Verticillium wilt resistance related GhBZR1 protein of Gossypium.hirsutum as well as coding gene and application of protein
A technology against verticillium wilt and upland cotton, applied in the field of molecular biology, can solve the problems that there are no research reports on BZR homologous genes
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Embodiment 1
[0050] Verticillium wilt disease resistance status of silent Verticillium wilt upland cotton variety 86-1 under Verticillium dahliae stress.
[0051] (1) Material
[0052] The wild-type Verticillium wilt-susceptible upland cotton variety Zhimian 86-1 and the Verticillium wilt-susceptible upland cotton variety Zhongzhimian 86-1 with GhBZR1 gene silenced were used as experimental materials.
[0053] (2) Culture medium preparation
[0054] Control group: wild type 86-1;
[0055] Experimental group: Zhimian 86-1, a Verticillium wilt-susceptible upland cotton variety with GhBZR1 gene silenced, and Zhimian 86-1 transformed with an empty vector (CLCrV-00) were inoculated with V991.
[0056] (3) Experimental design and trait determination
[0057] After the seeds were delinted with concentrated sulfuric acid before sowing, the seeds with the same plumpness and size were selected for subsequent experiments. The seeds were soaked in 70% ethanol for 5 minutes for surface sterilization...
Embodiment 2
[0063] Detection of Verticillium dahliae resistance in Arabidopsis thaliana by overexpressing GhBZR1 gene under stress of Verticillium dahliae.
[0064] (1) Material
[0065] The second-generation Arabidopsis thaliana and wild-type Arabidopsis thaliana overexpressing the GhBZR1 gene were used as experimental materials.
[0066] (2) Cultivation substrate
[0067] Medium: 1 / 2MS medium; 1 / 2MS+50mg / L Kanamycin medium
[0068] Cultivation substrate: vermiculite + 1 / 4MS nutrient solution
[0069] (3) Experimental Design and Determination of Disease Resistance
[0070] Wild-type Arabidopsis was cultured in 1 / 2MS medium, and transgenic Arabidopsis was cultured in 1 / 2MS+50mg / L kanamycin medium. After the seedlings grow to a certain size, the kanamycin-resistant transgenic Arabidopsis is transplanted into a nutrient pot. Verticillium dahliae stress treatment was carried out 7 days after transplanting, and the overexpressed plants were inoculated with Verticillium dahliae V991 by ro...
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