Molecular marker of rice major gene bph22 (t) resistant to brown planthoppers and application thereof
A technology for resisting brown planthopper and main gene, which is applied in the field of molecular genetics, can solve problems such as the complexity and instability of insect resistance identification, difficulty in effectively introducing and aggregating insect resistance genes, etc., and achieves convenient identification, convenient and fast detection, and cost saving effect
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Embodiment 1
[0043] (1) White hair / W2183BC 1 f 2 Population construction and phenotyping
[0044] (1) Li et al. (2006) identified resistance to BPH from the transgenic offspring of common wild rice in Guangxi and screened out BPH biotypes 1 and 2, Bangladesh, Mekong River (Vietnam), Jiulongjiang (Vietnam), The six biotypes in Pantnagar (India) all had broad-spectrum high-resistance W2183. BC constructed by crossing, backcrossing and selfing this resistance source with susceptible variety TN1 2 f 2 The population was mapped to the resistance gene, which was initially located between the molecular markers RM6506 and RM273. Reconstruction of BC using the sensual variety Baimao 1 f 2 Fine mapping of resistance genes in populations. Therefore, the present invention screens and develops SSR molecular markers based on PCR technology. In order to find simple and effective molecular markers that are closely linked to bph22(t), the present invention uses the susceptible variety Baimao as the...
Embodiment 2
[0067] Example 2 Verification of Molecular Markers
[0068] 1. Materials and methods
[0069] 1.1 Negative varieties: 20 copies, 14 copies of non-insect-resistant materials from the susceptible varieties Baimao, Bai R54, Teqing (materials preserved by our laboratory), 9311, Nipponbare, TN1, and Baimao×W2183 hybrids.
[0070] Positive varieties: resistant line W2183, Baimao × W2183, Bai R54 × W2183 hybrid offspring 19 insect-resistant materials.
[0071] Molecular marker primers: RM16846, RM16852, RM16853, RM16858, RM16874 and RM16888, the nucleotide sequences of which are respectively shown in SEQ ID No.1-12.
[0072] 1.2 Method
[0073] 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 RM16846, RM16852, RM16853, RM16858, RM16874 and RM16888, respectively. 10 μl system. The 10μl reaction system includes: 10×PCR buffer, 1.0μl; 10mMdNTPs, 0.1μl; 10μM primer, 0.4μl; 5U / μl Taq DN...
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