茶树CsbHLH2基因克隆及表达分析

doi:10.13305/j.cnki.jts.2015.05.012

Abstract

Abstract: The bHLH (basic Helix-Loop-Helix) transcription factor is one of the most important transcription factor in plants, which play important roles in plant growth and stress regulation. The bHLH transcription factor CsbHLH2 was cloned from tea plant (Camellia sinensis) cultivars ‘Shanchayihao’ by homologous cloning technology using cDNA template. Bioinformatics analysis showed that the CsbHLH2 contains 714 bp ORF and was predicted encoding 297 amino acid, the deduced protein molecular weight was 58.4 kD and its theoretical isoelectric point was 5.14. Amino acid sequence alignment revealed that CsbHLH2 was highly homologous to other higher plant bHLH proteins. Transient expression of recombinant plasmid CsbHLH2/PBI221-GFP in Arabidopsis protoplasts showed that CsbHLH2 was located in cell nuclei. Quantitative real-time PCR analysis of the expression profiles showed that the CsbHLH2 gene was expressed in bud, leaf, stem and root. The highest expression level of the CsbHLH2 was found in the young leaf. Different hormone treatments results showed that the CsbHLH2 was induced by ETH, MEJA and SA treatment, respectively.

Key words: Camellia sinensis (L.), bHLH transcription factor, subcelleular locatlization, phylogenetic analysis, expression analysis

CLC Number:

S571.1
Q52

HAN Yongtao, XIAO Bin, QIAN Wenjun, LIANG Shaoru. Cloning and Expression Analysis of CsbHLH2 in Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2015, 35(5): 481-490.

References 25

[1]	Ludwig S R, Habera L F, Dellaporta S L, et al. Lc, a member of the maize R gene family responsible for tissue-specific anthocyanin production, encodes a protein similar to transcriptional activators and contains the myc-homology region[J]. Proceedings of the National Academy of Sciences of the United States of America, 1989, 86(18): 7092-7096.
[2]	Carretero-Paulet L, Galstyan A, Roig-Villanova I, et al.Genome-wide classification and evolutionary analysis of the bHLH family of transcription factors in Arabidopsis, poplar, rice, moss, and algae[J]. Plant Physiol, 2010, 153: 1398-1412.
[3]	Gabriela Toledo-Ortiz, Enamul Huq, Peter H Quail.The Arabidopsis basic helix-loop-helix transcription factor family[J]. Plant Cell, 2003, 15(8): 1749-l770.
[4]	Li X X, Duan X P, Jiang H X, et al. Genome-wide analysis of basic helix-loop-helix transcription factor fam in rice and ArabMopsk[J]. Plant PhysioL, 2006, 141(4): l167-l184.
[5]	Yi Song, Chuangwei Yang, Shan Gao, et al. Age-Triggered and Dark-Induced Leaf Senescence Require the bHLH Transcription Factors PIF3, 4 and 5[J]. Mol Plant, 2014, 7(12): 1776-1787.
[6]	Miho Ikeda, Nobutaka Mitsuda, Masaru Ohme-Takagi, et a1. ATBS1 INTERACTING FACTORs negatively regulate Arabidopsis cell elongation in the triantagonistic bHLH system[J]. Plant Signaling & Behavior, 2013, 8(3): e23448.
[7]	Terri A Long, Hironaka Tsukagoshi, Wolfgang Busch, et al. The bHLH Transcription Factor POPEYE Regulates Response to Iron Deficiency in Arabidopsis Roots[J]. The Plant Cell, 2010(22): 2219-2236.
[8]	Seo J S, Joo J S, Kim M J, et al. OsbHLH148, a basic helix-loop-helix protein, interacts with OsJAZ proteins in a jasmonate signaling pathway leading to drought tolerance in rice[J]. Plant J, 2011, 65: 907-921.
[9]	Kiribuchi K, Sugimori M, Takeda M, et al. RERJ1, a jasmonic acid-responsive gene from rice, encodes a basic helix-loop-helix protein[J]. Biochem Biophys Res Comm, 2004, 325: 857-863.
[10]	Kiribuchi K, Jikummu Y, Kaku H, et al. Involvement of the basic helix-loop-helix transcription factor RERJ1 in wounding and drought stress responses in rice plants[J]. Biosei Biotechnol Biochem, 2005, 69: 1042-1044.
[11]	Ling H Q, Baure P, Bereczky Z, et al. The tomato fer gene encoding a bHLH protein controls iron uptake responses in roots[J]. Proc Natl Acad Sci, 2002, 99: 13938-13943.
[12]	Kanaoka M M, Pillitteri L J, Fujii H, et al. SCREAM/ICE1 and SCREAM2 specify three cell-state transitional steps leading to Arabidopsis stomatal differentiation[J]. Plant Cell, 2008, 11(20): 1775-1785.
[13]	Miura K, Jin J B, Lee J, et al. SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis[J]. Plant Cell, 2007, 19: 1403-1414.
[14]	Min Fan, Ming-Yi Bai, Jung-Gun Kim, et al. The bHLH Transcription Factor HBI1 Mediates the Trade-Off between Growth and Pathogen-Associated Molecular Pattern- Triggered Immunity in Arabidopsis[J]. Plant Cell, 26(2): 828-841.
[15]	Susheng Song, Tiancong Qi, Meng Fan, et al. The bHLH Subgroup IIId Factors Negatively Regulate Jasmonate-Mediated Plant Defense and Development[J]. PLOS Genetics, 2013, 9(7): 523-528.
[16]	Sang-Dong Yoo, Young-Hee Cho, Jen Sheen.Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis[J]. Nature Protocol, 2007, 2(7): 1565-1572.
[17]	徐刚, 姚银安. 水杨酸、茉莉酸和乙烯介导的防卫信号途径相互作用的研究进展[J]. 生物学杂志, 2009(1): 48-51.
[18]	Liu Q, Zhang G Y, Chen S Y.Structure and regulatory function of plant transcription factors[J]. Chin Sci Bull, 2001, 45(4): 271-278.
[19]	K C Babitha, S V Ramu, V Pruthvi, et al. Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress in Arabidopsis[J]. Transgenic Res, 2013(22): 327-341.
[20]	Wenwen Liu1, Huanhuan Tai1, Songsong Li, et al. bHLH122 is important for drought and osmotic stress resistance in Arabidopsis and in the repression of ABA catabolism [J]. New Phytologist, 2014, 201: 1192-1204.
[21]	Hongmei Li, Jiaqiang Sun, Yingxiu Xu, et al. The bHLH-type transcription factor AtAIB positively regulates ABA response in Arabidopsis[J]. Plant Mol Biol, 2007(65): 655-665.
[22]	Yajie Niu, Pablo Figueroa, John Browse.Characterization of JAZ-interacting bHLH transcription factors that regulate jasmonate responses in Arabidopsis[J]. Journal of Experimental Botany, 2011, 62(6): 2143-2154.
[23]	曹爱忠, 李巧, 陈雅平, 等. 利用大麦基因芯片筛选簇毛麦抗白粉病相关基因及其抗病机制的初步研究[J]. 作物学报, 2006(10): 1444-1452.
[24]	Sung-Hyun Kim, Tetsuo Oikawa, Junko Kyozuka, et al. The bHLH Rac Immunity1 (RAI1) Is Activated by OsRac1 via OsMAPK3 and OsMAPK6 in Rice Immunity[J]. Plant and Cell Physiology, 2012, 53(4): 740-754.
[25]	Lintao WU, Caifu DU, Zhangli ZENG, et al. Study on the Signal Transduction Pathway of Plant Defense to Pathogens[J]. Agricultural Science & Technology, 2014, 15(4): 517-519, 549.

Cloning and Expression Analysis of CsbHLH2 in Tea Plant (Camellia sinensis)

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 25

Related Articles 15

Metrics

Comments

Recommended 10

[1]	ZHOU Hanchen, YANG Jihong, XU Yujie, WU Qiong, LEI Pandeng. Phylogenetic Analysis of NUDX1 Gene Involved in Geraniol Biosynthesis [J]. Journal of Tea Science, 2022, 42(5): 638-648.
[2]	YAN Yuting, LI Yujie, WANG Qian, TANG Meijun, GUO Huawei, LI Hongliang, SUN Liang. The Expression Profiles of Chemosensory Protein 8 Orthologs in Two Closely Related Tea Geometrid Species, Ectropis obliqua Prout and Ectropis grisescens Warren [J]. Journal of Tea Science, 2022, 42(2): 200-210.
[3]	ZHOU Hanchen, LEI Pandeng. The Functional Identification of Two Alternative Splicing Transcripts of CsNES [J]. Journal of Tea Science, 2021, 41(6): 753-760.
[4]	LIU Miaomiao, ZANG Liansheng, SUN Xiaoling, ZHOU Zhongshi, YE Meng. Cloning and Expression Analysis of CsWRKY17 Transcription Factor in Tea Plants [J]. Journal of Tea Science, 2021, 41(5): 631-642.
[5]	LIN Xinying, WANG Pengjie, CHEN Xuejin, GUO Yongchun, GU Mengya, ZHENG Yucheng, YE Naixing. Identification of LOX Gene Family in Camellia sinensis and Expression Analysis in the Process of White Tea Withering [J]. Journal of Tea Science, 2021, 41(4): 482-496.
[6]	WANG Yanding, WANG Huan, LI Nana, WANG Lu, HAO Xinyuan, WANG Yuchun, DING Changqing, YANG Yajun, WANG Xinchao, QIAN Wenjun. Identification and Expression Analysis of Glucose-6-hosphate Dehydrogenase Gene (CsG6PDHs) in Camellia sinensis [J]. Journal of Tea Science, 2021, 41(4): 497-510.
[7]	GU Mengya, WANG Pengjie, CHEN Xuejin, ZHENG Yucheng, GUO Yongchun, LIN Xinying, GAO Ting, HOU Binghao, YE Naixing. Identification of Alcohol Dehydrogenase Gene Family and Their Expression Analysis in the Withering Process of White Tea [J]. Journal of Tea Science, 2021, 41(3): 302-314.
[8]	CHEN Siwen, KANG Rui, GUO Zhiyuan, ZHOU Qiongqiong, FENG Jiancan. Cloning and Expression Analysis of CsCML16 in Tea Plants (Camellia sinensis) under Low Temperature Stress [J]. Journal of Tea Science, 2021, 41(3): 315-326.
[9]	CHEN Yao, ZHANG Weifu, REN Hengze, XIONG Fei, ZHANG Haojie, YAO Lina, LIU ying, WANG Lu, WANG Xinchao, YANG Yajun, HAO Xinyuan. Genome-wide Investigation and Expression Analysis of DNA Demethylase Genes in Tea Plant (Camellia Sinensis) [J]. Journal of Tea Science, 2021, 41(1): 28-39.
[10]	JIANG Junmei, FANG Yuanpeng, NING Na, CHEN Meiqing, YANG Zaifu, WANG Yong, LI Xiangyang, XIE Xin. Cloning and Expression Analysis of CssHSP18.1 Gene in Camellia Sinensis [J]. Journal of Tea Science, 2020, 40(3): 328-340.
[11]	YU Shuping, XU Liyi, WU Rongmei, WANG Liyuan, WU Liyun, WEI Kang, CHENG Hao, WANG Yongqi. Genetic and Phylogenetic Analysis for Resources of Camellia Sinensis from Kaihua County in Zhejiang Province [J]. Journal of Tea Science, 2020, 40(3): 341-351.
[12]	CHEN Xuejin, WANG Pengjie, LIN Xinying, GU Mengya, ZHENG Yucheng, ZHENG Zhilin, YE Naixing. Identification and Expression Analysis of Terpene Synthesis Related Genes during the Withering of White Tea [J]. Journal of Tea Science, 2020, 40(3): 363-374.
[13]	LEI Lei, WANG Lu, YAO Lina, HAO Xinyuan, ZENG Jianming, DING Changqing, WANG Xinchao, YANG Yajun. Identification and Expression Analysis of Calcium-dependent Protein Kinase CsCDPK17 in Tea Plant (Camellia sinensis) [J]. Journal of Tea Science, 2019, 39(3): 267-279.
[14]	CHEN Jiangfei, YU Jinming, YANG Jiankun, YU Youben, XIAO Bin, YANG Yajun, WANG Weidong. Cloning and Expression Analysis of Na⁺/H⁺ Antiporter Gene CsNHX1 and CsNHX2 in Tea Plant (Camellia sinensis) [J]. Journal of Tea Science, 2018, 38(6): 559-568.
[15]	YE Xiaoli, PAN Junting, ZHU Jiaojiao, SHU Zaifa, CUI Chuanlei, XING Anqi, NONG Shouhua, ZHU Xujun, FANG Wanping, WANG Yuhua. Cloning and Expression Analysis of Small GTPase (CsRAC5) under Cold Stress in Tea Plant (Camellia sinensis) [J]. Journal of Tea Science, 2018, 38(2): 146-154.