Loofah gummy stem blight-resistant gene segments or gene markers and application thereof
A gene marker and gene fragment technology, applied in the field of plant biotechnology science, can solve problems such as economic loss, affecting the growth and development process of loofah plants, and achieve the effects of shortening breeding years, improving breeding efficiency, and improving identification efficiency.
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[0017] 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 the loofah resistance gene. The specific implementation process is as follows:
[0018] 1. DNA extraction: Take an appropriate amount of young leaves of loofah, 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.
[0019] 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 loofah genomic DNA 50ng as a template, and ca...
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