春大豆种子田间劣变性和劣变抗性的差异蛋白质组学研究

doi:10.3864/j.issn.0578-1752.2015.01.03

摘要/Abstract

摘要： 【目的】南方春大豆种子在生理成熟（R6或R7期）过程中易受高温高湿胁迫影响常会发生种子田间劣变，已经严重制约了中国南方春大豆生产和应用的发展。应用比较蛋白质组学技术，在蛋白表达水平上揭示高温高湿下南方春大豆种子田间劣变性和劣变抗性的机制，为遗传育种改良和新品种选育奠定种质基础。【方法】利用抗种子田间劣变种质湘豆3号和不抗种质宁镇1号为材料，在种子发育到生理成熟期时模拟田间高温高湿胁迫处理，运用双向电泳（2-DE）和MALDI-TOF/TOF鉴定技术研究春大豆种子蛋白质表达谱的变化。【结果】高温高湿胁迫处理和对照条件下（1、5、10、16和24 h），湘豆3号和宁镇1号大豆种子可溶性蛋白的每张2-DE重复胶上都可以检测到700多个可重复蛋白点，50个蛋白质点在处理与对照之间表达量上存在显著差异。其中有33个差异蛋白点经质谱分析成功鉴定；功能分类表明，这些成功鉴定的差异蛋白分别涉及细胞修复及防御（9%）、氧化还原平衡（12%）、蛋白合成（3%）、能量代谢（15%）、转运过程（15%）以及贮藏蛋白（31%）等代谢途径和细胞过程。此外，还有5个差异蛋白为未知功能蛋白。【结论】高温高湿胁迫下，抗性种质湘豆3号较不抗种质宁镇1号具有较强的抗氧化和细胞修复及防御能力，可能是其具有较强的抗种子田间劣变性的关键原因。

关键词: 春大豆种子, 种子田间劣变, 劣变抗性, 高温高湿, 蛋白质组学

Abstract: 【Objective】 Soybean seed is susceptible to high temperature and humidity (HTH) stress resulting in pre-harvest deterioration during its development and maturation (R6 or R7), which limits the production and utilization of soybean in south China. The aim of the present study is to reveal the pre-harvest seed deterioration and deterioration resistance mechanism of spring soybean under HTH stress at proteomic level, thus laying a foundation for breeding new cultivars.【Method】When the deterioration-sensitive cultivar Ningzhen No.1 and deterioration-tolerant cultivar Xiangdou No.3 developed to physiological period (R7), their plants were treated under HTH stress and control for 1, 5, 10, 16 and 24 h, respectively. The extracted proteins from the developing seed of treatment and control were analyzed by two-dimensional electrophoresis (2-DE), and the differentially expressed proteins were identified by MALDI-TOF/TOF. 【Result】Approximately 700 protein spots were detected on each 2-DE gel. Of them, 50 were found to be significantly changed in abundance, with 33 being successfully identified by MALDI-TOF/TOF. The identified differentially expressed proteins were involved in cell rescue and defense (9%), redox homeostasis (12%), protein synthesis (3%), energy metabolism (15%), transport pathway (15%), protein destination and storage (31%). Moreover, the function of 5 identified proteins is unknown. 【Conclusion】Pre-harvest deterioration-resistant cultivar Xiangdou No.3 possessed the greater ability of ROS scavenging and cell rescue and defense than deterioration-sensitive cultivar Ningzhen No.1 under HTH tress, which may be the major reasons why it is more deterioration-resistant than the latter.

Key words: spring soybean seed, pre-harvest seed deterioration, deterioration resistance, high temperature and humidity, proteomics

宋利茹，王爽，牛娟，马洪雨，舒英杰，杨艳，顾卫红，麻浩. 春大豆种子田间劣变性和劣变抗性的差异蛋白质组学研究[J]. 中国农业科学, 2015, 48(1): 23-32.

SONG Li-ru, WANG Shuang, NIU Juan, MA Hong-yu, SHU Ying-jie, YANG Yan, GU Wei-hong, MA Hao. Differentially Proteomics Analysis of Pre-Harvest Seed Deterioration and Deterioration Resistance in Spring Soybean[J]. Scientia Agricultura Sinica, 2015, 48(1): 23-32.

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参考文献

[1] 韩天富, 盖钧镒. 世界菜用大豆生产、贸易和研究的进展. 大豆科学, 2002, 21(4): 278-284.

Han T F, Gai J Y. Advance in production, trade and research of vegetable soybean in the world. Soyben Scinece, 2002, 21(4): 278-284. (in Chinese)

[2] Gibson L R, Mullen R E.Soybean seed quality reductions by high day and night temperature. Crop Science, 1996, 36: 1615-1619.

[3] 乔燕祥, 周建萍, 田齐建, 穆志新. 大豆种子老化过程中生理特性变化的研究. 植物遗传资源学报, 2010, 11(5) : 616-620.

Qiao Y X, Zhou J P, Tian Q J, Mu Z X. Changing of physiological characteristics of soybean seeds in aging course. Journal of Plant Genetic Resources, 2010, 11(5): 616-620. (in Chinese)

[4] 王芳, 王丽群, 田鑫, 顾卫红, 麻浩. 中国南方春大豆收获前后种子劣变的抗性研究.中国农业科学, 2007, 40(11): 2637-2647.

Wang F, Wang L Q, Tian X, Gu W H, Ma H. Pre-harvest and post-harvest seed deterioration resistance of spring soybean germplasm in South China. Scientia Agricultura Sinica, 2007, 40(11): 2637-2647. (in Chinese)

[5] Saha R R, Sultana W. Influence of seed ageing on growth and yield of soybean. Bangladesh Journal of Botany, 2008, 37: 21-26.

[6] Ren C, Bilyeu K D, Beuselinck P R. Composition, vigor, and proteome of mature soybean seeds developed under high temperature. Crop Science, 2009, 49: 1010-1022.

[7] 唐桂香, 汪自强, 董明远. 春播和秋播对南方春大豆种子活力的影响. 作物学报, 1998, 21(2):243-247.

Tang G X, Wang Z Q, Dong M Y. Effects of plantings in spring and fall on the vigor of spring soybean seed in southern region. Acta Agronomica Sinica, 1998, 21(2): 243-247. (in Chinese)

[8] McDonald M B. Seed deterioration: Physiology, repair and assessment. Seed Science and Technology, 1999, 27: 177-237.

[9] Egli D B, TeKrony D M, Heitholt J J. Air temperature during seed filling and soybean seed germination and vigor. Crop Science, 2005, 45: 1329-1335.

[10] 唐善德, 黄敏珍, 成金莲. 春大豆种子劣变的研究. 大豆科学, 1994, 13(3): 230-236.

Tang S D, Huang M Z, Cheng J L. A study on seed deterioration in spring soybeans. Soybean Science, 1994, 13(3): 230-236. (in Chinese)

[11] Wien H, Kueneman E. Soybean seed deterioration in the tropics: II. Varietal differences and techniques for screening. Field Crops Research, 1981, 4: 123-132.

[12] 马国荣. 国际热带农业研究所(IITA)的大豆育种. 大豆科学, 1989, 8(2): 203-205.

Ma G R. Soybean breeding program in the International Institute of Tropical Agriculture (IITA). Soybean Science, 1989, 8(2): 203-205. (in Chinese)

[13] 孟祥栋. 菜用大豆种子劣变与生理生化变化的关系. 大豆科学, 1993, 12: 259-264.

Meng X D. Relationship between seed deterioration and physiological and biochemical changes of vegetable soybeans. Soybean Science, 1993, 12: 259-264. (in Chinese)

[14] 赵垦田, 李立华. 人工老化过程红松种胚细胞物质外渗和超微结构变化. 东北林业大学学报, 2000, 28(3): 5-7.

Zhao K T, Li L H. Cellular substance exudation and ultrastructural change of seed embryos of pinus koraiensis artificially accelerated aging. Journal of Northeast Forestry University, 2000, 28(3): 5-7. (in Chinese)

[15] 王慧超, 何士敏, 李昌满. PEG渗调处理对植物种子的影响. 安徽农业科学, 2008, 36(6) :2224-2226.

Wang H C, He S M, Li C M. Effect of PEG osmotic conditioning treatment on plant seed. Journal of Anhui Agriculture Science, 2008, 36(6): 2224-2226. (in Chinese)

[16] Wang L Q, Ma H, Song L R, Shu Y J, Gu W H. Comparative proteomics analysis reveals the mechanism of pre-harvest seed deterioration of soybean under high temperature and humidity stress. Journal of Proteomics, 2012, 75(7): 2109-2127.

[17] 舒英杰, 周玉丽, 陶源, 王爽, 杨艳, 张国敏, 麻浩. 模拟田间劣变对生理成熟期春大豆植株生长及种子活力的影响. 大豆科学, 2013, 32(5): 635-639.

Shu Y J, Zhou Y L, Tao Y, Wang S, Yang Y, Zhang G M, Ma H. Effect of simulated pre-harvest deterioration stress on plant growth and seed vigor of spring soybean at physiological maturity stage. Soybean Science, 2013, 32(5): 635-639. (in Chinese)

[18] Parrish D J, Leopold A C. On the mechanism of aging in soybean seeds. Plant Physiology, 1978, 61: 365-369.

[19] Jain A, Shivanna K R. Loss of viability during storage is associated with changes in membrane phosphohpid. Physology Chemistry, 1989, 28 : 999-1002.

[20] Clarke N A, James P E. The effects of priming and accelerated ageing upon the nucleic acid content of leek seeds and their embryos. Journal of Experimental Botany, 1991, 42: 261-268.

[21] Krober V A, Collins F I. Effects of weather damage on the chemical composition of soybeans. Journal of American Oil Chemists Society, 1948, 25: 296-298.

[22] Ching T M. Adenosine tnphosphate and seed vigor// Khan A A. ed. The Physiology and Biochemistry of Seed Development, Dormancy, and Germination. Elsevier New York, 1982: 487-505.

[23] Duke S H, Kakefuda G, Henson C A, Loeffler N L. Role of the testa epidermis in the leakage of intracellular substances from imbibing soybean seeds and its implications for seedling survival. Physiologia Plantarum, 1986, 68: 625-631.

[24] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 1976, 72: 248-254.
[25] Blum H, Beier H, Gross H J. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis, 1987, 8: 93-99.

[26] Yao Y, Yang Y W, Liu J Y. An efficient protein preparation for proteomic analysis of developing cotton fibers by two-dimensional gel electrophoresis. Electrophoresis, 2006, 27: 4559-4569.

[27] Ma H Y, Song L R, Shu Y J, Wang S, Niu J, Wang Z K, Yu T, Gu W H, Ma H. Comparative proteomic analysis of seedling leaves of different salt tolerant soybean genotypes. Journal of Proteomics, 2012, 75(6): 1529-1546.

[28] Zhang L, Yu Z F, Jiang L, Jiang J, Luo H B, Fu L R. Effect of post-harvest heat treatment on proteome change of peach fruit during ripening. Journal of Proteomics, 2011, 74: 1135-1149.

[29] Huang H, Møller I M, Song S Q. Proteomics of desiccation tolerance during development and germination of maize embryos. Journal of Proteomics, 2012, 75: 1247-1262.

[30] Hashiguchi A, Sakata K, Komatsu S. Proteome analysis of early-stage soybean seedlings under flooding stress. Journal of Proteome Research, 2009, 8: 2058-2069.

[31] Wan X Y, Liu J Y. Comparative proteomics analysis reveals an intimate protein network provoked by hydrogen peroxide stress in rice seedling leaves. Molecular and Cellular Proteomics, 2008, 10: 1469-1488.

[32] Kim Y J, Lee S, Lee H M. Comparative proteomics analysis of seed coat from two black colored soybean cultivars during seed development. Plant Omics Journal, 2013, 6(6): 456-463.

[33] Major I T, Constabel C P. Functional analysis of the Kunitz trypsin inhibitor family in poplar reveals biochemical diversity and multiplicity in defense against herbivores. Plant Physiology, 2008, 146: 888-903.

[34] Wang J Q, Suen P K, Xu Z F, Jiang L W. A 64 kDa sucrose binding protein is membrane-associated and tonoplast-localized in developing mung bean seeds. Journal of Experimental Botany, 2009, 60(2): 629-639.

[35] Salekdeh G H, Siopongco J, Wade L J, Ghareyazie B, Bennett J. Proteomic analysis of rice leaves during drought stress and recovery. Proteomics, 2002, 2(9): 1131-1145.

[36] Pacold I, Anderson L E. Chloroplast and cytoplasmic enzymes: VI. Pea leaf 3-phosphoglycerate kinases. Plant Physiology, 1975, 55: 168-171.