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Upland cotton ghebf1 gene, protein and application

An upland cotton, gene technology, applied in the field of molecular biology, can solve the problem that there is no EBF homologous gene research report and so on

Active Publication Date: 2021-09-14
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Upland cotton, as the main planting species in my country, has important economic value, but there is no research report on EBF homologous genes

Method used

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  • Upland cotton ghebf1 gene, protein and application
  • Upland cotton ghebf1 gene, protein and application
  • Upland cotton ghebf1 gene, protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Cloning of Example 1 Upland Cotton GhEBF1 Gene

[0034] 1. Extraction of RNA

[0035] RNAprep Pure plant polyphenol polysaccharide total RNA extraction kit was used to extract RNA from cotton KV3 leaf samples in cotton varieties.

[0036] 2. Synthesis of cDNA

[0037] 2.1 Synthesis of intermediate fragment cDNA

[0038] The cDNA of the intermediate fragment was synthesized using the FastQuant cDNA First Strand Synthesis Kit to reverse transcribe the cDNA.

[0039] 2.2 Synthesis of cDNA at the 3' end

[0040] The synthesis system of cDNA at the 3' end is as follows:

[0041] Table 1

[0042]

[0043] Mix according to the above system, after centrifugation, place on PCR instrument at 42°C for 60 minutes, and at 70°C for 15 minutes, cool on ice after the reaction, and store at -20°C.

[0044] 3.3 Synthesis of cDNA at the 5' end

[0045] The cDNA synthesis system of 5'RACE is as follows:

[0046] first step:

[0047] Table 2

[0048]

[0049] Mix well, centrif...

Embodiment 2

[0116] Example 2 Effect of Silencing Verticillium Wilt-resistant Upland Cotton Variety KV3 on Verticillium Wilt Disease Resistance under the Stress of Verticillium dahliae

[0117] (1) Material

[0118] The wild-type Verticillium wilt-resistant upland cotton variety Zhiqian KV3 and the Verticillium wilt-resistant upland cotton variety Zhiqian KV3 with GhEBF1 gene silenced were used as experimental materials.

[0119] (2) Experimental grouping

[0120] Control group: Zhongzhi Cotton KV3 (provided by the Institute of Plant Protection, Chinese Academy of Agricultural Sciences);

[0121] Experimental group: GhEBF1 gene-silenced GhEBF1-resistant upland cotton planting KV3, transforming empty vector (CLCrV-00) planting cotton KV3 (refer to Zhang Huachong's master's thesis "VIGS technology analysis of genes related to Verticillium wilt resistance in upland cotton Preliminary identification of GHB2 function", Graduate School of Chinese Academy of Agricultural Sciences, 2016; and Ren...

Embodiment 3

[0131] Example 3 Verticillium dahliae stress transfection overexpression GhEBF1 Arabidopsis Verticillium dahlia resistance detection

[0132] (1) Material

[0133] The second-generation Arabidopsis thaliana and wild-type Arabidopsis transfected with the GhEBF1 gene were used as experimental materials (see Zhang Huachong's master's thesis "Preliminary identification of the function of GHB2, a gene related to resistance to Verticillium wilt in upland cotton by VIGS technology", China Graduate School of the Chinese Academy of Agricultural Sciences, 2016; and Ren Yuhong's master's thesis "Cotton Verticillium wilt control technology and the research on the function of two resistance-related genes", Graduate School of the Chinese Academy of Agricultural Sciences, 2018).

[0134] (2) Cultivation substrate

[0135] Medium: 1 / 2MS medium; 1 / 2MS+50mg / L Kanamycin medium;

[0136] Cultivation substrate: vermiculite + 1 / 4MS nutrient solution

[0137] (3) Experimental Design and Determina...

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Abstract

The invention discloses an upland cotton GhEBF1 gene, protein and application thereof, belonging to the technical field of molecular biology. The gene includes: 1) a DNA molecule as shown in SEQ ID NO: 1; 2) a DNA molecule as shown in the sequence shown in SEQ ID NO: 1 from the 270th nucleotide sequence at the 5' end; 3) with The DNA sequence defined in 1) or 2) has at least 70% homology and encodes a DNA molecule that encodes a protein related to plant Verticillium wilt resistance. Also disclosed are the protein encoded by the gene; and the application of the gene or the protein, including: 1) promoting the coordinated expression of plant leaves and roots; 2) improving the plant's ability to resist Verticillium wilt. Through gene silencing and overexpression analysis, the invention confirms its function in the upland cotton's resistance to Verticillium wilt, and provides candidate genes for the next step of cultivating disease-resistant varieties.

Description

technical field [0001] The invention relates to the technical field of molecular biology, relates to an upland cotton GhEBF1 gene, protein and application thereof, in particular to a GhEBF1 gene, protein and application thereof related to upland cotton Verticillium wilt resistance. 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 occurred and caused damage in a few areas of my country’s cotton provinces in the 1950s. With the transfer of bacteria-carrying cotton seeds, it spread very...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/11C12N15/82A01H5/06A01H5/12A01H6/20A01H6/60
CPCC07K14/415C12N15/8282
Inventor 简桂良卢美光杨武贺浪张华崇司宁
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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