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Application of GLK7 protein and coding gene thereof in drought resisting of plants

A technology for encoding genes and proteins, applied in the field of plant genetic engineering, to achieve the effects of increasing content, shortening breeding time, and improving efficiency

Active Publication Date: 2020-12-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are only a few reports on the function of GLK family proteins in plants, and the mechanism of its participation in drought resistance of monocot crop maize has not been reported yet

Method used

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  • Application of GLK7 protein and coding gene thereof in drought resisting of plants
  • Application of GLK7 protein and coding gene thereof in drought resisting of plants
  • Application of GLK7 protein and coding gene thereof in drought resisting of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the construction and detection of GLK7 gene overexpression vector

[0040] In order to study the molecular mechanism of GLK7 family proteins in plant drought resistance, total RNA was extracted from B73 maize (Zea mays L.), and cDNA was obtained by reverse transcription. The GLK7 gene (nucleoside The acid sequence is shown in SEQ ID NO.1, the amino acid sequence is shown in SEQ ID NO.2), and the primer has a restriction site, which is connected to the overexpression vector after digestion.

[0041] Carrier construction method:

[0042] (1) The total RNA of B73 corn was extracted with the RNA extraction kit of Magen Company, and the specific steps were referred to the instructions of the kit.

[0043] (2) Use the reverse transcription kit from Thermo Company to reverse transcribe the RNA into cDNA, and refer to the kit instructions for specific steps.

[0044] (3) Use cDNA as a template and F and R as primers to amplify the cDNA of GLK7 gene, run electropho...

Embodiment 2

[0052] Embodiment 2, construction and detection of GLK7 gene overexpression plant

[0053] The pBCXUN-GLK7 overexpression plasmid constructed in Example 1 was transformed into competent Agrobacterium strain EHA105 by heat shock method, and positive clones were identified by colony PCR. Inoculate a single colony of Agrobacterium correctly identified in 2-3 mL of liquid medium containing 100 μg / mL kanamycin and 50 μg / mL rifampicin, culture with shaking at 28 °C overnight, and transfer a large amount of liquid containing antibiotics the next day Shake culture in the medium, collect the cells after several transfers, and resuspend to OD 600 Between 0.8-1.0. The obtained recombinant Agrobacterium suspension was used to infect the young B73 maize embryos picked out under aseptic conditions, and the callus was induced to form seedlings. The transgenic plants were self-propagated to obtain the T3 generation for subsequent experiments. The RNA of the wild type and different transgen...

Embodiment 3

[0054] Example 3, phenotype detection of GLK7 gene overexpression maize drought treatment

[0055] Add 140g of soil to each small pot, add water to the tray, put 4 seeds in each small pot, cover with 50ml of soil, pour out the remaining water in the tray after absorbing the water, and put a seedling with uneven growth after emergence Remove it, add 1L of water in the tray, drain the water after it is full, start the drought treatment, and continue to observe the growth of the wild-type and transgenic plants after the drought treatment. Wild-type and transgenic plants were duplicated in 2 pots each. figure 2 It showed that the growth condition of the transgenic plants overexpressing GLK7 was better than that of the wild type, and the leaf wilting degree was lower than that of the wild type, which indicated that the transgenic plants were more drought-resistant than the wild type.

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular to application of GLK7 protein and a coding gene thereof in drought resisting of plants. The invention finds that, after overexpression of a GLK7 gene, the wilting degree of leaves of a transgenic plant is lower than that of a wild type plant under the drought treatment condition, and the chlorophyll content is higher than that of the wild type plant, which indicates that plant drought resistance can be remarkably improved through the overexpression of the gene, a gene resource is provided for cultivationand improvement of new varieties of drought-resistant plants, and a theoretical basis is provided for illustrating the molecular mechanism of a GLK7 transcription factor in plant drought stress response.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the application of GLK7 protein and its coding gene in plant drought resistance. Background technique [0002] As an important food crop and economic crop in the world, the growth and yield of corn are affected by various environmental factors. Among them, drought is one of the factors that have the greatest impact on maize yield, and the crop loss caused by it ranks first among abiotic stresses. Therefore, cultivating new drought-resistant varieties can effectively cope with the yield loss caused by drought stress and improve water use efficiency. Compared with traditional breeding, molecular breeding has some unique advantages. First, it can improve breeding efficiency without the need for years of breeding processes such as hybridization and screening of excellent traits. Second, it is highly directional and the results can be predicted. Editing or overexpr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/12A01H6/46
CPCC07K14/415C12N15/8273
Inventor 王瑜巩志忠杨欣欣
Owner CHINA AGRI UNIV
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