Genome-wide Identification of Gro/Tup1 Gene Family in Camellia sinensis and Their Expression Analysis Under Exogenous Phytohormones and Abiotic Stress Treatments

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

Abstract

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

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.

Figures/Tables 9

References 41

[1]	Courey AJ, Jia S. Transcriptional repression:the long and the short of it[J]. Genes Dev, 2001, 15(21):2786-2796. doi: 10.1101/gad.939601 URL
[2]	Krogan NT, Long JA. Why so repressed? Turning off transcription during plant growth and development[J]. Curr Opin Plant Biol, 2009, 12(5):628-636. doi: 10.1016/j.pbi.2009.07.011 pmid: 19700365
[3]	Mannervik M, Nibu Y, Zhang H, et al. Transcriptional coregulators in development[J]. Science, 1999, 284(5414):606-609. pmid: 10213677
[4]	Chen G, Courey AJ. Groucho/TLE family proteins and transcriptional repression[J]. Gene, 2000, 249(1/2):1-16. doi: 10.1016/S0378-1119(00)00161-X URL
[5]	Conner J, Liu Z. LEUNIG, a putative transcriptional corepressor that regulates AGAMOUS expression during flower development[J]. PNAS, 2000, 97(23):12902-12907. pmid: 11058164
[6]	Long JA, Ohno C, Smith ZR, et al. TOPLESS regulates apical embryonic fate in Arabidopsis[J]. Science, 2006, 312(5779):1520-1523. doi: 10.1126/science.1123841 URL
[7]	Kieffer M, Stern Y, Cook H, et al. Analysis of the transcription factor WUSCHEL and its functional homologue in Antirrhinum reveals a potential mechanism for their roles in meristem maintenance[J]. Plant Cell, 2006, 18(3):560-573. pmid: 16461579
[8]	Liu ZC, Karmarkar V. Groucho/Tup1 family co-repressors in plant development[J]. Trends Plant Sci, 2008, 13(3):137-144. doi: 10.1016/j.tplants.2007.12.005 URL
[9]	Hao YW, Wang XY, Li X, et al. Genome-wide identification, phylogenetic analysis, expression profiling, and protein-protein interaction properties of TOPLESS gene family members in tomato[J]. J Exp Bot, 2014, 65(4):1013-1023. doi: 10.1093/jxb/ert440 URL
[10]	Li HY, Huang KF, Du HM, et al. Genome-wide analysis of Gro/Tup1 family corepressors and their responses to hormones and abiotic stresses in maize[J]. J Plant Biol, 2016, 59(6):603-615. doi: 10.1007/s12374-016-0333-8 URL
[11]	Szemenyei H, Hannon M, Long JA. TOPLESS mediates auxin-dependent transcriptional repression during Arabidopsis embryogenesis[J]. Science, 2008, 319(5868):1384-1386. doi: 10.1126/science.1151461 pmid: 18258861
[12]	Nguyen D, Rieu I, Mariani C, et al. How plants handle multiple stresses:hormonal interactions underlying responses to abiotic stress and insect herbivory[J]. Plant Mol Biol, 2016, 91(6):727-740. doi: 10.1007/s11103-016-0481-8 pmid: 27095445
[13]	Pauwels L, Barbero GF, Geerinck J, et al. NINJA connects the co-repressor TOPLESS to jasmonate signalling[J]. Nature, 2010, 464(7289):788-791. doi: 10.1038/nature08854 URL
[14]	You Y, Zhai Q, An C, et al. LEUNIG_HOMOLOG mediates MYC2-dependent transcriptional activation in cooperation with the coactivators HAC1 and MED25[J]. Plant Cell, 2019, 31(9):2187-2205. doi: 10.1105/tpc.19.00115 URL
[15]	Causier B, Ashworth M, Guo W, et al. The TOPLESS interactome:a framework for gene repression in Arabidopsis[J]. Plant Physiol, 2012, 158(1):423-438. doi: 10.1104/pp.111.186999 pmid: 22065421
[16]	Gonzalez D, Bowen AJ, Carroll TS, et al. The transcription corepressor LEUNIG interacts with the histone deacetylase HDA19 and mediator components MED14(SWP)and CDK8(HEN3)to repress transcription[J]. Mol Cell Biol, 2007, 27(15):5306-5315. doi: 10.1128/MCB.01912-06 URL
[17]	Zhu J, Jeong JC, Zhu Y, et al. Involvement of Arabidopsis HOS15 in histone deacetylation and cold tolerance[J]. PNAS, 2008, 105(12):4945-4950. doi: 10.1073/pnas.0801029105 URL
[18]	Zhu Z, Xu F, Zhang Y, et al. Arabidopsis resistance protein SNC1 activates immune responses through association with a transcriptional corepressor[J]. PNAS, 2010, 107(31):13960-13965. doi: 10.1073/pnas.1002828107 URL
[19]	Shrestha B, Guragain B, Sridhar VV. Involvement of co-repressor LUH and the adapter proteins SLK1 and SLK2 in the regulation of abiotic stress response genes in Arabidopsis[J]. BMC Plant Biol, 2014, 14:54. doi: 10.1186/1471-2229-14-54 pmid: 24564815
[20]	芦梅. 中国茶叶产业发展的管理模式与经济学分析[J]. 福建茶叶, 2016, 38(9):99-100.
	Lu M. Management mode and economic analysis of tea industry development in China[J]. Tea Fujian, 2016, 38(9):99-100.
[21]	Liu SC, Jin JQ, Ma JQ, et al. Transcriptomic analysis of tea plant responding to drought stress and recovery[J]. PLoS One, 2016, 11(1):e0147306. doi: 10.1371/journal.pone.0147306 URL
[22]	Cheruiyot EK, Mumera LM, Ng’Etich WK, et al. High fertilizer rates increase susceptibility of tea to water stress[J]. J Plant Nutr, 2009, 33(1):115-129. doi: 10.1080/01904160903392659 URL
[23]	Wei CL, Yang H, Wang SB, et al. Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality[J]. Proc Natl Acad Sci USA, 2018, 115(18):E4151-E4158. doi: 10.1073/pnas.1719622115 URL
[24]	Xia EH, Zhang HB, Sheng J, et al. The tea tree genome provides insights into tea flavor and independent evolution of caffeine biosynjournal[J]. Mol Plant, 2017, 10(6):866-877. doi: 10.1016/j.molp.2017.04.002 URL
[25]	Xia EH, Tong W, Hou Y, et al. The reference genome of tea plant and resequencing of 81 diverse accessions provide insights into its genome evolution and adaptation[J]. Mol Plant, 2020, 13(7):1013-1026. doi: 10.1016/j.molp.2020.04.010 URL
[26]	Wang X, Feng H, Chang Y, et al. Population sequencing enhances understanding of tea plant evolution[J]. Nat Commun, 2020, 11(1):4447. doi: 10.1038/s41467-020-18228-8 URL
[27]	Zhang QJ, Li W, Li K, et al. The chromosome-level reference genome of tea tree unveils recent bursts of non-autonomous LTR retrotransposons in driving genome size evolution[J]. Mol Plant, 2020, 13(7):935-938. doi: 10.1016/j.molp.2020.04.009 URL
[28]	项丽慧, 陈林, 余文权, 等. 茶树GH1基因家族鉴定及其在茶鲜叶萎凋过程的表达[J]. 应用与环境生物学报, 2020, 26(4):878-885.
	Xiang LH, Chen L, Yu WQ, et al. Identification of the GH1 gene family in Camellia sinensis and expression analysis during the withering process of fresh tea leaves[J]. Chin J Appl Environ Biol, 2020, 26(4):878-885.
[29]	叶一隽, 李佳敏, 曹红利, 等. 茶树CsBBX基因家族的鉴定与表达[J]. 应用与环境生物学报, 2020, 26(6):1508-1516.
	Ye YJ, Li JM, Cao HL, et al. Identification and expression analysis of the CsBBX gene family in tea plants[J]. Chin J Appl Environ Biol, 2020, 26(6):1508-1516.
[30]	Xia EH, Li FD, Tong W, et al. Tea Plant Information Archive:a comprehensive genomics and bioinformatics platform for tea plant[J]. Plant Biotechnol J, 2019, 17(10):1938-1953. doi: 10.1111/pbi.v17.10 URL
[31]	Zhou CZ, Zhu C, Fu H, et al. Genome-wide investigation of superoxide dismutase(SOD)gene family and their regulatory miRNAs reveal the involvement in abiotic stress and hormone response in tea plant(Camellia sinensis)[J]. PLoS One, 2019, 14(10):e0223609. doi: 10.1371/journal.pone.0223609 URL
[32]	Ouyang S, Zhu W, Hamilton J, et al. The TIGR Rice Genome Annotation Resource:improvements and new features[J]. Nucleic Acids Res, 2007, 35(database issue):D883-D887. doi: 10.1093/nar/gkl976 URL
[33]	马洪磊. 植物辅抑制因子TPL/TPR蛋白结构与EAR基序相互作用分子机理研究[D]. 上海:中国科学院上海药物研究所, 2016.
	Ma HL. Structure of plant co-repressor TPL/TPR protein provides insights into mechanism of EAR motif binding[D]. Shanghai:Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 2016.
[34]	Anderson P. Post-transcriptional regulons coordinate the initiation and resolution of inflammation[J]. Nat Rev Immunol, 2010, 10(1):24-35. doi: 10.1038/nri2685 pmid: 20029446
[35]	Stahle MI, Kuehlich J, Staron L, et al. YABBYs and the transcriptional corepressors LEUNIG and LEUNIG_HOMOLOG maintain leaf polarity and meristem activity in Arabidopsis[J]. Plant Cell, 2009, 21(10):3105-3118. doi: 10.1105/tpc.109.070458 URL
[36]	Thines B, Katsir L, Melotto M, et al. JAZ repressor proteins are targets of the SCF(COI1)complex during jasmonate signalling[J]. Nature, 2007, 448(7154):661-665. doi: 10.1038/nature05960 URL
[37]	Chini A, Fonseca S, Fernández G, et al. The JAZ family of repressors is the missing link in jasmonate signalling[J]. Nature, 2007, 448(7154):666-671. doi: 10.1038/nature06006 URL
[38]	彭蕾. DNA序列的Exon-Intron结构的研究[D]. 北京:北京工业大学, 2001.
	Peng L. Research of exon-intron structures in the DNA sequences[D]. Beijing:Beijing University of Technology, 2001.
[39]	王晓斌, 刘国仰. 有关内含子功能研究的新进展[J]. 中华医学遗传学杂志, 2000(3):211-212
	Wang XB, Liu GY. New research progress on the function of intron[J]. Chin J Med Genet, 2000(3):211-212
[40]	单耀军. 内含子与基因间隔序列长度及基因表达量间关系的研究[D]. 保定:河北大学, 2007.
	Shan YJ. Investigating the relation of introns to the length of intergenic sequences and gene expression levels[D]. Baoding:Hebei University, 2007.
[41]	Lorenzo O, Chico JM, Sánchez-Serrano JJ, et al. JASMONATE-INSENSITIVE1 encodes a MYC transcription factor essential to discriminate between different jasmonate-regulated defense responses in Arabidopsis[J]. Plant Cell, 2004, 16(7):1938-1950. pmid: 15208388

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