Primers and method for improving resistance of susceptible rice varieties to black streaked dwarf viral disease
A black-streaked dwarf disease and rice technology, applied in the field of molecular genetics, can solve problems such as environmental pollution and difficulty in achieving expected results, and achieve the effects of clear location, convenient identification, and accelerated cultivation process
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[0028] 1) Under the genetic background of Huaidao 5, the carrier " qRBSDV-6 The construction of the near isogenic line of "
[0029] F 1 , and then use "Huaidao No. 5" as the recurrent parent for continuous backcrossing, from BC 1 f 1 From the beginning of the segregation generation, for different rice single plants, use the target resistance QTL provided by the present invention" qRBSDV-6 The SSR markers RM7158 and RM587 on both sides were used for genotype detection. The DNA was extracted using the conventional CTAB method.
[0030] PCR amplification system: 0.2μmolL -1 1.5 μL each of the forward and reverse primers, 200 μmolL -1 dNTPs 2μL, 10×PCRbuffer 2μL, 50~100ng DNA template 2μL, 1UμL -1 of tag Enzyme 0.3 μL, ddH 2 O10.7 μL.
[0031] The PCR reaction conditions were: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 1 min, renaturation at 55°C for 1 min, extension at 72°C for 1 min, 35 cycles, and finally extension at 72°C for 5 min.
[0032] Selec...
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