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Application of CBL-interacting protein kinases (CIPK)3 protein and encoding genes thereof to plant drought resistance

A technology for transgenic plants and plants, applied in applications, plant products, genetic engineering, etc., can solve the problems of unpredictable results, long breeding cycle, etc., and achieve the effects of reducing water loss, short breeding time, and strong purpose.

Inactive Publication Date: 2020-01-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional breeding methods need to identify and combine excellent traits. Accuracy and efficiency determine the success or failure of breeding. Although the biological safety and stability are high, the breeding cycle is long and the results are unpredictable

Method used

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  • Application of CBL-interacting protein kinases (CIPK)3 protein and encoding genes thereof to plant drought resistance
  • Application of CBL-interacting protein kinases (CIPK)3 protein and encoding genes thereof to plant drought resistance
  • Application of CBL-interacting protein kinases (CIPK)3 protein and encoding genes thereof to plant drought resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Construction and detection of embodiment 1 CIPK3 gene overexpression vector

[0041] Total RNA was extracted from B73 corn (Zea mays L.), and cDNA was obtained by reverse transcription. The cDNA was used as a template, and F and R were used as primers to amplify the CIPK3 gene. on the carrier.

[0042] CIPK3 gene overexpression vector construction method is as follows:

[0043] (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.

[0044] (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.

[0045] (3) Using cDNA as a template, F and R as primers, amplify the cDNA of the CIPK3 gene (as shown in SEQ ID NO.3, its encoded amino acid sequence is as shown in SEQ ID NO.1), and run electrophoresis on the amplified product The gel was cut and recovered, and the recovery m...

Embodiment 2

[0053] Construction and detection of embodiment 2 CIPK3 gene overexpression plant

[0054] The pBCXUN-CIPK3 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 different transgenic inbred lines w...

Embodiment 3

[0055] Example 3 CIPK3 gene overexpression maize drought treatment phenotype detection

[0056] Add 140g of soil to each small pot, add water to the tray, put 4 corn seeds with CIPK3 gene overexpression and the seeds of non-transgenic control plant B73 in each small pot, cover 50mL of soil, absorb the water and remove the remaining Pour off the water, remove a seedling with uneven growth after emergence, add 1L of water in the tray, pour off the water after it is full, start the drought treatment, and observe the drought treatment phenotype of the control and transgenic plants. Three pots of control and transgenic plants were replicated. The result is as figure 2 As shown, the growth condition of the transgenic plants overexpressing CIPK3 was significantly better than that of the control plants, and the leaf wilting degree was significantly lower than that of the control plants, indicating that the transgenic plants overexpressing the CIPK3 gene had stronger drought resistan...

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Abstract

The invention relates to application of a CBL-interacting protein kinases (CIPK)3 protein and encoding genes thereof to plant drought resistance. The CIPK3 protein has the amino acid sequence shown asSEQ ID NO.1. The CIPK3 genes are cloned, transgenic plants for overexpressing the CIPK3 genes are constructed to analyze characters of the obtained transgenic plants, thus it is found that by increasing the expression quantity of the CIPK3 protein in the plants, the transpiration rate of the plants can be significantly decreased, stomatal conductance of the plants can be significantly reduced, the water utilization rate of the plants is increased, and the growth conditions of the plants under the drought condition are improved. Therefore, the CIPK3 protein and the encoding genes thereof can be used for enhancing the drought resistance ability of the plants in production practice, the yield of the plants under the drought conditions is increased, and water resources are saved.

Description

technical field [0001] The invention relates to the field of genetic engineering and genetic breeding, in particular to the application of CIPK3 protein and its coding gene in plant drought resistance. Background technique [0002] As the global climate warms, the population continues to grow, fresh water resources are scarce, and the problem of drought is becoming increasingly prominent. Drought stress affects the growth and yield of crops and brings serious disasters to agricultural production. It has become a worldwide problem restricting agricultural production. The crop loss caused by it ranks first among abiotic stresses. Most important crops are sensitive to drought, so cultivating new drought-resistant varieties can effectively cope with the yield loss caused by drought stress and improve water use efficiency. Traditional breeding methods need to identify and combine excellent traits. Accuracy and efficiency determine the success or failure of breeding. Although the...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N9/12C12N15/54A01H5/00A01H6/46A01H6/54A01H6/60
CPCC12N9/1205C12N15/8261C12N15/8273
Inventor 巩志忠王瑜孙志慧秦少川
Owner CHINA AGRI UNIV
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