Extraction of Winter Turnip Rape Leaf Apoplast and Analysis of Protein Expression in Response to Low Temperature Stress

doi:10.7668/hbnxb.2016.04.008

Abstract

Abstract: In order to explore the mechanism of super cold resistant varieties of Winter turnip rape,select super cold resistant varieties of Winter turnip rape Longyou No.7 leaves as materials,using two different extraction liquids(Extraction A:0.1 mol/L Tris-HCl (pH=7.6),0.2 mol/L KCl,1 mmol/L PMSF,Extraction B:0.05 mol/L Tris-HCl (pH=7.6),0.01 mol/L EDTA-Na₂,0.02 mol/L Vc,1 mmol/L PMSF) to compare the extracted effect.The experiment showed that the extraction A liquid protein yield reached (0.073±0.004 6) mg/g,and the extraction ratio increased by 42.88% compared to liquid B.Through the Glucose-6-phosphate dehydrogenize (G₆PDH) activity detection and the extraction A was lower than B 0.91 percent;not only the extraction A extract protein was shorter than B 4.5 hours in the first oelectric focusing electrophoresis of two-dimensional electrophoresis,but also was get clear gel map;through the calculation of abundance,there were total 339 differentially expressed proteins discovered in Longyou No.7 under low temperature stress(4℃,48 h)later,changes in expression levels of more than two times the protein spots of 46,which 18 up-regulated protein expression spots,21 down-regulated protein expression spots,7 specific protein expression spots,these differential protein spots were suggested to be related to low temperature stress.This laid the foundation for the research of cold resistant varieties in Winter turnip rape Longyou No.7 proteomics aspect.

Key words: Apoplast, Longyou No.7, Protein, Two-dimensional electrophoresis

CLC Number:

Q945.78
Q946

YUAN Jinhai, LIU Zigang, SUN Wancang, ZENG Xiucun, MA Li, FANG Yuan, LIU Haiqing, GUO Rendi, WANG Zhijiang, CHEN Qi, WANG Kaiyin, LIU Linbo. Extraction of Winter Turnip Rape Leaf Apoplast and Analysis of Protein Expression in Response to Low Temperature Stress[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(4): 44-50. doi: 10.7668/hbnxb.2016.04.008.

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