喷施不同形态锰肥对叶用油菜镉累积及亚细胞分布的影响
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摘要
为探明喷施锰肥对叶用油菜累积镉的调控作用,采用盆栽试验,研究喷施不同浓度的MnSO_4、Mn(CH_3COO)_2和EDTA·Na_2Mn对两种叶用油菜(Cd低积累品种华骏和普通品种寒绿)生物量以及Cd和Mn含量的影响,并通过分析Cd在叶用油菜体内的吸收转运规律以及亚细胞分布特征揭示其作用机理。结果表明,喷施3种形态Mn肥对Cd低积累叶用油菜地上部和根部生物量无显著影响,喷施高浓度Mn(CH_3COO)_2和EDTA·Na_2Mn显著降低普通叶用油菜地上部生物量。喷施MnSO_4和Mn(CH_3COO)_2处理降低叶用油菜Cd含量、提高Mn含量的效果明显优于EDTA·Na_2Mn处理,其中,0.4 g·L~(-1)Mn的MnSO_4和0.2 g·L~(-1)Mn的Mn(CH_3COO)_2处理降低Cd低积累叶用油菜地上部Cd含量的效果最佳,可使其分别降低29.2%和31.0%。喷施Mn肥显著降低叶用油菜总Cd累积量,使Cd转运系数显著升高,叶用油菜地上部Cd含量与总Cd累积量呈极显著的相关性(P<0.01),而与Cd转运系数则无显著相关性。Cd和Mn在叶用油菜地上部主要分布在细胞可溶组分中,其次为细胞壁。喷施Mn肥后,Cd和Mn在细胞壁组分的分配比例降低,在可溶组分的分配比例升高。总之,喷施0.4 g·L~(-1)Mn的MnSO_4和0.2 g·L~(-1)Mn的Mn(CH_3COO)_2可以显著降低低积累叶用油菜地上部Cd含量,提高其Mn含量,同时不影响叶用油菜生长,是保障Cd低积累叶用油菜安全优质生产的较好措施。
In order to test the effect of foliar application of manganese(Mn)fertilizers on cadmium(Cd)accumulation in pak choi(Brassica chinensis), three different forms of Mn, MnSO_4, Mn(CH_3COO)_2 and EDTA·Na_2Mn, were sprayed among two pak choi cultivars(Huajun and Hanlv)in a pot experiment. We examined the biomass, Cd and Mn concentrations, Cd uptake and transport, and Cd and Mn subcellular distribution in pak choi. The results show that foliar application of Mn had no significant effect on the shoot and root biomasses of the low Cd-accumulating cultivar(Huajun); however, the foliar application of high dosages of Mn(CH_3COO)_2 and EDTA·Na_2Mn significantly reduced shoot biomass in the common cultivar of pak choi(Hanlv). Foliar application with MnSO_4 and Mn(CH_3COO)_2 resulted in greater alleviation of shoot Cd concentrations and a more notable enhancement of shoot Mn concentrations in pak choi than by spraying with EDTA·Na_2Mn. Shoot Cd concentrations in Huajun were the lowest under foliar feeding with 0.4 g·L~(-1) Mn(MnSO_4)and 0.2 g·L~(-1) Mn [Mn(CH_3COO)_2],which were decreased by 29.2% and 31.0%, respectively. Foliar Mn application reduced the total Cd accumulation significantly and increased the Cd transfer factor of pak choi markedly. The shoot Cd concentration of pak choi was positively correlated(P<0.01)with total Cd accumulation; however, it was not significantly correlated with the Cd transfer factor. In shoots of pak choi, the majority of Cd and Mn were associated with the soluble fraction, followed by the cell wall fraction. After foliar Mn application, the proportions of Cd and Mn in the cell wall fraction decreased and those in the soluble fraction increased. In summary, foliar feeding with 0.4 g·L~(-1) Mn(MnSO_4)and 0.2 g·L~(-1) Mn [Mn(CH_3COO)_2] significantly reduced Cd concentrations and increased Mn concentrations in shoots, and it did not affect the growth of Huajun, which is suitable for the safe and high-quality production of pak choi.
In order to test the effect of foliar application of manganese(Mn)fertilizers on cadmium(Cd)accumulation in pak choi(Brassica chinensis), three different forms of Mn, MnSO_4, Mn(CH_3COO)_2 and EDTA·Na_2Mn, were sprayed among two pak choi cultivars(Huajun and Hanlv)in a pot experiment. We examined the biomass, Cd and Mn concentrations, Cd uptake and transport, and Cd and Mn subcellular distribution in pak choi. The results show that foliar application of Mn had no significant effect on the shoot and root biomasses of the low Cd-accumulating cultivar(Huajun); however, the foliar application of high dosages of Mn(CH_3COO)_2 and EDTA·Na_2Mn significantly reduced shoot biomass in the common cultivar of pak choi(Hanlv). Foliar application with MnSO_4 and Mn(CH_3COO)_2 resulted in greater alleviation of shoot Cd concentrations and a more notable enhancement of shoot Mn concentrations in pak choi than by spraying with EDTA·Na_2Mn. Shoot Cd concentrations in Huajun were the lowest under foliar feeding with 0.4 g·L~(-1) Mn(MnSO_4)and 0.2 g·L~(-1) Mn [Mn(CH_3COO)_2],which were decreased by 29.2% and 31.0%, respectively. Foliar Mn application reduced the total Cd accumulation significantly and increased the Cd transfer factor of pak choi markedly. The shoot Cd concentration of pak choi was positively correlated(P<0.01)with total Cd accumulation; however, it was not significantly correlated with the Cd transfer factor. In shoots of pak choi, the majority of Cd and Mn were associated with the soluble fraction, followed by the cell wall fraction. After foliar Mn application, the proportions of Cd and Mn in the cell wall fraction decreased and those in the soluble fraction increased. In summary, foliar feeding with 0.4 g·L~(-1) Mn(MnSO_4)and 0.2 g·L~(-1) Mn [Mn(CH_3COO)_2] significantly reduced Cd concentrations and increased Mn concentrations in shoots, and it did not affect the growth of Huajun, which is suitable for the safe and high-quality production of pak choi.
引文
[1] Huang Y Y, He C T, Shen C, et al. Toxicity of cadmium and its health risks from leafy vegetable consumption[J]. Food&Function, 2017, 8(4):1373-1401.
[2]李霞,张慧鸣,徐震,等.农田Cd和Hg污染的来源解析与风险评价研究[J].农业环境科学学报, 2016, 35(7):1314-1320.LI Xia, ZHANG Hui-ming, XU Zhen, et al. Source apportionment and risk assessment of Cd and Hg pollution in farmland[J]. Journal of AgroEnvironment Science, 2016, 35(7):1314-1320.
[3] Chen H P, Yang X P, Wang P, et al. Dietary cadmium intake from rice and vegetables and potential health risk:A case study in Xiangtan,southern China[J]. Science of the Total Environment, 2018, 639:271–277.
[4]黄道友,朱奇宏,朱捍华,等.重金属污染耕地农业安全利用研究进展与展望[J].农业现代化研究, 2018, 39(6):1030-1043.HUANG Dao-you, ZHU Qi-hong, ZHU Han-hua, et al. Advances and prospects of safety agro-utilization of heavy metal contaminated farmland soil[J]. Research of Agricultural Modernization, 2018, 39(6):1030-1043.
[5]温娜,王景安,刘仲齐.利用AMMI模型分析稻米镉含量的基因型与环境互作效应[J].农业环境科学学报, 2015, 34(5):817-823.WEN Na, WANG Jing-an, LIU Zhong-qi. Analysis of genotypic and environmental effects on cadmium content in rice by AMMI model[J].Journal of Agro-Environment Science, 2015, 34(5):817-823.
[6] Socha A L, Guerinot M L. Mn-euvering manganese:The role of transporter gene family members in manganese uptake and mobilization in plants[J]. Frontiers in Plant Science, 2014, 5:106-122.
[7] Ullah I, Wang Y, Eide D J, et al. Evolution, and functional analysis of Natural Resistance-Associated Macrophage Proteins(NRAMPs)from Theobroma cacao and their role in cadmium accumulation[J]. Scientific Reports, 2018, 8(1):14412.
[8] Huang Q N, An H, Yang Y J, et al. Effects of Mn-Cd antagonistic interaction on Cd accumulation and major agronomic traits in rice genotypes by different Mn forms[J]. Plant Growth Regulation, 2017, 82(2):317-331.
[9]尹晓辉,邹慧玲,方雅瑜,等.施锰方式对水稻吸收积累镉的影响研究[J].环境科学与技术, 2017, 40(8):8-12.YIN Xiao-hui, ZOU Hui-ling, FANG Ya-yu, et al. Effects of manganese fertilizer on absorption and accumulation of Cd in rice[J]. Environmental Science&Technology, 2017, 40(8):8-12.
[10] Wang L, Xu Y M, Sun Y B, et al. Identification of pakchoi cultivars with low cadmium accumulation and soil factors that affect their cadmium uptake and translocation[J]. Frontiers of Environmental Science and Engineering, 2014, 8(6):877-887.
[11] Liang X F, Qin X, Huang Q Q, et al. Remediation mechanisms of mercapto-grafted palygorskite for cadmium pollutant in paddy soil[J]. Environmental Science and Pollution Research, 2017, 24(30):23783-23793.
[12]鲍士旦.土壤农化分析[M].三版.北京：中国农业出版社, 2000.BAO Shi-dan. Soil and agricultural chemistry analysis[M]. 3th. Edition. Beijing:China Agriculture Press, 2000.
[13] Xin J L, Huang B F. Subcellular distribution and chemical forms of cadmium in two hot pepper cultivars differing in cadmium accumulation[J]. Journal of Agricultural and Food Chemistry, 2014, 62(2):508-515.
[14] Xin J L, Huang B F, Dai H W, et al. Characterization of cadmium uptake, translocation, and distribution in young seedlings of two hot pepper cultivars that differ in fruit cadmium concentration[J]. Environmental Science and Pollution Research, 2014, 21(12):7449-7456.
[15]董如茵,徐应明,王林,等.土施和喷施锌肥对镉低积累油菜吸收镉的影响[J].环境科学学报, 2015, 35(8):2589-2596.DONG Ru-yin, XU Ying-ming, WANG Lin, et al. Effects of soil application and foliar spray of zinc fertilizer on cadmium uptake in a pakchoi cultivar with low cadmium accumulation[J]. Acta Scientiae Circumstantiae, 2015, 35(8):2589-2596.
[16] Rahman A, Nahar K, Hasanuzzaman M, et al. Manganese-induced cadmium stress tolerance in rice seedlings:Coordinated action of antioxidant defense, glyoxalase system and nutrient homeostasis[J].Comptes Rendus Biologies, 2016, 339(11/12):462-474.
[17]安振锋,方正.植物锰营养研究进展[J].河北农业科学, 2002, 6(4):35-41.AN Zhen-feng, FANG Zheng. Research progress on plant manganese nutrition[J]. Journal of Hebei Agricultural Sciences, 2002, 6(4):35-41.
[18]李光远,王凤华,蒋燕.叶面喷施锰对生菜生长和品质的影响[J].贵州农业科学, 2015, 43(3):54-57.LI Guang-yuan, WANG Feng-hua, JIANG Yan. Effects of foliage spraying Mn on growth and quality of lettuce[J]. Guizhou Agricultural Sciences, 2015, 43(3):54-57.
[19]张玉秀,李林峰,柴团耀,等.锰对植物毒害及植物耐锰机理研究进展[J].植物学报, 2010, 45(4):506-520.ZHANG Yu-xiu, LI Lin-feng, CHAI Tuan-yao, et al. Advances in research on manganese toxicity to plants and mechanism of manganese tolerance in plants[J]. Chinese Bulletin of Botany, 2010, 45(4):506–520.
[20] Zhang H, Yang H, Wang Y, et al. The response of ginseng grown on farmland to foliar-applied iron, zinc, manganese and copper[J]. Industrial Crops and Products, 2013, 45:388-394.
[21]张玉先,祁倩倩,罗奥,等.锰对大豆氮代谢相关指标及产量品质的影响[J].中国油料作物学报, 2009, 31(4):486-491.ZHANG Yu-xian, QI Qian-qian, LUO Ao, et al. Effects of manganese on soybean nitrogen metabolism yield and quality[J]. Chinese Journal of Oil Crop Sciences, 2009, 31(4):486-491.
[22] Yang M, Zhang Y Y, Zhang L J, et al. OsNRAMP5 contributes to manganese translocation and distribution in rice shoots[J]. Journal of Experimental Botany, 2014, 65(17):4849-4861.
[23] Sasaki A, Yamaji N, Yokosho K, et al. Nramp5 is a major transporter responsible for manganese and cadmium uptake in rice[J]. The Plant Cell, 2012, 24(5):2155-2167.
[24] Shi G R, Zhang Z, Liu C F. Silicon influences cadmium translocation by altering subcellular distribution and chemical forms of cadmium in peanut roots[J]. Archives of Agronomy and Soil Science, 2017, 63(1):117-123.
[25]王俊丽,梁娟,任建国.土壤镉污染对艾纳香生长及体内镉积累和分布特征的影响[J].西南师范大学学报（自然科学版）, 2017, 42(9):84-89.WANG Jun-Li, LIANG Juan, REN Jian-guo. Effects of soil cadmium contamination on growth of Blumea balsamifera and cadmium accumulation and subcellular distribution in plant[J]. Journal of Southwest China Normal University(Natural Science), 2017, 42(9):84-89.
[26]何蔚,陈永华,梁希,等.改良锰矿渣中木本植物筛选及锰的亚细胞分布和化学形态[J].环境工程, 2018, 36(9):154-160.HE Wei, CHEN Yong-hua, LIANG Xi, et al. Screening for tolerant woody plants for improved manganese slag and research on subcellular distribution and chemical form of manganese[J]. Environmental Engineering, 2018, 36(9):154-160.
[27]邓华,李明顺,陈英旭,等.锰在短毛蓼不同器官中的亚细胞分布及化学形态[J].广西师范大学学报（自然科学版）, 2010, 28(1):58-62.DENG Hua, LI Ming-shun, CHEN Ying-xu, et al. Subcellular distribution and chemical fractions of manganese in hyperaccumulator Polygonum pubescens Blume[J]. Journal of Guangxi Normal University(Natural Science Edition), 2010, 28(1):58-62.
[28]徐向华,施积炎,陈新才,等.锰在商陆叶片的细胞分布及化学形态分析[J].农业环境科学学报, 2008, 27(2):515-520.XU Xiang-hua, SHI Ji-yan, CHEN Xin-cai, et al. Subcellular distribution and chemical fractions of manganese in leaves of hyperaccumulator Phytolacca acinosa Roxb.[J]. Journal of Agro-Environment Science, 2008, 27(2):515-520.
[29]徐莜,杨益新,李文华,等.锰离子浓度及其转运通道对水稻幼苗镉吸收转运特性的影响[J].农业环境科学学报, 2016, 35(8):1429-1435.XU You, YANG Yi-xin, LI Wen-hua, et al. Effects of manganese concentrations and transporters on uptake and translocation of cadmium in rice seedlings[J]. Journal of Agro-Environment Science, 2016, 35(8):1429-1435.
[30] Thomine S, Lelièvre F, Debarbieux E, et al. AtNRAMP3, a multispecific vacuolar metal transporter involved in plant responses to iron deficiency[J]. The Plant Journal, 2003, 34(5):685-695.
[31] Koren′kov V, Park S, Cheng N H, et al. Enhanced Cd2+-selective root-tonoplast-transport in tobaccos expressing Arabidopsis cation exchangers[J]. Planta, 2006, 225(2):403-411.
[2]李霞,张慧鸣,徐震,等.农田Cd和Hg污染的来源解析与风险评价研究[J].农业环境科学学报, 2016, 35(7):1314-1320.LI Xia, ZHANG Hui-ming, XU Zhen, et al. Source apportionment and risk assessment of Cd and Hg pollution in farmland[J]. Journal of AgroEnvironment Science, 2016, 35(7):1314-1320.
[3] Chen H P, Yang X P, Wang P, et al. Dietary cadmium intake from rice and vegetables and potential health risk:A case study in Xiangtan,southern China[J]. Science of the Total Environment, 2018, 639:271–277.
[4]黄道友,朱奇宏,朱捍华,等.重金属污染耕地农业安全利用研究进展与展望[J].农业现代化研究, 2018, 39(6):1030-1043.HUANG Dao-you, ZHU Qi-hong, ZHU Han-hua, et al. Advances and prospects of safety agro-utilization of heavy metal contaminated farmland soil[J]. Research of Agricultural Modernization, 2018, 39(6):1030-1043.
[5]温娜,王景安,刘仲齐.利用AMMI模型分析稻米镉含量的基因型与环境互作效应[J].农业环境科学学报, 2015, 34(5):817-823.WEN Na, WANG Jing-an, LIU Zhong-qi. Analysis of genotypic and environmental effects on cadmium content in rice by AMMI model[J].Journal of Agro-Environment Science, 2015, 34(5):817-823.
[6] Socha A L, Guerinot M L. Mn-euvering manganese:The role of transporter gene family members in manganese uptake and mobilization in plants[J]. Frontiers in Plant Science, 2014, 5:106-122.
[7] Ullah I, Wang Y, Eide D J, et al. Evolution, and functional analysis of Natural Resistance-Associated Macrophage Proteins(NRAMPs)from Theobroma cacao and their role in cadmium accumulation[J]. Scientific Reports, 2018, 8(1):14412.
[8] Huang Q N, An H, Yang Y J, et al. Effects of Mn-Cd antagonistic interaction on Cd accumulation and major agronomic traits in rice genotypes by different Mn forms[J]. Plant Growth Regulation, 2017, 82(2):317-331.
[9]尹晓辉,邹慧玲,方雅瑜,等.施锰方式对水稻吸收积累镉的影响研究[J].环境科学与技术, 2017, 40(8):8-12.YIN Xiao-hui, ZOU Hui-ling, FANG Ya-yu, et al. Effects of manganese fertilizer on absorption and accumulation of Cd in rice[J]. Environmental Science&Technology, 2017, 40(8):8-12.
[10] Wang L, Xu Y M, Sun Y B, et al. Identification of pakchoi cultivars with low cadmium accumulation and soil factors that affect their cadmium uptake and translocation[J]. Frontiers of Environmental Science and Engineering, 2014, 8(6):877-887.
[11] Liang X F, Qin X, Huang Q Q, et al. Remediation mechanisms of mercapto-grafted palygorskite for cadmium pollutant in paddy soil[J]. Environmental Science and Pollution Research, 2017, 24(30):23783-23793.
[12]鲍士旦.土壤农化分析[M].三版.北京：中国农业出版社, 2000.BAO Shi-dan. Soil and agricultural chemistry analysis[M]. 3th. Edition. Beijing:China Agriculture Press, 2000.
[13] Xin J L, Huang B F. Subcellular distribution and chemical forms of cadmium in two hot pepper cultivars differing in cadmium accumulation[J]. Journal of Agricultural and Food Chemistry, 2014, 62(2):508-515.
[14] Xin J L, Huang B F, Dai H W, et al. Characterization of cadmium uptake, translocation, and distribution in young seedlings of two hot pepper cultivars that differ in fruit cadmium concentration[J]. Environmental Science and Pollution Research, 2014, 21(12):7449-7456.
[15]董如茵,徐应明,王林,等.土施和喷施锌肥对镉低积累油菜吸收镉的影响[J].环境科学学报, 2015, 35(8):2589-2596.DONG Ru-yin, XU Ying-ming, WANG Lin, et al. Effects of soil application and foliar spray of zinc fertilizer on cadmium uptake in a pakchoi cultivar with low cadmium accumulation[J]. Acta Scientiae Circumstantiae, 2015, 35(8):2589-2596.
[16] Rahman A, Nahar K, Hasanuzzaman M, et al. Manganese-induced cadmium stress tolerance in rice seedlings:Coordinated action of antioxidant defense, glyoxalase system and nutrient homeostasis[J].Comptes Rendus Biologies, 2016, 339(11/12):462-474.
[17]安振锋,方正.植物锰营养研究进展[J].河北农业科学, 2002, 6(4):35-41.AN Zhen-feng, FANG Zheng. Research progress on plant manganese nutrition[J]. Journal of Hebei Agricultural Sciences, 2002, 6(4):35-41.
[18]李光远,王凤华,蒋燕.叶面喷施锰对生菜生长和品质的影响[J].贵州农业科学, 2015, 43(3):54-57.LI Guang-yuan, WANG Feng-hua, JIANG Yan. Effects of foliage spraying Mn on growth and quality of lettuce[J]. Guizhou Agricultural Sciences, 2015, 43(3):54-57.
[19]张玉秀,李林峰,柴团耀,等.锰对植物毒害及植物耐锰机理研究进展[J].植物学报, 2010, 45(4):506-520.ZHANG Yu-xiu, LI Lin-feng, CHAI Tuan-yao, et al. Advances in research on manganese toxicity to plants and mechanism of manganese tolerance in plants[J]. Chinese Bulletin of Botany, 2010, 45(4):506–520.
[20] Zhang H, Yang H, Wang Y, et al. The response of ginseng grown on farmland to foliar-applied iron, zinc, manganese and copper[J]. Industrial Crops and Products, 2013, 45:388-394.
[21]张玉先,祁倩倩,罗奥,等.锰对大豆氮代谢相关指标及产量品质的影响[J].中国油料作物学报, 2009, 31(4):486-491.ZHANG Yu-xian, QI Qian-qian, LUO Ao, et al. Effects of manganese on soybean nitrogen metabolism yield and quality[J]. Chinese Journal of Oil Crop Sciences, 2009, 31(4):486-491.
[22] Yang M, Zhang Y Y, Zhang L J, et al. OsNRAMP5 contributes to manganese translocation and distribution in rice shoots[J]. Journal of Experimental Botany, 2014, 65(17):4849-4861.
[23] Sasaki A, Yamaji N, Yokosho K, et al. Nramp5 is a major transporter responsible for manganese and cadmium uptake in rice[J]. The Plant Cell, 2012, 24(5):2155-2167.
[24] Shi G R, Zhang Z, Liu C F. Silicon influences cadmium translocation by altering subcellular distribution and chemical forms of cadmium in peanut roots[J]. Archives of Agronomy and Soil Science, 2017, 63(1):117-123.
[25]王俊丽,梁娟,任建国.土壤镉污染对艾纳香生长及体内镉积累和分布特征的影响[J].西南师范大学学报（自然科学版）, 2017, 42(9):84-89.WANG Jun-Li, LIANG Juan, REN Jian-guo. Effects of soil cadmium contamination on growth of Blumea balsamifera and cadmium accumulation and subcellular distribution in plant[J]. Journal of Southwest China Normal University(Natural Science), 2017, 42(9):84-89.
[26]何蔚,陈永华,梁希,等.改良锰矿渣中木本植物筛选及锰的亚细胞分布和化学形态[J].环境工程, 2018, 36(9):154-160.HE Wei, CHEN Yong-hua, LIANG Xi, et al. Screening for tolerant woody plants for improved manganese slag and research on subcellular distribution and chemical form of manganese[J]. Environmental Engineering, 2018, 36(9):154-160.
[27]邓华,李明顺,陈英旭,等.锰在短毛蓼不同器官中的亚细胞分布及化学形态[J].广西师范大学学报（自然科学版）, 2010, 28(1):58-62.DENG Hua, LI Ming-shun, CHEN Ying-xu, et al. Subcellular distribution and chemical fractions of manganese in hyperaccumulator Polygonum pubescens Blume[J]. Journal of Guangxi Normal University(Natural Science Edition), 2010, 28(1):58-62.
[28]徐向华,施积炎,陈新才,等.锰在商陆叶片的细胞分布及化学形态分析[J].农业环境科学学报, 2008, 27(2):515-520.XU Xiang-hua, SHI Ji-yan, CHEN Xin-cai, et al. Subcellular distribution and chemical fractions of manganese in leaves of hyperaccumulator Phytolacca acinosa Roxb.[J]. Journal of Agro-Environment Science, 2008, 27(2):515-520.
[29]徐莜,杨益新,李文华,等.锰离子浓度及其转运通道对水稻幼苗镉吸收转运特性的影响[J].农业环境科学学报, 2016, 35(8):1429-1435.XU You, YANG Yi-xin, LI Wen-hua, et al. Effects of manganese concentrations and transporters on uptake and translocation of cadmium in rice seedlings[J]. Journal of Agro-Environment Science, 2016, 35(8):1429-1435.
[30] Thomine S, Lelièvre F, Debarbieux E, et al. AtNRAMP3, a multispecific vacuolar metal transporter involved in plant responses to iron deficiency[J]. The Plant Journal, 2003, 34(5):685-695.
[31] Koren′kov V, Park S, Cheng N H, et al. Enhanced Cd2+-selective root-tonoplast-transport in tobaccos expressing Arabidopsis cation exchangers[J]. Planta, 2006, 225(2):403-411.
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