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作物杂志,2020, 第6期: 8–16 doi: 10.16035/j.issn.1001-7283.2020.06.002

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

大豆G位点近等基因系叶片类囊体蛋白质组比较分析

赵宇杨1(), 宋健2, 邱丽娟1()   

  1. 1中国农业科学院作物科学研究所/国家农作物基因资源与遗传改良重大科学工程/农业农村部作物种质资源与生物技术重点开放实验室,100081,北京
    2长江大学生命科学学院,434025,湖北荆州
  • 收稿日期:2020-04-20 修回日期:2020-10-26 出版日期:2020-12-15 发布日期:2020-12-09
  • 通讯作者: 邱丽娟
  • 作者简介:赵宇杨,主要从事大豆基因资源挖掘与功能研究,E-mail： 398884889@qq.com
  • 基金资助:
    国家自然科学基金(3163005);中国农业科学院科技创新工程

Proteomic Comparation Analysis of Thylakoid in Leaves of G-Locus Near Isogenic Line in Soybean

Zhao Yuyang1(), Song Jian2, Qiu Lijuan1()   

  1. 1Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/The National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Germplasm & Biotechnology (Ministry of Agriculture and Rural Affairs), Beijing 100081, China
    2College of Life Sciences, Yangtze University, Jingzhou 434025, Hubei, China
  • Received:2020-04-20 Revised:2020-10-26 Online:2020-12-15 Published:2020-12-09
  • Contact: Qiu Lijuan

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摘要：

类囊体是光合作用和电子传递关键载体,是叶绿体核心组分,在植物亚细胞器蛋白质组研究中尤为重要。为了探究大豆叶片类囊体的蛋白质组,以1对遗传背景相似度为97.6%的大豆种皮颜色G位点近等基因系（NIL-G和NIL-Y）为材料,利用SDS-PAGE与分级质谱相结合的方法对大豆叶片类囊体蛋白进行分析。结果表明,在NIL-G和NIL-Y中鉴定到的总蛋白分别为2 170种和1 730种,特异蛋白分别为1 140种和700种,总蛋白和特异蛋白数量在NIL-G中均高于NIL-Y。特异蛋白的GO注释及KEGG通路分析表明,尽管这对近等基因系遗传背景相似度很高,但其叶片类囊体蛋白在生物途径、细胞组分和分子功能方面均存在差异。

关键词: 大豆, 质谱, 类囊体, 蛋白质组

Abstract:

Thylakoid is the key carrier of photosynthesis and electron transport and also the core component of chloroplast, which is particularly important in the study of plant subcellular organelle proteome. In order to explore the proteome of thylakoid in soybean leaves, a pair of G-locus near-isogenic line (NIL-G and NIL-Y) with 97.6% similarity in the genetic background was used as materials. SDS-PAGE and mass spectrometry were used to isolate the thylakoid protein from soybean leaves. The results showed that the number of total protein and specific protein in NIL-G was higher than that in NIL-Y, respectively. The numbers of total protein and specific protein in NIL-G were 2 170 and 1 140, while 1 730 and 700 in NIL-Y. GO annotation and KEGG pathway analysis of these specific proteins showed that although the high similarity among genetic background, there are differences in the biological pathway, cell components, and molecular functions of thylakoid proteins in soybean. This provide a new clue to the future research on thylakoid proteins in soybean.

Key words: Soybean, Mass spectrometry, Thylakoid, Proteome

图1

近等基因系叶片类囊体蛋白SDS-PAGE检测

表1

分段质谱鉴定近等基因系NIL-G和NIL-Y大豆叶片类囊体蛋白数目

材料
Material		 < 25kDa 25~45kDa > 45kDa 特异
Specific		 共有
Same		 总计
Total
NIL-G 485 656 1 029 1 140 1 030 2 170
NIL-Y 373 458 899 700 1 730

图2

近等基因系NIL-G和NIL-Y特异蛋白的生物学途径水平GO注释

图3

近等基因系NIL-G和NIL-Y特异蛋白的细胞组分水平GO注释

图4

近等基因系NIL-G和NIL-Y特异蛋白的分子功能水平GO注释

图5

近等基因系NIL-G中特异蛋白的KEGG通路分析

图6

近等基因系NIL-Y中特异蛋白的KEGG通路分析

表2

近等基因系NIL-G和NIL-Y特异蛋白中与光合作用和天线蛋白相关蛋白

蛋白通路
Pathway of protein		 NIL-G NIL-Y
基因编号Gene accession 功能Function 基因编号Gene accession 功能Function
光合作用 A5Z2K3 PSⅠ亚基PsaD C6SWI3 PSⅡR类亚基
Photosynthesis C6SX81 未知蛋白 C6SXP3 未知蛋白
C6SW43 未知蛋白 C6T308 假设的ATP合成酶亚单位b
C6T2Z2 未知蛋白 C6TFR5 未知蛋白
I1MNK0 放氧增强蛋白1 I1M3V1 PSⅡ修复蛋白PSB27-H1
I1M100 ATP合酶γ链
I1JCG8 铁氧还蛋白—NADP还原酶 -
I1NGD2 PSⅠ反应中心亚基II
A0A0R4J4B4 铁氧还蛋白—NADP还原酶
A0A0R4J389 PSⅠ反应中心亚基II
C6TC81 未知蛋白
I1JJ05 放氧增强蛋白2
I1K498 铁氧还蛋白-A
光合作用-天线蛋白 I1KR46 叶绿素a/b结合蛋白CP24 I1MNM3 叶绿素a/b结合蛋白6A
Photosynthesis- C6T8S8 未知蛋白 A0A0R0ERF9 叶绿素a/b结合蛋白CP26
antenna protein A0A0R0JQ91 叶绿素a/b结合蛋白P4
C6TM32 LHCII 1型叶绿素a/b结合蛋白
Q43437 PSⅡ的I型叶绿素a/b结合蛋白
A0A0R0K962 叶绿素a/b结合蛋白P4
I1MZ32 叶绿素a/b结合蛋白CP29

图7

近等基因系NIL-G和NIL-Y表达存在差异的蛋白质层次聚类分析

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