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Application of maize mitogen-activated protein kinase gene zmmpk20 in regulation of stomatal movement and plant heat tolerance

A technology for activating protein kinase and mitogen, applied in the application field of maize mitogen-activated protein kinase gene

Active Publication Date: 2021-05-25
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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After searching, the application of maize mitogen-activated protein kinase gene ZmMPK20 in regulating stomatal movement and plant heat resistance has not been reported yet.

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  • Application of maize mitogen-activated protein kinase gene zmmpk20 in regulation of stomatal movement and plant heat tolerance
  • Application of maize mitogen-activated protein kinase gene zmmpk20 in regulation of stomatal movement and plant heat tolerance
  • Application of maize mitogen-activated protein kinase gene zmmpk20 in regulation of stomatal movement and plant heat tolerance

Examples

Experimental program
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Embodiment 1

[0019] Example 1 Obtainment and molecular identification of a loss-of-function mutant of the maize mitogen-activated protein kinase gene ZmMPK20

[0020] 1. EMS mutagenesis to obtain leaf curl mutants under heat stress in maize

[0021] After EMS mutagenesis, the three-leaf stage plants were heat-treated at 42°C, and it was found that 1c5 showed a phenotype sensitive to heat stress, and the leaves were severely curled. There was no difference between 1c5 and wild-type B73 leaves under normal conditions.

[0022] 2. Use the F2 generation for resequencing to determine effective SNPs

[0023] The F1 generation B73 / 1c5 was obtained by backcrossing the mutant 1c5 with the wild type, and the F2 generation B73 / 1c5 was obtained after self-crossing. The segregation ratio of the wild type trait and the mutant trait was 3:1 in the F2 generation, indicating that this trait is a recessive trait And in line with Mendel's laws of inheritance. A population of 50 plants with mutant traits i...

Embodiment 2

[0026] Example 2 Maize Mitogen-Activated Protein Kinase Gene ZmMPK20 Transgenic Plant Obtained

[0027] 1. PCR method to clone ZmMPK20 gene fragment

[0028] Using the cDNA of maize B73 as a template, PCR amplification was performed with the following upstream and downstream primer combinations:

[0029] ZmMPK20-OE-F: 5'-GCTCTAGAATGTCACGTTTCATGTTCCTTC-3'

[0030] ZmMPK20-OE-R: 5'-GGGGTACCCTAATACATCCTTGACATGCCAG-3'

[0031] After electrophoresis of the PCR product, the gel was cut and recovered, and the ZmMPK20 gene fragment was purified.

[0032] 2. Vector Ligation and Transformation

[0033] First, connect the purified ZmMPK20 gene fragment to the intermediate cloning vector Blunt (product of Beijing Quanshijin Company), transform Escherichia coli DH5α, and obtain positive clones through antibiotic screening and colony PCR; The nucleotide sequence is exactly the same as that of SEQ ID NO.1, and the next step of digestion is carried out after the sequencing is correct.

...

Embodiment 3

[0044] Example 3 Application of Maize Mitogen-Activated Protein Kinase Gene ZmMPK20 in Regulating Stomatal Movement and Plant Heat Resistance

[0045] Using ZmMPK20 overexpression plants (OE1, OE2), the following physiological trait experiments were carried out to analyze the application of ZmMPK20 in the regulation of stomatal movement in Arabidopsis or other economic crops.

[0046] 1. Stomatal Movement Experiment in Arabidopsis

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Abstract

The invention discloses the application of a corn mitogen-activated protein kinase base ZmMPK20 in regulating stomatal movement and plant heat resistance. Comprehensive experiments confirmed that ZmMPK20 plays a positive regulatory role in high temperature regulation of stomatal movement, and also plays a regulatory role in plant response to high temperature stress. It is predicted that the application of the present invention will help improve the ability of plants to resist high temperature stress, have important theoretical guiding significance for cultivating new varieties of crops resistant to high temperature and high yield, and have great application value for agricultural production in my country.

Description

technical field [0001] The invention relates to the application of a maize mitogen-activated protein kinase gene, in particular to the application of a maize mitogen-activated protein kinase gene ZmMPK20 (Gene ID: 100191429) in regulating stomatal movement and plant heat resistance, belonging to molecular biology science and genetic engineering. Background technique [0002] The published ZmMPK20 gene sequence (Gene ID: 100191429) was analyzed by bioinformatics, indicating that it encodes a mitogen-activated protein kinase (Mitogen-activated protein kinase, MPK). It belongs to the MAPK kinase family and participates in the MAPKKK cascade reaction. [0003] According to the Corn Research website ( http: / / www.maizegdb.org ) provided by the eFP Blowser results showed that the ZmMPK20 gene was expressed in large quantities in plant leaves and other parts. [0004] The stomata of plant epidermis are the main channel for plants to transpiration and water loss. The opening and c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/12C12N15/82A01H5/00A01H6/46A01H6/20
CPCC12N9/12C12N15/8261C12N15/8271C12Y207/11024
Inventor 张伟吴琪琪陈冬花王美沈建霖程闯
Owner SHANDONG UNIV
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