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Application of glycine max cuticular wax synthetic gene or protein thereof

A protein and vegetable wax technology, applied in the field of genetics and breeding, can solve the problems of changing the proportion of wax components and affecting horny development, etc., and achieve the effects of improving drought stress resistance, disease resistance, and defense against mechanical damage.

Inactive Publication Date: 2020-01-14
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have shown that the mutation of the Glyma.07G028600 gene leads to a great change in the proportion of waxy components in the cuticle, and at the same time affects the development of cutin

Method used

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  • Application of glycine max cuticular wax synthetic gene or protein thereof
  • Application of glycine max cuticular wax synthetic gene or protein thereof
  • Application of glycine max cuticular wax synthetic gene or protein thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Isolation and structural analysis of GmGK1 gene in soybean

[0057] (1) Isolation of genes

[0058] The mRNA was isolated from the young leaves of the soybean variety - Hedou 12, and the first strand of cDNA was synthesized using the mRNA as a template and Oligo(T)17 as a primer. Then, using the first strand of the cDNA as a template, primers (5'-ATGTCGAAAGAGGATGTGTTTG-3') and primers (5'-TCAAAGAGCAAGATCAGCTAAGTC-3') were used for PCR amplification, and a 1560bp cDNA fragment of a GmGK1 gene was obtained , cloned into pGEM T Easy vector (TaKaRa Company) and named pGEM T Easy-GmGK1.

[0059] The GmGK1 CDS sequence contained in pGEM T Easy-GmGK1 is shown in SEQ ID NO: 1, with a total of 1560 bp, which encodes a protein of 519 amino acids (SEQ ID NO: 2).

[0060] (2) Gene structure analysis

[0061] The young leaves of Williams82 were used as materials to extract the DNA therein, and then the genomic DNA was used as a template to amplify the fragment of the G...

Embodiment 2

[0062] Example 2. Construction of soybean GmGK1 gene sense expression vector

[0063] The CDS of GmGK1 and the CDS of green fluorescent protein GFP were fused forwardly into the plant expression vector pCAMBIA3301H (CAMBIA Research Center) to obtain the positive expression vector of GmGK1 with the GFP tag. The full length of the vector is 11.2Kbp (such as figure 2 ), the resistance in Escherichia coli is kanamycin resistance, and the resistance in plants is glufosinate-ammonium resistance.

Embodiment 3

[0064] Example 3. Agrobacterium-mediated transformation of leguminous plants

[0065] In this example, the explants of the gmgk1 mutant containing the positive-sense expression vector of the GmGK1 regulatory gene were obtained by transforming soybean embryo tips through Agrobacterium-mediated transformation.

[0066] (A) Obtaining of soybean explants

[0067] 10,000 grains of the soybean cultivar Hedou 12 widely planted in the Huanghuaihai region were mutagenized by 250Gy gamma rays. A total of 1298 mutants of more than 900 lines were identified as M2 mutants, and the mutants with shrunken leaves were screened from them. material, phenotypically stable gmgk1 mutants were obtained after planting for 4 generations. The Glyma.07G028600 gene that controls the phenotype of the gmgk1 mutant was successfully identified by crossing the gmgk1 mutant with the distant species Williams82. The gene has a deletion of ten bases of TCTTTTATCC and an insertion of one in the exon A base. The...

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Abstract

The invention relates to the technical field of genetic breeding, in particular to application of a glycine max cuticular wax synthetic gene or a protein thereof. The research shows that a GmGk1 gene,derived from glycine max, for encoding glycerol triphosphokinase can regulate development of cuticular wax on the surfaces of glycine max leaves and thus can be used for realizing improvement of thedrought resistance and the disease resistance of the glycine max. The research shows that crinkles leaves of a gMgk1 mutant plant for overexpressing the GmGK1 gene are recovered normally, and the morphology of the leaves are consistent with the morphology of leaves of wild type glycine max. For the application, by using the GmGK1 as a marker, identification of plant germplasm, particularly the drought resistance of the glycine max, can be realized.

Description

technical field [0001] The invention relates to the technical field of genetic breeding, in particular to the application of soybean cuticle wax synthesis gene or protein thereof. Background technique [0002] As an ancient crop, soybean (Glycine max) has important and special economic value. It is sensitive to water shortage and has weak high temperature resistance. Abiotic stresses such as drought, high temperature, and salinity during the growth period of soybean are important factors that cause soybean yield reduction. Therefore, improving soybean drought resistance has become one of the key issues in modern breeding. [0003] The most important function of the cuticle for plants is to respond to the threat of drought, and it also has the functions of defending against mechanical damage, resisting the invasion of germs and insects, and blocking ultraviolet rays. The microstructure of the stratum corneum is divided into two layers, the stratum corneum and the wax. The w...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/54C12N15/82A01H5/12C12Q1/6895G01N33/68
CPCC12N9/1217C12N15/8273C12N15/8279C12Q1/6895C12Q2600/158C12Y207/02003G01N33/68
Inventor 冯献忠杨素欣高金珊宋晓峰
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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