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Gene GmWRKY148 capable of improving resistance to phytophthora sojae and application of gene

A soybean Phytophthora, gene technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of soybean yield and economic loss, resistant varieties cannot maintain continuous resistance, etc.

Active Publication Date: 2019-11-05
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Caused huge yield and economic losses to soybean production
However, due to the high genetic variability of P. sojae, resistant varieties often cannot maintain continuous resistance and are quickly overcome by new virulent races

Method used

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  • Gene GmWRKY148 capable of improving resistance to phytophthora sojae and application of gene
  • Gene GmWRKY148 capable of improving resistance to phytophthora sojae and application of gene
  • Gene GmWRKY148 capable of improving resistance to phytophthora sojae and application of gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1) Cloning and bioinformatics analysis of GmWRKY148 gene

[0042] Phytozome (http: / / www.phytozome.net / ) was used to find genes homologous to Arabidopsis AtWRKY44 in soybean with AtWRKY44 (AT2G37260.1) as a probe, and BLAST comparison was performed. As a result, it was found that the number of homologous genes was large, and after the induction of Phytophthora soybean P6497, the gene with a large expression change was selected, namely GmWRKY148, and it was determined as the research object ( figure 1 ). Primers were designed, and the primer sequences are listed in Table 1. The results of three-dimensional structure analysis of GmWRKY148 protein showed that ( figure 2 ), which consists of 7 α-helices and 4 β-sheets. Four β-sheets are typical features of WRKY domain, and the β-strand containing WRKYGQK motif can interact with the 6bp W-box (TTGACY) region. Evolutionary analysis showed that GmWRKY148 belongs to soybean WRKY transcription factor family gene, and is very...

Embodiment 2

[0051] The genetic engineering application of embodiment 2 GmWRKY148

[0052] According to the cDNA sequence of GmWRKY148, primers were designed to amplify the complete coding open reading frame (ORF), and adapter primers were introduced into the primers. For the DNA sequence, the vector is constructed by double restriction digestion, and the restriction sites are Kpn1 and BamH1. The complete ORF was inserted into the expression vector pBINGFP2, and the plant overexpression vector 35S-WRKY148::GFP ( Figure 7 ). It and the empty load were respectively transformed into Agrobacterium rhizogenes K599 by electric shock transformation method, and then the transgenic soybean root hairs were obtained by Agrobacterium-mediated method. Then according to the marker protein GFP on the carrier, the positive soybean root hairs obtained are initially screened using a fluorescent stereomicroscope, and the positive hairy roots are expanded and cultivated on the White medium ( Figure 8 , a...

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Abstract

The invention discloses a gene GmWRKY148 capable of improving resistance to phytophthora sojae and application of the gene, wherein the gene can enhance resistance of soybean to the phytophthora sojae. According to the disclosed gene GmWRKY148, the nucleotide sequence of the gene is shown as SEQ ID NO.1. Phenotype and phytophthora accumulation indexes of soybeans Williams with GmWRKY148 overexpression under the phytophthora stress condition indicate that the GmWRKY148 plays an important regulatory role in improving plant resistance to phytophthora. The gene is used for genetic engineering improvement of plant resistance to phytophthora root rot, it is found that the gene has the certain effect on breeding plant varieties with high resistance to phytophthora root rot, and by improving plantresistance to the phytophthora, the plant yield and quality service are improved.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to a gene GmWRKY148 for improving the resistance of Phytophthora soybean and its application. [0002] technical background [0003] Plants are often subjected to multiple abiotic and / or biotic stresses, such as drought and heat, drought and cold, salinity and heat, or any major abiotic stress combined with pathogen infection. Phytophthora sojae is an oomycete that can infect soybeans under favorable conditions. It has many physiological races and is difficult to control. The comprehensive control technology combining multiple control measures is a fundamental and effective control approach, among which the application of genetic engineering technology to cultivate disease-resistant varieties is the most effective in controlling soybean Phytophthora root rot. [0004] WRKY transcription factors are one of the largest transcriptional regulator families in plants, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C12N15/82C12N1/21C07K14/415C12R1/01C12R1/19
CPCC07K14/415C12N15/8282
Inventor 邢邯王莎莎崔晓霞黄颜众轩慧冬郭娜
Owner NANJING AGRICULTURAL UNIVERSITY
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