Lettuce plant

Abstract

To provide a new variety of lettuce.SOLUTION: A lettuce plant seed according to the present invention is deposited with a lettuce plant identified by either accession number FERM P-22453 or accession number FERM P-22454, or with a progeny line thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to lettuce plants. [Background technology]

[0002] Lettuce plants prefer cool climates. In recent years, they have been subject to diseases such as root rot and downy mildew, as well as poor growth, which are thought to be caused by high temperatures and humidity due to continuous cultivation and abnormal weather, and producers are looking for varieties that can be produced stably. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] M. FUJINAGA, “Studies on physiological races and phylogenetic analysis of lettuce root rot pathogen, Fusarium oxysporum f. sp. Lactucae”, Journal of General Plant Pathology volume 71, page457 (2005) [Non-Patent Document 2] T. SHIMIZU et.al., “Development Trend of Diseases on Lettuce (Lactuca sativa) of Organic Cultivation in Highlands of Nagano Prefecture and Their Control”, Annual report of the Kanto-Tosan Plant Protection Society 64:2017.12 p.41-46 Summary of the Invention [Problem to be solved by the invention]

[0004] Therefore, an object of the present invention is to provide a new lettuce plant. [Means for solving the problem]

[0005] To achieve the above objectives, a seed of the lettuce plant of the present invention has been deposited, which is a lettuce plant identified by accession number FERM P-22453 or accession number FERM P-22454 or a progeny thereof.

[0006] Lettuce plants of the present invention include the lettuce plants identified by accession number FERM P-22453 or accession number FERM P-22454.

[0007] The method for producing a lettuce plant of the present invention includes a self-pollination step of self-pollinating the lettuce plant of the present invention.

[0008] The method for producing a lettuce plant of the present invention includes a crossbreeding step of crossbreeding the lettuce plant of the present invention with another lettuce plant. Effect of the Invention

[0009] According to the present invention, a new lettuce plant can be provided. [Brief description of the drawings]

[0010] [Figure 1] FIG. 1 is a schematic diagram showing examples of leaf top overlap, leaf posture, and leaf lobe number of lettuce plant stems. [Diagram 2] FIG. 2 is a schematic diagram showing examples of leaf shapes of lettuce plants. [Diagram 3] FIG. 3 is a schematic diagram showing examples of the shape of the tip of a leaf of a lettuce plant, a longitudinal section of a leaf, and the size of the irregularities of the leaf. [Figure 4] FIG. 4 is a schematic diagram showing examples of the leaf marginal lobe type, leaf marginal lobe depth, and leaf vein type of lettuce plants. [Diagram 5] FIG. 5 is a schematic diagram showing examples of the size of a bulb of a lettuce plant and the shape of the bulb in longitudinal section. [Figure 6] FIG. 6 is a schematic diagram showing an example of axillary bud development and fasciation development in lettuce plants.

BEST MODE FOR CARRYING OUT THE INVENTION

[0011] <Lettuce plant> The lettuce plant of the present invention includes a lettuce plant identified (deposited) by accession number FERM P-22453 or accession number FERM P-22454 (hereinafter collectively also referred to as the "deposited strain") or its progeny strain. The lettuce plant of the present invention is characterized by including a lettuce plant identified by accession number FERM P-22453 or accession number FERM P-22454 or its progeny strain, and other configurations and conditions are not particularly limited.

[0012] In the present invention, the "lettuce plant" is classified into the genus Lactuca of the family Asteraceae, the species Lactuca sativa L., with the scientific name Lactuca sativa and the Japanese name chisha (lettuce, Lactuca, chisa). The lettuce plant may be, for example, a hybrid with a related species or a wild species.

[0013] In the present invention, the "cultivated lettuce plant", "cultivated lettuce variety", or "cultivated lettuce" is a lettuce plant or its variety, breeding line, or cultivated variety that is cultivated by humans and is excellent in cultivation science. The "cultivated lettuce plant", "cultivated lettuce variety", or "cultivated lettuce" may be their hybrids or hybrids with other lettuce species.

[0014] In the present invention, the "plant" means a plant individual indicating the whole plant.

[0015] In the present invention, the term "plant part" refers to a part of an individual plant. Examples of the "plant part" include plant cells, plant protoplasts, plant cell cultures or tissue cultures capable of regenerating plants, plant callus, plant clumps, plant cells isolated from plants or plant parts, leaves, pollen, embryos, cotyledons, hypocotyls, roots, root tips (root tips), anthers, pistils, flowers, ovaries, ovules, seeds, fruits, stems, seedlings, etc. Examples of the part of the individual plant include organs, tissues, cells, vegetative propagules, etc., and any of these may be used. Examples of the organs include petals, corollas, flowers, leaves, seeds, fruits, stems, roots, etc. Examples of the tissues are parts of the organs. Specific examples of the part of the plant individual include microspores, flowers, flower buds, pistils, anthers, pollen, ovaries, embryos, ovules, hypocotyls, embryo sacs, egg cells, cuttings, roots, root tips, stems, stalks, leaves, petioles, leaf pith, cotyledons, cells, meristematic cells, protoplasts, and seeds. The pollen may be mature or immature pollen. The part of the plant individual may be derived from any growth stage of a plant, for example, before rooting, after rooting, seedlings, cuttings, mature individuals, and the like. The part of the plant individual may be, for example, one type of organ, tissue, and / or cell, or two or more types of organs, tissues, and / or cells.

[0016] In the present invention, "root rot (lettuce root rot)" is a disease caused by a filamentous fungus. Examples of the pathogenic fungus causing the root rot include Fusarium oxysporum f. sp. Lactucae. The "race" refers to a fungus lineage with different pathogenicity, more specifically, a lineage that shows different pathogenicity against varieties with different resistance genes or resistance loci.

[0017] In the present invention, "downy mildew (lettuce downy mildew)" refers to a disease caused by a filamentous fungus. The pathogenic fungus of the downy mildew is, for example, Bremia lactea ( Bremia lactucaeThe term "race" refers to a bacterial strain having a different pathogenicity, more specifically, a strain exhibiting a different pathogenicity against varieties having different resistance genes or resistance gene loci.

[0018] In the present invention, "resistance" is also referred to as, for example, "disease resistance." The resistance means, for example, the ability to inhibit or suppress the occurrence and progression of a disease caused by infection with a pathogen, and specifically may mean, for example, the absence of disease, the halting of the progression of an already occurring disease, and the suppression (also referred to as "inhibition") of the progression of an already occurring disease.

[0019] In the present invention, the "root rot resistance" can be evaluated according to the measurement methods of the trait numbers 59 and 60 described below. The "downy mildew resistance" can be evaluated according to the measurement method of the trait number 52 described below.

[0020] <Deposited strain> An example of the lettuce plant of the present invention is a lettuce plant identified by accession number FERM P-22453 or accession number FERM P-22454, or a progeny thereof. Hereinafter, accession number FERM P-22453 and accession number FERM P-22454 are also referred to as lettuce cultivars Takii32 and Takii33, respectively. Information regarding the deposit of each cultivar is shown below.

[0021] (Takii32) Type of deposit: Domestic deposit Name of depository institution: National Institute of Technology and Evaluation, Patent Biological Deposit Center Address: Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818 Accession number: FERM P-22453 Identification mark: Takii32 Received: May 23, 2022

[0022] (Takii33) Type of deposit: Domestic deposit Name of depository institution: National Institute of Technology and Evaluation, Patent Biological Deposit Center Address: Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818 Accession number: FERM P-22454 Identification mark: Takii33 Received: May 23, 2022

[0023] The deposited line exhibits, for example, the morphological and physiological characteristics described in Table 1 below. In Table 1 below, the morphological and physiological characteristics are based on a trial production in Japan in 2022. In Table 1 below, the morphological and physiological characteristics are evaluated based on the Lettuce Species Examination Criteria (issued in June 2021, http: / / www.hinshu2.maff.go.jp / info / sinsakijun / kijun / 1361.pdf) issued by the Ministry of Agriculture, Forestry and Fisheries of Japan (MAFF). In addition, in Table 1 below, the morphological and physiological characteristics are evaluated based on the criteria described below. The morphological and physiological characteristics can be seen in Figures 1 to 6. In addition, the evaluation of the morphological and physiological characteristics may refer to the DUS test guidelines in UPOV. As the DUS test guidelines, for example, the guidelines published on April 5, 2017 (https: / / www.upov.int / edocs / tgdocs / en / tg013.pdf) can be referred to.

[0024] [Table 1A]

[0025] [Table 1B]

[0026] [Table 1C]

[0027] (Trait number 1) The "seed color" can be evaluated by visual observation. The "seed color" can be evaluated based on the following criteria: Class 1 (white, standard variety: Olympia), Class 2 (yellow), Class 3 (brown, standard variety: Cisco V lettuce), and Class 4 (black, Logic).

[0028] (Trait number 2) The "cotyledon size" means the length from the base to the tip of the cotyledon when fully expanded x the maximum width (cm), and can be evaluated by measuring it. The "cotyledon size" can be evaluated based on class 3 (small, standard variety: Romance), class 5 (medium, standard variety: Olympia), and class 7 (large).

[0029] (Trait number 3) The "cotyledon shape" refers to the shape of the cotyledon at the fully expanded stage, and can be evaluated by visual observation. The "cotyledon shape" can be evaluated based on the following criteria: Class 1 (broad ellipse, standard variety: Calmar), Class 2 (ellipse, standard variety: Olympia), and Class 3 (narrow ellipse, standard variety: Black Seeded Simson).

[0030] (Trait number 4) The "plant width" refers to the maximum diameter (cm) of the plant, and can be evaluated by measurement. The "plant width" can be evaluated based on the following criteria: Class 1 (very small), Class 3 (small, standard variety: Okayama salad greens), Class 5 (medium, standard variety: Olympia), Class 7 (large), and Class 9 (very large).

[0031] (Trait number 5) The "overlap of the upper leaves of the plant" refers to the degree of overlap of the upper leaves of the plant (degree of head formation) and can be evaluated by visual observation. The "overlap of the upper leaves of the plant" can be evaluated based on the following criteria: Class 1 (none or weak, standard variety: Prizehead), Class 2 (medium, standard variety: Costa Rica No. 4), and Class 3 (strong, standard varieties: Olympia and Vanguard 75).

[0032] (Trait number 7) The "leaf posture" refers to the posture of the leaf and can be evaluated by visual observation. The "leaf posture" can be evaluated based on the following criteria: Class 1 (upright, standard variety: Costa Rica No. 4), Class 3 (obliquely upward, standard varieties: Olympia and Cisco), and Class 5 (horizontal).

[0033] (Trait number 8) The "number of leaf lobes" refers to the number of leaf lobes, and can be evaluated by visual observation. The "number of leaf lobes" can be evaluated based on the following criteria: class 1 (none or very small, standard variety: Olympia), class 3 (few), class 5 (medium), class 7 (many), and class 9 (very many).

[0034] (Trait number 9) The "leaf shape" refers to the shape of the leaves of varieties with no or very few leaf lobes (limited to varieties with no or very few leaf lobes), and can be evaluated by visual observation. Class 1 (triangular), Class 2 (lanceolate), Class 3 (oblate, standard variety Olympia), Class 4 (narrow oblate), Class 5 (round, standard variety: The criteria for evaluation are as follows: Class 1 (White Boston), Class 2 (Broad oval), Class 3 (Oval, standard variety: Costa Rica No. 4), Class 4 (Narrow oval, standard variety: Red Kakichisha), Class 5 (Linear), Class 6 (Broad obovate), Class 7 (Oval, standard variety: Prizehead), Class 8 (Narrow ovate, standard variety: Red Kakichisha), Class 9 (Linear), Class 10 (Broad obovate), Class 11 (Oval, standard variety: Prizehead) and Class 12 (Olanceolate).

[0035] (Trait number 10) The "shape of the leaf tip" refers to the shape of the leaf tip of a variety with no or very few leaf lobes (limited to varieties with no or very few leaf lobes), and can be evaluated by visual observation. The evaluation can be performed based on the following criteria: Class 1 (acute, standard variety: Celtas), Class 2 (obtuse), Class 3 (circular), and Class 4 (oblate heart).

[0036] (Trait number 11) The "longitudinal section of the leaf" refers to the shape of the longitudinal section of the leaf of a variety with no or very few leaf lobes (limited to varieties with no or very few leaf lobes), and can be evaluated by visual observation. The evaluation can be performed based on the following criteria: Class 1 (inwardly curved), Class 3 (flat), and Class 5 (outwardly curved).

[0037] (Trait number 13) The "anthocyanin coloration of the leaves" refers to the intensity of the anthocyanin coloration of the leaves, and can be evaluated by visual observation. The "anthocyanin coloration of the leaves" can be evaluated based on the following criteria: Class 1 (none or very weak), Class 3 (weak), Class 5 (medium, standard variety: Prizehead), Class 7 (strong), and Class 9 (very strong).

[0038] (Trait number 16) The "leaf color" refers to the color of the leaves and can be evaluated by visual observation. The "leaf color" can be evaluated based on the standards of grade 1 (green), grade 2 (yellow-green), and grade (gray-green, standard variety: Celtas).

[0039] (Trait number 17) The "green shade of the leaves" refers to the green shade of the leaves and can be evaluated by visual observation. The "green shade of the leaves" can be evaluated based on the following criteria: class 1 (very light), class 3 (light), class 5 (medium), class 7 (dark), and class 9 (very dark).

[0040] (Trait number 18) The "leaf gloss" refers to the gloss of the leaf surface, and can be evaluated by visual observation. The "leaf gloss" can be evaluated based on the following criteria: class 1 (none or weak), class 3 (weak, standard variety: Okayama salad greens), class 5 (medium, standard variety: Olympia), class 7 (strong, standard variety: Nishinabeni), and class 9 (very strong).

[0041] (Trait number 19) The "leaf length" refers to the length of the leaf, and can be evaluated by gently spreading the leaf and measuring it. The "leaf length" can be evaluated based on the following criteria: Class 3 (short, standard variety: Okayama lettuce), Class 5 (medium, standard variety: Prizehead), and Class 7 (long, standard variety: Celtas).

[0042] (Trait number 20) The "leaf width" means the maximum width of the leaf, and can be evaluated by gently spreading the leaf and measuring it. The "leaf width" can be evaluated based on the following criteria: class 3 (narrow, standard variety: Celtas), class 5 (medium, standard variety: Prizehead), and class 7 (wide, standard variety: Vanguard).

[0043] (Trait number 21) The "leaf thickness" refers to the thickness of the leaf and can be evaluated by visual observation. The "leaf thickness" can be evaluated based on the following criteria: Class 1 (very thin), Class 2 (thin, standard variety: Prize Head), Class 3 (medium, standard variety: Olympia), Class 4 (thick), and Class 5 (very thick, Frill Ice).

[0044] (Trait number 22) The "leaf unevenness" refers to the strength of the unevenness (shrinkage) of the leaf surface, and can be evaluated by visual observation. The "leaf unevenness" can be evaluated based on the following criteria: Class 1 (none or weak, standard variety: Cisco), Class 3 (weak, standard variety: Olympia), Class 5 (medium, standard variety: Early Impulse), Class 7 (strong, standard variety: Nishinabeni), and Class 9 (very strong, standard variety: Black Seeded Simson).

[0045] (Trait number 23) The "size of leaf unevenness" refers to the size of the unevenness (shrinkage) on the leaf surface, and can be evaluated by visual observation. The "size of leaf unevenness" can be evaluated based on class 3 (small, standard variety: Black Seeded Simson), class 5 (medium, standard variety: Early Impulse), and class 7 (large, standard variety: Okayama lettuce).

[0046] (Trait number 24) The "waviness of the leaf margin" refers to the strength of the waviness of the leaf margin, and can be evaluated by visual observation. The "waviness of the leaf margin" can be evaluated as grade 1 (none or very weak), grade 3 (weak), grade 5 (medium, standard variety: Mikado Great 3204), grade 7 (strong, standard variety: Calmar, Grand Rapid), and grade 9 (very strong).

[0047] (Trait number 25) The "type of notches in the marginal area of ​​the leaves" refers to the type of notches in the marginal area of ​​the leaves, and can be evaluated by visual observation. The "type of notches in the marginal area of ​​the leaves" can be evaluated based on the following criteria: Class 1 (circular sawtooth), Class 2 (regular dentition), Class 3 (irregular dentition), Class 4 (double dentition), and Class 5 (triple dentition).

[0048] (Trait number 26) The "depth of the incision at the edge of the leaf" refers to the depth of the incision at the edge of the leaf, and can be evaluated by visual observation. The "depth of the incision at the edge of the leaf" can be evaluated based on the following criteria: Class 1 (none or very shallow), Class 3 (shallow), Class 5 (medium, standard variety: Olympia), Class 7 (deep), and Class 9 (very deep).

[0049] (Trait number 27) The "depth of the secondary notches on the leaf margin" means the depth of the secondary notches on the leaf margin of a variety having irregularly dentate, double-dentate or triple-dentate notches on the leaf margin (limited to varieties having irregularly dentate, double-dentate or triple-dentate notches on the leaf margin), and can be evaluated by visual observation. The "depth of secondary incisions at the margin of the leaf" can be evaluated based on the following criteria: class 3 (shallow), class 5 (medium), and class 7 (deep).

[0050] (Trait number 28) The "denseness of the notches at the edge of the leaf" refers to the denseness of the notches at the edge of the leaf, and can be evaluated by visual observation. The "denseness of the notches at the edge of the leaf" can be evaluated based on the following criteria: class 1 (very coarse), class 3 (coarse), class 5 (medium), standard variety: Calmar), class 7 (dense), and class 9 (very dense).

[0051] (Trait number 29) The "vein type" refers to the type of the veins, and can be evaluated by visual observation. The "vein type" can be evaluated based on the following criteria: Class 1 (non-fan-shaped, standard variety: Costa Rica No. 4), Class 2 (slightly fan-shaped, standard variety: Early Impulse), and Class 3 (fan-shaped, standard varieties: Olympia and Cisco).

[0052] (Trait number 30) The "bulb size" refers to the size of the bulb, and means the bulb size (bulb diameter x bulb height) of varieties with medium or strong overlap of the upper leaves of the plant (limited to varieties with medium or strong overlap of the upper leaves of the plant), which can be evaluated by measurement and visual observation. The evaluation can be performed based on the following criteria: Class 1 (very small), Class 3 (small), Class 5 (medium, standard variety: Olympia), Class 7 (large), and Class 9 (very large).

[0053] (Trait number 31) The "shape of the bulb in longitudinal section" refers to the shape of the bulb in longitudinal section, and refers to the shape of the bulb in longitudinal section of a variety with medium or strong overlap of the upper leaves of the plant (limited to varieties with medium or strong overlap of the upper leaves of the plant), and can be evaluated by visual observation. The evaluation can be performed based on the following criteria: Class 1 (narrow oval), Class 2 (wide oval), Class 3 (circular, standard variety: Excel Headgrass), Class 4 (oblate, standard variety: Cisco).

[0054] (Trait number 32) The "bulb firmness" refers to the firmness of the bulb of a variety with medium or strong overlap of the upper leaves of the plant (limited to varieties with medium or strong overlap of the upper leaves of the plant), and can be evaluated by visual observation. It can be evaluated based on the following standards: Class 1 (very loose), Class 2 (fairly loose), Class 3 (loose), Class 4 (slightly loose), Class 5 (medium, standard variety: Cisco), Class 6 (slightly firm), Class 7 (firm, standard variety: Fine), Class 8 (fairly firm, standard variety: Olympia), and Class 9 (very firm).

[0055] (Trait number 33) The "core size" refers to the maximum diameter of the core, and can be evaluated by measurement. It can be evaluated based on the following standards: Class 1 (extra thin), Class 2 (thin), Class 3 (medium, standard variety: Olympia), Class 4 (thick), and Class 5 (extra thick).

[0056] (Trait number 39) The "harvest time" refers to the time of harvest (the number of days from the sowing date) for varieties with medium or strong overlap of the upper leaves of the plant (limited to varieties with medium or strong overlap of the upper leaves of the plant), and can be evaluated by measurement and visual observation. It can be evaluated based on class 1 (very early), class 3 (early), class 5 (medium, standard variety: Olympia), class 7 (late, standard variety: Calmar), and class 9 (very late).

[0057] (Trait number 40) The "initiation of bolting" refers to the time when bolting starts under long-day conditions (the time when 50% of the test plants have bolted is the number of days from the sowing date), and can be evaluated by measurement and visual observation. The "initiation of bolting" can be evaluated based on class 1 (very early, standard variety: Green Leaf), class 3 (early, standard variety: Prize Head), class 5 (medium, standard variety: Gett, Falcon), class 7 (late, standard variety: Olympia), and class 9 (very late).

[0058] (Trait number 41) The "occurrence of fasciation" refers to the degree of fasciation of flower stalks under long-day conditions, and can be evaluated by visual observation. The "occurrence of fasciation" can be evaluated based on the following criteria: rank 1 (none or weak), rank 3 (weak), rank 5 (medium), rank (strong), and rank 9 (very strong).

[0059] (Trait number 42) The "presence or absence of male sterility" refers to the presence or absence of male sterility, and can be evaluated by visual observation. The "presence or absence of male sterility" can be evaluated based on grade 1 (absence) and grade 9 (presence).

[0060] (Trait number 52) The "mildew race Bl:27EU resistance" means the presence or absence of resistance to mildew race Bl:27EU. The "mildew race Bl:27EU resistance" can be evaluated based on grade 1 (absent, standard variety: Colorado) and grade 9 (present, standard varieties: Balesta, Bedford). Mildew race Bl:27EU is available from Naktuinbouw (Sotaweg 22, 2371 GD, Roelofarendsveen, The Netherlands; https: / / www.naktuinbouw.com / ).

[0061] The mildew resistance can be confirmed by the following test method. (Testing method for mildew resistance) -Maintenance of inoculum Downy mildew races are maintained in cultivars with no known or unknown resistance genes, e.g. Cobham Green, Lobjoits Green Cos, Hilde (DM12), Olof, etc. Alternatively, resistant cultivars or breeding lines are used to select specific isolates. Seed purity and quality of these cultivars are important, so seed producers can be contacted directly to produce quality seed, if necessary. · Conducting the test Temperature setting: 15~18℃ Lighting: Provide adequate lighting for the plants to grow well. The seedlings to be tested should be ones with fully developed cotyledons and should not be allowed to blanch. Inoculation concentration: The optimal spore concentration is 1 × 10 5 / ml, but at least 3 × 10 4 When using seedlings as the inoculum source, inoculation should be performed before the first true leaf appears. Reference varieties: Reference varieties carrying the resistance gene under test are included in each trial for checking. These reference varieties are available from GEVES (25 rue Georges Morel, CS 90024, 49071 Beaucouze cedex, France; https: / / www.geves.fr / geves / ) or from Naktuinbouw, the Netherlands. Number of test specimens: 20 or more Criteria for determining symptoms: Symptoms are determined at the following times: 1st time: When spore formation of the susceptible standard variety reaches its maximum (approximately 7 days after inoculation) 2nd vaccination: 3-4 days after the 1st vaccination (approximately 10 days after vaccination) 14 days after the third vaccination (Resistance may be indicated by leaf necrosis at the first evaluation stage.) For details, please refer to the UPOV test guideline (LETTUCE; TG / 13 / 11 Rev.).

[0062] (Trait number 59) The "root rot race 1 resistance" means the strength of resistance to root rot race 1. The "root rot race 1 resistance" can be evaluated based on the following criteria: class 3 (weak, standard varieties: Patriot, late-flowering Red Fire, Waldman's Green), class 5 (medium, standard variety: Salinas 88), and class 7 (strong, standard variety: Costa Rica No. 4).

[0063] The resistance to race 1 of root rot fungus can be confirmed by the following test method. (Testing method for resistance to race 1 of root rot fungus) -Maintenance of inoculum Pathogen: Lettuce root rot race 1 (Fusarium oxysporum f. sp. lactucae (Fol) Race 1) was used. Confirmation of pathogenicity: Confirm pathogenicity using a susceptible variety prior to testing. · Conducting the test Temperature: Keep within the range of approximately 20 to 28°C. Lighting: Natural light or a minimum of 15,000lx with a 14-hour light period. Inoculation preparation: After inoculating the test strain cultured in a nutrient medium (PSA medium, etc.) into the bran medium, mix and culture at approximately 25°C for 10 to 14 days. The medium with sufficient bacteria growth throughout is used as the inoculum source. Inoculation method: Add the inoculum to sterilized soil (or commercially available culture soil) at a volume ratio of 20:1 and mix well. Fill pots with this as the disease soil, and sow the germinated seeds. Test period: The severity of the disease is observed approximately 20 to 30 days after inoculation. Number of test specimens: At least 30 specimens per variety should be tested. Symptoms criteria The percentage of diseased plants above ground and the extent of the disease are investigated, and a disease index is calculated and judged.

[0064] (Trait number 60) The "root rot race 2 resistance" refers to the strength of resistance to root rot race 2. The "root rot race 1 resistance" can be evaluated based on class 3 (weak, standard varieties: Patriot, Salinas 88, Costa Rica No. 4), class 5 (medium), and class 7 (strong, standard varieties: Waldman's Green, late-flowering Red Fire).

[0065] The resistance to race 2 of Root Rot Fungus can be confirmed by the following test method. (Testing method for resistance to race 2 of root rot fungus) -Maintenance of inoculum Pathogen: Lettuce root rot race 2 (Fusarium oxysporum f. sp. lactucae Race 2) is used. Confirmation of pathogenicity: Confirm pathogenicity using a susceptible variety prior to testing. · Conducting the test Temperature: Keep within the range of approximately 20 to 28°C. Lighting: Natural light or a minimum of 15,000lx with a 14-hour light period. Inoculation preparation: After inoculating the test strain cultured in a nutrient medium (PSA medium, etc.) into the bran medium, mix and culture at approximately 25°C for 10 to 14 days. The medium with sufficient bacteria growth throughout is used as the inoculum source. Inoculation method: Add the inoculum to sterilized soil (or commercially available culture soil) at a volume ratio of 20:1 and mix well. Fill pots with this as the disease soil, and sow the germinated seeds. Test period: The severity of the disease is observed approximately 20 to 30 days after inoculation. Number of test specimens: At least 20 specimens per variety should be tested. Symptoms criteria The percentage of diseased plants above ground and the extent of the disease are investigated, and a disease index is calculated and judged.

[0066] In the present invention, a plant having "essentially all physiological and morphological characteristics of the deposited line" means a plant having the major traits of the deposited line (corresponding deposited line) from which the target plant is derived, when grown in the same environment. The major traits are traits (1) to (11) below, i.e., trait numbers 59, 60, 52, 40, 4, 30, 32, 24, 26, 28, and 22 in Table 1. The major traits are preferably traits of trait numbers 59, 60, and 52 in Table 1, i.e., traits (1) to (3) below. The major traits are preferably traits of trait numbers 40, 4, 30, and 32 in Table 1, i.e., traits (4) to (7) below. The main traits are preferably traits of trait numbers 24, 26, 28 and 22 in Table 1, i.e., traits (8) to (11) below. The plant having "essentially all physiological and morphological characteristics of the deposited line" may be a plant having the same traits as the deposited line except for, for example, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 trait, i.e., 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 trait may be traits different from the deposited line. The "traits different from the deposited line" may be the main traits of the deposited line or traits other than the main traits of the deposited line, but traits other than the main traits of the deposited line are preferred. The "traits different from those of the deposited line" can be implemented, for example, by introducing the traits and / or genes described below. In the plant having "essentially all physiological and morphological characteristics of the deposited line", all of the traits of trait numbers 59, 60, 52, 40, 4, 30, 32, 24, 26, 28, and 22 may be the same as those of the deposited line. (1) Strong resistance to race 1 of root rot fungus; (2) Strong resistance to race 2 of root rot fungi; (3) Resistance to downy mildew race Bl:27EU is positive; (4) The start of bolting is in the late period; (5) The width of the stock is "slightly medium"; (6) The size of the ball is "slightly large" or "large"; (7) The ball has a firm or medium firmness; (8) The leaf margins are moderately or somewhat strongly waviness; (9) The depth of the incisions on the margins of the leaves is shallow; (10) The density of the incisions on the edge of the leaf is "slightly medium"; (11) The leaf unevenness is “medium” or “slightly strong.”

[0067] Among the deposited lines, Takii32 has the same or different polymorphisms as the reference lettuce variety "Souther" (variety registration number 10884) for, for example, 14 SSR (simple sequence repeat) markers listed in Table 3A below. In Table 3A below, [Z] indicates the genotype of Souther, a genotype with a larger number of bases than that genotype is indicated as [Z+ (increased number of bases)], and a genotype with a smaller number of bases than that genotype is indicated as [Z- (decreased number of bases)]. For example, the genotype of Takii32 at the SSR markers SML15, SML26, SML22, SML42, SML45, SML60, SML3, LS_WGS_10, TK_39, LS_WGS_15, and TK_11 is "Z", indicating that it is the same genotype as the reference variety Souther. The genotype of Takii32 in TK_37 and TK_20 is "Z+15", which indicates that the genotype has 15 more bases than Souther. The genotype of Takii32 in TK_42 is "Z-12", which indicates that the genotype has 12 fewer bases than Souther. For the analysis using the SSR markers, see, for example, Reference 1 and Table 2 below. Reference 1: Lettuce DNA Variety Identification Manual (Ministry of Agriculture, Forestry and Fisheries website, published March 10, 2016, https: / / www.maff.go.jp / j / kanbo / tizai / brand / b_syokubut / attach / pdf / index-23.pdf)

[0068] [Table 2]

[0069] [Table 3A]

[0070] Among the deposited lines, Takii33 has the same or different polymorphisms as the reference lettuce variety "Souther" (variety registration number 10884) for the 14 SSR markers listed in Table 3B below. In Table 3B below, [S] indicates the genotype of Souther, a genotype with a larger number of bases than that genotype is indicated as [S+ (increased number of bases)], and a genotype with a smaller number of bases than that genotype is indicated as [S- (decreased number of bases)]. For example, the genotype of Takii33 at SSR markers SML22, SML42, SML45, SML60, SML3, LS_WGS_10, LS_WGS_15, and TK_11 is "Z", indicating that it is the same genotype as the reference variety Souther. The genotype of Takii33 in SML_15, SML_26, TK_39, TK_37 and TK_20 is "Z+10" or "Z+15", which indicates that the genotype has 10 or 15 more bases than Souther. The genotype of Takii33 in TK_42 is "Z-12", which indicates that the genotype has 12 fewer bases than Souther. For the analysis using the SSR markers, see, for example, Reference 1 and Table 2 above.

[0071] [Table 3B]

[0072] <Progeny lineage> The lettuce plant of the present invention may be a progeny of the deposited line. The progeny may be an individual plant of the progeny, or a part of an individual plant of the progeny, or a seed of the progeny.

[0073] In the present invention, a "progeny line" or a "progeny lettuce plant" (hereinafter collectively referred to as "progeny line") is a plant obtained from a deposited line of lettuce or its progeny. In the present invention, the progeny line may be a plant obtained by crossing the deposited line with another deposited line or another lettuce plant, or by crossing the deposited line with a wild lettuce plant. The progeny line may be obtained, obtainable, or derived directly or indirectly by selfing and / or cross-pollinating the deposited line or its progeny, or may be derived from a parent line obtained from the deposited line using traditional breeding methods such as selfing and / or cross-pollination. Examples of the progeny line include a selfed progeny line, a first generation hybrid F1 (first generation hybrid line, F1 hybrid), and the like. In obtaining the progeny line, the deposited line may be used as a female parent, a male parent, or both parents.

[0074] In the present invention, "crossing" refers to the crossing of two parental lines. The crossing may be "cross-pollination" or "self-pollination". The cross-pollination refers to fertilization by the union of two gametes originating from different plants. The self-pollination refers to the transfer of pollen from anthers to the stigma of the same plant. The self-pollination may also be referred to as, for example, self-fertilization. The crossing may include backcrossing, which is one of the traditional breeding methods.

[0075] The "backcrossing" is one of the traditional breeding techniques, in which a breeder repeatedly backcrosses a progeny line of a hybrid to one of the parent lines to introduce a trait into a plant or variety. The plant containing the trait to be introduced can be called, for example, a donor plant. The plant into which the trait is introduced can be called, for example, a recurrent parent. The backcrossing can be performed by crossing a donor plant with a recurrent parent, thereby obtaining a first generation hybrid F1 (first generation hybrid line, F1 hybrid). The progeny line having the trait is then crossed with the recurrent parent. Then, by backcrossing and / or selfing for several generations, the trait of the donor plant can be introduced into the recurrent parent.

[0076] In the present invention, the progeny line may be regenerated from a cell culture or tissue culture, a protoplast or a part of a plant individual derived from the deposited line, or may be obtained by self-pollination of the deposited line, or may be obtained by producing seeds from a plant individual of the deposited line.

[0077] In the present invention, the term "regeneration" refers to the generation or vegetative propagation of plants from cell culture, tissue culture or protoplasts.

[0078] The "tissue culture" or "cell culture" may be a composition containing isolated cells of the same or different types, or may be a collection of cells organized into a plant part. Methods for tissue culture of various tissues of lettuce plants and for regenerating plants from said tissue cultures are well known, see, for example, References 2 to 4 below. Reference 2: Pink DA and Carter PJ, “Propagation of lettuce (Lactuca sativa) breeding material by tissue culture”, Ann. Appl. Biol., 1987, vol. 110, pages 611-616 Reference 3: Ampomah-Dwamena C., Conner AJ and Fautrier AG, “Genotypic response of lettuce cotyledons to regeneration in vitro”, Sci. Hort., 1997, vol. 71, pages 137-145 Reference 4: Takayuki Mizutani, Takayuki Tanaka, "Nodal culture of flower stalks of lettuce and closely related wild species Lactuca serriola", Horticultural Research, 2008, Vol. 7, No. 1, pp. 17-21

[0079] The progeny line may have a desired trait. The progeny line may have "essentially all physiological and morphological characteristics of the deposited line" when cultivated under the same cultivation conditions. Specifically, the progeny line may have traits in common with the corresponding deposited line (derived deposited line). As a specific example, the progeny line may have one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, or eleven or more traits that match those of the deposited line. The progeny line may be a plant having the main traits of the deposited line. The main traits include traits of trait numbers 59, 60, and 52 in Table 1C, i.e., the above traits (1) to (3). Moreover, the main traits include traits of trait numbers 40, 4, 30, and 32 in Tables 1A to 1C, i.e., traits (4) to (7) above. Moreover, the main traits include traits of trait numbers 24, 26, 28, and 22 in Table 1B, i.e., traits (8) to (11) above. The progeny line may be a plant having the same traits as the deposited line except for, for example, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 trait, i.e., 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 trait may be different from the deposited line. The "characteristics different from the deposited line" may be the main character of the deposited line or a character other than the main character of the deposited line, but a character other than the main character of the deposited line is preferable. The "characteristics different from the deposited line" can be implemented, for example, by introducing the character and / or gene described below. In the progeny line, all of the character traits of character numbers 1 to 5, 7 to 11, 16 to 33, 39 to 42, 52, 59, and 60 may be the same as those of the deposited line. Examples of the characteristics different from the deposited line include resistance to races other than downy mildew race BI:27EU (Bl16 to 26, 28 to 37, and Bl5 to 11US), bacterial rot disease resistance, corky root disease resistance, lettuce mosaic virus disease resistance, aphid resistance, leafminer resistance, and resistance to browning of cut surfaces.Each trait can be introduced, for example, by crossing with a plant known to have a genetic locus associated with each trait.

[0080] The progeny line may include cells that contain at least one set of chromosomes derived from the deposited line. The progeny line may, for example, have at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of its alleles derived from the deposited line. That is, the progeny line may have at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% genetic complement with the deposited line.

[0081] The "allele" is any one or more genes, all of which are associated with a characteristic or trait in a lettuce plant. In a diploid cell or organism, the pair of alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

[0082] The genetic complementarity can be calculated, for example, by deciphering a molecular marker or a base sequence, comparing it with the molecular marker or base sequence of Takii32 or Takii33, and calculating the coincidence rate. Examples of the molecular marker include SNP (single nucleotide polymorphism) markers, amplified fragment length polymorphism (AFLP) markers, restriction fragment length polymorphism (RFLP) markers, microsatellite markers, sequence-characterized amplified region (SCAR) markers, and cleaved amplified polymorphic sequence (CAPS) markers. Genome analysis methods using the molecular markers are well known and widely published (for example, References 5 and 6 below). The base sequence can be deciphered, for example, by extracting a chromosome from the progeny line and sequencing the chromosome. The proportion of alleles derived from the deposited line and the proportion of genetic complementarity may be estimated, for example, by the number of crosses. In this case, the proportion can be estimated from the number of crosses from the deposited line. As a specific example, when the number of crosses from the deposited line is n, the proportion can be estimated as, for example, (1 / 2)n×100%. Reference 5: Sinchan Adhikari et.al, “Application of molecular markers in plant genome analysis: a review”, The Nucleus, 2017, Volume 60, Issue 3, pp. 283-297 Reference 6: Elcio P. Guimaraes et.al., “MARKER-ASSISTED SELECTION Current status and future perspectives in crops, livestock, forestry and fish”, 2007, Springer, 29-49

[0083] The ratio of alleles and genetic complementarity derived from the deposited line is preferably, for example, an average value of the ratios of a plurality of progeny lines. The plurality is, for example, the number of individuals that can be statistically examined, specifically, 200 or more individuals, and preferably 200 to 1000 individuals.

[0084] The progeny line may have SSR polymorphisms derived from the deposited line. The SSR polymorphisms of the deposited line are those shown in Tables 3A and 3B. For example, the progeny line may have at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen SSR polymorphisms that match those of the deposited line. In the present invention, when the target lettuce variety has, for example, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen SSR polymorphisms that match those of the deposited line, the target lettuce plant can be determined (distinguished, presumed, identified, or differentiated) to be a progeny line of the deposited line.

[0085] The progeny line may have, for example, a mutation or an introduced gene. In this case, the progeny line has, for example, one or more traits modified. The progeny line can be produced, for example, by introducing a mutation or an introduced gene into the deposited line or its progeny line. The mutation may be artificially introduced or naturally introduced. The mutation may be, for example, a chemical substance-induced mutation or a radiation-induced mutation. The mutation may also be introduced, for example, by a molecular biological technique or a genome editing technique (for example, Reference 7 below). The gene may be introduced, for example, by a method using Agrobacterium tumefaciens. Reference 7: Yanfei Mao et.al., “Gene editing in plants: progress and challenges”, National Science Review, 2019, vol. 6, pp. 421-437

[0086] The one or more traits include, for example, resistance to races other than downy mildew race BI:27EU (Bl16-26, 28-37, Bl5-11US), bacterial rot resistance, corky root disease resistance, lettuce mosaic virus resistance, aphid resistance, leafminer resistance, and resistance to browning of cut surfaces.

[0087] The transgene refers to a desired gene introduced into the genome of a plant, for example, by genetic engineering techniques or traditional breeding methods. The transgene may be derived from, for example, the same species or a different species. The transgene may be a gene containing the same base sequence as that of the species of origin or a gene containing a different base sequence. In the latter case, the different base sequence can be prepared, for example, by performing codon optimization, addition of a transcriptional regulator such as a promoter, or the like, on the same base sequence. The transgene may include a translation region and a non-translation region.

[0088] <Haploid and doubled haploid plants> The lettuce plants of the invention may be haploid and / or doubled haploid plants obtained, obtainable or derived from said deposited line. The haploid and / or doubled haploid plants of said deposited line may be used in a method of producing a parent line of said deposited line. In one embodiment, the invention may provide a plant of a haploid and / or doubled haploid plant, a plant part of a haploid and / or doubled haploid plant, or a seed of a haploid and / or doubled haploid plant.

[0089] The doubled haploid plant can be produced by doubling chromosomes in a haploid plant or cell (see, for example, Reference 8 below). As a specific example, haploid pollen or ovules are cultured under specific conditions to form plantlets with 1n chromosomes. The plantlets are then treated with a chemical substance such as colchicine to double the chromosomes. As a result, the cells of the plantlets have 2n chromosomes (doubled haploid). The doubled haploid plant and its progeny can be obtained by growing the plantlets after the treatment. Reference 8: Jim M. Dunwell, “Haploids in flowering plants: origins and exploitation”, Plant Biotechnology Journal, 2010, volume 8, pp. 377-424

[0090] <How to make lettuce plants> As described above, the method for producing a lettuce plant of the present invention includes a crossbreeding step of crossbreeding a first lettuce plant with a second lettuce plant, the first lettuce plant being the lettuce plant of the present invention. The production method of the present invention is characterized in that the lettuce plant of the present invention is used as at least one of the parents in the crossbreeding step, and other steps and conditions are not particularly limited.

[0091] The method for producing a lettuce plant of the present invention further comprises a selfing step of self-pollinating the lettuce plant of the present invention. The production method of the present invention is characterized by self-pollinating the lettuce plant of the present invention, and other steps and conditions are not particularly limited.

[0092] According to the production method of the present invention, progeny lines of the deposited line can be produced. The explanation of the production method of the present invention can be applied to the lettuce plant of the present invention.

[0093] In the present invention, the cross between the first lettuce plant (first parent line) and the second lettuce plant (second parent line) may be, for example, a cross between the same individuals (normal self-pollination), a cross between individuals of the same clone or between individuals of a line maintained as an inbred line (quasi-self-pollination), or a cross between different individuals (cross-pollination). In the case of normal self-pollination, for example, one of the first parent line and the second parent line is a female organ of the same individual, and the other is pollen of the same individual.

[0094] In the present invention, the first parent line is the lettuce plant of the present invention, for example, the lettuce plant deposited under the above-mentioned accession number FERM P-22453 or accession number FERM P-22454, or a progeny line thereof.

[0095] The second parent line is not particularly limited, and any lettuce plant can be used. The second parent line may be, for example, a lettuce plant of the same taxonomic species as the first parent line, or a lettuce plant of a different species. The second parent line may be, for example, the deposited line or a progeny line, or another lettuce plant.

[0096] The production method of the present invention may further include, for example, a growing step of growing a progeny line obtained in the crossbreeding step after the crossbreeding step. Examples of the growing conditions in the growing step include general growing conditions for lettuce plants.

[0097] The lettuce plant of the present invention can be obtained, for example, by the production method of the present invention.

[0098] <How to produce lettuce seeds> The present invention provides a method for producing lettuce seeds. The method for producing lettuce seeds of the present invention comprises the steps of selfing or crossing a lettuce plant of the deposited line with another lettuce plant, and optionally producing (collecting or harvesting) the resulting seeds. The method for producing seeds of the present invention may also provide a plant, plant part or seed by growing a seed of the lettuce plant.

[0099] The seed production method of the present invention may be a method for producing seeds derived from the deposited line. In this case, the seed production method of the present invention may include a step of (a) crossing the plant of the deposited line with another lettuce plant to produce seeds. The seed production method of the present invention may further include a step of (b) cultivating a lettuce plant from the seed of the step (a) to produce a lettuce plant derived from the deposited line, and a step of (c) self-pollinating or crossing the lettuce plant of the step (b) with another lettuce plant to produce an additional lettuce plant derived from the deposited line. Furthermore, the seed production method of the present invention may (d) optionally repeat the steps (b) and (c) one or more times to produce an additional lettuce plant derived from the deposited line. In this case, the additional lettuce plant obtained in the step (c) can be used as the lettuce plant cultivated from the seed of the step (a) in the step (b). The "one or more times" may be, for example, 1 to 10 times, 3 to 7 times, or 3 to 5 times. The seed production method of the present invention may further include a step of collecting or harvesting the seeds. The seed production method of the present invention may provide seeds produced by the above method, and plants or plant parts obtained by growing the seeds.

[0100] <Method of producing hybrid lettuce plants> The present invention provides a method for producing a hybrid lettuce plant. The method for producing a hybrid plant of the present invention comprises the step of crossing a lettuce plant of the present invention with another lettuce plant. The method for producing a hybrid plant of the present invention may comprise the step of collecting or harvesting seeds obtained by the cross. The method for producing a hybrid plant of the present invention may also provide seeds and hybrid plants or parts of hybrid plant individuals produced by the above methods.

[0101] <Method of introducing new traits> The present invention provides a method for introducing at least one new characteristic or trait (hereinafter, collectively referred to as "trait") into the deposited line. The method for introducing a trait of the present invention can also be referred to as, for example, a method for producing a lettuce plant into which a new trait has been introduced. The method for introducing a trait of the present invention includes, for example, (a) a step of crossing a plant of the deposited line with a lettuce plant containing at least one new trait to produce a progeny line, and (b) a step of selecting a progeny line containing at least one new trait. The method for introducing a trait of the present invention includes, for example, a step of (c) a step of crossing the progeny line with the deposited line to produce backcross progeny seeds, and (d) a step of selecting a backcross progeny containing at least one new trait and essentially all physiological and morphological characteristics of the deposited line. In the steps (b) and (d), the selection of a progeny line having a new trait may be carried out, for example, by detecting the trait, or by detecting a gene or molecular marker associated (linked) with the trait. The new traits include resistance to races other than downy mildew race BI:27EU (Bl16-26, 28-37, Bl5-11US), bacterial rot resistance, corky root disease resistance, lettuce mosaic virus resistance, aphid resistance, leaf miner resistance, and resistance to browning on cut surfaces.

[0102] The method for introducing a trait of the present invention may (e) optionally repeat the steps (c) and (d) one or more times to produce a lettuce plant containing at least one new trait. In this case, the progeny line in the step (c) of the method for introducing a trait of the present invention may be the backcross progeny selected in the step (d). The lettuce plant obtained or obtainable in the step (e) may have essentially all of the physiological and morphological characteristics of the deposited line. The "essentially all of the physiological and morphological characteristics" may be used in the explanation of the progeny line by replacing "progeny line" with "lettuce plant obtained or obtainable in the step (e)". The "one or more times" may be, for example, 1 to 10 times, 3 to 7 times, or 3 to 5 times. The method for introducing a trait of the present invention may include a step of collecting or harvesting seeds. The method for introducing a trait of the present invention may provide seeds produced by the above method, and parts of plants or plant individuals obtained by growing the seeds.

[0103] <Method of introducing transgene> The present invention provides a method for producing a plant derived from the deposited line, the plant containing at least one new characteristic or trait. The method for introducing a transgene of the present invention can also be referred to as a method for producing a lettuce plant having a new trait introduced therein.

[0104] The method for introducing a transgene of the present invention includes, for example, a step of introducing a mutation or transgene that confers at least one new trait into a plant of a deposited line. The introduction of the mutation or transgene can be carried out, for example, in the same manner as the introduction of a mutation or transgene in the progeny line. The lettuce plant obtained or obtainable by the introduction step may have essentially all of the physiological and morphological characteristics of the deposited line. The "essentially all of the physiological and morphological characteristics" can be used in the description of the progeny line by replacing "progeny line" with "lettuce plant obtained or obtainable by the introduction step". The method for introducing a transgene of the present invention may include a step of collecting or harvesting seeds. The method for introducing a transgene of the present invention may provide seeds produced by the above method, and parts of plants or plant individuals obtained by growing the seeds. Examples of the new traits include resistance to races other than downy mildew race BI:27EU (Bl16-26, 28-37, Bl5-11US), bacterial rot resistance, corky root disease resistance, lettuce mosaic virus resistance, aphid resistance, leaf miner resistance, and resistance to browning of cut surfaces.

[0105] <Lettuce plant regeneration and regeneration method> The invention provides lettuce plants regenerated from cell cultures, tissue cultures, or protoplasts of the deposited line (hereinafter referred to as "regenerates"). The invention may also provide cell or tissue cultures of regenerable cells, or protoplasts derived from the deposited line of lettuce plants. The cells, tissues, or protoplasts may be derived from tissues including leaves, pollen, embryos, cotyledons, hypocotyls, meristematic cells, roots, root tips, anthers, flowers, seeds, or stems.

[0106] The present invention provides a method for the propagation or multiplication of a lettuce plant of the deposited line. The propagation of the lettuce plant of the deposited line may be vegetative propagation of the lettuce plant of the deposited line. In this case, the regeneration method of the lettuce plant of the present invention may, for example, comprise the steps of (a) collecting tissue capable of propagation from the plant of the deposited line, (b) culturing the tissue to obtain a propagated shoot, and (c) rooting the propagated shoot to obtain a rooted plantlet. The regeneration method of the lettuce plant of the present invention may further comprise the step of (d) optionally growing a plant from the rooted plantlet. For the vegetative propagation method, see, for example, References 9 and 10 below. The regeneration method of the present invention may, for example, provide a plantlet, plant or part of a plant individual regenerated (produced) by the above method. The plant may have essentially all of the physiological and morphological characteristics of the deposited line. The above-mentioned "essentially all physiological and morphological characteristics" can be used in the description of the progeny line by replacing "progeny line" with "regenerated plant." Reference 9: Habtamu Gudisa Megersa, “Propagation Methods of Selected Horticultural Crops by Specialized Organs: Review”, Journal of Horticulture, 2017, Volume 4, Issue 2, 1000198 Reference 10: Nitish Kumar et.al., “In vitro Plant Propagation: A Review”, Journal of Forest Science, 2011, Vol. 21, No. 2, pp. 61-72

[0107] <Harvested and processed products of lettuce plants> The present invention provides harvested and / or processed products of the deposited line or progeny line, the harvested products being whole plants or parts of individual plants, preferably including leaves (e.g., petioles and blades), leaves and roots, or seeds.

[0108] The processed product includes any product obtained by processing the deposited line or progeny line. The processing is not particularly limited, and examples thereof include cutting, slicing, grounding, pureeing, drying, canning, bottling, washing, packaging, freezing and / or heat treatment. In the deposited line or progeny line, the part of the plant or plant individual used for the processed product is, for example, a leaf. The processed product may be, for example, the deposited line or progeny line that has been washed and packaged. The processed product may be, for example, housed in a container of any size or shape. Specific examples of the container include a bag, a box, a carton, and the like.

[0109] The invention may provide a container containing one or more lettuce plants, said container containing a whole plant or parts of an individual plant.

[0110] The present invention may provide a method for producing a lettuce plant as a food product (a method for producing a food product). The method for producing a food product of the present invention may, for example, comprise a step of collecting or harvesting a whole plant or a part of a plant individual of the deposited line or a progeny line, preferably a leaf of the deposited line or a progeny line. The method for producing a food product of the present invention may also comprise a step of cultivating a lettuce plant of the deposited line or a progeny line until it matures.

[0111] <Method of determining genotype> The present invention provides a method for determining or detecting the genotype of a deposited line or a progeny line. The genotype determination method of the present invention includes, for example, (a) a step of obtaining a nucleic acid sample from the deposited line or a progeny line, and (b) a step of detecting a genome in the nucleic acid sample. In the step (a), the method for preparing nucleic acid from the deposited line or the progeny line can be performed using a general method for preparing nucleic acid from tissue. In the step (b), for example, polymorphisms and / or alleles in the genome in the nucleic acid sample are detected. The detection of the polymorphism and / or allele can be carried out using, for example, SNP (single nucleotide polymorphism) genotyping, amplified fragment length polymorphism detection (AFLP), restriction fragment length polymorphism discrimination (RFLP) of genomic DNA, sequence-characterized amplified region detection (SCAR) of genomic DNA, cleaved amplified polymorphic sequence detection (CAPS) of genomic DNA, random amplified polymorphic detection (RAPD) of genomic DNA, polymerase chain reaction (PCR), DNA sequence, allele specific oligonucleotide (ASO) probe, or DNA microarray. The detection of the polymorphism and / or allele may be carried out, for example, by sequencing the base sequence of the genome, or may be carried out with reference to the SNP of the deposited lineage as described above. In the step (b), one polymorphism and / or allele in the genomic DNA may be detected, or two or more polymorphisms and / or alleles may be detected. The genotyping method of the present invention may include a step of storing the detection results of the polymorphisms and / or alleles in a computer readable medium. The present invention may provide a computer readable medium produced by such a method.

[0112] The genotype determination method of the present invention may be carried out on any lettuce plant (target lettuce plant) instead of the deposited line or the progeny line. In this case, the genotype determination method of the present invention may further include, for example, a step of determining whether the target lettuce plant is the progeny line based on the result of step (b). The determination may also be referred to as, for example, discrimination, estimation, appraisal, or identification. The determination may be made based on, for example, the rate of agreement between the result of step (b) and the genotype of the deposited line. EXAMPLES

[0113] The present invention will be described in detail below using examples, but the present invention is not limited to the embodiments described in the examples.

[0114] (1) Breeding of the deposited strain In the spring of 2017, a late-sprouting breeding line with strong plant vigor, excellent head formation in high temperatures, and resistance to multiple diseases such as downy mildew and root rot was crossed with a selected line with excellent heat and rain resistance to root rot to obtain the F1 generation. In the fall of the same year, the F1 generation was self-fertilized, and from the resulting F2 generation, individuals with plant vigor, large heads, late spouting, and resistance to root rot races 1 and 2 were selected and harvested. Selection and harvesting were repeated in the same manner until the F7 generation was reached in 2022, when it was determined that the target traits had been fixed and breeding was terminated.

[0115] The F6 generation was cultivated in a highland experimental field (Kawakami Village, Nagano Prefecture) in 2021 in two cropping types, June to July, with 320 plants each, to confirm that there was no variation in characteristics within the lineage and that the bred lettuce varieties had uniformity and stability. The F6 generation was then self-fertilized, and the resulting F7 generation seeds were deposited under accession numbers FERM P-22453 and FERM P-22454.

[0116] (2) Characteristics of the deposited strain The characteristics and traits of the plant individuals of the deposited lines were evaluated according to the Ministry of Agriculture, Forestry and Fisheries' examination criteria for variety registration. The results are shown in Table 4.

[0117] [Table 4A]

[0118] [Table 4B]

[0119] [Table 4C]

[0120] (3) Identification of the deposited strain and its progeny The deposited plant and the following commercially available lettuce varieties were subjected to the SSR marker analysis described in Tables 3A and 3B. In addition, an inoculation test was performed using the downy mildew race BI:27EU according to the evaluation criteria for lettuce varieties issued by MAFF, and the resistance to downy mildew against the race was evaluated. The results are shown in Table 5 below. Here, the number of polymorphisms in the table indicates the number of polymorphisms obtained by the marker. Z indicates the genotype of the reference variety Souther, and the polymorphism of each variety is indicated by the number of bases increased (+) or decreased (-) relative to the genotype. R indicates resistance to downy mildew race BI:27EU, and S indicates no resistance. As shown in Table 5 below, it was found that the deposited line and its progeny can be distinguished from other lettuce varieties by combining the multiple markers described in the table with the presence or absence of resistance to downy mildew race BI:27EU. (Developed by Takii Seeds) Souther, Red Fire, Dancing, Summer Surge, Lancer, Tough Ora 0566, Tough Ora 0567, Romalia, Oasis, Aristo, Summer Guy (Developed at Nagano Prefecture Vegetable and Flower Experiment Station) Shinano Green, Shinano Star (Developed by Sakata Seed) Bull Rush

[0121] [Table 5]

[0122] Although the present invention has been described above with reference to the embodiments and examples, the present invention is not limited to the above-mentioned embodiments and examples. Various modifications that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention. [Industrial Applicability]

[0123] As described above, the present invention provides a novel lettuce plant, which is extremely useful in the agricultural field, for example, in breeding.

Claims

1. Seeds of lettuce plants of the lettuce line Takii 32 or Takii 33, wherein said seeds are deposited under accession number FERM P-22453 or accession number FERM P-22454.

2. A lettuce plant or a part thereof grown from the seeds according to Claim 1.

3. including an inbred progeny line of the lettuce plant according to Claim 2, wherein said inbred progeny line has the following traits (1), (2) and (3): a lettuce plant or a part thereof: (1) having "strong" resistance to race 1 of Rhizoctonia solani; (2) having "strong" resistance to race 2 of Rhizoctonia solani; (3) having "present" resistance to downy mildew race Bl:27EU.

4. A first-generation hybrid lettuce plant or a part thereof, using the lettuce plant according to Claim 2 or 3 as at least one of the crossing parents.

5. A method for producing a lettuce plant, comprising an inbreeding step of inbreeding the lettuce plant according to Claim 2 or 3.

6. A method for producing a lettuce plant, comprising a crossing step of crossing the lettuce plant according to Claim 2 or 3 with another lettuce plant.

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