KR102342057B1 - Cold Stress and Clubroot Disease Resistant Novel Chinese Cabbage Variety - Google Patents

Abstract

The present invention relates to a new variety of cabbage resistant to low-temperature stress and root nodules, which has strong resistance to low-temperature stress, exhibits resistance to root nodules, and has excellent joint shape, so it can be usefully used as a breeding material and a material for genetic research have.

Description

Cold Stress and Clubroot Disease Resistant Novel Chinese Cabbage Variety

The present invention relates to a new variety of cabbage resistant to low temperature stress and root lump disease.

Chinese cabbage ( Brassica rapa L.) is a vegetable traditionally used in Korea as a source of vitamins that are easily scarce in winter because it is processed in the form of kimchi. Chinese cabbage is currently cultivated as a major vegetable in all regions of Korea from spring, summer, autumn, and winter. In 2017, the production amount of Chinese cabbage was 890 billion won, one of the top 10 vegetables in Korea, and the cultivated area was 32,000 ha, accounting for about 13% of the total vegetables. Due to the recent tendency to avoid salty and spicy foods such as westernization of diet, the cultivated area is showing a decreasing trend of 3% per year on average (2019, Agricultural Prospects).

Cabbage is one of the cruciferous vegetables that grow well in cool climates. Since it was introduced into Korea, it was mainly cultivated in autumn. Since the temperature is relatively low in spring, a spring cultivation type using a vinyl house or tunnel was developed in the southern region using late-planting varieties. Afterwards, a variety that can withstand the summer heat was developed, and cultivation was successful in high altitude regions over 700 m above sea level, such as Gangwon-do, where summer is relatively cool. Even cultivation became possible (2008, Korea Vegetable Seed Industry Development Company).

In 2010, the supply of Chinese cabbage was disrupted due to a rapid decrease in yield due to heavy rains during the harvest season in the high-altitude region of Gangwon-do, the main production area of cabbage in summer, and a late typhoon that hit the whole country in autumn. Since then, various severe weather conditions such as poor survival due to repeated low and high temperatures in the spring cabbage planting season, and the occurrence of the East Sea due to prolonged low temperatures outside the weather pattern of 3 to 4 degrees during the winter cabbage cultivation season have occurred. has grown

In addition, as the weather deteriorates in recent years, the growth of crops becomes poor, and environmental conditions favorable for the occurrence of pests and pests are formed, and the mass generation of specific pests is a problem. In particular, in the case of summer cultivation in high altitudes, root nodules frequently occur when the rainy season or rain frequently falls during a high temperature period. Root nodules are caused by Plasmodiophora brassicae , an obligate active parasitic bacterium that survives in the soil and forms nodules on the roots of Brassica vegetables (Braselton, 1995). The root nodule pathogen was classified as a fungi at the beginning of the study, but as a result of subsequent sequencing, it was found not to be related to the fungus (Castlebury & Domier, 1998, Down et al. 2002), and as a result of embryological classification using rRNA It was found to be a protozoa (Van de Peer et al. 2000).

Root nodules are occurring in all cabbage growing regions in Korea, including Ulleungdo, and when they occur in the early stages of growth, they pose a great threat to the supply of Chinese cabbage to such an extent that harvesting must be abandoned. Methods using crop protection agents and antagonists have been developed to control root nodules (Datnoff et al. 1987, Cheah et al. 2000, Yeoung et al. 2003), but in the case of crop protection agents, treatment costs are high, and treatment is carried out at every crop season. There is a difficulty that needs to be done, and the use of antagonists has a low treatment effect. Therefore, the development of root-knot-resistant varieties is the most fundamental countermeasure, and research related to the development of resistant varieties is in progress, but the development of Chinese cabbage exhibiting both low-temperature stress resistance and root-knot resistance has not been made yet.

Accordingly, the present inventors conducted a study to develop a new variety of Chinese cabbage having resistance to severe weather conditions and resistance to root nodules by using a haploid breeding method through spore culture to develop a genetically high-purity line in a short period of time. carried out to complete the present invention.

Korean Patent Publication No. 10-2017-0070343

One object of the present invention is to provide a Chinese cabbage plant having cold stress resistance (Brassica rapa . L) plant, wherein the plant is deposited with the seed accession number KCTC 13971BP.

Another object of the present invention is to provide a Chinese cabbage ( Brassica rapa . L) plant having cold stress and root nodule resistance, the plant being deposited with the seed accession number KCTC 13971BP It is to provide a part (plant part) of the Chinese cabbage plant .

Another object of the present invention is a Chinese cabbage ( Brassica rapa . L) plant having cold stress and root nodule resistance, wherein the plant is deposited with the seed accession number KCTC 13971BP. It includes the step of crossing the cabbage plant with another Brassica plant. It is to provide a method for producing a Chinese cabbage plant seed having increased resistance to cold stress and root nodules.

One aspect of the present invention provides a Chinese cabbage plant having cold stress and root-knot resistance (Brassica rapa . L) plant, wherein the plant is deposited with seed accession number KCTC 13971BP.

Since the Chinese cabbage deposited with the seed accession number KCTC 13971BP of the present invention exhibits resistance to low temperature stress, it has little effect on the growth of Chinese cabbage even with a sudden temperature drop. In addition, the Chinese cabbage of the present invention not only exhibits resistance to root nodules, one of the most serious diseases of Chinese cabbage, but also has an excellent shape. Therefore, the Chinese cabbage deposited with the seed accession number KCTC 13971BP of the present invention can be usefully utilized as a breeding material such as an intermediate model.

The Chinese cabbage of the present invention may be cultivated in a temperature range of -3 ℃ to 30 ℃, and may be sown from July to August, for example, from August 1 to August 31, in the weather of Korea. It is not limited.

According to one embodiment of the present invention, there is provided a Chinese cabbage plant having the following characteristics more than the control cultivar Bulam No. 3: (1) The maturation of the applied cultivar is slower than that of the control cultivar. (2) The green tea of the applied variety is smaller than that of the reference variety. (3) The color of the outer leaf is dark green for the applied variety, and green for the control variety. (4) The color of the inner leaf is yellow for the applied variety, and dark yellow for the control variety. (5) The outer leaf shape of the applied variety is an upside down egg shape, and the control variety is an egg shape. (6) The cultivar of this application is semi-gulgu, and the control cultivar is semi-conjugated.

As used herein, the term 'ripe' refers to a phase in which the growth of Chinese cabbage is completed, and the Chinese cabbage that has reached maturity includes being in a harvestable state.

As used in the present invention, the term 'growth' refers to the formation of a sphere when the leaf is differentiated and grows, and the leaf shrinks and fills the inside.

As used herein, the term 'conjugated' refers to a form in which the outer leaf covers the sphere, and 'half fusion' refers to a form in which the outer leaf does not completely cover the sphere.

The Chinese cabbage of the present invention may be derived from seeds deposited with accession number KCTC 13971BP. The new variety of Chinese cabbage of the present invention was named 'Wonkyo 20051', and the seeds were deposited with the Korea Research Institute of Bioscience and Biotechnology Microbial Resource Center, an international depository organization according to the Budapest Treaty, as of October 8, 2019 under the accession number KCTC 13971BP.

On the other hand, the Chinese cabbage of the present invention has a high fertilizer requirement as a middle seed, is sensitive to boron deficiency, and may cause browning of the outer leaves in the late growth period. The yield of Chinese cabbage of the present invention is about 5,268 kg/10a, which is lower than that of Bulam No. 3, which is a control variety, 6,538/10a.

Another aspect of the present invention is a cabbage ( Brassica rapa . L) plant having cold stress and root nodule resistance, wherein the plant is deposited with seed accession number KCTC 13971BP It provides a part (plant part) of the Chinese cabbage plant.

The leaves and/or stems obtained from the Chinese cabbage of the present invention can be used in various ways as ingredients such as kimchi, greens, and salads, and the parts of the Chinese cabbage such as seeds and/or cells obtained from the Chinese cabbage are different from other cabbage individuals, strains, Or it can be used to cultivate a variety, in particular, a cabbage having cold stress and root-knot traits.

According to one embodiment of the present invention, the part is a protoplast, an ovule, a cell, a pollen grain, an embryo, a cotyledon, a hypocotyledon, a root, an anther ( anther), stems, leaves and seeds.

Another aspect of the present invention is a Chinese cabbage ( Brassica rapa . L) plant having cold stress and root nodule resistance, wherein the plant is deposited with seed accession number KCTC 13971BP. It includes the step of crossing a Chinese cabbage plant with another Brassica plant. It provides a method for producing a Chinese cabbage plant seed having increased resistance to cold stress and root lump disease.

The method for producing a cabbage plant seed having increased resistance to low temperature stress and root nodule according to the present invention can be carried out through a breeding method generally performed in the art, etc. in order to introduce the trait of the parent variety to future individuals.

According to the new variety of cabbage resistant to low temperature stress and root nodules, it has strong resistance to low temperature stress, exhibits resistance to root nodules, and has an excellent joint shape, so it can be usefully used as a breeding material and a material for genetic research.

1 shows (A) 14-FS1 (Chunkwang), (B) 14-FS2 (Arari), (C) 14-FS14 (derived from Bulam Plus) and ( D) 14-FS15 (Chinese cultivar) It is a half cut form of Chinese cabbage material.
2 is a photograph showing the shape of Wonkyo 20051 cultivated in (A) 2017 and (B) 2018.

Hereinafter, the present invention will be described in more detail through one or more embodiments. However, these examples are for illustrative purposes of the present invention, and the scope of the present invention is not limited to these examples.

Example 1. Low-temperature stress and root-knot resistant cabbage line cultivation method

1-1. Early summer cultivation for selection of cultured specimens

By 2014, the breeding material developed by the Vegetable Department of the National Institute of Horticultural and Herbal Science and 37 major hybrid varieties sold in Korea were sown on April 16, 2014 and managed for a month, after which the seedlings were managed for a month. Planted in the vegetable department of the Academy of Sciences (located in Suwon-si, Gyeonggi-do), and growth survey was conducted on July 17 to investigate the horticultural characteristics (Table 1). Each variety was grown according to the standard Chinese cabbage cultivation method, covered with a black mulching film, and drip irrigation was performed. The characterization was conducted in accordance with the guidelines of the National Seed Resources for cabbage characterization (UPOV, 2003) and the investigation criteria of the Rural Development Administration's trial study design. Investigation could not be carried out on some subjects that could not be irradiated due to necrosis.

label designation origin year of seed production Seed production
(dog) 14-FS01 spring Korean commercial varieties - - 14-FS02 Arari Korean commercial varieties - - 14-FS03 ssam yellow-i Korean commercial varieties - - 14-FS04 Wonkyo No.20034 NIHHS 2013 528 14-FS05 Wonkyo No.20035 NIHHS 2013 631 14-FS06 Wonkyo No.20036 NIHHS 2013 533 14-FS07 Wonkyo No.20037 NIHHS 2013 538 14-FS08 Wonkyo No.20038 NIHHS 2013 541 14-FS09 Wonkyo No. 20039 NIHHS 2013 545 14-FS10 Wonkyo No.20040 NIHHS 2013 549 14-FS11 Wonkyo 20041 NIHHS 2013 553 14-FS12 Wonkyo 20042 NIHHS 2013 557 14-FS13 Wonkyo No.20043 NIHHS 2013 715 14-FS14 12-BK09 NIHHS (derived from 'Bullam Plus') 2012 290 14-FS15 12-BK10 NIHHS (derived from commercial varieties) 2012 291 14-FS16 12-BK11 NIHHS (derived from 'Hwimori') 2012 293 14-FS17 12-BK15 NIHHS (derived from 'Bullam Plus') 2012 299 14-FS18 12-BK7-11 NIHHS (derived from 'girl taste') 2013 455 14-FS19 12-BK7-12 NIHHS (derived from 'girl taste') 2013 456 14-FS20 12-BK9-2 NIHHS (derived from 'Bullam Plus') 2013 460 14-FS21 12-BK9-3 NIHHS (derived from 'Bullam Plus') 2013 461 14-FS22 12-BK11-2 NIHHS (derived from 'Hwimori') 2013 466 14-FS23 12-BK11-3 NIHHS (derived from 'Hwimori') 2013 467 14-FS24 12-BK15-1 NIHHS (derived from 'Bullam Plus') 2013 468 14-FS25 12-BK15-5 NIHHS (derived from 'Bullam Plus') 2013 472 14-FS26 12-BK7A-2x12-FA10-5 NIHHS (derived from commercial varieties) 2013 478 14-FS27 12-BK7A-3x12-FA10-1 NIHHS (derived from commercial varieties)r 2013 480 14-FS28 12-BK8A-1 NIHHS (from 'strong') 2013 485 14-FS29 12-BK8A-1x12-FA10-5 NIHHS (derived from commercial varieties) 2013 487 14-FS30 12-BK8A-5 NIHHS (from 'strong') 2013 496 14-FS31 12-BK8A-5x12-FA10-4 NIHHS (derived from commercial varieties) 2013 498 14-FS32 12-BK10A-3 NIHHS (derived from commercial varieties) 2013 505 14-FS33 12-BK10A-3x12-FA10-5 NIHHS (derived from Korean and Chinese varieties) 2013 507 14-FS34 12-BK10A-4 NIHHS (derived from Chinese varieties) 2013 508 14-FS35 12-BK10A-4x12-F10-3 NIHHS (derived from Korean and Chinese varieties) 2013 510 14-FS36 12-BK14A-4 NIHHS (from 'Handongji') 2013 522 14-FS37 12-BK14A-4x12-FA10-2 NIHHS (from Korean variety) 2013 524

1-2. Chinese cabbage spore culture

Selected specimens for spore culture were sown in late August (26th), 2014 to secure germination rate, and after 5 or more true leaves were developed, they were treated for 2 months in a low temperature treatment room (maintained at 5 to 10℃) on September 25th and then plastic The young buds of an individual that bloomed after being transferred to a pot with a diameter of 20 cm in the house were used as a material for culture.

Young buds with a size of 2 to 3 mm were selected and sterilized by wrapping 30 each by resource with gauze. It was sterilized by immersion in sodium hypochlorite (2%) solution for 15 minutes, then immersed in sterile water for 3 minutes and then taken out and immersed in fresh sterile water for 3 minutes, followed by washing three times.

For the preparation of pyrotechnics for culture, pour a small amount of 1X NLN (Nitsch and Nitsch 1967; Lichter 1982) medium, grind with a mortar and sieve through a 45㎛ sieve, adjust the volume to 30 ml with NLN medium, and centrifuge to remove the supernatant The washing process was performed three times.

After the last supernatant was drained, an experimental medium was added, and 2.5 ml each was dispensed into a 6 cm diameter Petri dish, sealed with parafilm and lap, and dented (Lee and Nam 1995). Then, in order to induce embryonic development, it was investigated whether or not embryos were developed by dark culture (30°C, 24 hours) and dark culture (25°C, 14 days), followed by light culture (25°C, 75-80 rmp shaking culture). As a medium, after mixing 1/2X NLN and 1/2X NLN with Vit., AgNO ₃ 1mg/L was added and basal medium adjusted to 13% of sucrose concentration, NAA 0.05mg/L and BAP 0.05mg/L Hormone medium to which was added was used.

When the embryoid body grows to about 7 mm in size, it is transferred to 1/2MS (Murashige and Skoog, Duchefa) solid medium containing 3% sucrose, and subcultured 2 to 4 times until it grows into a normal plant, and then sterilized soil It was planted in a plant and acclimatized until it grew into a normal plant.

1-3. seed propagation

Plants that have been acclimatized in Example 1-2 were managed for about 3 months in a low-temperature treatment room (maintained at 5 to 10° C.) depending on the growth situation, and after growing strong, they were transferred to a greenhouse and planted in a pot with a diameter of 20 cm to induce flowering. The formation of geodesic was induced by removing the first stem of the individual in which the stem was formed, and 3 to 5 stems were grown per individual and then brain pollination was carried out. For brain pollination, artificial insemination was performed by opening the top of a bud of about 5 mm that had not yet bloomed in the pollen of the same individual that had bloomed in the bag, and then covered the bag for a week to prevent polluted moisture. After 50 to 60 days after brain pollination, when the seeds ripen to yellow, the seeds were harvested before the pods opened.

1-4. Root lump inoculation and evaluation

Artificial inoculation of root nodules was performed by irrigating an inoculum of a certain concentration to inoculate. The seeds sown in Examples 1-3 were sown for each test material in a 50-hole plug cell tray (50). When the cotyledons were fully developed after about 7 days, each individual was drenched with 5 ml of the inoculum (Seosan strain (root nodule pathogen) distributed by the Korea Research Institute of Chemical Technology, 1×10 ⁶ concentration) (Kim et al., 2016). . Because the Seosan strain is highly pathogenic and infects all domestic varieties known to be resistant, selection of resistant breeding materials is difficult (data not presented). Seosan strain inoculum was treated at a corresponding low concentration. After irrigation, it was irrigated by immersion using a water tray and managed for 6 weeks.

For the evaluation of root nodule resistance, the disease index (DI) was investigated according to the degree of occurrence of symptoms after digging up all the plants and cleaning the roots. DI values were calculated according to the method of Werner et al. (2008). The abnormally deformed severity, from 0 to the main muscle, which showed no symptoms, was divided into 5 sections and evaluated according to the evaluation criteria shown in Table 2 (Kuginuki et al., 1999; Suwabe et al., 2003).

score disease 0 No root lumps One There is little degree of hypertrophy due to an epigenetic lump on the entourage, and they exist independently of each other 2 Humps are formed on the lateral roots, and the degree of hypertrophy is relatively large. 3 Humps grow on the main roots, they are joined to each other, and the degree of hypertrophy is large. 4 Humps grow on the main roots, they are joined together, and the degree of hypertrophy is very large.

1-5. Horticultural Characteristics Assessment

The spore culture-derived line in which the seeds were sufficiently propagated by carrying out Examples 1-3 were cultivated in the autumn at the vegetable department of the National Academy of Horticultural and Herbal Science in Iseo-myeon, Wanju-gun, Jeollabuk-do for horticultural property evaluation. It was cultivated according to the standard Chinese cabbage cultivation method, covered with a black mulching film, and drip irrigation was performed. Sowing was carried out on August 9, 2018 at the nursery of the National Institute of Horticultural and Herbal Science in Iseo-myeon, Wanju-gun, and planting was carried out on September 6, 2018 in a nearby open field. The characterization survey was conducted on November 5, 2018 in accordance with the National Seed Resources' Guidelines for Characteristics Investigation of Chinese cabbage (UPOV, 2003) and the survey criteria of the Rural Development Administration's trial study design.

Example 2. Selection of environmental resistant specimens and spore culture

In Korea, the four seasons are distinct, and the menstrual disorders of Chinese cabbage occur during hot summer and cold winter, and the supply disruption problem tends to recur every year. In order to develop a variety resistant to such temperature stress, it was attempted to select a model for culturing small spores through hot season cultivation. It was sown on April 16, 2014 in the test field of the National Institute of Horticultural and Herbal Science (Suwon) located in Suwon, the central region of the Republic of Korea. After that, it was cultivated according to the standard cultivation guidelines and harvested on July 17th, and major horticultural characteristics such as cohesiveness, number of inner leaves, and weight were investigated.

As a result of quantitatively surveying weekdays and weights by performing statistical analysis for each characteristic, significance was shown for each variety, but 11 resources for culturing small spores in consideration of the occurrence of physiological disorders such as internal rot and the shape of the bulbs. was selected (Fig. 1 and Table 3).

label Weekday
(g) nine leaf weight (g) Height (cm) Width (cm) Length (cm) Width (cm) Count 14-FS1 2291.0 1147.0 29.5 12.5 45.0 28.0 77.0 14-FS2 2978.5 2324.5 29.0 16.0 42.0 28.5 74.0 14-FS3 2423.5 1459.5 27.5 13.5 52.5 28.5 66.5 14-FS4 2051.5 911.0 33.0 11.5 53.0 30.0 47.5 14-FS5 875.0 436.5 33.0 7.0 41.5 21.5 27.5 14-FS6 2085.5 1061.0 37.0 13.5 49.0 30.5 61.0 14-FS7 1883.0 862.5 27.0 10.5 34.5 25.5 55.0 14-FS8 1615.5 430.5 23.0 9.5 37.5 16.5 43.5 14-FS9 1470.5 733.5 25.0 11.5 38.5 24.5 64.0 14-FS10 1014.0 601.5 35.0 8.0 50.5 23.5 34.5 14-FS11 2576.0 780.5 24.0 12.5 48.5 32.0 69.0 14-FS12 2540.5 989.5 26.0 12.5 42.5 25.5 73.5 14-FS13 2701.0 1809.5 33.0 16.0 47.5 30.0 79.5 14-FS14 2545.5 1587.5 30.0 17.0 51.0 31.0 83.5 14-FS15 1953.0 252.0 18.0 7.0 39.5 25.5 27.0 14-FS16 1766.5 712.0 21.0 11.5 38.5 26.0 59.0 14-FS17 1260.5 573.5 26.0 12.5 38.5 24.0 57.0 14-FS18 1962.0 591.5 26.5 10.5 55.0 28.5 60.5 14-FS19 2298.0 1438.0 36.0 16.0 54.0 33.5 66.0 14-FS20 2513.0 1362.5 28.0 16.5 45.0 26.5 78.5 14-FS21 2460.0 1304.5 25.0 14.5 46.5 26.5 89.5 14-FS22 1065.0 481.0 20.0 11.0 36.0 29.0 54.0 14-FS23 838.5 249.0 18.5 8.5 35.5 25.0 44.5 14-FS24 1885.0 679.5 24.0 12.0 43.5 27.0 65.0 14-FS25 837.0 195.0 24.0 7.0 38.0 28.0 11.0 14-FS26 2579.5 1230 29.5 12.5 49.5 34.5 75.0 14-FS27 2123.0 1035.5 30.0 14.0 45.5 33.5 67.0 14-FS28 1574.5 781.5 31.5 13.0 45.0 26.0 58.5 14-FS29 2204.5 1165.5 30.5 14.0 47.0 30.0 69.5 14-FS30 1660.0 921.0 31.0 14.5 52.5 27.5 62.5 14-FS31 3027.0 1603.5 35.5 13.5 47.5 32.0 67.0 14-FS32 1268.5 244.5 21.5 6.5 41.5 23.5 1.5 14-FS33 3126.0 1438.5 32.0 13.0 46.0 32.5 54.5 14-FS34 1763.0 362.0 22.5 8.5 36.0 19.5 26.0 14-FS35 3433.0 1807.5 35.0 16.0 48.5 34.0 55.5 14-FS36 531.5 110.5 16.5 3.5 33.0 26.5 17.5 14-FS37 2495 1397.5 37.0 13.5 45.5 32.5 63.5

Specifically, it was confirmed that the cultivation environment was not good enough to cause rot symptoms in '14-FS1 (Chunkwang, Sakata Korea)', the main variety of high-altitude summer cabbage (FIG. 1A). '14-FS2 (Arari, Nonghyup seedling, Fig. 1B))', developed as a summer cultivar in Jeongseon, '14-FS13', a vegetable family breeding line, and '14-FS35', a segregated generation of imported resources from China, Spheres over 1.8 kg were formed, but most of them were found to be very small or rotten. '14-FS2' grew well even in the summer cultivation conditions of flat land, forming 2.3 kg spheres. The inner leaf color was yellow and there was no sign of rot, so it was selected as a culture model. In addition, 11 varieties were selected, including '14-FS14' (FIG. 1C), '14-FS28', and '14-FS15' (FIG. 1D) with dark leaves, with dark leaves and small bulbs. did '14-FS13' and '14-FS35' formed relatively heavy spheres, but were culled due to poor horticultural properties such as poor shape and stiff leaves.

For 11 selected resources, from 2014 to 2015, spore culture was performed according to Example 1-2. For culturing small spores, 90 to 120 flower buds were cultured using NLN-13 as a basic medium. With the exception of '14-FS18 (the descendants of 'Winter Taste', a Korean collection variety)', which did not develop buds smoothly, 200 or more were cultivated for 10 resources, and 167 points from 4 resources including '14-FS02' of haploid-derived embryoid bodies were obtained. The embryoid bodies were acclimatized using 1/2MS solid medium, and the individuals with normally formed roots were transplanted into sterilized soil and acclimatized. Individuals well acclimatized in the soil and successfully grown as plants were planted in pots with a diameter of 20 cm after low-temperature treatment (maintained at 5 to 10° C., for about 3 months) to induce flowering, and then seeds were obtained by brain pollination. Finally, seeds were obtained from a total of 13 doubled haploids, including 2 points from '14-FS02', 1 point from '14-FS15', 6 points from '14-FS21' and 4 points from '14-FS31' ( Table 4).

label number of buds number of embryoid bodies double haploid
number of plants 14-FS01 210 - - 14-FS02 390 78 2 14-FS03 240 - - 14-FS14 200 - - 14-FS15 350 One One 14-FS17 210 - - 14-FS18 90 - - 14-FS19 300 - - 14-FS21 270 56 6 14-FS28 210 - - 14-FS31 330 32 4

In the case of '14-FS02', the highest number of haploid-derived embryoid bodies were generated by culturing 390 immature pyrophyllites, 78, but the number of plants that were able to successfully form roots and bear seeds was 2, and the efficiency of obtaining doubled haploids was 2.6. % was found to be low. On the other hand, in the case of '14-FS15', only one embryoid body was obtained, but it was successfully acclimatized and harvested. In particular, in the case of '14-FS02', it was confirmed that too many embryoid bodies were generated at once, so that they grew competitively in a small culture vessel, making it difficult to develop into a normal plant.

Example 3. Root-knot resistance test of nurturing resources

In Example 2, artificial inoculation of root nodules was carried out in the method of Examples 1-4 using 13 doubled haploids in which seeds were normally propagated and 'Bulam 3', a major domestic Chinese cabbage variety, and root-knot disease of cultivation resources. Resistance was assayed.

As a result, among the cultivated lines, '14-FS31-C1', '14-FS31-C3', '14-FS21-C2', '14- Seven intermediate resistance resources with a DI of 10 or less, such as FS21-C3', '14-FS21-C4', '14-FS21-C6' and '14-FS02-C1', were selected. The selected resources were propagated by brain pollination for horticultural characteristics test in the next year (Table 5).

label Disease Index label Disease Index Bulam No. 3 25.00 14-FS21-C3 0.00 14-FS31-C1 9.62 14-FS21-C4 0.00 14-FS31-C2 23.21 14-FS21-C5 11.54 14-FS31-C3 0.00 14-FS21-C6 0.00 14-FS31-C4 30.77 14-FS15-C1 25.00 14-FS21-C1 25.00 14-FS02-C2 32.50 14-FS21-C2 5.00 14-FS02-C1 0.00

Example 4. Horticultural Characteristics Test and Selection of Breeding Lines

For testing of horticultural properties, brain pollination was performed on 7 root-knot resistance selection resources in Example 3, '14-FS21-C3', '14-FS21-C4', '14-FS21-C6', More than 100 seeds were obtained at 6 points of '14-FS15-C1', '14-FS31-C3', and '14-FS02-C1'. After that, in order to investigate the horticultural characteristics of the propagated pear haploid line and 83 points of vegetables and resources possessed, autumn cultivation was carried out in the open field in Wanju area in 2018. 83 resources are 17 points ('18-FH10' to '18-), including 9 commercially available hybrid varieties ('18-FH01' to '18-FH09') and 'Wongyo 20034', a line cultivated in the vegetable department. FH26'), 5 points of spore culture-derived lines of root-knot-resistant cabbage selection resources ('18-FH44' to '18-FH48'), 41 points of crossbreeding of cabbage resources that were resistant to root-knot disease Pyeongchang strains owned by the vegetable family ('18-FH49' to '18-FH89'), and 11 points ('18-FH90' to '18-FH100') of the breeding and culturing of resources that were resistant to the low concentration of the western acid strain. Due to the unfavorable weather conditions, 4 of the hybrid varieties could not form a sphere. The weight of the entire plant, the weight of the bulb, and the number of leaves were investigated for each resource, and resources with excellent bulb characteristics were selected so that they could enter the domestic market with high demand for high quality.

As a result, the resources used in this experiment were high-purity strains that were either brain pollinated for more than 3 generations or were cultivated through spore culture, and the number of leaves varied from 13 to 194, with an average of 42. It was smaller than the average of 71. This was confirmed to be consistent with the report that the yield was high due to high stress and disease resistance due to the phenomenon of hybrid strength in one-to-one hybrid varieties (Kazutaka et al. 2016). Among the resources used in this experiment, the number of leaves was around 50, which is more than the average number of leaves of 42 plants and resources cultivated together. The quality of yellow '18-FH98' was evaluated as excellent ('18. Packaging Fair).

Meanwhile, looking at the autumn climate of Jeonju in 2018, the average daily temperature for 7 consecutive days from October 8 to 14 was 2.9℃ lower than the average temperature for the past 20 years on average, and from October 31 to November 1 The average temperature was 3.1°C lower for 3 days as well. This period was about a month after planting on September 6, and the growth of Chinese cabbage was very vigorous. However, in the case of the '18-FH98' resource, it was not a one-to-one hybrid, and showed resistance to low-temperature stress even without hybridization, and its excellence was recognized. The selected '18-FH98' forms small, hard spheres and has excellent leaf characteristics, so it was also selected by 4 private companies including Samsung Seedling.

label cross breed name Weekdays (g) weight (g) number of leaves 18-FH001 Chu Kang 2453.3 1320.0 69.3 18-FH002 whistle Sphere not formed - - 18-FH003 Bulam No. 3 2346.7 1160.0 71.7 18-FH004 yellow mini 3050.0 2158.3 74.3 18-FH005 yellow Sphere not formed - - 18-FH006 fan Sphere not formed - - 18-FH007 chuno 2351.7 1321.7 65.0 18-FH008 bulam plus 2340.0 1146.7 75.0 18-FH009 yellow castle Sphere not formed - - 18-FH010 Wonkyo No.20034 1888.3 940.0 38.3 18-FH011 Wonkyo No.20035 1645.0 540.0 35.3 18-FH012 Wonkyo No.20036 1035.0 475.0 33.3 18-FH013 Wonkyo No.20037 1961.3 806.7 61.7 18-FH014 Wonkyo No.20038 2444.0 1023.3 43.7 18-FH015 Wonkyo No. 20039 1211.7 545.0 64.0 18-FH016 Wonkyo No.20040 1328.3 406.7 24.7 18-FH017 Wonkyo 20041 1163.3 450.0 56.3 18-FH018 Wonkyo 20042 2558.3 1313.3 67.7 18-FH019 Wonkyo No.20043 3166.7 1161.7 58.7 18-FH020 Wonkyo No.20044 1166.7 676.7 43.7 18-FH021 Wonkyo No.20045 2130.0 1305.0 76.0 18-FH022 Wonkyo No.20046 Sphere not formed - - 18-FH023 Wonkyo No. 20047 Sphere not formed - - 18-FH024 Wonkyo No.20048 Sphere not formed - - 18-FH025 Wonkyo No.20049 Sphere not formed - - 18-FH026 Wonkyo No.20050 1861.7 901.7 43.7 18-FH044 17-RFE36 Sphere not formed - - 18-FH045 17-RFE81 3326.7 1385.0 50.0 18-FH046 17-RFE33 1361.7 915.0 39.0 18-FH047 17-RFE108 2300.0 840.0 57.3 18-FH048 17-RFE98 3751.7 1551.7 51.0 18-FH049 17-RGW03 Sphere not formed - - 18-FH050 17-RGW04 2036.7 746.7 27.0 18-FH051 17-RGW06 19966.7 498.3 27.0 18-FH052 17-RGW08 2131.7 150.0 12.7 18-FH053 17-RGW19 Sphere not formed - - 18-FH054 17-RGW34 2348.3 508.3 25.3 18-FH055 17-RGW35 2021.7 703.3 30.3 18-FH056 17-RGW46 995.0 256.7 26.0 18-FH057 17-RGW47 1846.7 357.7 23.0 18-FH058 17-RGW51 Sphere not formed - - 18-FH059 17-RGW60-4 1658.3 328.3 12.7 18-FH060 17-RGW61-5 2006.7 256.7 23.0 18-FH061 17-RGW63 1093.3 225.0 12.7 18-FH062 17-RGW132 Sphere not formed - - 18-FH063 17-RGW133 2078.3 1046.7 49.7 18-FH064 17-RGW137 1598.3 153.3 15.3 18-FH065 17-RGW202 Sphere not formed - - 18-FH066 17-RGW203-4 Sphere not formed - - 18-FH067 17-RGW205-3 2645.0 410.0 21.7 18-FH068 17-RGW206 3003.3 400.0 26.3 18-FH069 17-RGW208 2463.3 323.3 24.7 18-FH070 17-RGW209 2843.3 416.7 18.7 18-FH071 18-RFN8 1943.3 703.3 32.0 18-FH072 18-RFN52 1628.3 765.0 57.3 18-FH073 18-RFN75 1700.0 351.7 23.0 18-FH074 18-RFN77 1830.0 506.7 27.7 18-FH075 18-RFN78 1865.0 321.7 13.7 18-FH076 18-RFN124 1608.3 400.0 34.7 18-FH077 16-GW01 2161.7 770.0 44.3 18-FH078 16-GW10 3148.3 1411.7 44.3 18-FH079 16-GW16 1280.0 306.7 27.0 18-FH080 16-GW18 1501.7 121.7 22.3 18-FH081 16-GW67 1161.7 376.7 30.0 18-FH082 16-GW36 2395.0 1375.0 194.3 18-FH083 16-GW74 2233.3 1166.7 48.7 18-FH084 16-GW82 2846.7 1451.7 77.0 18-FH085 16-GW85 1566.7 586.7 51.3 18-FH086 16-GW86 1208.3 510.0 46.7 18-FH087 16-GW102 1870.0 741.7 62.3 18-FH088 16-GW107 2351.7 875.0 38.7 18-FH089 16-GW112 3516.0 1366.7 83.7 18-FH090 Wonkyo 20042 2081.7 830.0 49.7 18-FH091 14-FS21-C3 1261.7 258.3 14.7 18-FH092 14-FS21-C4 2200.0 796.7 44.0 18-FH093 14-FS21-C6 2460.0 873.3 47.0 18-FH094 17-RFC127 3203.3 1408.3 60.7 18-FH095 17-RFC148 2566.7 1380.0 34.7 18-FH096 14-FS15-C1 2556.7 1010.0 42.0 18-FH097 14-FS31-C3 2088.3 898.3 43.0 18-FH098 14-FS02-C1 2248.3 1255.0 50.7 18-FH099 16-RB7 2000.0 430.0 26.7 18-FH100 16-RB8 1845.0 296.7 24.3

'18-FH98' was named 'Wongyo 20051' after passing the job cultivating variety review meeting in November 2018, and the seeds of 'Wongyo 20051' were transferred to the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center as of October 8, 2019. It was deposited (Accession No. KCTC 13971BP). It was confirmed that 'Wongyo 20051' has a medium herbaceous character, a semi-conjugated type, and the inner leaf color is yellow similar to that of the control variety, 'Bulam No. 3', and the outer leaf color is darker green than the control variety. and variable characteristics are shown in Tables 7 and 8, respectively.

varieties and
genealogy rudimentary conspicuous outer leaf color inner leaf color leaf type mock up sleep Bulam No. 3 middle semiconjugation green dark yellow oval middle middle 18-FH98 small type semi-determined dark green yellow big honey egg middle just

varieties and
genealogy Weekday
(g) custody
(g) mouth width
(cm) gogo
(cm) postcard
(cm) leaf width
(cm) number of leaves
(Hawk) inner leaf water
(Hawk) Bulam No. 3 2,153.3a 1,018.3b 151.7a 230.0a 33.0b 21.3b 16.0a 58.0a 18-FH98 2,248.3a 1,255.0a 123.3b 263.3b 35.0a 25.7a 12.0b 38.7b

As a result of cultivation in 2018, the total weight of the plant was 2,248.3 g, which was similar to 2,340.0 g of the control group. 6). In addition, 'Wongyo 20051' forms a hard and large ball even in the autumn cultivation environment in 2018 when it rains frequently and the low temperature comes early (FIG. 2), so it is useful as a breeding material such as intermediate specimens that are resistant to low temperature stress and root nodules. It was confirmed that it can be used.

Up to now, the present invention has been looked at focusing on the embodiments thereof. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (5)

As a Chinese cabbage ( Brassica rapa . L) plant with cold stress and root nodule resistance,
The plant is a Chinese cabbage plant that was deposited with the seed accession number KCTC 13971BP.
[Claim 2] The Chinese cabbage plant according to claim 1, which has the following characteristics more than that of the control variety, Bulam No. 3:
(1) The maturation of the applied cultivar is slower than that of the control cultivar.
(2) The green tea of the applied variety is smaller than that of the reference variety.
(3) The color of the outer leaf is dark green for the applied variety, and green for the control variety.
(4) The color of the inner leaf is yellow for the applied variety, and dark yellow for the control variety.
(5) The outer leaf shape of the applied variety is an upside-down egg-shaped, and the control variety is an egg-shaped.
(6) The cultivar of this application is semi-gyle-gu, and the control cultivar is semi-conjugated.
A plant part of a Chinese cabbage plant according to claim 1 .
4. The method according to claim 3, wherein the part is a protoplast, an ovule, a cell, a pollen grain, an embryo, a cotyledon, a hypocotyledon, a root, an anther, A part of a Chinese cabbage plant which is selected from the group consisting of stems, leaves and seeds.
A method for producing a Chinese cabbage plant seed having increased resistance to cold stress and root lump disease, comprising the step of crossing the Chinese cabbage plant according to claim 1 with another Brassica plant.

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