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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (10): 1417-1425.doi: 10.3724/SP.J.1006.2017.01417

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS •     Next Articles

Cloning of β-amylase Gene (CsBAM3) and ItsExpression ModelResponseto Cold Stress in Tea Plant

HAO Xin-Yuan1,**,YUEChuan1,2,**,TANG Hu1,QIAN Wen-Jun1,WANG Yu-Chun1,WANG Lu1, WANG Xin-Chao1,YANG Ya-Jun1,*   

  1. 1 Tea Research Institute, Chinese Academy of Agricultural Sciences / National Center for Tea Improvement / Key Laboratory of Tea Plant Biology and Resources Utilization, Ministry of Agriculture, Hangzhou 310008, China; 2College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002
  • Received:2017-02-13 Revised:2017-05-10 Online:2017-10-12 Published:2017-05-23
  • Contact: 王新超, E-mail: xcw75@tricaas.com Tel: 0571-86653162; 杨亚军: E-mail: yjyang@tricaas.com E-mail:haoxy@tricaas.com
  • Supported by:

    This study was supported by the Zhejiang Provincial Natural Science Foundation (LY16C160001), the Earmarked Fund for China Agriculture Research System (CARS-23), and the Chinese Academy of Agricultural Sciences through an Innovation Project for Agricultural Sciences and Technology (CAAS-ASTIP-2014-TRICAAS).

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Abstract:

A major enzyme of starch breakdown in plant leaf, β-amylase (BAM), plays important roles in response to abiotic stresses. In tea plant, a differentially expressed gene involved in starch metabolism was identified from previous RNA-Seq analysis of cold acclimation. Its full-length cDNA was cloned and sequenced, showing that it is a homolog of BAM3 in Arabidopsis (named CsBAM3). CsBAM3 contains 548 amino acid residues and is grouped into subfamily II together with BAM1 and BAM3 from Arabidopsis. According to protein sequence and 3D structure analyses, CsBAM3 was considered as a chloroplast-targeted protein with β-amylase activity. Promoter cloning and sequence analysis showed a couple of responsive elements related to circadian, light, cold and phytohormone, indicating its complex regulating mechanism. CsBAM3 showed the expression higher in leaf, relative lower in stem and flower and extremely low in root. The expression of CsBAM3 in mature leaf was significantly up-regulated during the early time courses of winter cold acclimation and maintained at relative high level during the following period. With the treatments of 4℃ and 0℃, the transcript levels of CsBAM3 in both mature leaves and young shoots were dramatically increased, interestingly, much higher level was detected in young shoots. The cold environment similar to spring cold spell was created using climate chamber. And the expression changes of CsBAM3 in shoots were determined then after the tea plants were moved into climatic chamber at different sprouting phrases. CsBAM3 was quickly up-regulated by cold once the first leaf unfolded. Our study illuminated that CsBAM3 is a key β-amylase coding gene involved in starch metabolism of tea plant, which can be stimulated in both mature leaves and young shoots by cold stress.

Key words: β-amylase, Tea plant, Gene clone, Cold stress, Expression regulation

[1]蒋跃林 , 李倬. 我国茶树栽培界限的气候划分. 生态农业研究, 2000, 8(1): 89–92
Jiang Y L, Li Z. Climate divison of tea tree cultivated limit in China. Eco-agric Res, 2000, 8(1):89–92 (in Chinese with English abstract)
[2]郭湘, 唐茜, 许燕, 陈玖琳, 王自琴. 早春霜冻对不同茶树品种芽叶的生化成分及制茶品质的影响. 云南大学学报, 2015, 37: 930–938
Guo X, Tang Q, Xu Y, Chen J L, Wang Z Q. Effect of frost in early spring on biochemical composition and sensory qulity of new shoots in different tea cultivars. J Yunnan Univ, 2015, 37: 930–938 (in Chinese with English abstract)
[3]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
[4]Benina M, Obata T, Mehterov N, Ivanov I, Petrov V, Toneva V, Fernie A R, Gechev T S. Comparative metabolic profiling of Haberlea rhodopensis, Thellungiella halophyla, and Arabidopsis thaliana exposed to low temperature. Front Plant Sci, 2013, 4: 499
[5]Lee J H, Yu D J, Kim S J, Choi D, Lee H J. Intraspecies differences in cold hardiness, carbohydrate content and beta-amylase gene expression of Vaccinium corymbosum during cold acclimation and deacclimation. Tree Physiol, 2012, 32: 1533–1540
[6]Janska A, Aprile A, Zamecnik J, Cattivelli L, Ovesna J. Transcriptional responses of winter barley to cold indicate nucleosome remodelling as a specific feature of crown tissues. Funct Integr Genom, 2011, 11: 307–325
[7]Gupta A K, Kaur N. Sugar signalling and gene expression in relation to carbohydrate metabolism under abiotic stresses in plants. J Biosci, 2005, 30: 761–76
[8]Baena-Gonzalez E, Sheen J. Convergent energy and stress signaling. Trends Plant Sci, 2008, 13: 474–482
[9]Smeekens S, Ma J, Hanson J, Rolland F. Sugar signals and molecular networks controlling plant growth. Curr Opin Plant Biol, 2010, 13: 274–279
[10]Stitt M, Zeeman S C. Starch turnover: pathways, regulation and role in growth. Curr Opin Plant Biol, 2012, 15: 282–292
[11]Yu T S, Zeeman S C, Thorneycroft D, Fulton D C, Dunstan H, Lue W L, Hegemann B, Tung S Y, Umemoto T, Chapple A, Tsai D L, Wang S M, Smith A M, Chen J, Smith S M. alpha-Amylase is not required for breakdown of transitory starch in Arabidopsis leaves. J Biol Chem, 2005, 280: 9773–9779
[12]Yue C, Cao H L, Wang L, Zhou Y H, Huang Y T, Hao X Y, Wang Y C, Wang B, Yang Y J. Wang X C. Effects of cold acclimation on sugar metabolism and sugar-related gene expression in tea plant during the winter season. Plant Mol Biol, 2015, 88: 591–608
[13]Fulton D C, Stettler M, Mettler T, Vaughan C K, Li J, Francisco P, Gil M, Reinhold H, Eicke S, Messerli G, Dorken G, Halliday K, Smith A M, Smith S M, Zeeman S C. Beta-AMYLASE4, a noncatalytic protein required for starch breakdown, acts upstream of three active beta-amylases in Arabidopsis chloroplasts. Plant Cell, 2008, 20: 1040–1058
[14]Li J, Francisco P, Zhou W, Edner C, Steup M, Ritte G, Bond C S, Smith S M. Catalytically-inactive beta-amylase BAM4 required for starch breakdown in Arabidopsis leaves is a starch-binding-protein. Arch Biochem Biophys, 2009, 489: 92–98
[15]Monroe J D, Storm A R, Badley E M, Lehman M D, Platt S M, Saunders L K, Schmitz J M, Torres C E. beta-Amylase1 and beta-amylase3 are plastidic starch hydrolases in Arabidopsis that seem to be adapted for different thermal, pH, and stress conditions. Plant Physiol, 2014, 166: 1748–1763
[16]Peng T, Zhu X, Duan N, Liu J H. PtrBAM1, a beta-amylase-coding gene of Poncirus trifoliata, is a CBF regulon member with function in cold tolerance by modulating soluble sugar levels. Plant Cell Environ, 2014, 37: 2754–2767
[17]Kaplan F, Guy C L. Beta-Amylase induction and the protective role of maltose during temperature shock. Plant Physiol, 2004, 135: 1674–1684
[18]Fowler S, Thomashow M F. Arabidopsis transcriptome profiling indicates that multiple regulatory pathways are activated during cold acclimation in addition to the CBF cold response pathway. Plant Cell, 2002. 14: 1675–1690
[19]Reinhold H, Soyk S, Simkova K, Hostettler C, Marafino J, Mainiero S, Vaughan C K, Monroe J D, Zeeman S C. Beta-amylase-like proteins function as transcription factors in Arabidopsis, controlling shoot growth and development. Plant Cell, 2011, 23: 1391–1403
[20]丁菲. 低温胁迫下与茶树糖代谢相关基因的克隆与表达.安徽农业大学硕士学位论文, 安徽合肥, 2012
Ding F. Cloning and Expression of Genes Related to Glyconmetabolism in Tea Plant (Camellia sinensis (L.) O. Kuntze) Under Low Temperature. MS Thesis of Anhui Agricultural  University,Hefei, China, 2012 (in Chinese with English abstract)
[21]岳川. 茶树糖类相关基因的挖掘及其在茶树冷驯化中的表达研究. 中国农业科学院博士学位论文,北京, 2015
Yue C. Cloning and Expression Analysis of Sugar-Related Genes during Cold Acclimation in Tea Plant. PhD Dissertation of Chinese Academy of Agricultural Sciences,Beijing, China, 2015 (in Chinese with English abstract)
[22]Chang S, Puryear J, Cairney J. A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep, 1993, 11: 113–116
[23]Hao X, Horvath D P, Chao W S, Yang Y, Wang X, Xiao B. Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant (Camellia sinensis (L.) O. Kuntze). Int J Mol Sci, 2014, 15: 22155–22172
[24]彭婷. 枳叶片抑制差减杂交cDNA文库构建及PtrBAM1基因抗寒功能鉴定. 华中农业大学博士学位论文,中国武汉, 2013
Peng T. Suppression Subtractive Hybridization cDNA Library Construction for Low Temperature-Treated Poncirus trifoliata and Characterization of PtrBAM1. PhD Dissertation of Huangzhong Agurcultural University, Wuhan, China, 2013 (in Chinese with English abstract)
[25]Sparla F, Costa A, Lo Schiavo F, Pupillo P, Trost P. Redox regulation of a novel plastid-targeted beta-amylase of Arabidopsis. Plant Physiol, 2006, 141: 840–850
[26]Kaplan F, Guy CL. RNA interference of Arabidopsis beta-amylase8 prevents maltose accumulation upon cold shock and increases sensitivity of PSII photochemical efficiency to freezing stress. Plant J, 2005, 44: 730–743
[27]Ingkasuwan P, Netrphan S, Prasitwattanaseree S, Tanticharoen M, Bhumiratana S, Meechai A, Chaijaruwanich J, Takahashi H, Cheevadhanarak S. Inferring transcriptional gene regulation network of starch metabolism in Arabidopsis thaliana leaves using graphical Gaussian model. BMC Syst Biol, 2012, 6: 100
[28]Davies P J. Plant Hormones. Netherlands: Springer-Verlag, XI, 2010, p 833
[29]Takeda S, Kowyama Y, Takeuchi Y, Matsuoka K, Nishimura M, Nakamura K. Spatial patterns of sucrose-inducible and polygalacturonic acid-inducible expression of genes that encode sporamin and β-amylase in sweet potato: gene structure and expression. Plant Cell Physiol, 1995, 36: 321–333
[30]Samojedny D, Orzechowski S. New look at starch degradation in Arabidopsis thaliana L. chloroplasts. Postepy Biochem, 2007, 53: 74–83
[31]Shin H, Oh Y, Kim D. Differences in cold hardiness, carbohydrates, dehydrins and related gene expressions under an experimental deacclimation and reacclimation in Prunus persica. Physiol Plant, 2015, 154: 485–499
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