作物学报 ›› 2017, Vol. 43 ›› Issue (05): 708-717.doi: 10.3724/SP.J.1006.2017.00708
袁连玉,陈应娟,魏旭,童华荣*
YUAN Lian-Yu,CHEN Ying-Juan,WEI Xu,TONG Hua-Rong*
摘要:
金属耐受蛋白MTP (metal tolerance protein)是阳离子转运蛋白(CDF)家族的重要成员, 在植物重金属转运过程中发挥重要调控作用。本研究以中茶108茶树为试验材料, 通过RT-PCR和RACE方法克隆到茶树重金属耐受蛋白基因CsMTP11 (GenBank登录号为KX450265), 其全长cDNA为1197 bp, 编码398个氨基酸残基, 其编码蛋白分子量为44.85 kD, 等电点为5.34。在线软件分析表明, CsMTP11蛋白具有5个跨膜结构域, 且含有CDF家族的其他保守结构域。系统进化树分析结果表明, 茶树CsMTP11与葡萄VvMTP11进化同源性最近, 其氨基酸序列相似度高达90%。基因表达模式分析表明, CsMTP11基因在茶树老叶中的表达量最高, 根中的表达量最低, 另外, CsMTP11基因受重金属Mn和Co离子胁迫诱导表达。CsMTP11-YFP融合蛋白在拟南芥原生质体共定位试验表明, CsMTP11-YFP融合蛋白定位于质膜。CsMTP11在酿酒酵母及其突变株中的异源表达可以提高其对重金属Mn和Co离子的耐受性。综上所述, 茶树CsMTP11属于Mn-CDF亚家族, 可能参与茶树对重金属锰和钴的转运过程。
| [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 |
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