Verticillium wilt resistance related GhBZR1 protein of Gossypium.hirsutum as well as coding gene and application of protein
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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
Pending Publication Date: 2021-01-12
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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Upland cotton, as the main planting species in my country, has important
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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
[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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Abstract
The invention discloses verticillium wilt resistance related GhBZR1 protein of Gossypium.hirsutum as well as a coding gene and application of the protein. A nucleotide sequence of the protein is as shown in SEQ ID NO.1. The invention provides the protein coded by the verticillium wilt resistance related gene GhBZR1 of the Gossypium.hirsutum. The verticillium wilt resistance related gene GhBZR1 isobtained by cloning from leaves of a verticillium wilt infected Gossypium.hirsutum variety 86-1, and RT-PCR proves that the gene GhBZR1 has synergistic expression in the leaves and root systems underthe stress of Verticillium dahliae. Disease resistance evaluation of arabidopsis thaliana for transformation and overexpression of the gene proves that the gene can improve the Verticillium dahliae resistance of the verticillium wilt infected Gossypium.hirsutum variety, so that the gene can be used for improving genetic improvement of the verticillium wilt resistance of the Gossypium.hirsutum or genetic improvement of the verticillium wilt resistance of other Verticillium dahliae host crops.
Description
technical field [0001] The invention relates to the field of molecular biology, in particular to the GhBZR1 protein related to Verticillium wilt resistance of upland cotton and its coding gene and application. Background technique [0002] Cotton is an important economic crop and occupies an important position in the national economy. It is not only an important raw material for textile, chemical, pharmaceutical and national defense industries, but also an important export commodity. Among them, upland cotton (Gossypium. hirsutum) is the main planting species, accounting for more than 99% of the cotton area in my country. Verticillium wilt is one of the most important diseases in cotton production. It was introduced into my country in the 1930s, and began to spread and cause damage in some areas of China's cotton-producing provinces in the early 1950s after liberation. At the beginning, the damage in the main cotton-producing areas of our country increased year by year, and ...
Claims
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