Pea downy mildew resistance gene fragment or gene marker and application
A technology of gene markers and gene fragments, applied in the field of plant biotechnology science, can solve problems such as economic loss and affecting the growth and development of pea plants, and achieve the effects of shortening the breeding period, improving breeding efficiency, and shortening the excavation cycle
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[0015] The identification of disease-resistant genes plays an important role in the theoretical research of crop disease-resistant genetics and the selection of disease-resistant varieties. This method can quickly identify downy mildew resistance genes in pea. The specific implementation process is as follows:
[0016] 1. DNA extraction: Take an appropriate amount of young leaves of pea, dry them with silica gel, grind them into powder, and extract the total genomic DNA by CTAB method. The extracted DNA was detected by an ultraviolet spectrophotometer, and samples with an OD260 / OD280 value of 1.8-2.0 were selected and stored at -20°C for future use.
[0017] 2. Amplification and cloning of homologous fragments of disease resistance genes: according to the amino acid sequences GG(L / I / V / M / S)GKTT and G(L / N ) PL (A / T / S) (L / V), designed a pair of degenerate primers: NBS15'-GKTT GGIGGIWSIGGIAARACIACG-3' and NBS25'-GCIGCIIARIGGRTTICC-3'. Take 50 ng of pea genomic DNA as a template...
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