[1]甄文轩,杨晓桐,郭歌扬,等.大豆叶片蛋白响应干旱胁迫的双向电泳分析[J].大豆科学,2019,38(01):35-40.[doi:10.11861/j.issn.1000-9841.2019.01.0035]
　ZHEN Wen-xuan,YANG Xiao-tong,GUO Ge-yang,et al.Two-dimensional Electrophoresis Pattern Analysis of Leaf Proteins from Soybean under Drought Stress[J].Soybean Science,2019,38(01):35-40.[doi:10.11861/j.issn.1000-9841.2019.01.0035]
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大豆叶片蛋白响应干旱胁迫的双向电泳分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第38卷 期数: 2019年01期 页码: 35-40 栏目: 出版日期: 2019-01-20
Title:
Two-dimensional Electrophoresis Pattern Analysis of Leaf Proteins from Soybean under Drought Stress
作者:
甄文轩; 杨晓桐; 郭歌扬; 宋颖; 赵雨薇; 臧新
(郑州大学 生命科学学院，河南 郑州 450001）
Author(s):
ZHEN Wen-xuan;  YANG Xiao-tong;  GUO Ge-yang;  SONG Ying;  ZHAO Yu-wei;  ZANG Xin
（School of Life Science, Zhengzhou University, Zhengzhou 450001, China）
关键词:
大豆; 干旱胁迫; 双向电泳; 表达图谱
Keywords:
Soybean;  Drought stress;  Two-dimensional electrophoresis;  Expression map
DOI:
10.11861/j.issn.1000-9841.2019.01.0035
文献标志码:
A
摘要:
大豆是我国重要的农作物之一，蛋白质组学技术是解析植物响应逆境胁迫的重要工具之一。为进一步推进对响应干旱胁迫的大豆叶片蛋白的鉴定工作，以大豆幼苗的叶片为研究对象，用甘露醇溶液模拟干旱胁迫，处理24 h后提取叶片蛋白粉进行双向电泳分离和图谱差异分析。结果表明：采取三氯醋酸（TCA）-丙酮沉淀法提取的叶片蛋白进行双向电泳分离能够得到稳定清晰的双向电泳蛋白图谱。采取PD-Quest软件对干旱胁迫下的大豆叶片蛋白表达图谱进行差异分析，共检测和匹配425个大豆叶片蛋白点，其中差异表达的蛋白点有101个，60个表达量上调，41个表达量下调。本研究结果表明有一部分大豆叶片蛋白响应了干旱胁迫。
Abstract:
Soybean is one of the important crops in China. Proteomics technology is one of the important tools to analyze plant response to stress. To lay the foundation for further identification of homologous proteins of soybean leaf protein in response to drought stress, the leaves of soybean seedlings were used as research objects. The mannitol solution was used to simulate drought stress. After 24 h treatment, the leaf protein powder was extracted for two-dimensional electrophoresis separation and spectral difference analysis. The results showed that the leaf protein was extracted by trichloroacetic acid-acetone precipitation method for two-dimensional electrophoresis analysis to obtain a stable and clear two-dimensional electrophoresis protein map. PD-Quest software was used to analyze the protein expression profiles of soybean leaves under drought stress. A total of 425 soybean leaf protein spots were detected and matched. There were 101 differentially expressed protein spots, among which 60 were up-regulated, and 41 were down-regulated. The results of this study indicated that there were some soybean leaf proteins responded to drought stress.

参考文献/References:

［1］王厚鑫,刘鸣达, 张惠, 等.施硅对草地早熟禾生长特性和抗旱性的影响［J］.北方园艺, 2007, 31 (9) :135-137. (Wang H X, Liu M D, Zhang H, et al. Effects of silicon application on growth characteristics and drought resistance of Kentucky bluegrass ［J］. Northern Horticulture, 2007, 31 (9): 135-137.)
［2］高亚梅,韩毅强,杜吉到,等. 干旱胁迫对大豆酶活性的影响［J］. 黑龙江八一农垦大学学报,2007(4):13-16.(Gao Y M, Han Y Q, Du J D, et al. Effects of drought stress on soybean enzyme activity［J］. Journal of Heilongjiang Bayi Agricultural University, 2007(4): 13-16.)
［3］高蕾,孙海龙.干旱胁迫对大豆幼苗生长及生理特性的影响［J］.黑龙江科学,2011,2(6):7-9.(Gao L, Sun H L. Effects of drought stress on growth and physiological characteristics of soybean seedlings［J］.Heilongjiang Science,2011,2(6):7-9.)
［4］董兴月,林浩,刘丽君,等.干旱胁迫对大豆生理指标的影响［J］.大豆科学,2011,30(1):83-88. (Dong X Y, Lin H, Liu L J, et al. Effects of drought stress on physiological indexes of soybean［J］.Soybean Science,2011,30(1):83-88.)
［5］王燕平,王晓梅,侯国强,等.干旱胁迫对不同生态型大豆生理生化特征的影响［J］.中国农学通报,2014,30(12):93-100.（Wang Y P, Wang X M, Hou G Q,et al. Effects of drought stress on physiological and biochemical characteristics of different ecotypes of soybean［J］. Chinese Agricultural Science Bulletin, 2014, 30(12): 93-100.）
［6］王春艳,庞艳梅,李茂松,等.干旱胁迫对大豆气孔特征和光合参数的影响［J］.中国农业科技导报,2013,15(1):109-115. (Wang C Y, Pang Y M, Li M S, et al.Effects of drought stress on stomatal characteristics and photosynthetic parameters of soybean［J］. China Agricultural Science and Technology Review, 2013, 15(1): 109-115.)
［7］Mohammadi P P，Moieni A，Hiraga S，et al. Organ-specific pro-teomic analysis of drought-stressed soybean seedlings［J］.Journal of Proteomics,2012,75: 1906-1923.
［8］Khatoon A，Rehman S，Hiraga S，et al．Organ-specific proteomics analysis for identification of response mechanism in soybean seedlings under flooding stress［J］.Journal of Proteomics,2012, 75: 5706-5723．
［9］Komatsu S, Yamamoto A, Nakamura T, et al. Comprehensive analysis of mitochondria in roots and hypocotyls of soybean under flooding stress using proteomics and metabolomics techniques［J］.Journal of Proteome Research, 2011, 10(9):3993-4004.
［10］Ma H Y，Song L R，Shu Y J，et al．Comparative proteomic analysis of seedling leaves of different salt tolerant soybean genotypes［J］．Journal of Proteomics，2012, 75: 1529-1546．
［11］王文军. 大豆种子冷害和PEG引发的蛋白质组学分析［D］. 呼和浩特：内蒙古农业大学, 2008.(Wang W J. Proteomic analysis of soybean seed chilling injury and PEG induced ［D］. Hohhot: Inner Mongolia Agricultural University, 2008.)
［12］Damerval C,de Vienne D,Zivy M.Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat seeding proteins［J］.Electrophories,1986,7(1):52-54.
［13］Braford M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding［J］.Analytical Biochemistry,1976,72:248-254.
［14］Park I, Ndjomou J, Wen Y, et al. Inhibition of HCV replication by oxysterol-binding protein-related protein 4 (ORP4) through interaction with HCV NS5B and Alteration of lipid droplet formation［J］. PLoS One, 2013, 8(9):e75648.
［15］Herman E M，Helm R M，Jung R，et al．Genetic modification removes an immune dominant allergen from soybean［J］.Plant Physiology,2003,132(1):36-43．
［16］Hurkman W J, Tanaka C K.Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis［J］.Plant Physiology,1986 ,81(3):802-806．
［17］Meyer Y, Grosset J, Chartier Y, et al. Preparation by two-dimensional electrophoresis of proteins for antibody production: Antibodies against proteins whose synthesisis reduced by auxin in tobacco mesophyll protoplasts［J］.Electrophoresis, 1988,9(11):704-712．
［18］Wang W，Vignani R，Scali M，et al．Removal of lipid contaminants by organic solvents from oilseed protein extract prior to electrophoresis［J］. Analytical Biochemistry,2004,329(1):139 -141．
［19］Hurkman W J,Tanaka C K.Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis［J］.Plant Physiology,1986,81(3):802-806．

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备注/Memo

收稿日期：2017-08-26

基金项目：河南省高等学校重点科研项目（18A180005）；大学生创新创业训练计划（201810459067）。
第一作者简介：甄文轩（1998-），女，学士，主要从事生物化学研究。E-mail:18838200120@163.com。
通讯作者：臧新（1968-），男，博士，副教授，主要从事植物细胞工程蛋白组学与植物细胞工程研究。E-mail: zangxin@zzu.edu.cn。

更新日期/Last Update: 2019-01-22