Plant low potassium stress resistant related protein GmWRKY50 as well as encoding gene and application thereof

A low-potassium tolerance gene technology, applied in the plant low-potassium stress-resistant protein GmWRKY50 and its coding gene and application field, can solve the problems of restricting crop yield, affecting the normal growth and development of plants, and large potassium consumption

Active Publication Date: 2014-09-17
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Crops, especially soybeans, consume a lot of potassium, and the potassium ion concentration in the soil available in the rhizosphere is generally 0.1-6.0mmol / L, which is often lower than the concentration required by the plant, causing the plant to encounter a low potassium environment and seriously Affect the normal growth and development of plants
When the content of potassium ions in the soil solution is low and the supply of potassium fertilizer is insufficient, it will seriously restrict the crop yield

Method used

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  • Plant low potassium stress resistant related protein GmWRKY50 as well as encoding gene and application thereof
  • Plant low potassium stress resistant related protein GmWRKY50 as well as encoding gene and application thereof
  • Plant low potassium stress resistant related protein GmWRKY50 as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1, acquisition of soybean protein GmWRKY50 and its coding gene and recombinant expression vector

[0041] Total RNA was extracted from seedling leaves of soybean line 06-71 (Glycine max(L.)Merr.) cultured under normal conditions, and cDNA was obtained by reverse transcription. Using the cDNA as a template, primers PF and PR Under the guidance, use the conventional PCR method to amplify. After the reaction, the PCR amplification product is subjected to 1% agarose gel electrophoresis detection, and a DNA fragment of about 1 kb is recovered and purified; the vector pCXSN (T-DNA in the vector pCXSN The fragment contains the lethal gene ccdB, flanked by XcmI restriction recognition sequences. The vector was purchased from the Arabidopsis Biological Resource Center (The Arabidopsis Biological Resource Center, ABRC; website: http: / / abrc.osu.edu / ) , the product catalog number is Vector: 5019471951) was digested with XcmⅠ, and the vector skeleton fragment was recover...

Embodiment 2

[0045] Embodiment 2, the acquisition of recombinant Agrobacterium tumefaciens

[0046] The recombinant vector pCXSN-GmWRKY50 obtained in Example 1 was transformed into Agrobacterium tumefaciens GV3101 by freeze-thaw method (document: Amanda M Davis, Anthony Hall, Andrew J Millar, Chiarina Darrah and Seth J Davis, Protocol: Streamlined sub-protocols for floral- dip transformation and selection of transformants in Arabidopsis thaliana, 2009, 5:310.1186 / 1746-4811-5-3; the public can obtain from the Institute of Oil Crops, Chinese Academy of Agricultural Sciences), and obtain Agrobacterium tumefaciens GV3101 containing the recombinant vector pCXSN-GmWRKY50 , and the recombinant Agrobacterium was named GV3101 / pCXSN-GmWRKY50;

[0047] The empty vector pCXSN was transformed into Agrobacterium tumefaciens GV3101 by freeze-thawing method to obtain Agrobacterium tumefaciens GV3101 containing the empty vector pCXSN, and the recombinant Agrobacterium was named GV3101 / pCXSN.

Embodiment 3

[0048] Embodiment 3, the acquisition and identification of transgenic Arabidopsis

[0049] 1. Obtaining transgenic Arabidopsis

[0050] Using the two recombinant Agrobacteria obtained in Example 2, 180 strains of Columbia ecotype Arabidopsis Columbia-0 (hereinafter referred to as wild-type Arabidopsis WT) were transformed by flower bud soaking method (document: Xia T, Xiao D, Liu D , Chai W, Gong Q, Wang NN. Heterologous expression of ATG8c from soybean confers tolerance to nitrogen deficiency and increases yield in Arabidopsis. PLoS One. 2012;7(5):e37217; publicly available from Institute of Oil Crops, Chinese Academy of Agricultural Sciences ), to obtain T 3 3 Arabidopsis lines homozygous transgenic GmWRKY50, T 3 Generation of 3 Arabidopsis lines that were homozygously transformed into empty vectors, the specific method is as follows:

[0051] 1. Take the recombinant Agrobacterium GV3101 / pCXSN-GmWRKY50 or GV3101 / pCXSN and culture it with YEP medium containing 50 μg / ml kan...

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Abstract

The invention discloses a plant low potassium stress resistant related protein GmWRKY50 as well as an encoding gene and application thereof. The protein GmWRKY50 comes from soybean (Glycine max(L.)Merr.), and is the protein of the following a) or b): a) a protein consisting of amino acid sequences shown as a sequence 1 in a sequence table; and b) a protein which is subjected to substitution and / or deletion and / or addition of one or more amino acid residues in the amino acid sequences of the sequence 1 in the sequence table, is related to the low potassium stress resistance and / or volume and number of organs and is derived from (a). The experiment proves that the low potassium stress resistance of the T3 generation homozygous transgenic plant obtained by the recombinant expression vector GmWRKY50 Arabidopsis containing the DNA molecules shown by the sequence 2 in the sequence table for encoding the protein is obviously higher than that of the wild Arabidopsis plant under the same condition. The protein has significance in breeding and research for improving the low potassium stress resistance.

Description

technical field [0001] The invention relates to a protein GmWRKY50 related to plant tolerance to low potassium stress and its coding gene and application. Background technique [0002] Potassium is the most abundant nonpolar cation contained in plants, and its total amount accounts for 2%-6% of the total dry weight of plants, and plays a key role in the growth and development of plants. Potassium ions participate in many important physiological and biochemical processes of plants, such as maintaining cell charge balance, regulating the activity of various enzymes, regulating cell turgor, participating in protein synthesis, affecting plant stomatal movement, photosynthesis, and promoting cell elongation. Plants with severe potassium deficiency show deficiency symptoms such as root shrinkage, fragile stems, poor leaf vitality, burnt edges of old leaves, small and shrunken seeds and fruits, short plants, slow growth, and are susceptible to pests and frost damage. Although pota...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N7/01C12N15/82C12N15/84A01H5/00
Inventor 朱晓玲陈海峰王程郝青南崔嘉成沙爱华陈李淼周蓉张婵娟张晓娟陈水莲周新安
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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