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Application of CIPK9 protein and coding gene of CIPK9 protein in drought resistance of plants

A technology of transgenic plants and plants, applied in applications, plant products, genetic engineering, etc., can solve the problems of long breeding cycle and unpredictability of results, and achieve the effects of short breeding time, reduced water loss, and enhanced drought resistance

Inactive Publication Date: 2020-01-17
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 CIPK9 protein and coding gene of CIPK9 protein in drought resistance of plants
  • Application of CIPK9 protein and coding gene of CIPK9 protein in drought resistance of plants
  • Application of CIPK9 protein and coding gene of CIPK9 protein in drought resistance of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040]Example 1 Construction and detection of CIPK9 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 CIPK9 gene. on the carrier.

[0042] CIPK9 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 CIPK9 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 metho...

Embodiment 2

[0053] Example 2 Construction and detection of CIPK9 gene overexpression plants

[0054] The pBCXUN-CIPK9 overexpression plasmid constructed in Example 2 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 was...

Embodiment 3

[0055] Example 3 Phenotype Detection of CIPK9 Gene Overexpression Maize under Drought Treatment

[0056] Add 140g of soil to each small pot, add water to the tray, put 4 seeds of CIPK9 gene overexpressed corn 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 status of the transgenic plants overexpressing CIPK9 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 CIPK9 gene had stronger drought resis...

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Abstract

The invention relates to application of CIPK9 protein and a coding gene of CIPK9 protein in drought resistance of plants. The CIPK9 protein has an amino acid sequence shown in SEQ ID NO.1. Cloning ofaCIPK9 gene, construction of transgenic plants for overexpression of the CIPK9 gene and analysis of characters of the obtained transgenic plants find that increase of the expression quantity of the CIPK9 protein in plants can significantly reduce the transpiration rate and stomatal conductance of plants and increase the moisture utilization rate of plants and growth conditions of plants under a drought condition. Therefore, the CIPK9 protein and the coding gene of the CIPK9 protein can be used for enhancing drought resistance of plants, increasingthe yield of plants under the drought conditionand savingwater resources in production practice.

Description

technical field [0001] The invention relates to the fields of genetic engineering and genetic breeding, in particular to the application of CIPK9 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 th...

Claims

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Application Information

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