姜GST基因家族成员鉴定与表达分析

doi:10.16420/j.issn.0513-353x.2023-0384

摘要/Abstract

摘要：

为解析姜（Zingiber officinale Roscoe）谷胱甘肽硫转移酶（glutathione-S-transferase，GST）基因家族的信息，基于姜基因组数据，分析鉴定了ZoGST家族成员及其表达模式，并进一步利用qRT-PCR检测了10个ZoGST在不同胁迫下的表达特性。在姜基因组中共鉴定到58个GST家族成员，其编码蛋白的氨基酸长度在110 ~ 675 aa之间，分子量12.21 ~ 72.51 kD，等电点4.55 ~ 9.51。系统进化树显示，ZoGST可分为Ⅰ~Ⅹ亚家族，第Ⅶ和Ⅹ亚家族成员居多数；ZoGST家族成员分布在11条染色体上，共线性分析表明该家族成员存在6对片段重复；顺式作用调控元件分析发现上游启动子区含有与光响应、逆境胁迫、激素调节相关的作用元件；转录组测序数据分析显示，ZoGST有一定的组织表达特异性且响应病害、低温等逆境胁迫，其中，ZoGSTDHAR5、ZoGSTF9、ZoGSTF13等在姜不同生长时期、不同部位、低温和病害胁迫下均有较高表达。qRT-PCR分析表明，盐胁迫下ZoGSTDHAR2（除根茎外）、ZoGSTEF1G2、ZoGSTF9在姜不同组织表达量均显著下调；淹水胁迫下，ZoGSTDHAR2等在叶和根茎中表达量显著上调；干旱胁迫下，ZoGSTF9等在根中表达量显著上调。

关键词: 姜, GST, 基因家族, 表达模式

Abstract:

To analyze the information of the glutathione-S-transferase（GST）gene family in ginger （Zingiber officinale Roscoe.），based on ginger genome data，the members and expression patterns of the ZoGST family were identified and analyzed. Furthermore，the expression characteristics of 10 ZoGST under different stress conditions were detected using qRT-PCR. A total of 58 members of the GST family were identified in the ginger genome，encoding proteins with amino acid lengths ranging from 110 to 675 aa，molecular weights ranging from 12.21 to 72.51 kD，isoelectric points ranging from 4.55 to 9.51. The phylogenetic tree shows that ZoGST can be divided intoⅠ-Ⅹsubfamilies，with members from the Ⅶ andⅩsubfamilies being the majority. The members of the ZoGST family are distributed on 11 chromosomes，and collinearity analysis shows that there are 6 pairs of segment duplications in this family member. Analysis of cis-acting regulatory elements revealed that the upstream promoter region contains regulatory elements related to light response，stress，and hormone regulation. Transcriptome sequencing data analysis shows that ZoGST has certain tissue expression specificity and responds to stress such as diseases and low temperatures. Among them，ZoGSTDHAR5，ZoGSTF9，ZoGSTF13，etc. are highly expressed in ginger at different growth stages，different parts，low temperatures，and disease stresses. qRT-PCR analysis showed that under salt stress，the expression levels of ZoGSTDHAR2（except for rhizomes），ZoGSTEF1G2，and ZoGSTF9 were significantly downregulated in different tissues of ginger. Under flooding stress，the expression levels of ZoGSTDHAR2 and others were significantly upregulated in leaves and rhizome. Under drought stress，the expression levels of ZoGSTF9 and others were significantly upregulated in roots.

Key words: Zingiber officinale Roscoe., GST, gene family, expression pattern

王艳红, 席克勇, 田野, 刘德麒, 周克贵, 尹军良, 刘奕清, 朱永兴. 姜GST基因家族成员鉴定与表达分析[J]. 园艺学报, 2024, 51(8): 1803-1822.

WANG Yanhong, XI Keyong, TIAN Ye, LIU Deqi, ZHOU Kegui, YIN Junliang, LIU Yiqing, ZHU Yongxing. Identification and Expression Pattern Analysis of GST in Zingiber officinale[J]. Acta Horticulturae Sinica, 2024, 51(8): 1803-1822.

图/表 12

参考文献 51

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登录号 Accession ID	处理样品描述 Description	数据大小 Data size	参考文献 Reference
PRJNA788194	不同发育阶段根茎、不同组织 Different developmental stages of rhizomes，different tissues	未知 Unknown	Li et al.,2021
PRJNA592215	红茎内部组织、绿茎、黄根茎、红茎表皮 Internal tissue of red stem，green stem，yellow rhizome，red stem epidermis	254 Mb	Li et al.,2022
PRJNA380847	姜根茎低/高土壤湿度下用无菌水和青枯菌接种 Inoculate ginger rhizomes with sterile water and Ralstonia solanacearum under low/high soil moisture	未知 Unknown	Jiang et al.,2018
PRJNA380972	高土壤湿度增加姜中青枯菌感染率 High-soil-moisture-elevated susceptibility to R. solanacearum infection in ginger	31.5 Gb	Huang et al.,2021
PRJNA911443	姜采收后26、10和2 ℃贮藏条件下的根茎 Rhizome of ginger harvested under storage conditions of 26，10，and 2 ℃	16.8 Gb	Zhang et al.,2023

登录号 Accession ID	处理样品描述 Description	数据大小 Data size	参考文献 Reference
PRJNA788194	不同发育阶段根茎、不同组织 Different developmental stages of rhizomes，different tissues	未知 Unknown	Li et al.,2021
PRJNA592215	红茎内部组织、绿茎、黄根茎、红茎表皮 Internal tissue of red stem，green stem，yellow rhizome，red stem epidermis	254 Mb	Li et al.,2022
PRJNA380847	姜根茎低/高土壤湿度下用无菌水和青枯菌接种 Inoculate ginger rhizomes with sterile water and Ralstonia solanacearum under low/high soil moisture	未知 Unknown	Jiang et al.,2018
PRJNA380972	高土壤湿度增加姜中青枯菌感染率 High-soil-moisture-elevated susceptibility to R. solanacearum infection in ginger	31.5 Gb	Huang et al.,2021
PRJNA911443	姜采收后26、10和2 ℃贮藏条件下的根茎 Rhizome of ginger harvested under storage conditions of 26，10，and 2 ℃	16.8 Gb	Zhang et al.,2023

基因名 Gene name	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer	长度/bp Length
ZoGSTDHAR2	CTCCTCCTGCCATTTCCTCG	GATTCACCTCCACGACTTTGTATG	228
ZoGSTEF1G2	AAGGTGTTAGGCGAGGTCAAG	TGGGTGCTTCTTCCTCTTCTTC	169
ZoGSTCHQ1	GGTCACAGAATGGGTGGAGAG	TGGACCATCCGAGCAATCAC	129
ZoGSTT1	AGAGTCATGCCATATTGAGCTACC	ATTGAACCACGGCGTAGATTG	140
ZoGSTL1	CTTCCTCCGTCCCTCGATTC	GTTATCCAGGTGCGTTGTGC	107
ZoGSTL2	TCCCGCTTTGTGAAGGAGG	CGCAAATGGGCAGGTGTATG	111
ZoGSTF9	GGTGGTACGACGGACGAGAAG	TGACGGGATAGTGGCAGAGG	145
ZoGSTU13	TTGCTGGATCTGTGGGTGAGTC	GTATTGCACGATGATGAGCGACT	201
ZoGSTU16	TGGATCTGTGGGTGAGTCCTTTC	TGTTGTCCAAGTCCTGCTCCTG	96
ZoGSTU27	GGTGTTACTGGATCTGTGGGTGAG	GCAACGGGCTCTTGTTGTCC	116
RBP	CCTATGAAGCGTAGAAACACAAG	GGAAGGACAACATCCCAAATC	123

基因名 Gene name	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer	长度/bp Length
ZoGSTDHAR2	CTCCTCCTGCCATTTCCTCG	GATTCACCTCCACGACTTTGTATG	228
ZoGSTEF1G2	AAGGTGTTAGGCGAGGTCAAG	TGGGTGCTTCTTCCTCTTCTTC	169
ZoGSTCHQ1	GGTCACAGAATGGGTGGAGAG	TGGACCATCCGAGCAATCAC	129
ZoGSTT1	AGAGTCATGCCATATTGAGCTACC	ATTGAACCACGGCGTAGATTG	140
ZoGSTL1	CTTCCTCCGTCCCTCGATTC	GTTATCCAGGTGCGTTGTGC	107
ZoGSTL2	TCCCGCTTTGTGAAGGAGG	CGCAAATGGGCAGGTGTATG	111
ZoGSTF9	GGTGGTACGACGGACGAGAAG	TGACGGGATAGTGGCAGAGG	145
ZoGSTU13	TTGCTGGATCTGTGGGTGAGTC	GTATTGCACGATGATGAGCGACT	201
ZoGSTU16	TGGATCTGTGGGTGAGTCCTTTC	TGTTGTCCAAGTCCTGCTCCTG	96
ZoGSTU27	GGTGTTACTGGATCTGTGGGTGAG	GCAACGGGCTCTTGTTGTCC	116
RBP	CCTATGAAGCGTAGAAACACAAG	GGAAGGACAACATCCCAAATC	123

基因1	基因2	异义替换频率（K_a）	同义替换频率（K_s）	异义替换/同义替换
Gene 1	Gene 2	Non-synonymous	Synonymous	K_a/K_s
ZoGSTL1	ZoGSTL3	0.141203891	0.527940985	0.267461505
ZoGSTL2	ZoGSTL3	0.096538384	0.315771474	0.305722310
ZoGSTDHAR2	ZoGSTDHAR3	0.059875159	0.365643255	0.163752943
ZoGSTF2	ZoGSTF4	0.389135669	0.971421814	0.400583622
ZoGSTU2	ZoGSTU3	0.113145323	0.259868329	0.435394813
ZoGSTU11	ZoGSTU14	0.073775974	0.133473189	0.552740031