中国春小麦肌醇磷脂依赖的磷脂酶C基因的全基因组鉴定及表达分析

doi:10.3864/j.issn.0578-1752.2020.24.001

中国农业科学 ›› 2020, Vol. 53 ›› Issue (24): 4969-4981.doi: 10.3864/j.issn.0578-1752.2020.24.001

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

中国春小麦肌醇磷脂依赖的磷脂酶C基因的全基因组鉴定及表达分析

司旭阳¹(),贾哓玮¹(),张洪艳¹,贾羊羊¹,田士军¹,张科²(),潘延云¹()

¹河北农业大学生命科学学院/河北省植物生理与分子病理学重点实验室,河北保定 071000
²河北农业大学农学院/省部共建华北作物改良与调控国家重点实验室/河北省作物生长调控实验室,河北保定 071001

收稿日期:2020-03-23 接受日期:2020-06-02 出版日期:2020-12-16 发布日期:2020-12-28
通讯作者: 张科,潘延云
作者简介:司旭阳,E-mail： sixuyanglove@163.com。|贾哓玮,E-mail： jia999666@126.com。
基金资助:
河北省自然科学基金(C2017204095);河北省高等学校科学技术研究项目(ZD2017039)

Genomic Profiling and Expression Analysis of Phosphatidylinositol- specific PLC Gene Families Among Chinese Spring Wheat

SI XuYang¹(),JIA XiaoWei¹(),ZHANG HongYan¹,JIA YangYang¹,TIAN ShiJun¹,ZHANG Ke²(),PAN YanYun¹()

¹College of Life Sciences, Hebei Agricultural University/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, Baoding 071000, Hebei
²College of Agronomy, Hebei Agricultural University/State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Crop Growth Regulation of Hebei Province, Baoding 071001, Hebei

Received:2020-03-23 Accepted:2020-06-02 Online:2020-12-16 Published:2020-12-28
Contact: Ke ZHANG,YanYun PAN

摘要/Abstract

摘要：

【目的】探知小麦基因组序列中肌醇磷脂依赖的磷脂酶C（PLC）的编码基因,解析小麦中PLC基因的结构与进化特征,揭示TaPLC基因在小麦各组织中的表达模式及其响应盐胁迫和干旱胁迫过程中的表达规律,以便深入分析小麦TaPLC基因调节小麦应答盐或干旱胁迫中的生理作用。【方法】基于Ensembl Plants全基因组数据库,以水稻和拟南芥PLC基因为参考序列,检索小麦TaPLC基因家族（http://plants.ensembl.org/index.html）,并利用在线蛋白结构分析工具（Pfam、CDD和SMART）对TaPLC基因进行结构分析和鉴定;运用ExPASy（http://cn.expasy.org/tools）分析TaPLC基因的分子和生化特征;使用WoLF PSORT（https://www.genscript.com/wolf-psort.html）预测TaPLC基因的亚细胞定位;运用MEGA 7.0软件进行系统进化树分析;并利用实时荧光定量PCR分析TaPLC基因在不同组织以及幼苗期在盐或干旱胁迫下的表达特征。【结果】在小麦基因组中鉴定了11个TaPLC基因序列,其编码蛋白与其他植物PLC结构相似,均具有X和Y保守催化结构域以及C2结构域;系统进化分析表明11个TaPLC基因可分为4组,即TaPLC1A/TaPLC1D、TaPLC2A/TaPLC2B/TaPLC2D、TaPLC3A/TaPLC3B/TaPLC3D、TaPLC4A/TaPLC4B/TaPLC4D,均为植物PLC基因家族的直系同源基因,在进化方面具有保守性,但也存在一定的分歧;通过与小麦祖先种比对分析,发现B亚基因组中的TaPLC1在异源六倍体形成前就已经缺失,进一步表明小麦亚基因组间进化的不对称性;亚细胞定位预测TaPLC基因定位于叶绿体、线粒体或细胞质中;TaPLC基因家族各成员的启动子普遍存在植物激素响应和应激响应相关的顺式调控元件;实时定量结果表明,不同的TaPLC基因在不同的组织中有不同的表达水平,其主要在根、叶、穗和籽粒中表达;且多数基因参与小麦对盐和干旱胁迫的响应,无论是在耐旱和耐盐品种中,还是在对照品种中,TaPLC基因均受到胁迫的诱导表达,并在12 h内迅速达到峰值。【结论】在小麦基因组中共鉴定11个TaPLC基因,编码典型的植物PLC蛋白,在进化上具有保守性,但亚基因组间也存在进化的不对称性。不同的TaPLC基因有各自的组织和细胞表达特征,在小麦生长发育过程中起调节作用;TaPLC基因在应答盐或干旱胁迫中起重要作用。

关键词: 小麦, 肌醇磷脂依赖的磷脂酶C, 盐胁迫, 干旱胁迫

Abstract:

【Objective】To characterize the gene family encoding phosphatidylinositol-specific phospholipase-C (PI-PLC), the gene number, structure, phylogenetic relationship and expression pattern of TaPLC genes were analyzed. In addition, we explored their relative transcription levels in various tissue and the expression pattern under drought and salt stress to further analysis the physiological role of TaPLC genes in response to abiotic stress.【Method】Based on the genome database (Ensembl Plants, http://plants.ensembl.org/index.html), TaPLC genes were identified from Triticum aestivum, and the gene structures were analyzed by some bioinformatics tools, such as Pfam, CDD and SMART. The protein sequence characteristics of the members in TaPLC family were analyzed with the online server ExPASy (http://cn.expasy.org/tools). The subcellular localization of TaPLC genes were predicted using the WoLF PSORT (https://www.genscript.com/wolf-psort.html). Phylogenic tree analysis was performed by software MEGA 7.0. The real-time PCR was used to detect the gene expression levels in different tissues under abiotic stress. 【Result】A total of 11 TaPLC genes members within the heterologous hexaploid wheat genome were identified and analyzed. All TaPLC genes have conserved X-Y catalytic domains and C2 domains, just like other plant PLC genes. Subcellular localization showed that these proteins were located in the chloroplast or mitochondria. Phylogenetic analysis revealed that all of the TaPLC genes were unevenly classified into four numbered, TaPLC1A/TaPLC1D, TaPLC2A/TaPLC2B/TaPLC2D, TaPLC3A/TaPLC3B/TaPLC3D, and TaPLC4A/TaPLC4B/TaPLC4D, which were all orthologous genes of plant PLC genes in plants, and highly conserved during evolution. However, the deletion of TaPLC1 in B subgenome indicates asymmetry evolution in different subgenomes. Further analysis of cis-regulatory elements elucidated that these members of TaPLC family shared similar elements, such as the phytohormone and the stress response cis-elements. Each PLC gene has a unique tissue expression pattern, and they are expressed mainly in roots, leaves, spikes, and gains. In addition, PLC genes mediate the response of wheat to salt and drought stresses, which are induced rapidly by salt or drought stress stimulation. 【Conclusion】The TaPLC family in Triticum aestivum contains 11 members which have conserved X-Y catalytic domains and C2 domains. Although the TaPLC genes were highly conserved, they were asymmetrically evolved in different subgenomes. They had specific expression pattern in different tissues, and were induced by salt or drought stress, which suggested TaPLC genes play a role in response to salt or drought in wheat.

Key words: Triticum aestivum, PI-PLC, salt stress, drought stress

司旭阳,贾哓玮,张洪艳,贾羊羊,田士军,张科,潘延云. 中国春小麦肌醇磷脂依赖的磷脂酶C基因的全基因组鉴定及表达分析[J]. 中国农业科学, 2020, 53(24): 4969-4981.

SI XuYang,JIA XiaoWei,ZHANG HongYan,JIA YangYang,TIAN ShiJun,ZHANG Ke,PAN YanYun. Genomic Profiling and Expression Analysis of Phosphatidylinositol- specific PLC Gene Families Among Chinese Spring Wheat[J]. Scientia Agricultura Sinica, 2020, 53(24): 4969-4981.

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基因名称 Gene name	基因ID Gene ID	染色体定位^a Chromosomal location	开放阅读框 Open reading frame (bp)	蛋白质^b Protein
基因名称 Gene name	基因ID Gene ID	染色体定位^a Chromosomal location	开放阅读框 Open reading frame (bp)	大小 Size (aa)	分子量 MW (kD)	等电点 pI
TaPLC1A	TraesCS1A02G069300	1A:51700021—51707185	1758	585	66.01	6.07
TaPLC1D	TraesCS1D02G071800	1D:52338417—52345805	1761	586	66.19	6.03
TaPLC2A	TraesCS2A02G084000	2A:38534860—38538480	1830	609	68.5	5.91
TaPLC2B	TraesCS2B02G098500	2B:58321242—58325230	1821	606	68.22	6.06
TaPLC2D	TraesCS2D02G082000	2D:35259471—35263395	1824	607	68.45	5.88
TaPLC3A	TraesCS4A02G109000	4A:129086595—129090166	1902	633	71.11	5.54
TaPLC3B	TraesCS4B02G195200	4B:420197892—420201623	1902	633	71.00	5.75
TaPLC3D	TraesCS4D02G195800	4D:340085748—340090648	1902	633	71.05	5.84
TaPLC4A	TraesCS5A02G155300	5A:333407514—333413175	1773	590	65.73	6.05
TaPLC4B	TraesCS5B02G153600	5B:283008744—283014326	1770	589	65.65	6.06
TaPLC4D	TraesCS5D02G160300	5D:250061407—250066699	1770	589	65.65	6.06

基因Gene	亚细胞定位Subcellular localization
TaPLC1A	线粒体：8,叶绿体：4,细胞核：1 Mito: 8, chlo: 4, nucl: 1
TaPLC1D	线粒体：8,叶绿体：4,细胞核：1 Mito: 8, chlo: 4, nucl: 1
TaPLC2A	叶绿体：7,细胞核：3,线粒体：2.5,线粒体基质：2 Chlo: 7, nucl: 3, mito: 2.5, cyto_mito: 2
TaPLC2B	叶绿体：6,线粒体：5.5,线粒体基质：3.5,细胞核：1 Chlo: 6, mito: 5.5, cyto_mito: 3.5, nucl: 1
TaPLC2D	叶绿体：7,线粒体：4.5,线粒体基质：3,细胞核：1 Chlo: 7, mito: 4.5, cyto_mito: 3, nucl: 1
TaPLC3A	细胞质：8,细胞核：4,叶绿体：1 Cyto: 8, nucl: 4, chlo: 1
TaPLC3B	细胞质：8,细胞核：3,叶绿体：1,线粒体：1 Cyto: 8, nucl: 3, chlo: 1, mito: 1
TaPLC3D	细胞质：6,细胞核：3,线粒体：2,过氧化物：2 Cyto: 6, nucl: 3, mito: 2, pero: 2
TaPLC4A	线粒体：10,叶绿体和线粒体：6.83333,线粒体基质：5.83333,叶绿体：2.5 Mito: 10, chlo_mito: 6.83333, cyto_mito: 5.83333, chlo: 2.5
TaPLC4B	线粒体：10,叶绿体和线粒体：6.83333,线粒体基质：5.83333,叶绿体：2.5 Mito: 10, chlo_mito: 6.83333, cyto_mito: 5.83333, chlo: 2.5
TaPLC4D	线粒体：10,叶绿体和线粒体：6.83333,线粒体基质：5.83333,叶绿体：2.5 Mito: 10, chlo_mito: 6.83333, cyto_mito: 5.83333, chlo: 2.5

中国春小麦肌醇磷脂依赖的磷脂酶C基因的全基因组鉴定及表达分析

Genomic Profiling and Expression Analysis of Phosphatidylinositol- specific PLC Gene Families Among Chinese Spring Wheat

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