NtCHA1 gene capable of regulating and controlling potassium content of tobacco leaves and application of NtCHA1 gene
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A technology of potassium content and genetics, applied in the field of genetic engineering
Pending Publication Date: 2022-01-18
YUNNAN ACAD OF TOBACCO AGRI SCI
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Embodiment 1
[0059] Example 1 Using CRISPR / cas9 to Knockout the NtCHA1 Gene to Reduce Potassium Content in Tobacco Leaf
[0060] 1.1 Experimental materials
[0061] 1.1.1 Tested strains
[0062] Escherichia coli DH5α. Agrobacterium (Agrobacterium) competent cells C58C1. The above biological materials are also preserved in this laboratory, and the applicant declares that they can be released to the public for verification experiments within 20 years from the date of application.
[0063] 1.1.2 Tobacco varieties tested
[0064] Tobacco: wild-type tobacco Yunyan 87, preserved in this laboratory, the applicant declares that it can be released to the public for verification experiments within 20 years from the date of application.
[0065] 1.1.3 Test reagents
[0066] pHSE401 vector, pK2GW7 vector, Trans-T1 competent cells, plasmid extraction kits, PCR product recovery kits, gel recovery kits, plant RNA extraction kits, and plant genome extraction kits were purchased from Beijing Quanshiji...
Embodiment 2
[0104] Example 2 Using overexpression of NtCHA1 to increase the content of potassium ions in tobacco leaves
[0105] 2.1 Experimental materials
[0106] 2.1.1 The tested bacterial strain is the same as the tested bacterial strain recorded in 1.1.1 in the examples.
[0107] 2.1.2 Tobacco varieties tested
[0108] Tobacco: Tobacco Yunyan 87, preserved in this laboratory, the applicant declares that it can be released to the public for verification experiments within 20 years from the date of application.
[0109] 2.1.3 Test reagents
[0110] PCR-Blunt II-TOPO cloning vector, pK2GW7 vector, plasmid extraction kit, PCR product recovery kit, gel recovery kit, plant RNA extraction kit, and plant genome extraction kit were purchased from Beijing Quanshijin Biotechnology Co., Ltd. ; 6-BA 6-benzylaminopurine; CRISPR / Cas9 plant expression vector preserved in Nanben Laboratory; LRClonaseTMⅡenzyme Mix, reverse transcriptase M.MLV, RNase inhibitor were purchased from Promega, USA; T4 DNA ...
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Abstract
The invention relates to the technical field of genetic engineering, and relates to a gene for regulating and controlling the potassium content of tobacco leaves, in particular to an NtCHA1 gene capable of regulating and controlling a potassium content of tobacco leaves and application of the NtCHA1 gene. The NtCHA1 gene has a nucleotide sequence shown as SEQ ID NO. 1. The invention further provides an application based on the NtCHA1 gene for regulating and controlling the potassium content of the tobacco leaves, the potassium content of the tobacco leaves can be remarkably increased by overexpressing the NtCHA1 gene in the tobacco leaves, and the potassium content of the tobacco leaves is remarkably reduced compared with that of a control group after the gene is knocked out through CRISPR/cas9. According to the invention, a regulation and control function of the NtCHA1 gene on the potassium content of the tobacco leaves is defined, a new evidence is provided for analyzing a molecular regulation and control mechanism of the potassium ion content of the tobacco leaves, and a basis is also provided for application of improving the potassium content of the tobacco leaves through overexpression of the gene.
Description
technical field [0001] The invention relates to the technical field of genetic engineering, and relates to a gene for regulating the potassium content of tobacco leaves, in particular to an NtCHA1 gene capable of regulating the potassium content of tobacco leaves and an application thereof. Background technique [0002] Tobacco is an important foliar economic crop in my country, and the potassium content of tobacco leaves seriously affects the economic value of tobacco leaves. Potassium ions in tobacco leaves participate in the physiological and biochemical reactions of tobacco leaves, and are closely related to the stress resistance of tobacco plants. For tobacco agriculture, it also affects the internal quality and industrial availability of tobacco leaves. Therefore, potassium content is usually used as a measure of tobacco leaf quality. Important indicators. Potassium has an important influence on the maturity, flavor, flammability, safety and other aspects of tobacco l...
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