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Application of phytophthora sojae gene PsIR1 capable of inducing plant disease resistance

A technology of Phytophthora sojae and gene, applied in the biological field, can solve the problems of time-consuming and labor-intensive, inability to have lasting broad-spectrum resistance, etc., and achieve the effects of enhancing plant resistance and slowing down the infection speed.

Inactive Publication Date: 2012-07-04
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to overcome the time-consuming and labor-intensive shortcomings of conventional breeding and the inability to have long-lasting broad-spectrum resistance, the present invention provides a Phytophthora soybean effector PsIR1 as a candidate material for breeding. This protein can induce the high expression of the plant PR1 gene and cause disease resistance in tobacco. sex

Method used

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  • Application of phytophthora sojae gene PsIR1 capable of inducing plant disease resistance
  • Application of phytophthora sojae gene PsIR1 capable of inducing plant disease resistance
  • Application of phytophthora sojae gene PsIR1 capable of inducing plant disease resistance

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 PsIR1 can inhibit the infection of Phytophthora tabacum

[0024] 1. Vector Construction

[0025] According to Liu, et al, Two host cytoplasmic effectors are required for pathogenesis of Phytophthora sojae by suppression of host defenses. Plant physiology, 2011, 155, 1: 490-501 article construction method to obtain PVX:PsIR1 and PVX:GFP plasmids.

[0026] 2. Agrobacterium-mediated transient expression

[0027] The PVX:PsIR1 and PVX:GFP plasmids were transferred into the GV3101 Agrobacterium (Biovector) bacterial strain by electroporation technology, and the positive transformant of Agrobacterium was added in 3ml of LB culture solution of kanamycin (50 μg / ml), 220rpm, 28- Cultivate at 30°C for 48h, centrifuge at 4000rpm for 4min, collect the bacteria, and wash with 10mM MgCl 2 Resuspended, repeated three times, with 10mM MgCl 2 Set the volume to OD600=0.4-0.6. Tobacco grown for 6-8 weeks was taken, and the third to sixth fully expanded true leaves from the t...

Embodiment 2

[0030] Example 2 PsIR1 Causes High Expression of Salicylic Acid Pathway Defense Response Genes

[0031] The PVX:PSIR1 and PVX:GFP obtained in Example 1 were transiently expressed on tobacco according to the method of Example 1, and samples were taken when expressing 0d and 5d, respectively, according to the Protocol provided by the Total RNA Purification System kit (Invitrogen). Fluorescence quantitative PCR operation was carried out according to ABI 7300 Sequence Detection System (Applied Biosystems, USA) software and guidelines. PCR reaction solution: use Real time PCR kit ( Premix Ex TaqTM, TaKaRa, DaLian, China) to carry out the reaction. The RNA extracted from transiently expressed tobacco leaves was reverse-transcribed, and the obtained cDNA was used as a template for relative quantitative PCR amplification. Mix 50ng of cDNA and 0.2μM primers, 0.4ul ROX Reference Dye and 10ul Premix Ex TaqTM was mixed in 20μl of PCR reaction system. The PCR program was: pre-denatur...

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Abstract

The invention belongs to the field of biotechnology and discloses application of phytophthora sojae gene PsIR1 capable of inducing plant disease resistance. The invention relates to application of the phytophthora sojae gene PsIR1 (GenBank: HQ231784.1) in constructing a transgenic plant with broad-spectrum resistance against plant pathogen. In the invention, the phytophthora sojae gene PsIR1 is inserted into a PVX expression vector, and the functions of the PsIR1 on tobacco are analyzed through the agrobacterium-mediated instant expression. The result shows that: PsIR1 can obviously improve the resistance of tobacco against phytophthora and can induce the salicylic acid channel to defense the high expression of the response genes PR1 and PR2. Therefore, a conclusion can be made that PsIR1 causes accumulation of salicylic acid, the increase of salicylic acid can improve the broad-spectrum resistance of the plant, and thus the PsIR1 can enhance the disease resistance of tobacco.

Description

technical field [0001] The invention belongs to the field of biotechnology and discloses the application of a soybean Phytophthora gene PsIR1 capable of inducing plant disease resistance. Background technique [0002] With the rapid growth of the world population and the reduction of arable land, the world's food problem has become increasingly serious. Cultivating more food on limited land resources is the top priority for human survival. Breeders not only want to increase the yield of crops, but also reduce the loss of crops. Among them, plant diseases have always been the biggest bottleneck hindering the increase in yield of fine varieties. Therefore, breeding high-yield and disease-resistant crops is the direction pursued by breeders in recent years. . Conventional breeding methods are time-consuming and labor-intensive, but with the development of biology, varieties bred by genetic engineering methods can not only shorten the breeding time, but also allow multiple exce...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 窦道龙刘廷利茹艳艳刘莉刘沛菡
Owner NANJING AGRICULTURAL UNIVERSITY
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