group consisting of: (a) the amino acid sequence shown in SEQ ID NO: 2, 14, 17, 20, 41, 42, 43, 44 or 45; and (b) an amino acid sequence encoded by the nucleotide sequence shown in SEQ ID NO: 1, 3, 13, 15, 16, 18, 19, 21, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40.

Images