Molecular marker of brown planthopper-resistant major gene bph7 of rice and application thereof
A technology of anti-BPH and main gene, applied in the field of molecular genetics, can solve complex and difficult problems such as effective introduction and aggregation of insect-resistant genes, and achieve the effects of difficulty, shortening the breeding cycle, and convenient identification
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Embodiment 1
[0040] (1) 9311 / T12 F 2 Population construction and phenotyping
[0041] (1) In 1988, Kabir & Khush used insect resistance screening to find that the rice variety T12 was resistant to the brown planthopper population (biotype 4). Genetic analysis showed that the variety carried a pair of recessive brown planthopper resistance genes and named it for bph7 (Kabir & Khush, 1988 Genetic analysis of resistance to brown planthopper in rice, Oryza sativa L. Plant Breeding 100, 54–58). The rice variety comes from the germplasm resource bank of the Chinese Academy of Agricultural Sciences, and T12 comes from the rice germplasm resource bank of the International Rice Research Institute. Insect resistance identification experiments showed that T12 had moderate resistance to the brown planthopper population in my country. In order to find simple and efficient and compatible with bph7 Tightly linked molecular markers, in the present invention, the insect-susceptible variety 9311 i...
Embodiment 2
[0064] Example 2 Verification of molecular markers
[0065] 1. Materials and methods
[0066] 1.1 Materials
[0067] Negative varieties: 10 copies, 8 copies of non-insect-resistant materials in the breeding combination of susceptible variety 9311, Taichung Local No. 1 (TN1), and 9311 X T12.
[0068] Positive varieties: 9 insect-resistant materials from the breeding combination of insect-resistant variety T12 and 9311 X T12.
[0069] Molecular marker primers: RM28295, RM519, RM28305, RM3448, RM313, RM28374.
[0070] 1.2 Method
[0071] Genomic DNA of rice samples was extracted by CTAB extraction method (the method is the same as in Example 1). Sample DNA was amplified with primers RM28295, RM519, RM28305, RM3448, RM313 and RM28374, respectively. The reaction system included 0.10 μM primers, 250 μM dNTPs, 1× PCR reaction buffer (50 mM KCl, 10 mM Tris-HCl pH8.3, 1.5 mM MgCl 2 ), 100ng DNA template, 1U Taq enzyme. The reaction program was: 94°C pre-denaturation for 5 minu...
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