KR101046511B1 - 'P ＆ B-AR5' Plant-Based Red Pepper Anthrax Resistant, Promoted Through Cross Species Between Capsicum annuum and C. baccatum - Google Patents

Abstract

The present invention is selected from the country, as well as India and Indonesia, according to Pepper anthrax being raised as a serious issue in many countries of Southeast Asia Susceptible Cultivars of Capsicum annuum and resistance allied species Capsicum baccatum between generations interspecific hybrids segregating population (BC ₁ F ₁₎ a resistance object (# 99) and Susceptible commercial inbred line (HN11) and the hybrid progeny selected, immobilized BC ₂ F through continuous child (selfing) and every generation resistance test of the resistance object, starting from (BC ₂ F _{1) It} is about the growth process and system characteristics of the _7th generation anthrax resistant plant system 'P &B-AR5'.

Peppers, which are grown all over the world, are suffering from severe yield loss and quality deterioration due to invasion of various pathogens throughout the growing season. In particular, pepper anthrax is known as one of the most damaging diseases in countries and regions including rainy season or rainy season during the growing season, as well as Korea and many countries in Southeast Asia. Until now, chemical control by disinfectant spraying has been mainly used. However, due to the difficulty in selecting the timing of drug control due to frequent rainfall during the cultivation period, and the emergence of new fungicide-resistant pathogens, the control value is not so high. .

The most economical, effective and environmentally friendly way to solve this problem is to cultivate resistant varieties, but there is no commercially grown single anthrax resistant strains and varieties worldwide. The reason for this is the absence of substantial resistant breeding efforts due to the absence of stable assay methods, which are essential prerequisites for resistant sarcoma, and the absence of commercially available highly anthrax resistant gene sources. Therefore, in the present invention, based on the previously registered Korean Patent No. 0504172, 'Culturing method for hybrid pepper by cross-breeding', which includes clues and solutions to the above prerequisites, a hybrid sub-separation group of two pathogenic cultivars and resistant myomas It is possible to select highly resistant individuals from them and to foster their descendants, and to develop a high anthrax resistant system with horticultural characteristics through continuous anthrax resistance test for each generation. Includes strategies for developing commercial anthrax resistant strains and breeds. That is, the present invention selects resistant individuals selected from the first generation of cross-female hybrids (BC ₁ F ₁ ) and resistant individuals from the BC ₂ F ₁ generations that hybridize the pathogenic commercial inbred line, 'HN11', which is resistant to persistent children and resistance. The screening is characterized by the development of a species-to-hybrid 6th generation strain with fixed anthrax resistance and horticultural characteristics.

Techniques for breeding pepper anthrax resistant strains using cross-crossing can be applied to almost all cross-breeding breeding methods that want to develop various pest resistant strains and varieties that damage peppers in addition to anthrax. In addition, the anthrax resistant strains fostered in the present invention are expected to make an absolute contribution to the development of strains and varieties suitable for all regions and countries where anthrax is afflicted with not only Korea, but also to further enhance the international competitiveness of the Korean seed industry. We think that we can provide electricity.

Pepper, Anthrax, Species Hybridization, Species-Cultivation, Disease Resistant, Resistant Black, Selection, System Development

Description

Pepper 'P & B-AR5' resistant to pepper anthracnose developed by interspecific hybridization between Capsicum annuum and C. baccatum}

The present invention is selected from the country, as well as India and Indonesia, according to Pepper anthrax being raised as a serious issue in many countries of Southeast Asia Susceptible Cultivars of Capsicum annuum and resistance allied species Capsicum baccatum between generations interspecific hybrids segregating population (BC ₁ F ₁₎ Highly resistant individuals were selected from the hybridization of one resistant entity (# 99) with the pathogenic commercial inbred line 'HN11' (BC ₂ F ₁ ), and selected and fixed through continuous selfing and generational resistance tests. BC ₂ F It is about the development of anthracnose-resistant plant family 'P &B-AR5' of ₇ generations.

Peppers grown all over the world are suffering from severe yield loss and quality deterioration due to invasion of various pathogens throughout the growing season. In particular, pepper anthrax is known as one of the most damaging diseases in countries and regions including rainy season or rainy season during the growing season, as well as late blight and cyanosis. It is estimated that the annual damage caused by anthrax is about 100 billion won, accounting for more than 10% of the total output of open field cultivation. In addition, the scale of damage to anthrax is increasing gradually in rainy season cultivation in southern China, India, Indonesia, Thailand and Taiwan. Until now, chemical control by disinfectant spraying has been mainly used. However, due to the difficulty in selecting the timing of drug control due to frequent rainfall during the cultivation period, and the emergence of new fungicide-resistant pathogens, the control value is not so high. . In Korea, Colletotrichum gloeosporioides was identified as 95% of the anthrax bacteria collected in the field in the early 1990s, but more than 99% have been identified as C. acutatum strains. This shows a variation in the distribution of pathogens due to continuous overspray of fungicides. In the latter case, resistance to fungicides developed to date has been reported to be much higher than that of other pathogens. Recent reports also show that in Indonesia and Taiwan, almost all pathogens present on the packaging are C. acutatum . In India and Thailand, the predominant pathogenic pathogens of C. capsici have been identified as C. capsici . In comparison with pathogen species or isolates, C. acutatum is generally the highest and other strains are relatively low. Therefore, in case of chemical control by continuous drug application, its effectiveness is highly doubted. Therefore, it can be sure that the best control measures are to cultivate and cultivate resistant varieties as the most economical, effective and environmentally friendly method. However, there are no examples of commercially grown single anthrax resistant strains and varieties worldwide. The reason for this is the absence of substantial resistant breeding efforts due to the absence of stable assay methods, which are essential prerequisites for resistant sarcoma, and the absence of commercially available highly anthrax resistant gene sources. There have been a number of assays for anthrax-resistant breeding reported so far, but recently developed by Taiwan's AVRDC, the microinjection method improved and established by the research team has been found to be the most stable. In the selection of anthrax resistant gene sources, the response could be very different depending on the inoculation method and pathogen isolates used. The recently developed microinjection method was used to inoculate with the most pathogenic C. acutatum strain. In case of cultivation, there is no single source of resistance gene yet, but some strains belonging to Capsicum baccatum species are resistant. Therefore, in order to introduce anthrax resistance to C. annuum , a species of C. annuum , species hybridization must be carried out. In this process, hybridization cannot be obtained due to the phenomenon that the embryo degenerates during the development. Was observed. In order to overcome this problem, cross-breeding was carried out using peppers of various cultivars. After the fertilization, the cultivation process of the pears was confirmed step by step. there was. Therefore, normal hybrid plants could be obtained by extracting and culturing cotyledons of the cotyledon in combinations showing partial compatibility. The growth of the hybrids between species was very normal and the fire structure was good, but the observation of pollen grains after the reorganization showed hybrid infertility. In order to overcome infertility, cross-hybridization was carried out using the hybrid plants as mothers and the parent seedlings as pot plants, and several hundred BC ₁ F ₁ generations were obtained. More than two-thirds of the plants in the first isolated population showed male sterility, resulting in normal fruiting. Therefore, when anthrax resistance test was performed on the fruit of 88 fruiting fruit, it was found that some individuals showed high resistance and overcome the problem of hybrid infertility. The results of the above study were published in some papers and almost all of them were included in the registered patent No. 0504172, 'By breeding method of hybrid pepper by cross hybridization'.

Therefore, the present invention fosters hybridization with commercial inbred lines used by seed companies using highly resistant individuals selected according to the above patents, and selects highly resistant individuals through resistance tests and then raises them. Each generation includes the development of anthrax resistant cultivars by continuously selecting and immobilizing highly resistant individuals through resistance tests.

Document 1 AVRDC. 1999. Off-season tomato, pepper, and eggplant. In: AVRDC 1998 Progress Report. p. 20-30. Tainan, Taiwan.

[Document 2] Yun Bok Yoon, Hyo Geun Park. 2001. Test Methods for Fruit Anthrax Resistance of Red Pepper: Spore Formation of Pathogens, Inoculation Method, Inoculation Concentration and Environmental Conditions after Inoculation. Korean Society for Horticultural Science 42: 389-393.

3 JB Yoon et al. 2004. Genetic resources resistant to anthracnose in the genus Capsicum . J. Kor. Soc. Hort. Sci. 45: 318-323.

4 JB Yoon et al. 2006. Overcoming two post-fertilization genetic barriers in interspecific hybridization between Capsicum annuum and C. baccatum for introgression of anthracnose resistance. Breed. Sci. 56: 31-38.

[Document 5] Yun Bok Yoon, Dong Cheol Yang, Hyo Geun Park. 2005. Breeding method of hybrid pepper by cross-breeding, Korean Patent No. 0504172.

The present invention has been started based on the registered patent No. 0504172 to introduce a new cultivar resistant strain by introducing pepper anthracnose resistance present in the recent species Capsicum baccatum into the cultivar C. annuum . Species - backcrossing BC ₁ F resistance object of a highly selected from _one separate group (# 99) is used as a planter friendly and BC ₂ F ₁ Separation created by crossing the 'HN11' systems that use a commercial inbred line in the seed company Highly resistant individuals were selected from the group, and the horticultural group was close to the cultivar, and the reared group was selected by rearing and re-producing the inferior hybrid infertility. Later generations tried to foster strains with horticultural characteristics and high resistance to anthrax by repeating selection and offspring.

The present invention is for fostering cultivated strains introduced pepper anthracnose resistance and using them to present strategies for commercial cultivated strains and breeding, and highly resistant individuals (# 99) selected according to the registered patent No. 0504172. Horticultural characteristics are close to cultivars and hybridized with resistance test by micro-injection to hybridized BC ₂ F ₁ group using parent seedlings and 'HN11', a commercial inbred line of seed companies. Selecting and regenerating individuals with high anthrax resistance that has recovered from infertility, fostering a later group BC ₂ F ₂ and then repeating selection and offspring to foster a system with fixed horticultural characteristics and anthrax resistance. It is characterized by consisting of steps.

The present invention relates to the development of pepper anthracnose resistant strains and will be recognized as the first and best technology worldwide. In addition, we can reduce the damage caused by the annual decrease of more than 100 billion won in Korea, and increase the international competitiveness of the Korean seed industry by providing an opportunity to export strains and varieties to several countries in Southeast Asia where the damage of anthrax is increasing day by day. I think it will greatly contribute.

The present invention, as in the registered patent No. 0504172 and Figure 1, through the cross-breeding (species F ₁ ) and female hybridization (species F ₁ × SP26) between two pathogenic cultivars (SP26) and resistant species (PBC81) Begin with anthrax resistant individuals (# 99) selected from BC ₁ F ₁ population, the first anthrax-resistant interstitial population that was created, foster isolates and perform resistance testing by microinjection to sustain highly resistant individuals from the subgroup. The selection includes the process of cultivating a fixed system with horticultural characteristics as well as high anthrax resistance. In other words, BC ₁ F ₁ 99 was used as a potted plant and hybridized with 'HN11', a commercial inbred line of seed companies, to create a BC ₂ F ₁ separation group. Anthrax bacteria predominate in Korea based on the micro-injection method. Highly resistant individuals were selected by performing a resistance assay using the KSCa-1 strain, one of ( C. acutatum ). BC ₂ F ₂ populations were raised by the offspring, and highly resistant individuals were selected and regenerated by the resistance test. As a result of repeating the process of selecting and substituting resistant individuals from the created BC ₂ F ₃ populations, the 6 generations of BC ₂ F ₃ generations were raised, resulting in a lineage that did not show individual variation in horticultural characteristics. As a result of anthrax resistance test on these strains, all individuals showed high resistance, and it was judged that they successfully introduced new strains by introducing resistance existing in cultivars. By investigating the horticultural characteristics of the cultivated strains, we could obtain distinct strains. Among them, we selected a strain that is considered to be highly commercialized because the horticultural characteristic is close to the cultivated species, and anthrax resistant strain 'P &B-AR5' And some important horticultural characteristics.

A. Nurturing later hybrid and resistance lines

Highly anthrax-resistant BC ₁ F ₁ 99 was used as a potted plant to hybridize with the 'HN11' strain, a commercial inbred line of seed companies, to foster BC ₂ F ₁ population and to test the next generation BC ₂ through resistance test. F ₂ populations were raised. In this group, individuals were selected and children were raised through the resistance test to foster BC ₂ F ₃ group. After that, the process of selecting individuals and children through the resistance test was repeated to develop 6 generations of BC ₂ F ₇ children. The process is shown in FIG.

2. The selfing of the selected resistant individual shall be covered with a small envelope made of folded paper with oil to prevent pollen contamination by placing a large amount of pollen from different flowers of the same plant on the front of the flower that has just blossomed. gave. One week after artificial pollination, fruit is growing and the bag is peeled off and labeled to indicate that it is a child.

3. When the fruit was completely colored red after artificial hybridization, it was harvested and received seeds. The received seeds were dried in the cold room to remove moisture, and then used as the next experimental material.

4. The harvested new generation of seeds were sterilized with disinfectant and trisodium phosphate, and placed in a petri dish coated with wet paper towels. Seeds germinated from the third day of cultivation were sown in 50-hole seedling trays and placed in pots or houses at the time of the formation of the first mines.

5. The degree of resistance of the grown resistant system is as shown in FIG.

B. Anthrax resistance assay using microinjection

1. After flowering, fruits were harvested for 35-45 days, washed with distilled water, and inoculated using microinjection.

2. As the pathogen, KSCa-1 strain, one of those identified as C. acutatum , was used, and the strain was stored in a refrigerator at 4 ° C., one week before inoculation, and cultured in PDA medium. In order to maintain the pathogenicity of the pathogen, strains were inoculated once every three months into two pathogenic red pepper fruits, and the pathogens were re-isolated from the developed lesions and cultured in PDA medium.

3. The concentration of inoculum was 1x10 ⁵ conidia / ml and inoculated at 3 places per fruit. The inoculated fruits were put in an acrylic box covered with wet paper towels, sealed with a commercial clean lab, incubated in a culture chamber at 25 ° C for 2 days, peeled off the sealed wrap, and cultured for another 5 days. It was. The degree of resistance was expressed as a percentage of diseased spots / total inoculated spots.

C. Horticultural Characterization

1. For the evaluation of horticultural characteristics of selected anthrax resistant strains, the characteristics were briefly examined visually according to the contents of the Breeding Bulletin of National Species.

2. The investigated characteristics were the degree of anthocyanin pigmentation of the hypocotyl and the plant type, the presence or absence of downy and leafy green leaves of the plant stem, the direction and flower color of the peduncle, the color of the green fruit and the fruit before maturation, and the shape of the cotton cut horizontally. , Texture and maturation of fruit surface, presence and absence of fruit stems, number of ventricle and thickness of flesh, shape of calyx and pungent taste of fruit.
The horticultural characteristics investigated were as follows.
-Anthocyanin pigmentation on the hypocotyls and plant primitives standing
-No or little downy stem of plant stem, leaves are dark green
-The peduncle's posture is slightly downward and the flower color is white.
-The color before fruit maturation is green and the fruit is thick and short
-The shape of the slice cut horizontally is circular
The texture of the fruit surface is slightly rough and the color of maturity is red.
-There is no depression of the fruit tap and the shape of the tip is sharp
-Fruit ventricular number 2 and medium flesh thickness
-Calyx is exposed and fruit is slightly hot

Figure 1 shows the development of anthrax resistant strain 'P & B-AR5'. After the hybridization, the separation group was created, and the individual selection and the children were repeated to proceed with six generations. The process of fostering a later group and line through the children of selected individuals after the selection of individuals through resistance test every generation.

2 shows the fruit characteristics and disease resistance of anthrax resistance system 'P & B-AR5'. Individual disease resistance evaluation of the fostered strain was expressed as a percentage and confirmed that the strain was fixed to a highly resistant strain without significant differences between individuals.

Claims (2)

a) obtaining species F ₁ by cross-breeding between C26 annuum SP26 and PBC81 resistant C. baccatum; b) obtaining the interstitial F ₁ and again forming an interstitial BC ₁ F ₁ population, which is an anthrax resistant interstitial segregation group through SP26 and female hybridization (species F ₁ × SP26); c) breeding inter-breeding BC ₂ F ₁ populations by mating with said interstitial BC ₁ F ₁ with a mother of commercial line HN11; d) a method for breeding P & B-AR5, characterized in that the anthrax resistant individual selection and offspring are repeated from BC ₂ F ₁ to obtain anthrax resistant strain in BC ₂ F ₇ The method of breeding 'P & B-AR5' according to claim 1, wherein the finally obtained anthrax resistant strain includes the following characteristics. -Anthocyanin pigmentation on the hypocotyls and plant primitives standing -No or little downy stem of plant stem, leaves are dark green -The peduncle's posture is slightly downward and the flower color is white. -The color before fruit maturation is green and the fruit is thick and short -The shape of the slice cut horizontally is circular The texture of the fruit surface is slightly rough and the color of maturity is red. -There is no depression of the fruit tap and the shape of the tip is sharp -Fruit ventricular number 2 and medium flesh thickness -Calyx is exposed and fruit is slightly hot

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