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Late Blight Resistance Gene From Solanum Americanum And Methods Of Use

A transgenic plant, resistance technology, applied in the direction of genetic engineering, biochemical equipment and methods, introduction of foreign genetic material using vectors, etc., can solve problems such as the inability to perform map-based cloning

Active Publication Date: 2018-02-16
TWO BLADES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Although traditional map-based cloning has been used to isolate resistance (R) genes from plants, many plant genomes carry large chromosomal segments that cannot be subjected to traditional map-based cloning due to recombination arrest (Gaut et al. (2007) NatureRev.Genet.8:77-84), and Solanaceae plants are no exception

Method used

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  • Late Blight Resistance Gene From Solanum Americanum And Methods Of Use
  • Late Blight Resistance Gene From Solanum Americanum And Methods Of Use
  • Late Blight Resistance Gene From Solanum Americanum And Methods Of Use

Examples

Experimental program
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Effect test

Embodiment 1

[0187] Example 1: Solanum nigrum is a rich source of late blight resistance genes

[0188] We set out to study the immune response to Phytophthora infestans in a panel of Solanum nigrum (2n) and Solanum nigrum (6n) lines obtained from three European seed collections (Table 1). Flow cytometry analysis identified lines 944750095, A54750014 and A14750006 originally listed as Solanum nigrum as diploid rather than hexaploid (data not shown). This is consistent with a previous report of frequent misidentifications between Solanum nigrum and Solanum nigrum (Manoko et al. (2007) Plant Syst. Evol. 267:1-11). For the purposes of the present invention, unless otherwise stated or apparent from the text of the examples, all plant lines will be assigned to the broadly defined Solanum nigrum group. It is furthermore recognized that the present invention is also not dependent on the R gene of the present invention being isolated from a particular plant species or even being said species of t...

Embodiment 2

[0195] Example 2: RenSeq gene mapping shows that Rpi-amr3 is located near R2 on potato chromosome 4

[0196] Inoculation of the pathogen on the leaves of young F2 plants (F1 954750186x 944750095) revealed 99 resistant and 6 susceptible plants (15:1 segregation), indicating the presence of two unlinked dominant Rpi genes. To segregate the gene, we self-pollinated 15 disease-resistant F2 plants and determined resistance segregation patterns in 30-200 F3 progeny. In the DLA test on leaves from young plants (8-10 weeks old), the four populations segregated 3:1. Interestingly, this result was inconsistent with that in older plants, suggesting the presence of additional resistance genes in adult plants (greater than 12 weeks old), which were assessed as susceptible when young.

[0197] We hypothesized that the underlying late blight resistance genes encode NB-LRR / NLR proteins. We applied RenSeq to the resistant (R) and susceptible (S) parents, and to pooled DNA from the 50 most su...

Embodiment 3

[0200] Example 3: Combination of RenSeq and PacBio sequencing enables assembly of full-length sequences of 14 co-segregated full-length NLR genes

[0201] One of the main motivations of this project is to establish an R gene cloning method that does not require the construction of a BAC library. We previously found that the high copy number and sequence similarity of NLR genes complicates de novo assembly of short Illumina RenSeq reads (Jupe et al (2013) Plant J. 76:530-544; Andolfo et al (2014) BMC Plant Biol. 14:120). We therefore developed NLR enrichment combined with the longer read technology provided by PacBio RSII (Eid et al. (2008) Science 323:133-138). We used our Solanum NLR bait library (Jupe et al. (2013) Plant J. 76:530-544) from two independent libraries (1.5 kb and 2.5 kb gDNA fragments) derived from the Rpi-amr3-carrying parental line 944750095 The complete NLR complement was captured (see Methods for details). The 1.5kb and 2.5kb fractions were sequenced on...

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Abstract

Compositions and methods and for enhancing the resistance of plants to a plant disease caused by a Phytophthora species are provided. The compositions comprise nucleic acid molecules encoding resistance (R) gene products and variants thereof and plants, seeds, and plant cells comprising such nucleic acid molecules. The methods for enhancing the resistance of a plant to a plant disease caused by aPhytophthora species comprise introducing a nucleic acid molecule encoding an R gene product into a plant cell. Additionally provided are methods for using the plants in agriculture to limit plant disease.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Patent Application No. 62 / 159,240, filed May 9, 2015, which is hereby incorporated by reference in its entirety. [0003] References to Sequence Listings submitted as text files [0004] The official text of the sequence listing is submitted electronically via EFS-Web as a sequence listing in ASCII format, file named 070294-0095.TXT, created on April 22, 2016, and has a size of 27.4 kilobytes, and submitted at the same time as this manual. The sequence listing contained in this document in ASCII format is part of this specification and is hereby incorporated by reference in its entirety. field of invention [0005] The present invention relates to the fields of gene separation and plant improvement, in particular to enhancing the resistance of plants to plant diseases by using disease resistance genes. Background of the invention [0006] Late blight, caused by t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
CPCC12N15/8261Y02A40/146A23K10/30A23L19/00A23V2002/00C12N15/8282C12Q1/6895C12Q2600/13C12Q2600/158
Inventor K·威特克J·琼斯
Owner TWO BLADES FOUND
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