Functional molecular marker of related genes of cell nuclear male sterility of peppers and application thereof
A technology with the same function as the nucleus, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve the problems of fine positioning of pepper GMS fertility genes and no molecular markers, etc.
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Embodiment 1
[0080] Example 1, Development of Molecular Marker InDel-12 and Polymorphism Detection
[0081] 1. Development of Molecular Marker InDel-12
[0082] 1. Statistically locate the pollen fertility of each individual plant in the population
[0083] Using the capsicum male sterile line 09C460AB as the test material (fertile plants and sterile plants are near-isogenic lines), a mapping population consisting of 1110 individual plants was obtained. The pollen fertility of each individual plant in the positioning population was counted separately.
[0084] 2. Development of molecular marker InDel-12
[0085] (1) Take the fertile individual plants of the positioning population obtained in step 1, and mix the pools to obtain a fertile pool. The number of individual plants in each fertile pool is 30.
[0086] (2) Take the sterile individual plants of the positioning population obtained in step 1, and mix the pools to obtain sterile pools. The number of single plants in each sterile p...
Embodiment 2
[0119] Embodiment 2, application of molecular marker InDel-12
[0120] 1. Application 1
[0121] The peppers to be tested were 46 pepper inbred lines and varieties whose genotype was known as MsMs (see Table 1). All pepper inbred lines and varieties are publicly available from the College of Horticulture, China Agricultural University.
[0122] Table 1
[0123]
[0124]
[0125] 1. Carry out genotyping of the pepper to be tested according to the method of polymorphism detection one in Example 2.
[0126] 2. Plant the pepper samples to be tested in the field, manage them in the field routinely, and observe the pollen fertility after maturity.
[0127] For the experimental results of step 1, see figure 2 (M is a DNA marker, F is a fertile pool, S is a sterile pool, and the rest are 46 pepper inbred lines and varieties). The results show that the band type of the PCR amplification product of the pepper to be tested is all band type A (showing a band, which is 176bp), ...
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