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Physiological Characteristics of Cold Resistance and Proteonmic Analysis of Proteins for Cold Stressed Winter Rapa (Brassica.Rapa) by Two-dimensional Electrophoresis

Author: PuZuoZuo
Tutor: SunWanCang
School: Gansu Agricultural University
Course: Crop Genetics and Breeding
Keywords: Brassica winter rape Low temperature stress Physiological and biochemical Two-dimensional electrophoresis
CLC: S565.4
Type: Master's thesis
Year: 2010
Downloads: 180
Quote: 0
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Abstract


Plants in the growth and development process will encounter various biotic and abiotic stress factors, especially chilling injury phenomenon, involving almost all plants, severely limit plant distribution area and affect biological production globally each year due to low temperature injury caused crop losses of up to several hundred billion dollars in China each year by the low temperature damage an area of ??more than 200 million square kilometers. Therefore, improving the hardiness of plants is of great practical significance and application value. In order to understand the relationship of of osmolytes Rape cold tolerance test to the two Brassica winter oilseed rape Gansu oil on the 6th day oil 4 test material by freezing the relative conductivity of rapeseed roots and leaves superoxo dismutase (SOD), hydrogen peroxide enzyme (CAT), soluble proteins, malondialdehyde (MDA) and other physiological and biochemical indices were detected. The results show that: Loong Oil No. 6 (DQW-1) strong resistance to cold in the days of the oil on the 4th, Gansu oil No. 6, and the days of oil on the 4th of SOD and CAT activity, soluble protein content, MDA content along with the decrease of temperature are showing the variation increased and then decreased. Low temperatures, strong cold resistance Gansu oil No. 6 can maintain higher SOD activity, CAT activity of soluble protein and lower MDA content, SOD and CAT activity, soluble protein and MDA content in the can to a certain extent reflect the hardiness of winter rape Brassica. 2 in order to establish a bidirectional electrophoresis system suitable for rape leaves, trial of winter oilseed rape Brassica material to explore some important technical aspects of the two-dimensional electrophoresis to determine the amount of sample, the sample preparation method of its protein, isoelectric focus, staining an important part of optimization. The results are as follows: the amount of TCA-acetone precipitation method, the protein precipitate obtained is high, but the impurities, after protein precipitation less purified by acetone precipitation method, but fewer impurities, and a high concentration of protein solution can be obtained after cleavage. Protein sample volume by the comparison determines unidirectional SDS-PAGE gel electrophoresis test infected like to 35μg 45μg; infected samples test strips for two-dimensional gel electrophoresis 17cm (linear, pH3-10) The amount available 900μg about focusing better, two-dimensional electrophoresis map with clear background. The dyeing method of the gel BBC-G250, before dyeing, containing 12% TCA fixative gum, the size of the adhesive surface does not change, and is conducive to the analysis software. To learn more about the rape hardiness proteome test using two-dimensional electrophoresis system optimized to after 12h cold treatment of winter rape Brassica materials, its leaves proteome preliminary analysis of the differences The results show that: the total of 56 protein spots changed in expression levels of 34 protein spots were up-regulated, the expression of 22 protein spots decreased. The trial initially established two-dimensional electrophoresis system suitable for rape leaf proteomics methods used in the research of rapeseed resilience mechanisms, and provide a reference for rape leaves proteomics research.

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CLC: > Agricultural Sciences > Crop > Economic crops > Oil crops > Rapeseed ( Brassica )
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