茶树Gro/Tup1基因家族鉴定及外源激素和非生物胁迫下表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2021-0184

摘要/Abstract

摘要：

明确Groucho/Thymidine uptake 1（Gro/Tup1）辅转录调控因子在茶树基因组中的数量、结构及其在非生物胁迫和外源激素处理下表达差异,为揭示Gro/Tup1在茶树激素信号转导途径及其在茶树逆境胁迫中的作用奠定理论基础。对茶树Gro/Tup1家族进行鉴定、分析,同时对其编码蛋白理化性质、进化关系、顺式作用元件及其在干旱、低温（4℃）、赤霉素（GA₃）、脱落酸（ABA）和茉莉酸甲酯（MeJA）处理下表达模式进行分析。结果显示:茶树Gro/Tup1家族具有13个成员,分为TOPLESS/TOPLESS-related（TPL/TPR）和LEUNIG/LEUNIG HOMOLOG（LUG/LUH）两个亚家族。顺式作用元件预测结果表明,茶树Gro/Tup1家族成员启动子区域有大量与GA、ABA、MeJA响应以及干旱和低温应答的顺式作用元件;RT-qPCR分析结果表明,不同处理下,茶树Gro/Tup1家族成员呈现不同表达模式。重度干旱时,CsLUG3和CsLUG4基因的表达量显著上调,CsTPL2表达水平在轻度干旱时就开始受到抑制,CsTPL8和CsTPL9不受干旱调控;低温处理下CsTPL2和CsTPL3表达量显著上调;GA₃处理下,多数基因表达量上调;ABA处理下多数基因表达量均呈现先显著增加后显著下降趋势;MeJA处理下,大部分基因表达水平都受到显著抑制。茶树含有13个Gro/ Tup1家族成员,其结构进化高度保守,能够响应非生物胁迫及外源激素,其基因应答呈现不同表达谱。

关键词: 茶树, Groucho/Thymidine uptake 1（Gro/Tup1）家族, 植物激素, 非生物胁迫

Abstract:

The objective of this work is to clarify the number and structure of the co-transcription regulatory factor Groucho/Thymidine uptake 1（CsGro/Tup1）in Camellia sinensis genome,and its expression differences under abiotic stresses and exogenous phytohormones treatments,which will lay a theoretical foundation for revealing the role of Gro/Tup1 in hormone signal transduction pathway and its role in C. sinensis tree stress. The CsGro/Tup1 family members were identified and analyzed. Meanwhile,the physicochemical properties of its encoded proteins,evolutionary relationships,and cis-acting elements of CsGro/Tup1s were investigated,and the relative expression levels of CsGro/Tup1 members were measured under drought,low temperature,GA₃,ABA and MeJA treatments. The results showed that the CsGro/Tup1 family had 13 members,which were divided into two subfamilies:TOPLESS/TOPLESS-related（TPL/TPR）and LEUNIG/LEUNIG HOMOLOG（LUG/LUH）. The prediction results of cis-acting elements showed that Gro/Tupls’ promoter region had a large number of cis-acting elements in response to GA,ABA,MeJA,drought and low temperature. RT-qPCR results demonstrated that Gro/Tup1s presented different expression patterns under different treatments. In severe drought,the expressions of CsLUG3 and CsLUG4 genes were significantly up-regulated,the expressions of CsTPL2 were inhibited at light drought,CsTPL8 and CsTPL9 were not regulated by drought. The expressions of CsTPL2 and CsTPL3 were significantly up-regulated under low temperature treatment. Under GA₃ treatment,most gene expressions were up-regulated. Under ABA treatment,the expressions of most genes significantly increased in the beginning and significantly decreased later. Most of the gene expressions were significantly inhibited under MeJA treatment. In conclusion,C. sinensis has 13 CsGro/Tup1 family members,and their structures are evolutionarily conservative,and CsGro/Tup1s are able to respond to different abiotic stresses and exogenous phytohormone,and their responses present various profiles.

Key words: Camellia sinensis, Groucho/Thymidine uptake 1（Gro/Tup1）family, phytohormone, abiotic stress

詹冬梅, 朱晨, 周承哲, 黄雪婷, 赖钟雄, 郭玉琼. 茶树Gro/Tup1基因家族鉴定及外源激素和非生物胁迫下表达分析[J]. 生物技术通报, 2021, 37(12): 1-12.

ZHAN Dong-mei, ZHU Chen, ZHOU Cheng-zhe, HUANG Xue-ting, LAI Zhong-xiong, GUO Yu-qiong. Genome-wide Identification of Gro/Tup1 Gene Family in Camellia sinensis and Their Expression Analysis Under Exogenous Phytohormones and Abiotic Stress Treatments[J]. Biotechnology Bulletin, 2021, 37(12): 1-12.

图/表 9

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基因名称 Gene name	上游引物 Upstream primer	下游引物 Downstream primer
CsLUG1	GCGAGTCATCTGGGTAGTAAGT	GTTGTTGGTGCTCTCTTGCT
CsLUG2	GCTGAGAGTGCTTTGGATGG	GCTGAAGGATGTTGAGTGCT
CsLUG3	TTGGAATGCGAGTGGTCAGT	TGGTGCTGTGGTGTCAACAT
CsLUG4	CGTGGCAAGCCCTTAAAG	TGGTGTCAGCATCTGGAGT
CsTPL1	CATGTGTTGAAGAGACTGAGG	TGGGCAAATCATCAGAAGAG
CsTPL2	GCTACCGCTGTTGAGATTCTTG	ATTTGCCAGGGTGAGGAGTAG
CsTPL3	CCTTCCAACCTACACCAGC	CACCAAGACCAATAGCTCCAC
CsTPL4	TCGGCTTTGTCAGTTCTACG	CCGGAAGTGTTATAGGAGGTG
CsTPL5	GCGACTTGAGCATCTCCTTG	GCTGCCACTTCAAAGCCT
CsTPL6	TGGCTCGCTCTCCATGTATG	TGCAAGAGGAACGTGATGGT
CsTPL7	AGATGTTGGTTTGGTTCAGGG	GCTTTGGTTAATGAGGTTCCG
CsTPL8	TTCAAGGTTGCGGACTCTCAT	GATTGTTTGCTGGTGATGGAG
CsTPL9	GTCGCCTGTAACTAACCCACT	TGAGGAACCGGAGATGGATTAG
Csβ-actin	GCCATCTTTGATTGGAATGG	GGTGCCACAACCTTGATCTT
CsSAND	CCAATTGCCCCCTTAATGAC	GCAATCATTTCCTTCGTGGAG

基因名称 Gene name	上游引物 Upstream primer	下游引物 Downstream primer
CsLUG1	GCGAGTCATCTGGGTAGTAAGT	GTTGTTGGTGCTCTCTTGCT
CsLUG2	GCTGAGAGTGCTTTGGATGG	GCTGAAGGATGTTGAGTGCT
CsLUG3	TTGGAATGCGAGTGGTCAGT	TGGTGCTGTGGTGTCAACAT
CsLUG4	CGTGGCAAGCCCTTAAAG	TGGTGTCAGCATCTGGAGT
CsTPL1	CATGTGTTGAAGAGACTGAGG	TGGGCAAATCATCAGAAGAG
CsTPL2	GCTACCGCTGTTGAGATTCTTG	ATTTGCCAGGGTGAGGAGTAG
CsTPL3	CCTTCCAACCTACACCAGC	CACCAAGACCAATAGCTCCAC
CsTPL4	TCGGCTTTGTCAGTTCTACG	CCGGAAGTGTTATAGGAGGTG
CsTPL5	GCGACTTGAGCATCTCCTTG	GCTGCCACTTCAAAGCCT
CsTPL6	TGGCTCGCTCTCCATGTATG	TGCAAGAGGAACGTGATGGT
CsTPL7	AGATGTTGGTTTGGTTCAGGG	GCTTTGGTTAATGAGGTTCCG
CsTPL8	TTCAAGGTTGCGGACTCTCAT	GATTGTTTGCTGGTGATGGAG
CsTPL9	GTCGCCTGTAACTAACCCACT	TGAGGAACCGGAGATGGATTAG
Csβ-actin	GCCATCTTTGATTGGAATGG	GGTGCCACAACCTTGATCTT
CsSAND	CCAATTGCCCCCTTAATGAC	GCAATCATTTCCTTCGTGGAG

基因名称 Gene name	基因ID Gene ID	大小 Size/aa	分子量 Molecular weight/kD	等电点 pI	亚细胞定位 Subcellular localization	不稳定系数 Instability index	脂溶指 Aliphatic index	亲水指数 GRAVY	外显子数 Number of exons
CsLUG1	TEA022439.1	864	94.51266	7.02	质体Plastid	46.11	77.78	0.342	17
CsLUG2	TEA004970.1	660	73.86218	8.04	细胞核Nucleus	45.09	67.7	0.566	16
CsLUG3	TEA020168.1	907	100.83071	6.48	细胞核Nucleus	53.22	67.42	0.648	19
CsLUG4	TEA025268.1	757	81.54457	6.08	细胞核Nucleus	40.86	40.86	0.352	17
CsTPL1	TEA031994.1	540	60.18248	8.07	细胞核Nucleus	37.53	79.65	0.368	14
CsTPL2	TEA019471.1	1196	131.4561	6.33	细胞核Nucleus	40.35	83.09	0.27	27
CsTPL3	TEA016267.1	1319	145.85055	6.87	细胞核Nucleus	39.33	77.87	0.317	30
CsTPL4	TEA030187.1	569	64.03845	6.27	叶绿体Chloroplast	39.24	91.92	0.193	6
CsTPL5	TEA024264.1	723	81.24096	8.17	细胞核Nucleus	43.53	85.01	0.244	20
CsTPL6	TEA022067.1	1069	118.97318	6.14	内质网Reticulum	39.47	86.43	0.222	24
CsTPL7	TEA022077.1	759	86.17475	6.72	细胞核Nucleus	43.25	85.03	0.181	18
CsTPL8	TEA020620.1	1195	131.50275	6.43	叶绿体Chloroplast	40.03	80.14	0.312	26
CsTPL9	TEA023424.1	1188	131.09747	6.3	细胞核Nucleus	38.29	79.55	0.317	26

基因名称 Gene name	基因ID Gene ID	大小 Size/aa	分子量 Molecular weight/kD	等电点 pI	亚细胞定位 Subcellular localization	不稳定系数 Instability index	脂溶指 Aliphatic index	亲水指数 GRAVY	外显子数 Number of exons
CsLUG1	TEA022439.1	864	94.51266	7.02	质体Plastid	46.11	77.78	0.342	17
CsLUG2	TEA004970.1	660	73.86218	8.04	细胞核Nucleus	45.09	67.7	0.566	16
CsLUG3	TEA020168.1	907	100.83071	6.48	细胞核Nucleus	53.22	67.42	0.648	19
CsLUG4	TEA025268.1	757	81.54457	6.08	细胞核Nucleus	40.86	40.86	0.352	17
CsTPL1	TEA031994.1	540	60.18248	8.07	细胞核Nucleus	37.53	79.65	0.368	14
CsTPL2	TEA019471.1	1196	131.4561	6.33	细胞核Nucleus	40.35	83.09	0.27	27
CsTPL3	TEA016267.1	1319	145.85055	6.87	细胞核Nucleus	39.33	77.87	0.317	30
CsTPL4	TEA030187.1	569	64.03845	6.27	叶绿体Chloroplast	39.24	91.92	0.193	6
CsTPL5	TEA024264.1	723	81.24096	8.17	细胞核Nucleus	43.53	85.01	0.244	20
CsTPL6	TEA022067.1	1069	118.97318	6.14	内质网Reticulum	39.47	86.43	0.222	24
CsTPL7	TEA022077.1	759	86.17475	6.72	细胞核Nucleus	43.25	85.03	0.181	18
CsTPL8	TEA020620.1	1195	131.50275	6.43	叶绿体Chloroplast	40.03	80.14	0.312	26
CsTPL9	TEA023424.1	1188	131.09747	6.3	细胞核Nucleus	38.29	79.55	0.317	26