缺Ca/Fe及盐分胁迫下番茄Pb吸收转运与关键基因的表达关系
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摘要
选取台湾黄圣女番茄为供试材料,通过水培试验研究缺Ca/Fe及盐分胁迫三种处理对Pb在番茄中富集的影响,并测定不同处理下根部CNGC1(环核苷酸门控非选择性阳离子通道)和HMA3(P1B-ATPase酶)基因的相对表达量,进而探究CNGC1和HMA3基因的表达对番茄吸收累积Pb的影响。结果表明:与对照相比,缺钙处理显著增强了根对Pb的吸收能力并抑制了Pb往地上部转运过程,缺Fe处理对根吸收能力无显著影响而增加了往地上部转运量,盐分处理同时增强了根部吸收及往地上部转运能力;不同处理下番茄根部单位质量Pb净吸收量与CNGC1离子通道的活性水平呈现出极显著正相关(P<0.01),说明番茄根部CNGC1离子通道在不同处理下Pb的吸收可能起着重要作用;番茄Pb转运系数与HMA3的表达量呈现极显著负相关(P<0.01),说明番茄HMA3的高表达抑制了番茄对Pb的地上转运能力。因此,缺Ca/Fe及盐分胁迫处理对Pb在番茄中富集有不同影响,CNGC1离子通道和HMA3的表达量在一定程度上决定了Pb在番茄中的分布。
A hydroponic experiment was conducted to investigate the characteristics of lead(Pb)accumulation in tomato(Lycopersicon esculentum Mill)exposed to Fe-deficiency, Ca-deficiency, and salinity stress. The genes HMA3 and CNGC1 were located by homology comparison, and their gene expression levels in roots were determined. These results were used to discuss the effect of these two gene expressions on Pb accumulation. The results showed that:(1)Compared with control groups, the Ca-deficiency treatment significantly increased Pb uptake by roots, but inhibited root-to-shoot Pb translocation. The Fe-deficiency treatment significantly increased the amount of Pb translocation from roots to shoots but did not significantly influence root Pb uptake. The salinity treatment significantly enhanced the uptake and transportation of Pb.(2)A very significant positive correlation between Pb uptake by per unit root mass and activity level of CNGC1 cation channel was observed in the three treatments, indicating that CNGC1 cation channel played an important role in Pb uptake. Furthermore,there was a negative correlation between the expression of HMA3 and Pb translocation from roots to shoots, implying that high expression of HMA3 restricted Pb transport to the shoots. Hence, Fe-deficiency, Ca-deficiency, and salinity stress have different effects on Pb accumulation in tomato, and expression levels of HMA3 and CNGC1 control the distribution of Pb in tomato.
A hydroponic experiment was conducted to investigate the characteristics of lead(Pb)accumulation in tomato(Lycopersicon esculentum Mill)exposed to Fe-deficiency, Ca-deficiency, and salinity stress. The genes HMA3 and CNGC1 were located by homology comparison, and their gene expression levels in roots were determined. These results were used to discuss the effect of these two gene expressions on Pb accumulation. The results showed that:(1)Compared with control groups, the Ca-deficiency treatment significantly increased Pb uptake by roots, but inhibited root-to-shoot Pb translocation. The Fe-deficiency treatment significantly increased the amount of Pb translocation from roots to shoots but did not significantly influence root Pb uptake. The salinity treatment significantly enhanced the uptake and transportation of Pb.(2)A very significant positive correlation between Pb uptake by per unit root mass and activity level of CNGC1 cation channel was observed in the three treatments, indicating that CNGC1 cation channel played an important role in Pb uptake. Furthermore,there was a negative correlation between the expression of HMA3 and Pb translocation from roots to shoots, implying that high expression of HMA3 restricted Pb transport to the shoots. Hence, Fe-deficiency, Ca-deficiency, and salinity stress have different effects on Pb accumulation in tomato, and expression levels of HMA3 and CNGC1 control the distribution of Pb in tomato.
引文
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[6]介明.盐碱地的危害与改良研究[J].绿色科技, 2016(7):122-124.JIE Ming. Effects of nitrogen-phosphate fertilization on camellia[J].Journal of Green Science and Technology, 2016(7):122-124.
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[8]胡妮,陈柯罕,李取生,等.盐胁迫下苋菜品种有机酸变化对Cd累积和耐盐性的影响[J].农业环境科学学报, 2016, 35(5):858-864.HU Ni, CHEN Ke-han, LI Qu-sheng, et al. Effects of salinity-inducted organic acid variation on Cd accumulation and salinity tolerance of edible amaranth[J]. Journal of Agro-Environment Science, 2016, 35(5):858-864.
[9]陈秀玲,张磊.小麦/花生不同间作方式对花生吸收积累Cd的影响[J].环境化学, 2014, 33(9):1469-1475.CHEN Xiu-ling, ZHANG Lei. Influence on cadmium uptake by peanut of wheat/peanut intercropping modes[J]. Environmental Chemistry, 2014,33(9):1469-1475.
[10] Kizhaerals S, Virgine T, Kaliyaperumal J, et al. Biochemical changes and zinc fractions in arbuscular mycorrhizal fungus(Glomus intraradices)inoculated and uninoculated soils under differential zinc fertilization[J]. Applied Soil Ecology, 2009, 43(1):32-39.
[11]陈惠君,谭玲,李取生,等. Cr/Pb低累积菜心品种筛选及其根际机理研究[J].农业环境科学学报, 2016, 35(7):1249-1256.CHEN Hui-jun, TAN Ling, LI Qu-sheng, et al. Screening and preliminary rhizosphere mechanisms of low Cr/Pb accumulation cultivars of Chinese flowering cabbages(Brassica parachinensis L.)[J]. Journal of Agro-Environment Science, 2016, 35(7):1249-1256.
[12] Cuypers A, Smeets K, Vangronsveld J. Heavy metal stress in plants[M]//Plant stress biology:From genomics to systems biology. WileyVCH Verlag GmbH&Co. KGaA, 1999:161-178.
[13]余丹萍,李取生,王立立,等.缺Fe/Zn及盐胁迫下苋菜对Cd及矿质元素的吸收与IRT1表达的关系[J].农业环境科学学报, 2016, 35(2):234-239.YU Dan-ping, LI Qu-sheng, WANG Li-li, et al. Cadmium and mineral element accumulation and IRT1 gene expression of Amaranth hybidus L. under iron/zinc deficiencies or salt stress[J]. Journal of AgroEnvironment Science, 2016, 35(2):234-239.
[14]李楠.大豆NRAMP家族基因的克隆及其功能研究[D].北京:中国科学院大学, 2016.LI Nan. Cloning and function study of soybean NRAMP family gene[D]. Beijing:University of Chinese Academy of Sciences, 2016.
[15] Morel M, Crouzet J, Gravot A, et al. AtHMA3, a P1B-ATPase allowing Cd/Zn/Co/Pb vacuolar storage in Arabidopsis[J]. Plant Physiology, 2009, 149(2):894-904.
[16] Wojas S, Ruszczyńska A, Bulska E, et al. Ca2+-dependent plant response to Pb2+, is regulated by LCT1[J]. Environmental Pollution,2007, 147(3):584-592.
[17] Lee S, Kim Y Y, Lee Y, et al. Rice P1B-type heavy-metal ATPase,OsHMA9, is a metal efflux protein[J]. Plant Physiology, 2007, 145(3):831-842.
[18] Lee Y. AtPDR12 contributes to lead resistance in Arabidopsis[J].Plant Physiology, 2005, 138(2):827-836.
[19] Sunkar R, Kaplan B, Bouch N, et al. Expression of a truncated tobacco NtCBP4 channel in transgenic plants and disruption of the homologous Arabidopsis CNGC1 gene confer Pb2+tolerance[J]. Plant Journal,2000, 24(4):533-542.
[20] Arazi T, Sunkar R, Kaplan B, et al. A tobacco plasma membrane calmodulin-binding transporter confers Ni2+, tolerance and Pb2+, hypersensitivity in transgenic plants[J]. The Plant Journal, 1999, 20(2):171-182.
[21] Mei X, Li S, Li Q, et al. Sodium chloride salinity reduces Cd uptake by edible amaranth(Amaranthus mangostanus L.)via competition for Ca channels[J]. Ecotoxicology&Environmental Safety, 2014, 105(7):59-64.
[22]李仁,吴新新,李蔚,等.番茄水通道蛋白基因SIAQP的克隆与序列分析[J].中国农业科学, 2012, 45(2):302-310.LI Ren, WU Xin-xin, LI Wei, et al. Cloning and sequence analysis of the aquaporins gene SlAQP in tomato[J]. Scientia Agricultura Sinica,2012, 45(2):302-310.
[23] Sterckeman T, Redjala T, Morel J L. Influence of exposure solution composition and of plant cadmium content on root cadmium shortterm uptake[J]. Environmental&Experimental Botany, 2011, 74(1):131-139.
[24] Ye J, Yan C, Liu J, et al. Effects of silicon on the distribution of cadmium compartmentation in root tips of Kandelia obovata(S. L.)Yong.[J].Environmental Pollution, 2012, 162(5):369-373.
[25] Marek V, Cornelia K, Ingrid L, et al. Root anatomy and element distribution vary between two Salix caprea isolates with different Cd accumulation capacities[J]. Environmental Pollution, 2012, 163(1):117-126.
[26]王月,侯和胜.植物环核苷酸门控通道(CNGC)基因家族的结构与功能[J].植物生理学报, 2007, 43(1):1-8.WANG Yue, HOU He-sheng. Structure and function of plant cyclic nucleotide-gated channel(CNGC)gene family[J]. Plant Physiology Journal, 2007, 43(1):1-8.
[27] Kaplan B, Sherman T, Fromm H. Cyclic nucleotide-gated channels in plants.[J]. Febs Letters, 2007, 581(12):2237-2246.
[28]刘海娇,杜立群,林金星,等.植物环核苷酸门控离子通道及其功能研究进展[J].植物学报, 2015, 50(6):779-789.LIU Hai-jiao, DU Li-qun, LIN Jin-xing, et al. Recent advances in cyclic nucleotide-gated ion channels with their functions in plants[J].Chinese Bulletin of Botany, 2015, 50(6):779-789.
[29] Argüello J M, Eren E, Gonzálezguerrero M. The structure and function of heavy metal transport P1B-ATPases[J]. Biometals, 2007, 20(3/4):233.
[30]余刚.四翅滨藜抗逆相关基因的筛选及其通道蛋白功能解析[D].长春:吉林大学, 2016.YU Gang. Functional isolation of stress-responsive genes from Atriplex canescens and functional characterization of multiple transporters[D]. Changchun:Jilin University, 2016.
[31]侯晓龙,陈加松,刘爱琴,等. Pb胁迫对金丝草体内Pb化学形态及细胞分布的影响[J].生态与农村环境学报, 2012, 28(3):271-276.HOU Xiao-long, CHEN Jia-song, LIU Ai-qin, et al. Effects of Pb stress on fractionation and distribution of Pb in Pogonatherum crinitum[J]. Journal of Ecology and Rural Environment, 2012, 28(3):271-276.
[32]徐劼,于明革,陈英旭,等.铅在茶树体内的分布及化学形态特征[J].应用生态学报, 2011, 22(4):891-896.XU Jie, YU Ming-ge, CHEN Ying-xu, et al. Characteristic of distribution and chemical forms of Pb in tea plant varieties[J]. Chinese Journal of Applied Ecology, 2011, 22(4):891-896.
[33]孙伟安,李以圭,陈新明.金属配合物溶液的热力学研究Ⅰ. Pb(Ⅱ)-Cl-配位体系[J].深圳大学学报, 1992(增刊2):37-45.SUN Wei-an, LI Yi-gui, CHEN Xin-ming. Thermodynamics of the metal complexes in aqueous solutionsⅠ. Pb(Ⅱ)-Cl-complex system[J]. Journal of Shenzhen University Science and Engineering, 1992(Suppl 2):37-45.
[2]陈梦妮,杜天庆,张泽宇,等.苗期甜高粱和玉米对土壤Pb胁迫的响应及抗性评价[J].中国农业大学学报, 2016, 21(11):17-23.CHEN Meng-ni, DU Tian-qing, ZHANG Ze-yu, et al. Resistance evaluation and response of sweet sorghum and corn during seedling stage under Pb stress[J]. Journal of China Agricultural University, 2016, 21(11):17-23
[3]蔡立梅,黄兰椿,周永章,等.东莞市农业土壤和蔬菜铅含量特征分析[J].环境科学与技术, 2010, 33(2):122-125.CAI Li-mei, HUANG Lan-chun, ZHOU Yong-zhang, et al. Characteristic analysis of lead concentration in agricultural topsoil and vegetables of Dongguan Guangdong Province[J]. Environmental Science and Technology, 2010, 33(2):122-125.
[4]雷琼,马文哲,王文光.我国蔬菜重金属铅污染研究进展与防治措施[J].农产品加工(学刊), 2012(9):105-107.LEI Qiong, MA Wen-zhe, WANG Wen-guang. The heavy metal lead pollution progress and control measures of the vegetables in China[J].Academic Periodical of Farm Products Processing, 2012(9):105-107.
[5] Gabrijel O, Davor R, Zed R, et al. Cadmium accumulation by muskmelon under salt stress in contaminated organic soil[J]. Science of the Total Environment, 2009, 407(7):2175-2182.
[6]介明.盐碱地的危害与改良研究[J].绿色科技, 2016(7):122-124.JIE Ming. Effects of nitrogen-phosphate fertilization on camellia[J].Journal of Green Science and Technology, 2016(7):122-124.
[7]李取生,楚蓓,石雷,等.珠江口滩涂湿地土壤重金属分布及其对围垦的影响[J].农业环境科学学报, 2007, 26(4):224-228.LI Qu-sheng, CHU Bei, SHI Lei, et al. Heavy metal distribution in tidal wetland soils and its effect on reclamation in the Pearl River Estuary[J]. Journal of Agro-Environment Science, 2007, 26(4):224-228.
[8]胡妮,陈柯罕,李取生,等.盐胁迫下苋菜品种有机酸变化对Cd累积和耐盐性的影响[J].农业环境科学学报, 2016, 35(5):858-864.HU Ni, CHEN Ke-han, LI Qu-sheng, et al. Effects of salinity-inducted organic acid variation on Cd accumulation and salinity tolerance of edible amaranth[J]. Journal of Agro-Environment Science, 2016, 35(5):858-864.
[9]陈秀玲,张磊.小麦/花生不同间作方式对花生吸收积累Cd的影响[J].环境化学, 2014, 33(9):1469-1475.CHEN Xiu-ling, ZHANG Lei. Influence on cadmium uptake by peanut of wheat/peanut intercropping modes[J]. Environmental Chemistry, 2014,33(9):1469-1475.
[10] Kizhaerals S, Virgine T, Kaliyaperumal J, et al. Biochemical changes and zinc fractions in arbuscular mycorrhizal fungus(Glomus intraradices)inoculated and uninoculated soils under differential zinc fertilization[J]. Applied Soil Ecology, 2009, 43(1):32-39.
[11]陈惠君,谭玲,李取生,等. Cr/Pb低累积菜心品种筛选及其根际机理研究[J].农业环境科学学报, 2016, 35(7):1249-1256.CHEN Hui-jun, TAN Ling, LI Qu-sheng, et al. Screening and preliminary rhizosphere mechanisms of low Cr/Pb accumulation cultivars of Chinese flowering cabbages(Brassica parachinensis L.)[J]. Journal of Agro-Environment Science, 2016, 35(7):1249-1256.
[12] Cuypers A, Smeets K, Vangronsveld J. Heavy metal stress in plants[M]//Plant stress biology:From genomics to systems biology. WileyVCH Verlag GmbH&Co. KGaA, 1999:161-178.
[13]余丹萍,李取生,王立立,等.缺Fe/Zn及盐胁迫下苋菜对Cd及矿质元素的吸收与IRT1表达的关系[J].农业环境科学学报, 2016, 35(2):234-239.YU Dan-ping, LI Qu-sheng, WANG Li-li, et al. Cadmium and mineral element accumulation and IRT1 gene expression of Amaranth hybidus L. under iron/zinc deficiencies or salt stress[J]. Journal of AgroEnvironment Science, 2016, 35(2):234-239.
[14]李楠.大豆NRAMP家族基因的克隆及其功能研究[D].北京:中国科学院大学, 2016.LI Nan. Cloning and function study of soybean NRAMP family gene[D]. Beijing:University of Chinese Academy of Sciences, 2016.
[15] Morel M, Crouzet J, Gravot A, et al. AtHMA3, a P1B-ATPase allowing Cd/Zn/Co/Pb vacuolar storage in Arabidopsis[J]. Plant Physiology, 2009, 149(2):894-904.
[16] Wojas S, Ruszczyńska A, Bulska E, et al. Ca2+-dependent plant response to Pb2+, is regulated by LCT1[J]. Environmental Pollution,2007, 147(3):584-592.
[17] Lee S, Kim Y Y, Lee Y, et al. Rice P1B-type heavy-metal ATPase,OsHMA9, is a metal efflux protein[J]. Plant Physiology, 2007, 145(3):831-842.
[18] Lee Y. AtPDR12 contributes to lead resistance in Arabidopsis[J].Plant Physiology, 2005, 138(2):827-836.
[19] Sunkar R, Kaplan B, Bouch N, et al. Expression of a truncated tobacco NtCBP4 channel in transgenic plants and disruption of the homologous Arabidopsis CNGC1 gene confer Pb2+tolerance[J]. Plant Journal,2000, 24(4):533-542.
[20] Arazi T, Sunkar R, Kaplan B, et al. A tobacco plasma membrane calmodulin-binding transporter confers Ni2+, tolerance and Pb2+, hypersensitivity in transgenic plants[J]. The Plant Journal, 1999, 20(2):171-182.
[21] Mei X, Li S, Li Q, et al. Sodium chloride salinity reduces Cd uptake by edible amaranth(Amaranthus mangostanus L.)via competition for Ca channels[J]. Ecotoxicology&Environmental Safety, 2014, 105(7):59-64.
[22]李仁,吴新新,李蔚,等.番茄水通道蛋白基因SIAQP的克隆与序列分析[J].中国农业科学, 2012, 45(2):302-310.LI Ren, WU Xin-xin, LI Wei, et al. Cloning and sequence analysis of the aquaporins gene SlAQP in tomato[J]. Scientia Agricultura Sinica,2012, 45(2):302-310.
[23] Sterckeman T, Redjala T, Morel J L. Influence of exposure solution composition and of plant cadmium content on root cadmium shortterm uptake[J]. Environmental&Experimental Botany, 2011, 74(1):131-139.
[24] Ye J, Yan C, Liu J, et al. Effects of silicon on the distribution of cadmium compartmentation in root tips of Kandelia obovata(S. L.)Yong.[J].Environmental Pollution, 2012, 162(5):369-373.
[25] Marek V, Cornelia K, Ingrid L, et al. Root anatomy and element distribution vary between two Salix caprea isolates with different Cd accumulation capacities[J]. Environmental Pollution, 2012, 163(1):117-126.
[26]王月,侯和胜.植物环核苷酸门控通道(CNGC)基因家族的结构与功能[J].植物生理学报, 2007, 43(1):1-8.WANG Yue, HOU He-sheng. Structure and function of plant cyclic nucleotide-gated channel(CNGC)gene family[J]. Plant Physiology Journal, 2007, 43(1):1-8.
[27] Kaplan B, Sherman T, Fromm H. Cyclic nucleotide-gated channels in plants.[J]. Febs Letters, 2007, 581(12):2237-2246.
[28]刘海娇,杜立群,林金星,等.植物环核苷酸门控离子通道及其功能研究进展[J].植物学报, 2015, 50(6):779-789.LIU Hai-jiao, DU Li-qun, LIN Jin-xing, et al. Recent advances in cyclic nucleotide-gated ion channels with their functions in plants[J].Chinese Bulletin of Botany, 2015, 50(6):779-789.
[29] Argüello J M, Eren E, Gonzálezguerrero M. The structure and function of heavy metal transport P1B-ATPases[J]. Biometals, 2007, 20(3/4):233.
[30]余刚.四翅滨藜抗逆相关基因的筛选及其通道蛋白功能解析[D].长春:吉林大学, 2016.YU Gang. Functional isolation of stress-responsive genes from Atriplex canescens and functional characterization of multiple transporters[D]. Changchun:Jilin University, 2016.
[31]侯晓龙,陈加松,刘爱琴,等. Pb胁迫对金丝草体内Pb化学形态及细胞分布的影响[J].生态与农村环境学报, 2012, 28(3):271-276.HOU Xiao-long, CHEN Jia-song, LIU Ai-qin, et al. Effects of Pb stress on fractionation and distribution of Pb in Pogonatherum crinitum[J]. Journal of Ecology and Rural Environment, 2012, 28(3):271-276.
[32]徐劼,于明革,陈英旭,等.铅在茶树体内的分布及化学形态特征[J].应用生态学报, 2011, 22(4):891-896.XU Jie, YU Ming-ge, CHEN Ying-xu, et al. Characteristic of distribution and chemical forms of Pb in tea plant varieties[J]. Chinese Journal of Applied Ecology, 2011, 22(4):891-896.
[33]孙伟安,李以圭,陈新明.金属配合物溶液的热力学研究Ⅰ. Pb(Ⅱ)-Cl-配位体系[J].深圳大学学报, 1992(增刊2):37-45.SUN Wei-an, LI Yi-gui, CHEN Xin-ming. Thermodynamics of the metal complexes in aqueous solutionsⅠ. Pb(Ⅱ)-Cl-complex system[J]. Journal of Shenzhen University Science and Engineering, 1992(Suppl 2):37-45.