Method for obtaining wheat root related drought resistant protein through utilizing dimensional electrophoresis and MALDI-TOF-MS technology

A MALDI-TOF-MS, two-dimensional electrophoresis technology, applied in the biological field, can solve the problems of cultivating drought-resistant varieties, time-consuming, cumbersome methods, etc.

Inactive Publication Date: 2013-09-04
QINGHAI UNIVERSITY
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Problems solved by technology

[0005] At present, the research on drought resistance mostly adopts physiological and biochemical methods. This method is cumbersome, and the drought resistance of plants can only be roughly identified by measuring multiple indicators, which takes a long time and cannot fundamentally solve

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  • Method for obtaining wheat root related drought resistant protein through utilizing dimensional electrophoresis and MALDI-TOF-MS technology
  • Method for obtaining wheat root related drought resistant protein through utilizing dimensional electrophoresis and MALDI-TOF-MS technology
  • Method for obtaining wheat root related drought resistant protein through utilizing dimensional electrophoresis and MALDI-TOF-MS technology

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Embodiment Construction

[0015] The present invention will be described in detail below in conjunction with specific embodiments.

[0016] (1) Material treatment: The spring wheat variety Qingqing 38 seedlings were subjected to PEG6000 drought stress and rewatering as treatments, and the corresponding normal water supply was used as a control, and root proteins were extracted for two-dimensional electrophoresis.

[0017] (2) Two-dimensional electrophoresis: mainly by et al. and the IPGphor isoelectric focusing system guidelines. A total of 350 μl of hydration solution containing 900 μg of sample protein (with a trace of 0.001% bromophenol blue added) was continuously added along the focusing tank of the PROTAIN IEF Cell electrophoresis instrument (BIO-RAD). Cover the sample with a 17cm IPG strip with the glue side down, and after passive absorption for 1 hour, add 2ml of mineral oil to cover the strip. Isoelectric focusing and second-dimension vertical plate SDS-PAGE were slightly modified accordin...

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Abstract

The invention discloses a method for obtaining a wheat root related drought resistant protein through utilizing dimensional electrophoresis and an MALDI-TOF-MS technology. The method comprises a step of material treatment, a step of dimensional electrophoresis, a step of atlas analysis, and a step of MALDI-TOF-MS, and the MALDI-TOF-MS step is characterized in that differential protein spots undergo in-gel enzymatic hydrolysis, ultraflex TOF/TOF is used to carry out MALDI-TOF mass spectrum fingerprint analysis, and obtained PMF data are identified through retrieving Protein Data Bank by Mascot software. In the invention, the combination of dimensional electrophoresis and the MALDI-TOF-MS technology is adopted, the protein expression difference between the drought stressed wheat root and the normal-water-supply wheat root is analyzed, and the identification is further carried out through the mass spectrum, so the root expression protein obtained through the method is directly linked to the property, and is of greater significance than the gene level and the other organs of crops.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for obtaining wheat root-related drought-resistant protein by using two-dimensional electrophoresis and MALDI-TOF-MS technology. Background technique [0002] At present, arid and semi-arid areas in the world account for 43% of the cultivated land area. Most countries are facing a water resource crisis, and my country is no exception. Drought is widely distributed in our country, especially in northern China. Drought has become a major natural disaster in northern my country. One, there are different degrees of drought every year that endanger agricultural production and lead to a reduction in grain production. Drought is a major threat to precision agriculture and the stability of agricultural yields. Qinghai Province is located on the Qinghai-Tibet Plateau in the northwest of my country. It has a plateau continental climate, dry and windy, with little precipitation, uneven...

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Application Information

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IPC IPC(8): G01N21/25
Inventor 叶景秀姚有华谢德庆
Owner QINGHAI UNIVERSITY
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