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作物学报 ›› 2019, Vol. 45 ›› Issue (3): 390-400.doi: 10.3724/SP.J.1006.2019.84056

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

花生膜联蛋白基因家族成员的结构和表达分析

王慧敏1,李新国2,万书波3,张智猛4,丁红4,李国卫2,高文伟1,*(),彭振英1,2,*()   

  1. 1 新疆农业大学, 新疆乌鲁木齐830052
    2 山东省农业科学院生物技术研究中心 / 山东省作物遗传改良与生态生理重点实验室, 山东济南250100
    3 山东省农业科学院, 山东济南250100
    4 山东省花生研究所, 山东青岛266100
  • 收稿日期:2018-04-19 接受日期:2018-10-08 出版日期:2019-03-12 网络出版日期:2018-11-05
  • 通讯作者: 高文伟,彭振英
  • 作者简介:E-mail: whuiminyspa@163.com
  • 基金资助:
    本研究由山东省自然科学基金重大基础研究项目(植物根系耐盐、抗旱、耐瘠薄的调控机理);山东省农业科学院农业科技创新工程项目(CXGC2018B05);山东省自然科学基金项目资助(ZR2014YL043)

Structure and expression analysis of the members of peanut annexin gene family

Hui-Min WANG1,Xin-Guo LI2,Shu-Bo WAN3,Zhi-Meng ZHANG4,Hong DING4,Guo-Wei LI2,Wen-Wei GAO1,*(),Zhen-Ying PENG1,2,*()   

  1. 1 Xinjiang Agricultural University, Wulumuqi 830052, Xinjang, China
    2 Research Center of Biotechnology, Shandong Academy of Agricultural Sciences / Shandong Key Laboratory of Crop Genetic Improvement and Ecology and Physiology, Jinan 250100, Shandong, China
    3 Shandong Academy of Agricultural Sciences, Jinan 250100, Shandong, China
    4 Shandong Peanut Research Institute, Qingdao 266100, Shandong, China
  • Received:2018-04-19 Accepted:2018-10-08 Published:2019-03-12 Published online:2018-11-05
  • Contact: Wen-Wei GAO,Zhen-Ying PENG
  • Supported by:
    This study was supported by the Major Basic Research Project of Shandong Natural Science Foundation;Shandong Academy of Agricultural Sciences Agricultural Science and Technology Innovation Project(CXGC2018B05);Shandong Natural Science Foundation(ZR2014YL043)

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摘要：

植物膜联蛋白(annexin)是一类钙依赖性磷脂结合蛋白, 参与调控植物代谢和生长发育并协同调节抗旱、耐盐等多种抗逆反应, 其结构在不同植物中具有物种特异性。为系统分析花生膜蛋白基因家族, 对30个花生annexin (annexin of Arachis hypogaea, AnnAh)基因进行了生物信息学分析。结果表明 30个AnnAhs不均匀分布在13条染色体上, 其中A、B基因组中各有13个和17个。AnnAhs含有2~8个内含子, 但大多数含有5~6个内含子。聚类分析表明, 植物annexin聚类关系比较复杂, 低等植物、单子叶、双子叶植物annexin间隔分布, AnnAhs穿插其中, 分布于各个分支中; 但在各个小分支中, AnnAhs 基本上都与双子叶植物annexin聚在一起, 其中与大豆、苜蓿、向日葵亲缘关系较近, 其次是拟南芥; 但个别AnnAhs与单子叶植物和低等植物annexin聚在一起。30个AnnAhs均无跨膜结构域, 其中有16个AnnAhs定位于细胞质, 其余定位不明确。对AnnAhs可变剪切分析显示, 仅有11个发生可变剪切事件, 占38%; 根中发生的最多, 其次是叶中, 种子中最少。根据表达谱数据分析发现, AnnAhs在seed2和根中表达量较高, 其次是seed1, 在叶中表达量较低。本文可为花生抗性育种提供一定的理论支撑。

关键词: 花生, 膜联蛋白, 基因结构分析, 可变剪切分析, 表达模式分析

Abstract:

Annexin is a kind of calcium-dependent phospholipid binding proteins involved in the regulation of plant metabolism, growth and development, drought resistance and salt tolerance, and its structure is species-specific in different plants. In order to have a systematic analysis of the annexin gene family of peanut, we identified 30 annexin genes from the peanut genome database, and analyzed their characteristics using bioinformatics method. Peanut annexin (annexin of Arachis hypogaea, AnnAh) genes were unevenly distributed on 13 chromosomes, with 13 in A genome and 17 in B genome. There were 2 to 8 introns in the AnnAhs, with 5 to 6 introns in most AnnAhs. Phylogenetic analysis showed that the clustering relationship was complex. The annexins of lower plant, monocotyledonous and dicotyledonous plants were distributed at interval, with AnnAhs inserting in each branch. However, in each small branch, AnnAhs were basically clustered with the dicotyledonous plant annexins, and close to soybean, alfalfa and sunflower, followed by Arabidopsis; but several AnnAhs were associated with monocotyledonous and lower plant annexins. All 30 AnnAhs had no transmembrane domain, and 16 of them were located in cytoplasm, and the others’ localization was uncertain. Results of alternative splicing (AS) analysis of AnnAhs showed that only 11 AnnAhs experienced AS, which accounted for about 38% of all AnnAhs; the AS events occurred most in roots, followed by leaves, and the least in seeds. The expression level of AnnAhs was high in seed2 and root, followed by seed1, and lower in leaf. The comprehensive analysis of AnnAhs can provide some theoretical support for peanut resistance breeding.

Key words: peanut, annexin, gene structure analysis, alternative splicing analysis, expression pattern analysis

表1

Anneixn家族基因"

基因名称
Gene name		 染色体位置
Chr. position		 氨基酸个数
Amino acid number		 亚细胞定位
Subcellular localization		 蛋白完整性
Protein integrity		 Annexin
结构域
Annexin
domain		 重复序列
Repetitive sequence		 其他结构域
Other domains
Aradu.V26BD Aradu.A03 279 C 完整 Complete 4 4 0
Aradu.86DER Aradu.A03 286 – 不完整 Incomplete 4 3 0
Aradu.KJ1YM Aradu.A04 274 – 不完整 Incomplete 3 2 0
Aradu.3L5NK Aradu.A05 315 C 完整 Complete 4 4 0
Aradu.WYZ5E Aradu.A05 323 C 不完整 Incomplete 4 4 0
Aradu.9BC7H Aradu.A07 342 C 不完整 Incomplete 4 4 0
Aradu.23XWK Aradu.A08 309 C 不完整 Incomplete 3 3 0
Aradu.N8MUP Aradu.A08 279 – 不完整 Incomplete 3 3 0
Aradu.4J11T Aradu.A08 339 – 不完整 Incomplete 3 3 0
Aradu.S16C5 Aradu.A08 321 – 完整 Complete 4 4 0
Aradu.SCC75 Aradu.A10 266 C 不完整 Incomplete 3 3 0
Aradu.IZQ3Z Aradu.A10 315 C 完整 Complete 4 4 0
Aradu.MBZ2M Aradu.A10 333 – 完整 Complete 2 2 0
Araip.1LR8I Araip.B02 89 C 不完整 Incomplete 2 2 0
Araip.KR6F4 Araip.B03 323 – 完整 Complete 2 2 2
Araip.X0F2S Araip.B03 317 – 完整 Complete 4 4 0
Araip.CCM9G Araip.B03 286 – 不完整 Incomplete 4 3 0
Araip.RGY04 Araip.B04 142 C 不完整 Incomplete 1 1 0
Araip.R8WRM Araip.B04 207 – 不完整 Incomplete 3 2 0
Araip.HD6QL Araip.B05 316 C 完整 Complete 4 4 0
Araip.0MR1X Araip.B05 315 C 完整 Complete 4 4 0
Araip.FX5SI Araip.B07 316 C 完整 Complete 4 4 0
Araip.J58EQ Araip.B08 135 C 不完整 Incomplete 2 2 0
Araip.YGP4J Araip.B08 315 – 完整 Complete 3 3 0
Araip.6KP6U Araip.B08 295 – 完整 Complete 3 3 0
Araip.Y8EDR Araip.B08 321 – 完整 Complete 4 4 0
Araip.X9BIG Araip.B10 317 C 完整 Complete 4 4 0
Araip.Z0Q6Q Araip.B10 311 C 完整 Complete 4 3 0
Araip.WA456 Araip.B10 315 C 完整 Complete 4 3 0
Araip.RPP1M Araip.B10 364 – 完整 Complete 2 2 0
Aradu.V26BD Aradu.A03 279 C 完整 Complete 4 4 0
Aradu.86DER Aradu.A03 286 – 不完整 Incomplete 4 3 0

图1

植物annexin蛋白聚类分析 红色: 双子叶植物; 绿色: 单子叶植物; 黑色: 低等植物; 黑色圆点: AnnAhs。"

图2

AnnAhs基因结构分析 因部分AnnAhs无UTR, 故均用ORF进行基因结构分析。"

图3

AnnAhs的保守结构域分析"

表2

AnnAhs可变剪切分析"

基因名称
Name		 种子前期
Seed1		 种子后期
Seed2		 根
Root		 叶
Leaf
Aradu.S16C5 — — TSS —
Aradu.N8MUP — — TSS —
Aradu.3L5NK — — — TTS
Araip.CCM9G — TTS, AE — TTS, AE
Araip.WA456 — — TTS, AE —
Aradu.IZQ3Z TTS, AE AE AE AE
Araip.HD6QL TSS, ES TSS, ES — —
Aradu.KJ1YM TTS — TTS TTS, AE, ES
Araip.Z0Q6Q — — ES —
Araip.R8WRM — — ES —
Araip.RPP1M — — ES —

图4

AnnAhs表达模式分析"

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