SNP molecular marker related to infertility of capsicum and application thereof

A molecular marker, nuclear sterility technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., which can solve the problems of restricting the large-scale application of GMS and unclear physical location and allelic relationship of pepper GMS genes. , to reduce the cost of seed production, accelerate the breeding process, and speed up the breeding process.

Active Publication Date: 2019-05-14
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, only a few genes have been identified as loci, such as ms1, ms3, ms8, ms w 、ms nw and msc-1 are located on pepper chromosomes 5, 1, 4, 5, 5 and 2, respectively, and the physical locations of the other pepper GMS genes and the allelic relationship between them are still unclear
[0005] In addition, when using the GMS dual-purpose line to produce the first-generation pepper hybrid seeds, the conventional method is still used, that is, from the early flowering stage, after the individual plant fertility is manually identified, about 50% of the fertile plants in the population are pulled out one by one. Greatly limit the large-scale application of GMS

Method used

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  • SNP molecular marker related to infertility of capsicum and application thereof
  • SNP molecular marker related to infertility of capsicum and application thereof
  • SNP molecular marker related to infertility of capsicum and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] (1) Plant material

[0057] Seeds (about 250 seeds) of pepper GMS dual-purpose line ("SC13AB") were soaked to accelerate germination and seedlings were raised in plug trays. When the seedlings grow 5 to 6 flat true leaves, they are planted in Zengcheng Experimental Base of South China Agricultural University, and they are managed with conventional fertilizer and water.

[0058] (2) Phenotypic identification

[0059] After the colony plants grow to the flowering stage, the fertility phenotypes of the individual plants are identified one by one. The presence or absence of pollen was first observed with the naked eye, and then the phenotype identification of pollen fertility was carried out one by one by the method of pollen in vitro culture and germination. Those with a large amount of pollen observed by naked eyes and able to germinate normally after in vitro pollen culture are judged as fertile plants; those without pollen observed by naked eyes and confirmed by micro...

Embodiment 2

[0111] Example 2 A method for molecular marker identification of capsicum fertility and seedling stage

[0112] (1) Nursing

[0113] (1) Soak the capsicum seeds in clear water for 10 hours, dry them and wrap them in cotton cloth;

[0114] (2) Accelerate germination in a 30°C incubator for 48 hours, and sow it in a nutrition cup after the buds appear;

[0115] (3) After the seedlings are unearthed and the cotyledons are flattened, the seedlings are falsely planted to the plug;

[0116] (4) After the seedlings grow to 2 to 3 true leaves, obtain the pepper seedling samples; then sample and extract DNA;

[0117] (2) Genotyping

[0118] The total volume of the PCR reaction system is 1 μL, including 0.36 μL DNA solution (50 ng), 0.5 μL 2×KASP Mastermix 1536, and 0.14 μL Primer mix. Gradient PCR reaction program includes: denaturation at 95°C for 15 minutes; denaturation at 94°C for 20s, annealing at 61°C to 55°C (0.6°C per cycle, 10 cycles in total) annealing for 60s; denaturati...

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Abstract

The invention belongs to the technical field of vegetable molecular breeding, and particularly relates to an SNP molecular marker related to infertility of capsicum and application thereof. The SNP molecular marker related to the infertility of the capsicum is SNP2, SNP3, SNP4, SNP7 or SNP8, the nucleotide sequence of an SNP site is shown in SEQ ID NO.1-5, and the molecular marker is co-separatedor closely linked to pepper fertility phenotype. The invention also provides a KASP primer for identifying the above SNP molecular marker; the primer can be used for rapidly identifying the pepper fertility phenotype, and can greatly accelerate the transfer process of a novel capsicum male infertility line. The SNP molecular marker related to the infertility of the capsicum developed belongs to acosegregation SNP marker of a nuclear infertility gene, and the plant fertility can be determined in the seedling stage of a pepper, thereby reducing the seed production cost. The marker can also accelerate the transfer process of the novel capsicum male infertility line.

Description

technical field [0001] The invention belongs to the technical field of vegetable molecular breeding, and in particular relates to a SNP molecular marker related to capsicum sterility and an application thereof. Background technique [0002] Capsicum is a vegetable crop of the Solanaceae family that is widely cultivated worldwide. The heterosis of pepper is obvious, and the first-generation hybrid seed is the main utilization form of pepper for current and future production. The use of male sterility to reproduce the first generation of hybrid seeds can avoid artificial emasculation, thereby reducing the cost of seed production, and is an ideal way to utilize heterosis in peppers. [0003] There are mainly two types of male sterility used in the production of pepper hybrid seeds, namely, cytoplasmic male sterility (CMS) and genic male sterility (GMS). Compared with the former, the latter usually has the advantages of more stable sterility and is not limited by the restorati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
Inventor 程蛟文吴智明董骥驰胡开林
Owner SOUTH CHINA AGRI UNIV
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