Method of researching change of proteome of rice responding rice blast bacterial infection through iTRAQ technology

A rice blast fungus and proteome technology, applied in the field of plant biology, can solve problems such as inaccurate research results

Inactive Publication Date: 2015-08-05
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

To overcome the problem of inaccurate research effects of existing technologies

Method used

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  • Method of researching change of proteome of rice responding rice blast bacterial infection through iTRAQ technology
  • Method of researching change of proteome of rice responding rice blast bacterial infection through iTRAQ technology
  • Method of researching change of proteome of rice responding rice blast bacterial infection through iTRAQ technology

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Embodiment 1

[0043] 1. Application of iTRAQ technology to study rice proteome identification method in response to blast fungus infection

[0044] (1) Sample preparation:

[0045] 1. Extraction of total protein from rice materials and leaves. The rice seedlings at the 3.5-4 leaf stage of the disease-resistant material (H4) and the disease-resistant material (Zhongeranzhan) were inoculated with Magnaporthe grisea, and samples were taken after 0 and 24 hours of inoculation, and quickly placed in liquid nitrogen to cool and then placed in Store at -80°C for later use; take rice tissue samples, grind them into powder with liquid nitrogen and transfer them to a 50ml centrifuge tube, add 25ml of extract solution (TCA / acetone) to precipitate at -20°C for 1 hour. Centrifuge at 10,000rpm for 45min, discard the supernatant. After air-drying, add SDT buffer (4% SDS, 1mM DTT, 150mM Tris-HCl pH8.0) at a volume of 10:1, vortex and mix well, and place in a boiling water bath for 5min. Ultrasonication ...

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Abstract

The invention discloses a method of researching the change of a proteome of rice responding rice blast bacterial infection through an iTRAQ technology and belongs to the field of plant biotechnology. The method includes following steps: A. extracting a plant materials and leaf protein; B. performing further FASP enzymolysis, peptide fragment marking, SCX classification and LC-MS analysis to the protein sample obtained in the step A; C. performing data query and quantitative analysis to the result after the LC-MS analysis to obtain the change situation of the whole proteome during an interaction process between rice and rice blast bacteria. The method can quickly and effectively screening a candidate gene relative to disease resistance of rice, can be used for researching the candidate gene relative to the disease resistance of the rice well with the proteomics researching method, iTRAQ, which is high-throughput and accurate, can be widely used for researching and developing the characters of plant disease resistance and breeding new plant varieties being strong in disease resistance with combination of the transgenic technology.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a method for studying rice proteome changes in response to blast fungus infection by using iTRAQ technology. Background technique [0002] The study of plant disease resistance and disease resistance mechanism is the hotspot and focus of plant pathology and plant disease resistance breeding research. In natural conditions, plants are often under biotic or abiotic stress. When infected by foreign pathogens, plants will resist the invasion of pathogens by changing the expression pattern of their own proteome or the activity of enzymes. The generation of plant disease resistance involves a series of complex signal transduction processes. Current studies have found and identified the activation of some disease resistance pathways related to pathogen infection from various plants. [0003] However, most of the current research focuses on the use of gene chip, gene expr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848
Inventor 王加峰黄明刘浩董双玉陈志强王慧
Owner SOUTH CHINA AGRI UNIV
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