Application of GrVe gene providing plant with verticillium wilt resistance
A verticillium wilt and gene technology, applied in the field of plant genetic engineering, can solve the problems of long growth period, low yield, late maturity, etc., and achieve the effects of delaying onset, good resistance effect, and reducing disease fingers
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[0046] The methods used in the following embodiments are conventional methods unless otherwise specified, and the primer sequences used are all synthesized by Shanghai Handsome Biotechnology Co., Ltd., and the stated percentages are all mass percentages. The gene in this experiment was derived from black peak grape (Vitis vinifera).
[0047] (1) Grape GrVe cloning and sequence analysis
[0048] Primers were designed according to a receptor-like protein gene (accession number: XM_002263197) published in the Heifeng grape genome
[0049] P1: 5'-ATGTATCGAATCCTTTGCTTC-3',
[0050] P2:5'-CTAACTTCTTCTTGCTCCATG-3'
[0051] To amplify the cDNA template of Heifeng grape, add 1 μl of cDNA template to the 25 μl reaction system, 5 nmol of each primer, 5 μl of 5×primeSTARbuffer (Mg 2+ plus) PCR buffer, 0.2mMdNTP, 1UprimeSTAR HS DNA Polymerase (TaKaRa company) for PCR amplification. The PCR amplification conditions were: 94°C 3', 94°C 45'', 56°C 45'', 72°C 3', cycle 36 times, and then e...
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