Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (05): 708-717.doi: 10.3724/SP.J.1006.2017.00708


Cloning and Function Analysis of Metal Tolerance Gene (CsMTP11) in Tea Plant (Camellia sinensis L. O. Kuntze)

YUAN Lian-Yu,CHEN Ying-Juan,WEI Xu,TONG Hua-Rong*   

  1. College of Food Science, Southwest University, Chongqing 400715, China
  • Received:2016-08-23 Revised:2017-01-21 Online:2017-05-12 Published:2017-02-17
  • Contact: Dong Huarong, E-mail: huart@swu.edu.cn, Tel: 023-68251298 E-mail:yuanlianyu88@163.com
  • Supported by:

    This work was supported by the National Natural Science Foundation of China (31400583), the Natural Science Foundation Project of CQ CSTC (cstc2014jcyjA80011), and Fundamental Research Funds for the Central Universities (XDJK2014C070).


As a member of cation diffusion facilitator family, metal tolerance protein (MTP) is involved in the regulation of heavy metal stress process in plant. Here, we cloned a MTP gene, CsMTP11 (GenBank accession number: KX450265) from Zhongcha 108 tea plant. The full-length cDNA of CsMTP11 is 1197 bp, and the gene encodes a novel protein of 398 amino acids, which shares significant sequence similarity with VvMTP11(90%). The molecular weight and theoretical pI of this protein are 44.85 kD and 5.34, respectively. The conserved domain analysis indicated that CsMTP11 contained five transmembrane domains and the conserved CDF domain. The phylogenetic tree indicated that CsMTP11 had sequence conservation among different species. The transcriptional level of CsMTP11 in old leaf was higher than that in root. Futher more, the heavy metal Mn and Co stresses induced the expression of CsMTP11. The subcellular localization assay using CsMTP11-YFP fusion gene expressed in the Arabidopsis protoplast showed that CsMTP11 was localized in the plasma membranes. CsMTP11 heterologous expression in wild-type yeast BY4741 and the mutants showed that it was able to increase tolerance to Mn and Co. Taken together , our results indicated that CsMTP11 is a member of the Mn-CDF family, and it may be involved in the regulation of heavy metal stress process in tea plant.

Key words: Camellia sinensis, Heavy metal tolerance, Gene clone, Mn

[1] Szymczycha-Madeja A, Welna M, Pohl P. Elemental analysis of teas and their infusions by spectrometric methods. Trac-Trends Anal Chem, 2012, 35: 165–181 [2] Brzezicha-Cirocka J, Grembecka M, Szefer P. Monitoring of essential and heavy metals in green tea from different geographical origins. Environ Monitor Assess, 2016, 188: 1–11 [3] Santos D, Batoreu C, Mateus L, Dos Santos A P M, Aschner M. Manganese in human parenteral nutrition: Considerations for toxicity and biomonitoring. Neurotoxicology, 2014, 43: 36–45 [4] Sloot W N, Gramsbergen J B P. Axonal-transport of manganese and its relevance to selective neurotoxicity in the rat basal ganglia. Brain Res, 1994, 657: 124–132 [5] 雷雨, 苏晓倩, 王芳, 李珍, 萧力争. 茶树重金属胁迫研究概况. 中国食物与营养, 2010, (5): 19–23 Lei Y, Su X Q, Wang F, Li Z, Xiao L Z. Research situation of heavy metal stress in tea plant. Food and Nutrition in China, 2010, (5): 19–23 [6] 常硕其, 彭克勤, 张亚莲, 刘红艳, 付海平. 加锰处理对茶树锰含量及生理变化的影响研究. 茶叶科学, 2008, (5): 331–338 Chang S Q, Peng K Q, Zhang Y L, Liu H Yan, Fu H P. Effect of Mn addition treatment on accumulation of Mn and physiological active substance in tea plants. Journal of Tea Science, 2008, (5): 331–338 [7] Ozdemir Y, Gucer S. Speciation of manganese in tea leaves and tea infusions. Anal Lett, 1998, 31: 679–689 [8] Yemane Michael, Chandravanshi B S, Wondimu Taddese. Levels of essential and non-essential metals in leaves of the tea plant (Camellia sinensis L.) and soil of Wushwush farms, Ethiopia. Food Chem, 2008, 107: 1236–1243 [9] Street R, Szakova J, Drabek O, Mladkova L. The status of micronutrients (Cu, Fe, Mn, Zn) in tea and tea infusions in selected samples imported to the Czech Republic. Czech J Food Sci, 2006, 24: 62–71 [10] 姚元涛, 张丽霞, 宋鲁彬, 田丽丽. 茶树锰素营养研究现状与展望. 中国茶叶, 2009, (3): 10–11 Yao Y T, Zhang L X, Song L B, Tian L L. Research status andprospect of manganese nutrition in tea plant. China Tea, 2009, (3): 10–11 [11] Memon A R, Chino M, Hidaka H, Hara K, Yatazawa M. Manganese toxicity in field-grown tea plants and the micro-distribution of manganese in the leaf tissues as revealed by electron-probe X-ray micrography. Soil Sci Plant Nutr, 1981, 27: 317–328 [12] 姚元涛, 陶吉寒, 宋鲁彬, 田丽丽, 刘腾飞, 贾厚振. 钙、锰、铝及与硼的协同胁迫对茶树的毒害效应. 植物生理学报, 2015, 11: 1867–1872 Yao Y T, Yao J H, Song L B, Tian L L, Liu T F, Jia H Z. Poison effects of synergistic stress of calcium, manganese, aluminum and boron on tea plant. Plant Physiol J, 2015, 11: 1867–1872 [13] Yuan L Y, Yang S G, Liu B X, Zhang M, Wu K Q. Molecular characterization of a rice metal tolerance protein, OsMTP1. Plant Cell Rep, 2012, 31: 67–79 [14] Lorraine W E, Pittman Jon K. Dissecting pathways involved in manganese homeostasis and stress in higher plant cells. In: Hell R, Mendel R R, Eds. Cell Biology of Metals and Nutrients. Berlin, Germany: Springer-Verlag Berlin, Heidelberger Platz 3, D-14197, 2010. 95–117 [15] Paulsen I T, Saier M H. A novel family of ubiquitous heavy metal ion transport proteins. J Membr Biol, 1997, 156: 99–103 [16] Wei Y N, Fu D. Binding and transport of metal ions at the dimer interface of the Escherichia coli metal transporter YiiP. J Biol Chem, 2006, 281: 23492–23502 [17] Montanini B, Blaudez D, Jeandroz S, Sanders D, Chalot M. Phylogenetic and functional analysis of the cation diffusion facilitator (CDF) family: improved signature and prediction of substrate specificity. BMC Genomics, 2007, 8: 107 [18] Becher M, Talke I N, Krall L, Kramer U. Cross-species microarray transcript profiling reveals high constitutive expression of metal homeostasis genes in shoots of the zinc hyperaccumulator Arabidopsis halleri. Plant J, 2004, 37: 251–268 [19] Cho M, Chardonnens A N, Dietz K J. Differential heavy metal tolerance of Arabidopsis halleri and Arabidopsis thaliana: a leaf slice test. New Phytol, 2003, 158: 287–293 [20] Migocka M, Papierniak Anna, Maciaszczyk-Dziubinska E, Pozdzik P, Posyniak E, Garbiec A, Filleur S. Cucumber metal transport protein MTP8 confers increased tolerance to manganese when expressed in yeast and Arabidopsis thaliana. J Exp Bot, 2014, 65: 5367–5384 [21] Chen Z H, Fujii Y M, Yamaji N, Masuda S, Takemoto Y, Kamiya T, Yusuyin Y, Iwasaki K, Kato S I, Maeshima M, Ma J F, Ueno D. Mn tolerance in rice is mediated by MTP8.1, a member of the cation diffusion facilitator family. J Exp Bot, 2013, 64: 4375–4387 [22] Delhaize E, Gruber B D, Pittman J K, White R G, Leung H, Miao Y S, Jiang L W, Ryan P R, Richardson A E. A role for the AtMTP11 gene of Arabidopsis in manganese transport and tolerance. Plant J, 2007, 51: 198–210 [23] Tanaka N, Fujiwara T, Tomioka R, Kraemer U, Kawachi M, Maeshima M. Characterization of the histidine-rich loop of arabidopsis vacuolar membrane zinc transporter AtMTP1 as a sensor of zinc level in the cytosol. Plant Cell Physiol, 2015, 56: 510–519 [24] Arrivault S, Senger T, Kramer U. The Arabidopsis metal tolerance protein AtMTP3 maintains metal homeostasis by mediating Zn exclusion from the shoot under Fe deficiency and Zn oversupply. Plant J, 2006, 46: 861–879 [25] Gaitan-Solis E, Taylor N J, Siritunga D, Stevens W, Schachtman D P. Overexpression of the transporters AtZIP1 and AtMTP1 in cassava changes zinc accumulation and partitioning. Front Plant Sci, 2015, 6: 492 [26] Delhaize E, Gruber B D, Pittman J K, White R G, Leung H, Miao Y S, Jiang L W, Ryan P R, Richardson A E. A role for the AtMTP11 gene of Arabidopsis in manganese transport and tolerance. Plant J, 2007, 51: 198–210 [27] Delhaize E, Kataoka T, Hebb D M, White R G, Ryan P R. Genes encoding proteins of the cation diffusion facilitator family that confer manganese tolerance. Plant Cell, 2003, 15: 1131–1142 [28] Nies D H, Silver S. Ion efflux systems involved in bacterial metal resistances. J Ind Microbiol, 1995, 14: 186–199 [29] Kambe T, Narita H, Yamaguchi-Iwai Y, Hirose J, Amano T, Sugiura N, Sasaki R, Mori K, Iwanaga T, Nagao M. Cloning and characterization of a novel mammalian zinc transporter, zinc transporter 5, abundantly expressed in pancreatic beta cells. J Biol Chem, 2002, 277: 19049–19055 [30] Singh S, Parihar P, Singh R, Singh V. P, Prasad S. M. Heavy metal tolerance in plants: role of transcriptomics, proteomics, metabolomics, and lonomics. Front Plant Sci, 2016, 6: 1143-1178 [31] Kobae Y, Uemura T, Sato M H, Ohnishi M, Mimura T, Nakagawa T, Maeshima M. Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis. Plant Cell Physiol, 2004, 45: 1749–1758 [32] Desbrosses-Fonrouge A G, Voigt K, Schroder A, Arrivault S, Thomine S, Kramer U. Arabidopsis thaliana MTP1 is a Zn transporter in the vacuolar membrane which mediates Zn detoxification and drives leaf Zn accumulation. Febs Lett, 2005, 579: 4165–4174

[1] ZHANG Ling-Xiao,JIAO Zhen-Zhen,BU Hua-Hu,WANG Yi-Ru,LI Jian,ZHENG Jun. Functional identification of maize cation/proton antiporter ZmNHX7 [J]. Acta Agronomica Sinica, 2020, 46(8): 1185-1194.
[2] JIANG Hui-Bing,YANG Sheng-Mei,LIU Yu-Fei,TIAN Yi-Ping,SUN Yun-Nan,CHEN Lin-Bo,TANG Yi-Chun. Biological characteristics and cytological studies on anther abortion of male sterile Camellia crassocolumna [J]. Acta Agronomica Sinica, 2020, 46(7): 1076-1086.
[3] Tao FENG,Chun-Yun GUAN. Cloning and characterization of phytochrome interacting factor 4 (BnaPIF4) gene from Brassica napus L. [J]. Acta Agronomica Sinica, 2019, 45(2): 204-213.
[4] Yu-Zhang CHEN,Shou-Xi CHAI,Hong-Bo CHENG,Yu-Wei CHAI,Chang-Gang YANG,Kai-Min TAN,Lei CHANG. Effects of straw-incorporation combined with autumn plastic mulching on soil water consumption characteristics and winter wheat yield in arid farming areas [J]. Acta Agronomica Sinica, 2019, 45(2): 256-266.
[5] Tao FENG,Chun-Yun GUAN. Cloning and Characterization of Brassinazole-resistant (BnaBZR1 and BnaBES1) CDS from Brassica napus L. [J]. Acta Agronomica Sinica, 2018, 44(12): 1793-1801.
[6] Zhang PENG, Hua-Rong TONG, Guo-Lu LIANG, Yi-Qi SHI, Lian-Yu YUAN. Protoplast Isolation and Fusion Induced by PEG with Leaves and Roots of Tea Plant (Camellia sinensis L. O. Kuntze) [J]. Acta Agronomica Sinica, 2018, 44(03): 463-470.
[7] HAO Xin-Yuan,YUEChuan,TANG Hu,QIAN Wen-Jun,WANG Yu-Chun,WANG Lu, WANG Xin-Chao,YANG Ya-Jun. Cloning of β-amylase Gene (CsBAM3) and ItsExpression ModelResponseto Cold Stress in Tea Plant [J]. Acta Agron Sin, 2017, 43(10): 1417-1425.
[8] ZHANG Yan-Wei,ZHANG Li-Feng,LI Wei,WANG Cai-Jie,ZHANG Jun,XU Ran. Identification, Expression and Variation Analysis of Salt Tolerance Related GmNAC Genes in Soybean [J]. Acta Agron Sin, 2016, 42(07): 990-999.
[9] TAN Tai-Long,FENG Tao,LUO Hai-Yan,PENG Ye,LIU Rui-Yang,GUAN Chun-Yun. Cloning and Characterization of Phospholipids:Diacylglycerol Acyltransferase (BnPDAT1) cDNA from Brassica napus L. [J]. Acta Agron Sin, 2016, 42(05): 658-666.
[10] QIAN Wen-Jun,YUE Chuan,CAO Hong-Li,HAO Xin-Yuan,WANG Lu,WANG Yu-Chun,HUANG Yu-Ting,WANG Bo,WANG Xin-Chao,XIAO Bin,YANG Ya-Jun. Cloning and Expression Analysis of a Neutral/alkaline Invertase Gene (CsINV10) in Tea Plant (Camellia sinensis L. O. Kuntze) [J]. Acta Agron Sin, 2016, 42(03): 376-388.
[11] WANG Bo,CAO Hong-Li,HUANG Yu-Ting,HU Yu-Rong,QIAN Wen-Jun,HAO Xin-Yuan, WANG Lu,YANG Ya-Jun,WANG Xin-Chao. Cloning and Expression Analysis of Auxin Efflux Carrier Gene CsPIN3 in Tea Plant (Camellia sinensis) [J]. Acta Agron Sin, 2016, 42(01): 58-69.
[12] MA Li-Gong,ZHANG Yun-Hua,MENG Qing-Lin,SHI Feng-Mei,LIU Jia,LI Yi-Chu,WANG Zhi-Ying. Cloning and Function Analysis of Pathogenesis Related Protein Gene HaPR1 from Sunflower (Helianthus annuus) [J]. Acta Agron Sin, 2015, 41(12): 1819-1827.
[13] MA Chun-Lei,YAO Ming-Zhe,WANG Xin-Chao,JIN Ji-Qiang,MA Jian-Qiang,CHEN Liang. Cloning and Expression of Three Genes Involved in the Biosynthesis of Chlorophyll in Different Albescent Stages of “Baiye 1” [J]. Acta Agron Sin, 2015, 41(02): 240-250.
[14] LIU Shao-Jie,CHI Lin,XIE Wen-Gang,HAN Nan,CHEN Ying-Hui,TANG Qian. Genetic Diversity of Gulin-niupicha (Camellia sinensis [L.] O. Kuntze) Germplasm Resources [J]. Acta Agron Sin, 2014, 40(12): 2118-2127.
[15] CAO Hong-Li,YUE Chuan,ZHOU Yan-Hua,WANG Lu,HAO Xin-Yuan,YANG Ya-Jun*,WANG Xin-Chao*. Molecular Cloning and Expression of a bZIP Transcription Factor Gene CsbZIP1 in Tea Plant (Camellia sinensis) [J]. Acta Agron Sin, 2014, 40(09): 1702-1709.
Full text



No Suggested Reading articles found!