小麦穗突变体SDA1与野生型叶片蛋白差异分析

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (5790 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要小麦(Triticum aestivum)穗突变体SDA1是由隐性多效性单基因小麦穗发育异常基因1(Triticum aestivum spike development atrophy 1, TaSDA1)控制的小麦突变体，表现为穗部发育萎缩不结实、叶片变窄、植株变矮等。为进一步了解TaSDA1基因发生突变的机制，本研究以小麦穗突变体SDA1与野生型为材料，进行主要农艺性状如抽穗期、株高、旗叶长宽、穗长、小穗数调查，并对小麦抽穗期叶片总蛋白进行双向电泳(two-dimensional electrophoresis, 2-DE)，对差异蛋白质点进行质谱分析和qRT-PCR验证。结果表明，SDA1的主要农艺性状存在显著性差异；利用PDQuest8.0.1 软件分析得到27个差异蛋白，有23个质谱检测成功，其中有6个蛋白在SDA1中缺失，17个蛋白在SDA1中表达下调；qRT-PCR检测结果与差异蛋白质谱检测结果一致。在SDA1中，23个质谱成功的差异蛋白主要包括5个光合作用中核酮糖-1,5-二磷酸羧化酶/加氧酶(ribulose-l,5-bisphosphate carboxylase/oxygenase, Rubisco)全酶的大小亚基，4个(SSP7101, SSP7110, SSP7112和SSP8418)表达下调，1个缺失(SSP213)；参与能量代谢的叶绿体中的甘油醛-3-磷酸脱氢酶A(glyceraldehyde-3-phosphate dehydrogenase A, GAPA, SSP8000)表达下调；具有抗氧化能力的过氧化物氧化还原酶(peroxiredoxin, Prx, SSP7320)表达下调；1个RNA结合蛋白(SSP1309)表达下调，导致2个翻译延伸因子(translation elongation factor, eEF-Tu, SSP4614和SSP4802)表达下调，最终导致蛋白合成受阻；参与抗逆境胁迫的蛋白(SSP3738和SSP5209)表达下调，SSP3719蛋白缺失。qRT-PCR验证结果表明，核酮糖1,5二磷酸羧化酶大亚基蛋白(SSP7101)、叶绿体PtrTox结合蛋白(SSP7303)、翻译延伸因子(SSP4614)、磷酸丙糖异构酶(SSP5209)在转录水平与蛋白表达结果一致。SDA1变异表型可能与旗叶的光合作用能力下降、能量代谢紊乱、抗氧胁迫能力下降、抗胁迫能力下降、mRNA编辑过程以及蛋白合成过程受阻有关。本研究结果表明，TaSDA1基因具有多效复杂的调控功能。穗部和叶片的发育状况决定了小麦的产量，阐明SDA1的遗传机理将为小麦穗部和叶片性状的遗传改良奠定理论基础。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	周丽敏
	连俊方
	马文洁
	宋晓朋
	武炳瑾
	张德强
	孙道杰

关键词 ：小麦, 穗发育突变体, 双向电泳(2-DE)

Abstract：SDA1 mutant of wheat (Triticum aestivum) which is incomplete development in spike, narrow development in leaves and dwarf development, is controlled by a recessive pleiotropic single gene Triticum aestivum spike development atrophy 1 (TaSDA1) and it is discovered in the experiment field, belonging to natural mutation. To further research the mutation mechanism of TaSDA1 gene, the present study investigated some major agronomic traits, such as the heading time, flag leaf width and length, the number and length of the spike, dissecting the differences of flag leaves expressed-protein through two-dimensional electrophoresis (2-DE) between SDA1 and wild-type during the heading time, and performed the qRT-PCR verification of the differentially expressed proteins. According to the t test, there were significant differences of the flag leaves' width and length between SDA1 and wild-type. Also the total height of the SDA1 was much lower than that of the wild-type. SDA1 was later in heading than the wild-type. Twenty-seven differentially expressed protein spots were selected by PDQuest (version 8.0.1) image software, among which 23 protein spots were successfully identified. The Mascot software was used to search against the NCBInr database and Uniprot database for protein annotation. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to explore further biological functions of the 2-DE proteins. Among these differentially expressed proteins in SDA1, SSP7101, SSP7110, SSP7112 and SSP8418, related to the Rubisco holoenzyme composition, which were the key enzymes of the Rubisco holoenzyme in photosynthesis, were down-regulated and one of the Rubisco holoenzyme (SSP213) composition showed deficiency. Glyceraldehyde-3-phosphate dehydrogenase A (GAPA, SSP8000), one of the important proteins of glyceraldehyde-3-phosphate dehydrogenase (GAPD), was down-regulated. Peroxiredoxin (Prx, SSP7320), which was important to the oxidation resistance, also showed down-expressed in SDA1. One RNA-binding protein (SSP1309) was down-regulated in SDA1, leading to the down-regulation of 2 translation elongation factors (eEF-Tu, SSP4614 and SSP4802), and finally made the protein biosynthesis blocked. Some of 23 proteins, participating in the adversity stress response, were down-regulated (SSP3738, SSP5209) and showed defiency (SSP3719) in SDA1. qRT-PCR was used to validate ribulose-1,5-bisphosphatecarboxylase/oxygenase large subunit (chloroplast) (SSP7101), chloroplast-localized PtrToxA-binding protein1 (SSP7303), translation elongation factor G(SSP4614) and triosephosphate isomerase (SSP5209) genes expression at the transcriptional level, which were consistent with the protein expressed amounts detected by 2-DE. The mutation phenotype of SDA1, might be related to the flag leaves' differentially-expressed proteins in the decline of the photosynthetic capacity, disorder of the energy metabolism, the decline of oxygen stress resistance and coercive capabilities, process of mRNA editing and the blocked protein biosynthesis. The development of spike and leaves is of great importance to the grain yield of wheat, thus a better understanding of TaSDA1 gene will lay foundation to the theory of genetic improvement of spike and leaf development.

Key words： Wheat Spike mutant Two-dimensional electrophoresis (2-DE)

收稿日期: 2015-10-26 出版日期: 2016-04-01

基金资助:国家重点基础研究计划;陕西省自然基金项目;陕西省重点科技创新团队

通讯作者: 孙道杰 E-mail: chinawheat@hotmail.com

引用本文:

周丽敏连俊方马文洁宋晓朋武炳瑾张德强孙道杰. 小麦穗突变体SDA1与野生型叶片蛋白差异分析[J]. , 2016, 24(5): 637-648.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I5/637

[1]单保山, 常文锁.小麦旗叶对产量性状及籽粒蛋白含量作用的研究[J].河北农业大学学报, , (2):5-9
[2]Shan Baoshan, Chang Wensuo.1991. Effects of Fla g Leaf on Grain Protein and Yield in Wheat[J]. Journal of Hebei Agticultural University, 2:5-9.(in Chinese)
[3]杜丽芬, 李明飞, 刘录祥, 等.一个化学诱变的小麦斑点叶突变体的生理和遗传分析[J].作物学报, 2014, 40(6):1020-1026
[4]Du Lifen, LI Mingfei, LIU Luxiang, et al.2014. Physiological Characteristics and Genetic Analysis on a Spotted-leaf Wheat Derived from Chemical Mutation[J]. ACTA AGRONOMICA SINICA, 40(6): 1020?1026.(in Chinese)
[5]封德顺, 徐勤迎, 王洪刚, 等.2007.白粉病菌侵染后小麦叶片蛋白质变化的研究[J].华北农学报, 22(3): 123-126.
[6]Feng Deshun, Xu Qinying, Wang Hong gang, et al.2007.Changes of Protein in Wheat Leaf After the Infection of Powdery Mildew[J]. Acta Agriculturae Boreali-Sinica, 22(3): 123-126.(in Chinese)
[7]黄修文, 张艳菊, 冯清.2012. UV-B辐射处理小麦苗期叶片的蛋白质组分析[J], 中国农业大学学报, 17(2): 31-36.
[8]Huang Xiuwen, Zhang Yanju, Feng Qing.2012.Chen Weixia, Zhang Wenhui, Li Baoyun. Analysis of wheat seeding leaf protiome after UV-B bradiation[J]. Journal of China Agricultural University, 17(2): 31-36.(in Chinese)
[9]李亚宁, 陈春, 李国东, 等.2013. 磺胺甲恶唑对小麦叶片蛋白和叶绿素含量及SOD酶活性的影响[J]. 生态毒理学报, 8(4): 543-548.
[10]Li Yangning, Chen Chun, Li Guodong, 2013.Effects of Sulfamethoxazole(SMZ) on the Content of Chlorophy(CHL) and Soluble Protein(SP), and the Superoxide Dismutases (SOD) Activity of Wheat, Triticum aestivum[J]. Asian Journal of Ecotoxicology, , 8(4): 543-548.(in Chinese)
[11]宋全昊刘路平李法计, 等.2013.小麦穗部发育多效基因的遗传分析与基因定位[J]. 西北植物学报, 33(4): 0643-0648.
[12]Song Quanhao, Liu Luping, Li Faji, et al.2013.Genitic Analysis and Gene Mapping of Spike Development Pleiotropic Gene in Wheat[J]. Acta Botanica Boreali-Occidentalia Sinica, 33(4): 0643-0648.(in Chinese)
[13]宋素洁, 古佳玉, 郭会君, 等.2012. 小麦叶绿素缺失突变体 Mt6172及其野生型叶片蛋白质组学双向差异凝胶电泳分析[J]. 作物学报., 38(9): 1592-1606.
[14]Somg Sujie, Gu Jiayu, Guo Huijun, et al.2012. Proteomic Analysis of Leaves of the Chlorophyll-Deficient Wheat Mutant Mt6172 and Its Wild-Type through 2D-Difference Gel Electrophoresis[J]. ACTA AGRONOMICA SINICA, 38(9): 1592-1606.(in Chinese)
[15]王德龙, 叶武威, 王俊娟, 等.2010. 干旱胁迫下棉花 SSH 文库构建及其抗旱相关基因分析[J]. 作物学报, 36(12): 2035?2044.
[16]Wang Delong, Ye Wuwei, Wang JunJuan, et al.2010. Construction of SSH Library and Its Analyses of Cotton Drought Associated Genes under Drought Stress[J] . ACTA AGRONOMICA SINICA, 36(12): 2035?2044. (in Chinese)
[17]吴同彦, 杨学举, 刘会, 等.2009. 小麦叶片性状与主要农艺性状的相关性研究.安徽农业科学, 37( 1): 94-96.
[18]Wu Tongyan, Yang Xueju, Liu Hui, et al.Correlation Analysis on Leaf Characters and Some Main Agronomic Traits of Wheat[J]. Journal of Anhui Agri. Sci. 2009, 37( 1): 94-96.(in Chinese)
[19]辛庆国, 刘录祥, 于元杰, 等.2007. 离子注入技术及其在小麦育种中的应用[J].麦类作物学报, 27( 2): 354-357.
[20]Xin Qingguo, Liu Luxiang, Yu Yuanjie, et al.2007. Ion Beam Implantation Technique and Its Application in Wheat Breeding[J]. Journal of Triticeae Crops, 27( 2): 354-357.(in Chinese)
[21]杨鹏, 韩锦峰, 黄晋玲.2014. 棉花晋 A 细胞质雄性不育系及其保持系差异蛋白质组分析[J].中国农业科学, 47(20):3929-3940.
[22]Yang Peng, Han Jinfeng, Huang Jinling.2014. Proteome Analysis of Cytoplasmic Male Sterility and Its Maintaince in JA-CMS Cotton[J]. Scientia Agricultura Sinica, 47(20):3929-3940. (in Chinese)
[23]赵天祥, 孔秀英, 周荣华, 等.2009. EMS诱变六倍体小麦堰展4110的形态突变体鉴定与分析[J].中国农业科学, 42:755-764.
[24]Zhao Tianxiang, Kong Xiuying, Zhou Ronghua, Gao Shuangcheng, Jia Jizeng.Morphological Identification and Analysis of EMS Mutants from Hexaploid Wheat Cultivar Yanzhan 4110[J].Scientia Agricultura Sinica, 2009, 42(3):755-764
[25]Andersson and Taylor.2003.Structural framework for catalysis and regulation in ribulose-1, 5- bisphosphate carboxylase/oxygenase[J]. Arch Biochem Biophys, 414⑵: 130-140.
[26]Avilan L, Maberly CS, Mekhalf M, et al.2012. Regulation of glyceraldeh-yde-3-phosphate dehydrogenase in the eustigmatophyte Pseudo characiopsis ovalis is intermediate between a chlorophyte and a diatom[J] . Eur J Phycol, 47(3): 207-215.
[27]Benndorf D, Loffhagen N, Babel W, et al.1999 Induction of heat shock proteins in response to primary alcohols in Acine-tobacter calcoaceticus[J]. Electrophoresis, 20(4-5): 781?789.
[28]Damerval C, de Vienne D, Zivy M, et al.1986.Thiellement H. Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins[J]. Electrophoresis, 7(1): 52-54.
[29]Demmig B, Winter K, Krger A, et al.1987. Photo inhibition and zeaxanthin formation in intact leaves: A possible role of the xanthophyll cycle in the dissipation of excess light energy[J]. Plant Physiol, 84: 218?224
[30]Esquivel MG, Anwamzzaman M, Spreitzerb R J.2000. Deletion of nine carboxyl-terminal residues of the Rubisco small subunit decreases thermal stability but does not eliminate f unction[J]. FEBS Leters, (520): 73-76.
[31]Forbes K.P., Addepalli B., Hunt A.G.J. et al.2006. An Arabidopsis Fip1 homolog interacts with RNA and provides conceptual links with a number of other polyadenylation factor subunits[J]. Acta Biophysica Sinica, 281:176-186.
[32]G?rg A, Obermaier C, Boguth G, et al.1999. Recent developments in two-dimensional gel electrophoresis with immobilized pH gradients: wide pH gradients up to pH 12, longer separation distances and simplified procedures[J]. Electrophoresis, 20(4-5): 712?717.
[33]Howard TP, Lloyd JC, Raines CA.2011. Inter-species variation in the oligomeric states of the higher plant Calvin cycle enzymes glyceral-dehyde-3-phosphate dehydrogenase and phosphor ribulo kinase[J]. Expe Bot, 62(11): 3799-3805.
[34]Hu H, Boisson-Dernier A, Israelsson-Nordstrom M, et al.2010. Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells[J]. Nat Cell Biol, 12: 87-93.
[35]Isermann sermann K, Liebau E, Roeder T et al.2004. A peroxiredoxin specifically expressed in two types of pharyngeal neurons is required for normal growth and egg production in Caenorhabditis elegans[J]. Journal of Molecular Biology, 338: 745-755.
[36]Ishida H, Nishimori Y, Sugisawa M, et al.1997. The large subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase is fragmented into 37kDa and 16kDa polypeptides by active oxygen in the lysates of chloroplasts from primary leaves of wheat[J]. Plant Cell Physiol, 38: 471–479
[37]Ito H, Iwabuchi M, Ogawa K.2003. The sugar-metabolic enzymes aldolase and triose-phosphate isomerase are targets of glutathionylation in Arabidopsis thaliana:Detection using biotinylated glutathione[J]. Plant and Cell Physiology, 44 (7): 655–660.
[38]Kaul T, Reddy PS, Mahanty S, et al.2011. Biochemical and molecular characterization of stress-induced β-carbonic anhydrase from a C4 plant Pennisetum glaucum[J]. Plant Physiol, 168: 601-610.
[39]Krause G H, Weis E.1991. Chlorophyll fluorescence and photosynthesis: The basics. Ann Review Plant Physiol[J]. Plant Mol Biol, 45: 633?652.
[40]Makino A, Mae T, Ohira K.1985. Enzymic properties of ribulose-1, 5-bisphosphate carboxylase/oxygenase purified from rice leaves[J]. Plant Physiol, 79: 57–61.
[41]Mayer K, Schuller C, Wambutt R, et al.1999. Sequence and analysis of chromosome 4 of the plant Arabidopsis thaliana. EU, CSHL and WU[J]. Arabidopsis Sequencing Project Nature, 402: 769-777.
[42]Nelson N, Ben-Shem A, et al.2004. The complex architecture of oxygenic photosynthesis[J]. Nat Rev Mol Cell Biol, 5: 971–982.
[43]Salekdeh G H, Siopongco J, Wade L J, et al.2002. Approach to analyzing drought and salt-responsiveness in rice[J]. Field Crops Research，76：199–219.
[44]Sirover MA.2011. On the functional diversity of glyceraldehyde-3-phosphate dehydrogenase ：Biochemical mechanisms and regulatorycontro[J]l. Biochimi Biophys Acta, 1080: 741-751.