Flue-cured tobacco germ plasm cultivation method for increasing potassium ion content of tobacco leaves

A cultivation method and technology of potassium ions, applied in the field of cultivation of flue-cured tobacco germplasm, to achieve the effect of improving potassium ion uptake capacity and content

Inactive Publication Date: 2012-07-18
GUIZHOU TOBACCO SCI INST
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Problems solved by technology

[0005] These studies mainly demonstrated that potassium ion channel genes play an important role in crop potassium metabolism by heterologously expressing potassium ion channel genes from different sources in different crops, but so far there has been no report of overexpression of tobacco's own potassium ion inward transporter in tobacco NtHAK1 gene, while antisense represses tobacco's own potassium ion export channel gene TORK1 Methods for genetically modified breeding

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  • Flue-cured tobacco germ plasm cultivation method for increasing potassium ion content of tobacco leaves
  • Flue-cured tobacco germ plasm cultivation method for increasing potassium ion content of tobacco leaves

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experiment example

[0050] Experimental Example: Rapid Detection of Potassium Ion Content in Transgenic Tobacco Leaves

[0051]The thin layer of transgenic tobacco seedling leaves was cut for β-glucuronidase (GUS) staining, and the transgenic tobacco seedlings stained blue were selected for potassium ion content analysis. From the transgenic tobacco seedlings and non-transgenic tobacco seedlings transplanted into flower pots, the tobacco plants with relatively consistent growth were selected for analysis. After 30 days of cultivation, the middle three leaves of each tobacco plant were collected, killed at 105°C for 15 minutes, and dried at 80°C. Dry, remove the midrib, crush through a 0.25 mm sieve, and weigh for analysis. The content of potassium ions in the sample was determined according to the general standard method: Determination of potassium in tobacco - flame photometry (YC / T 173-2003). Through comparative analysis with non-transgenic tobacco seedlings, it was found that the potassiu...

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Abstract

The invention discloses a flue-cured tobacco germ plasm cultivation method for increasing the potassium ion content of tobacco leaves. The flue-cured tobacco germ plasm cultivation method comprises the following steps: firstly, cloning a tobacco auxin inducible promoter Pcel7 according to a conventional method; secondly, artificially synthesizing gene sequences of a tobacco potassium ion outward channel TORK1, a potassium ion inward transporter NtHAK1 and a terminator Tnos; thirdly, constructing a binary plant expression vector pSH-C-T-35S-H of a PCEL7 drive negative-sense TORK1 and a CaMV35Spositive-sense NtHAK1; fourthly, transforming soil agrobacterium and a tobacco explant; and fifthly, differentiating, transplanting and detecting a transgenic tobacco plant. According to the flue-cured tobacco germ plasm cultivation method disclosed by the invention, the drain of tobacco potassium ions is reduced by inducing an auxin signal; and meanwhile, the intake of the tobacco potassium ionsis increased, so as to facilitate the increase of the potassium ion content of the tobacco leaves.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for cultivating flue-cured tobacco germplasm capable of increasing the potassium ion content of tobacco leaves. Background technique [0002] Potassium is a recognized quality element of tobacco, and increasing the potassium content of tobacco is very important to reduce tar, reduce harmful substances, and increase safety. The potassium content of tobacco leaves in the United States and Zimbabwe is mostly 4-6%, while the potassium content of Chinese tobacco leaves is 1-2% on average, rarely exceeding 2%, and even less than 1%. The low content of potassium in tobacco seriously limits the quality and development of tobacco in my country. [0003] Potassium deficiency in tobacco leaves can be divided into exogenous potassium deficiency and qualitative potassium deficiency. Among them, the ability and level of tobacco's own uptake, transport, fixation and discharge ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 赵杰宏王轶韩洁
Owner GUIZHOU TOBACCO SCI INST
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