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Application of AtMPK6 in plant drought combatting aspect

A technology of Arabidopsis, gene, applied in the field of genetic engineering

Inactive Publication Date: 2013-09-18
HENAN UNIVERSITY
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  • Abstract
  • Description
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  • Application Information

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  • Application of AtMPK6 in plant drought combatting aspect
  • Application of AtMPK6 in plant drought combatting aspect
  • Application of AtMPK6 in plant drought combatting aspect

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Experimental program
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Effect test

Embodiment 1

[0022] Analysis of Drought Resistance of At2g43790 Gene

[0023] Such as figure 1 As shown, the seedlings were grown for 4 weeks under the conditions of light / dark cycle of 8 / 16h, temperature of 22°C, dark temperature of 18°C, relative humidity of 80%, and light intensity of 90mol·m-2·s-1, and group A was normal Watering, group B stopped watering for 10 days. The results of multiple experiments showed that: under normal growth conditions, WT and Atmpk6 plants grew well without significant difference (Figure A); after 10 days of continuous drought, the growth status of WT and Atmpk6 plants was significantly different, and 95% of the WT plants were drought-dead. However, 98% of Atmpk6 plants were still alive (Panel B). The results of these experiments indicated that Arabidopsis thaliana acquired stronger drought resistance after loss of AtMPK6 function.

Embodiment 2

[0025] Detection of ROS changes in WT and Atmpk6 protoplasts under stress treatment

[0026] Such as figure 2 As shown, the ROS fluorescent probe H2DCF-DA was used to load WT and Atmpk6 root protoplasts. On the laser confocal fluorescence microscope, 488nm, Time-Course scanning, the dynamic changes of ROS after ABA treatment were detected. After adding 10 μM ABA, Atmpk6 protoplasts The fluorescence intensity of the protoplasts increased rapidly, and reached the maximum at about 10 min, indicating that a large amount of ROS was produced and accumulated in the protoplasts at this time, while the fluorescence intensity of WT protoplasts did not change much.

Embodiment 3

[0028] Ca in WT and Atmpk6 plants under osmotic stress 2+ Kinetic Change Detection

[0029] The aequorin gene was transformed into Arabidopsis thaliana, and the seeds of the first-generation transgenic plants were obtained, which were spread on MS medium containing 30 μg L-1 of hygromycin B, and the transferred aequorin gene was screened out. plants were analyzed.

[0030] Such as image 3 As shown, under the simulated drought treatment of 330mM mannitol, Ca 2+ There is a significant difference in the changes: WT Ca 2+ The highest concentration peak was 0.5978 μM (n=15), and the highest peak concentration of Atmpk6 calcium ion was 1.868647 μM (n=12).

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Abstract

The increasingly serious water resource shortage phenomena make the drought problems become the focus of the world. Droughts limit the growth and development of plants, which results in the yield reduction and even complete crop failure. The grain security in China is also threatened. A mitogen-activated protein (MAP) kinase of the gene code AtMPK6 (Arabidopsis thaliana MAP kinase 6, No: At2g43790) has the function for adjusting the generation and accumulation of active oxygen in plant bodies threatened by droughts. Mutant plants can still survive under the condition of 10 days' successive dry soil, but wild plants die for the droughts under the same conditions. The contents of active oxygen, Ca2+ and water in mutant plants are higher than those in wild plants under stress conditions. In the present invention, the drought combatting function and the adjusting mechanism of AtMPK6 for plants are analyzed, and the metabolic mechanism of active oxygen induced by abscisic acid (ABA) hormone in stress plants with AtMPK6 is established, which lays down theory and production practice bases for the culture of novel drought combatting transgenic crop species.

Description

technical field [0001] The invention relates to the application of the gene At2g43790 encoding Arabidopsis thaliana AtMPK6 in the analysis of the mechanism of plant drought resistance, the cultivation of drought-resistant transgenic plant varieties, etc., and belongs to the field of genetic engineering. Background technique [0002] Drought and drought have been the main natural disasters faced by human beings since ancient times. The shortage of water resources is increasing day by day, seriously restricting the growth and development of plants, and threatening food security worldwide. With the deepening of molecular biology and genetic engineering research, many laboratories in the world are also trying to find genes related to plant drought resistance, and plan to improve the drought resistance of crops through genetic engineering technology, but the research progress in this area is still very slow. [0003] Existing studies have shown that when encountering drought str...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/12A01H5/00
Inventor 江静王文乐张华王淑静
Owner HENAN UNIVERSITY
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