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.
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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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