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2025年8月17日 星期日
  2016, Vol. 24 Issue (5): 637-648    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
小麦穗突变体SDA1与野生型叶片蛋白差异分析
周丽敏1,连俊方1,马文洁2,宋晓朋1,武炳瑾2,张德强1,孙道杰1
1. 西北农林科技大学南校区
2.
Analysis of Differentially Expressed Proteomes of Wheat (Triticum aestivum) Leaves Between SDA1 Spike Mutant and Its Wild-type
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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。
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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 wordsWheat    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
 
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