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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (07): 1047-1055.doi: 10.3724/SP.J.1006.2015.01047


Changes of DNA Methylation Levels and Patterns in Tea Plant (Camellia sinensis) during Cold Acclimation

ZHOU Yan-Hua,CAO Hong-Li,YUE Chuan,WANG Lu,HAO Xin-Yuan,WANG Xin-Chao*,YANG Ya-Jun*   

  1. Tea Research Institute of Chinese Academy of Agricultural Sciences / Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture / National Center for Tea Improvement, Hangzhou 310008, China
  • Received:2015-01-15 Revised:2015-04-02 Online:2015-07-12 Published:2015-05-04
  • Contact: 王新超, E-mail: wangxinchao@caas.cn; 杨亚军, E-mail: yjyang@tricaas.com E-mail:zhouyh@tricaas.com


Low temperature is one of the most critical environmental factors that limit tea plant growth, survival and geographical distribution. Tea plant can enhance its cold tolerance after undergoing a period of cold acclimation. DNA methylation is one of the epigenetic phenomena, and can be altered by environmental stress in plant. In order to explore the relationship between DNA methylation and low temperature stress response in tea plant, methylation sensitive amplified polymorphism (MSAP) and high performance liquid chromatography (HPLC) were used to analyze the changes of DNA methylation level and pattern in this study. The MSAP results showed that 905, 968, and 970 methylated bands were amplified with 50 selected primers in non-acclimated sample (CK), fully acclimated and de-acclimated samples, with the methylation levels of 50.6%, 54.1%, and 54.2%, respectively. DNA methylation levels in fully acclimated and de-acclimated samples were increased compared with CK. HPLC results were similar with MSAP results. In addition, DNA demethylation and methylation were both occurred during cold acclimation, but the DNA methylation was increased more than the

Key words: Tea plant, Cold acclimation, DNA methylation, MSAP, HPLC

[1]虞富莲. 论茶树原产地和起源中心. 茶叶科学, 1986, 6: 1–8

Yu F L. Discussion on the originating place and the originating centre of tea plant. J Tea Sci, 1986, 6: 1–8 (in Chinese with English abstract)

[2]王新超, 杨亚军. 茶树抗性育种研究现状. 茶叶科学, 2003, 23: 94–98

Wang X C, Yang Y J. Research progress on resistance breeding of tea plant. J Tea Sci, 2003, 23: 94–98 (in Chinese with English abstract)

[3]杨亚军, 郑英雷, 王新超. 冷驯化和ABA对茶树抗寒力及其体内脯氨酸含量的影响. 茶叶科学, 2004, 24: 177–182

Yang Y J, Zheng L Y, Wang X C. Effect of cold acclimation and ABA on cold hardiness, contents of proline of tea plants [Camellia sinensis (L.) O. Kuntze]. J Tea Sci, 2004, 24: 177–182 (in Chinese with English abstract)

[4]杨亚军, 郑雷英, 王新超. 低温对茶树叶片膜脂脂肪酸和蛋白质的影响. 亚热带植物科学, 2005, 34(1): 5–9

Yang Y J, Zheng L Y, Wang X C. Changes of membrane fatty acid composition and protein in tea leaves at low temperature. Subtrop Plant Sci, 2005, 34(1): 5–9 (in Chinese with English abstract)

[5]Wang X C, Zhao Q Y, Ma C L, Zhang Z H, Cao H L, Kong Y M, Yue C, Hao X Y, Chen L, Ma J Q, Jin J Q, Li X, Yang Y J. Global transcriptome profiles of Camellia sinensis during cold acclimation. BMC Genom, 2013, 14: 415

[6]曹红利, 岳川, 郝心愿, 王新超, 杨亚军. 茶树胆碱单加氧酶CsCMO的克隆及甜菜碱合成关键基因的表达分析. 中国农业科学, 2013, 46: 3087–3096

Cao H L, Yue C, Hao X Y, Wang X C, Yang Y J. Cloning of choline monooxygenase (CMO) gene and expression analysis of the key glycine betaine biosynthesis-related genes in tea plant (Camellia sinensis). Sci Agric Sin, 2013, 46: 3087–3096 (in Chinese with English abstract)

[7]曹红利, 岳川, 周艳华, 王璐, 郝心愿, 杨亚军, 王新超. 茶树bZIP 转录因子基因CsbZIP1 的克隆与表达定位. 作物学报, 2014, 40: 1702–1709

Cao H L, Yue C, Zhou Y H, Wang L, Hao X Y, Yang Y J, Wang X C. Molecular cloning and expression of a bZIP transcription factor gene CsbZIP1 in tea plant (Camellia sinensis). Acta Agron Sin, 2014, 40: 1702–1709 (in Chinese with English abstract)

[8]岳川, 曹红利, 周艳华, 王璐, 郝心愿, 王新超, 杨亚军. 茶树谷胱甘肽还原酶基因CsGRs的克隆与表达分析. 中国农业科学, 2014, 47: 3277–3289

Yue C, Cao H L, Zhou Y H, Wang L, Hao X Y, Wang X C, Yang Y J. Cloning and expression analysis of glutathione reductase genes (CsGRs) in tea plant (Camellia sinensis). Sci Agric Sin, 2014, 47: 3277–3289 (in Chinese with English abstract)

[9]Bender J. DNA methylation and epigenetics. Annu Rev Plant Biol, 2004, 55: 41–68

[10]Vanyushin B F, Ashapkin V V. DNA methylation in higher plants: past, present and future. Biochim Biophys Acta, 2011, 1809: 360–368

[11]Chan S W, Henderson I R, Jacobsen S E. Gardening the genome: DNA methylation in Arabidopsis thaliana. Nat Rev Genet, 2005, 6: 351–360

[12]Wang W, Zhao X, Pan Y, Zhu L, Fu B, Li Z. DNA methylation changes detected by methylation-sensitive amplified polymorphism in two contrasting rice genotypes under salt stress. J Genet Genom, 2011, 38: 419–424

[13]Jullien P E, Kinoshita T, Ohad N, Berger F. Maintenance of DNA methylation during the Arabidopsis life cycle is essential for parental imprinting. Plant Cell, 2006, 18: 1360–1372

[14]Adams K L, Percifield R, Wendel J F. Organ-specific silencing of duplicated genes in a newly synthesized cotton allotetraploid. Genetics, 2004, 168: 2217–2226

[15]潘雅姣, 傅彬英, 王迪, 朱苓华, 黎志康. 水稻干旱胁迫诱导DNA甲基化时空变化特征分析. 中国农业科学, 2009, 42: 3009–3018

Pan Y J, Fu B Y, Wang D, Zhu L H, Li Z K. Spatial and temporal profiling of DNA methylation induced by drought stress in rice. Sci Agric Sin, 2009, 42: 3009–3018 (in Chinese with English abstract)

[16]Steward N, Ito M, Yamaguchi Y, Koizumi N, Sano H. Periodic DNA methylation in maize nucleosomes and demethylation by environmental stress. J Biol Chem, 2002, 277(40): 37741–37746

[17]高桂珍, 应菲, 陈碧云, 李浩, 吕晓丹, 闫贵欣, 许鲲, 伍晓明. 热胁迫过程中白菜型油菜种子DNA的甲基化. 作物学报, 2011, 37: 1597–1604

Gao G Z, Ying F, Chen B Y, Li H, Lv X D, Yan G X, Xu K, Wu X M. DNA methylation of seed in response to heat stress in Brassica rapa L. Acta Agron Sin, 2011, 37: 1597–1604 (in Chinese with English abstract)

[18]Dowen R H, Pelizzola M, Schmitz R J, Lister R, Dowen J M, Nery J R, Dixon J E, Ecker J R. Widespread dynamic DNA methylation in response to biotic stress. Proc Natl Acad Sci USA, 2012, 109: E2183–2191

[19]Portis E, Acquadro A, Comino C, Lanteri S. Analysis of DNA methylation during germination of pepper (Capsicum annuum L.) seeds using methylation-sensitive amplification polymorphism (MSAP). Plant Sci, 2004, 166: 169–178

[20]Banaei Moghaddam A M, Fuchs J, Czauderna T, Houben A, Mette M F. Intraspecific hybrids of Arabidopsis thaliana revealed no gross alterations in endopolyploidy, DNA methylation, histone modifications and transcript levels. Theor Appl Genet, 2010, 120: 215–226

[21]Sha A H, Lin X H, Huang J B, Zhang D P. Analysis of DNA methylation related to rice adult plant resistance to bacterial blight based on methylation-sensitive AFLP (MSAP) analysis. Mol Genet Genomics, 2005, 273: 484–490

[22]Tan M P. Analysis of DNA methylation of maize in response to osmotic and salt stress based on methylation-sensitive amplified polymorphism. Plant Physiol Biochem, 2010, 48: 21–26

[23]Osabe K, Clement J D, Bedon F, Pettolino F A, Ziolkowski L, Llewellyn D J, Finnegan E J, Wilson I W. Genetic and DNA methylation changes in cotton (Gossypium) genotypes and tissues. PLoS One, 2014, 9(1): e86049

[24]Yaish M W, Peng M, Rothstein S J. Global DNA methylation analysis using methyl-sensitive amplification polymorphism (MSAP). Methods Mol Biol, 2014, 1062: 285–298

[25]Johnston J W, Harding K, Bremner D H, Souch G, Green J, Lynch P T, Grout B, Benson E E. HPLC analysis of plant DNA methylation: a study of critical methodological factors. Plant Physiol Biochem, 2005, 43: 844–853

[26]Zhao Y L, Yu S X, Ye W W, Wang H M, Wang J J, Fang B X. Study on DNA cytosine methylation of cotton (Gossypium hirsutum L.) genome and its implication for salt tolerance. Agric Sci China, 2010, 9: 783–791

[27]Chakrabarty D, Yu K W, Paek K Y. Detection of DNA methylation changes during somatic embryogenesis of Siberian ginseng (Eleuterococcus senticosus). Plant Sci, 2003, 165: 61–68

[28]Richards E J. DNA methylation and plant development. Trends Genet, 1997, 13: 319–323

[29]李雪林, 林忠旭, 聂以春, 郭小平, 张献龙. 盐胁迫下棉花基因组DNA表观遗传变化的MSAP分析. 作物学报, 2009, 35: 588–596

Li X L, Li Z X, Nie Y C, Guo X P, Zhang X L. MSAP analysis of epigenetic changes in cotton (Gossypium hirsutum L.) under salt stress. Acta Agron Sin, 2009, 35: 588–596 (in Chinese with English abstract)

[30]Cervera M T, Ruiz-Garcia L, Martinez-Zapater J M. Analysis of DNA methylation in Arabidopsis thaliana based on methylation-sensitive AFLP markers. Mol Genet Genom, 2002, 268: 543–552

[31]盖树鹏, 张风, 张玉喜, 郑国生. 低温解除牡丹休眠进程中基因组DNA甲基化敏感扩增多态性(MSAP)分析. 农业生物技术学报, 2012, 20: 261–267

Gai S P, Zhang F, Zhang Y X, Zheng G S. Analysis of genomic DNA methylation during chilling induced endo-dormancy release by methylation sensitive amplified polymorphism (MSAP) technology in tree peony (Paeonia suffruticosa). J Agric Biotech, 2012, 20: 261–267 (in Chinese with English abstract)

[32]张勇, 邓科君, 张韬, 彭金华, 周建平, 任正隆. 黑麦基因组DNA甲基化修饰位点的MSAP分析. 麦类作物学报, 2009, 29: 559–564

Zhang Y, Deng K J, Zhang T, Peng J H, Zhou J P, Ren Z L. Analysis on genomic DNA methylation modification of rye by methylation-sensitive ampl ification polymorphism. J Triticeae Crops, 2009, 29: 559–564 (in Chinese with English abstract)

[33]Kalberer S R, Wisniewski M, Arora R. Deacclimation and reacclimation of cold-hardy plants: current understanding and emerging concepts. Plant Sci, 2006, 171: 3–16

[34]Thomashow M F. Plant cold acclimation:Freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol, 1999, 50: 571–599

[35]Christov N.K, Yoneyama S, Shimamoto Y, Imai R. Differential expression of wheat genes during cold acclimation. Cytol Genet, 2007, 41: 142–150

[36]Burn J E, Bagnall D J, Metzger J D, Dennis E S, Peacock W J. DNA methylation, vernalization, and the initiation of flowering. Proc Natl Acad Sci USA, 1993, 90: 287–291

[37]Zhang L, Wang Y, Zhang X H, Zhang M, Han D G, Qiu C P, Han Z H. Dynamics of phytohormone and DNA methylation patterns changes during dormancy induction in strawberry (Fragaria x ananassa Duch.). Plant Cell Rep, 2012, 31: 155–165

[38]Pan Y J Wang W S, Zhao X Q, Zhu L H, Fu B Y, Li Z K. DNA methylation alterations of rice in response to cold stress. Plant Omics J, 2011, 4: 364–369

[39]Mayer B F, Ali-Benali M A, Demone J, Bertrand A, Charron J. Cold acclimation induces distinctive changes in the chromatin state and transcript levels of COR genes in Cannabis sativa varieties with contrasting cold acclimation capacities. Physiol Plant, 2014: DOI: 10.1111/ppl.12318

[40]徐青, 余云舟, 赵萌, 孙东晓. DNA甲基化在动植物遗传育种中的研究进展. 生物技术通讯, 2011, 2: 113–117

Xu Q, Yu Y Z, Zhao M, Sun D X. Progress of DNA methylation in genetics and breeding of plant and animal. Lett Biotech, 2011, 2: 113–117 (in Chinese with English abstract)

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