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Sclerotia oxalic acid decarboxylase gene SsOXDC2 and application thereof in improvement in soybeans for disease resistance

A technology of acid decarboxylase and gene, which is applied in the field of isolation and cloning and functional verification, and can solve the problems of activity reduction and other problems

Inactive Publication Date: 2013-04-24
HUAZHONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the enzyme-catalyzed oxalate degradation reaction occurs in the acidic environment created by S. sclerotiorum infection, and the efficiency of the enzyme is greatly affected by the environment, the activity of plant-derived oxalate oxidase is likely to be affected by the environment and reduce

Method used

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  • Sclerotia oxalic acid decarboxylase gene SsOXDC2 and application thereof in improvement in soybeans for disease resistance
  • Sclerotia oxalic acid decarboxylase gene SsOXDC2 and application thereof in improvement in soybeans for disease resistance
  • Sclerotia oxalic acid decarboxylase gene SsOXDC2 and application thereof in improvement in soybeans for disease resistance

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Embodiment 1

[0024] Embodiment 1: the isolation clone of sclerotinia oxalate decarboxylase gene SsOXDC2

[0025] Obtain some information of the sclerotinia oxalate decarboxylase gene from the fungal genome database (http: / / www.broadinstitute.org / scientific-community / data), containing two oxalate decarboxylase gene sites in the sclerotinia genome, respectively are SS1G_08814 and SS1G_10796, located on different chromosome segments. The SS1G_08814 transcribed region contains 4 exons, 3 introns, the full length is 1575bp, the cDNA coding region contains 1368bp, encoding 455 amino acids; the SS1G_10796 transcribed region contains 3 exons, 2 introns, the full length is 1665bp , the cDNA coding region contains 1515bp, encoding 504 amino acids. Through the analysis of the two amino acid sequences by TargetP 1.1Server, it was found that both amino-terminals contained signal peptide sequences.

[0026] The present invention shows that the Solexa expression profile analysis of Sclerotinia strain E...

Embodiment 3

[0038] Example 3: Functional verification of the oxalate decarboxylase gene SsOXDC2 of Sclerotinia sclerotiorum

[0039] pGR106 is an infection vector transformed from a plant single-stranded RNA virus-Potato Virus X (PVX) (see image 3 B), the foreign gene is inserted into the vector PVX coat protein (CP) promoter, and expressed as a fusion protein with the CP protein. After pGR106 is integrated into the plant genome through Agrobacterium-mediated transformation, it will express the coding potato X virus, and the foreign gene will be expressed at the same time. The exogenous gene can be packaged together with the viral genome, and expressed in other plant tissues along with the replication and transmission of the virus. The specific steps are as follows:

[0040] 1. Transformation vector construction

[0041] The expression vector of virus-mediated sclerotinia oxalate decarboxylase gene SsOXDC2 is plasmid pGR106 (see image 3 B) Build on the basis. First, T-OXDC was used...

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Abstract

The invention belongs to the technical field of plant pathology and molecular biology thereof, and relates to the plant transgene technology. A sclerotinia oxalic acid decarboxylase gene SsOXDC2 capable of resisting sclerotiniose is obtained by utilizing the plant gene cloning technology, wherein the nucleotide sequence of the gene SsOXDC2 is shown in SEQ ID NO: 1 in a sequence table; and the coded protein sequence of the gene SsOXDC2 is shown inSEQ ID NO: 3 in the sequence table. The invention further discloses a method for obtaining a transgenic plant by the clone, the vector construction and the screening of the sclerotinia oxalic acid decarboxylase gene SsOXDC2. The gene SsOXDC2 provided by the invention is suitable for the genetic improvement in enhancement of the capability of the soybeans for resisting the sclerotiniose.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. It specifically relates to the isolation, cloning and functional verification of an oxalate decarboxylase gene SsOXDC2 derived from Sclerotinia sclerotiorum. The oxalate decarboxylase gene SsOXDC2 can endow soybean with the ability to resist soybean sclerotinia caused by Sclerotinia sclerotiorum. Background technique [0002] Sclerotinia sclerotiorum is an important plant pathogenic fungus with a wide range of hosts, which can infect more than 450 kinds of plants in 75 families, mainly herbaceous and succulent woody plants, causing plant sclerotinia (Purdy , 1979; Boland et al, 1994; Bolton et al, 2006), causing huge losses in the production of a variety of commercial crops. In my country, Sclerotinia has become a huge hazard to soybean production. [0003] Sclerotinia can secrete a large amount of oxalic acid during the pathogenic process, and most of the studies in recent y...

Claims

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Application Information

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IPC IPC(8): C12N15/60C12N9/88A01P3/00C12N15/82A01H5/00C12R1/645
Inventor 姜道宏付艳苹程家森谢甲涛岳吉成
Owner HUAZHONG AGRICULTURAL UNIVERSITY