烟草幼苗根系分泌自毒物质种类及PAEs对根系抗氧化性能的影响
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摘要
采用GC-MS技术鉴定水培烟草Burley及K326在幼苗期不同生长阶段的根系分泌物;并用不同浓度邻苯二甲酸二丁酯(DBP)、邻苯二甲酸二异辛酯(DIOP)溶液浇灌盆栽烟草幼苗,研究其根系抗氧化性能变化。结果如下:(1)Burley根系分泌物主要有3类化合物,其中自毒物质邻苯二甲酸酯(PAEs)在二叶龄期、四叶龄期、六叶龄期的相对含量分别为7.6%、0.3%、未检出;而K326根系分泌物主要有9类化合物,PAEs在二叶龄期、四叶龄期、六叶龄期的相对含量分别为35.6%、51.3%、2.2%。(2)浓度高于0.1 mmol/L的PAEs使根中超氧阴离子自由基产生的速率显著(P<0.05)增加;随着DIOP及DBP浓度的增加,超氧化物歧化酶、过氧化氢酶活性增加,在0.5 mmol/L时达到最大,然后随着处理浓度的增加而下降。丙二醛的浓度随着这两种PAEs处理浓度的增加而增大。结果表明:烟草根系分泌的自毒物质PAEs达到0.5 mmol/L时,能降低根系的抗氧化性能,造成根尖细胞膜系统的氧化损伤,引起根吸收功能等一系列生理生化变化,并最终表现出自毒作用。
Root exudates of two varieties(Burley and K326)of tobacco seedlings cultured in a hydroponic medium were collected,and identified using gas chromatography-mass spectrometry(GC-MS). Further,the effects of different concentrations of dibutyl phthalate(DBP)and diisooctyl phthalate(DIOP)on root antioxidant capacity were assessed using pot culture experiments. The results revealed three main compounds in Burley root exudates. The relative content of autotoxin phthalate esters(PAEs)at the two-,four-,and six-leaf stages were 7.6%,0.3%,and not detected,respectively.However,there were nine main compounds in K326 root exudates. The relative PAE content at the two-,four-,and six-leaf stages were 35.6%,51.3%,and 2.2%,respectively. PAEs such as DIOP and DBP were identified as the major autotoxins in root exudates of both tobacco seedlings. The rate of superoxide anion radical generation in roots significantly increased at concentrations greater than 0. 1 mmol/L DIOP and DBP(P < 0. 05). Both superoxide dismutase and catalase activities increased with increasing DIOP concentrations,with a peak at 0. 5 mmol/L,and subsequently decreased at higher concentrations. Accumulated malondialdehyde concentrations increased with increasing DIOP and DBP concentrations,and the magnitude of malondialdehyde content was DIOP > DBP,which indicated the order of their toxic effect. Finally,our findings also revealed that when PAEs reached 0. 5 mmol/L,they decreased the antioxidant capacity of the root system,initiated oxidative damage of the root cell membrane system,and caused a further decrease in root absorption and mineral metabolism,and thereby led to autotoxicity in the tobacco plants.
Root exudates of two varieties(Burley and K326)of tobacco seedlings cultured in a hydroponic medium were collected,and identified using gas chromatography-mass spectrometry(GC-MS). Further,the effects of different concentrations of dibutyl phthalate(DBP)and diisooctyl phthalate(DIOP)on root antioxidant capacity were assessed using pot culture experiments. The results revealed three main compounds in Burley root exudates. The relative content of autotoxin phthalate esters(PAEs)at the two-,four-,and six-leaf stages were 7.6%,0.3%,and not detected,respectively.However,there were nine main compounds in K326 root exudates. The relative PAE content at the two-,four-,and six-leaf stages were 35.6%,51.3%,and 2.2%,respectively. PAEs such as DIOP and DBP were identified as the major autotoxins in root exudates of both tobacco seedlings. The rate of superoxide anion radical generation in roots significantly increased at concentrations greater than 0. 1 mmol/L DIOP and DBP(P < 0. 05). Both superoxide dismutase and catalase activities increased with increasing DIOP concentrations,with a peak at 0. 5 mmol/L,and subsequently decreased at higher concentrations. Accumulated malondialdehyde concentrations increased with increasing DIOP and DBP concentrations,and the magnitude of malondialdehyde content was DIOP > DBP,which indicated the order of their toxic effect. Finally,our findings also revealed that when PAEs reached 0. 5 mmol/L,they decreased the antioxidant capacity of the root system,initiated oxidative damage of the root cell membrane system,and caused a further decrease in root absorption and mineral metabolism,and thereby led to autotoxicity in the tobacco plants.
引文
[1]Walker T S,Bais H P,Grotewold E,Vivanco J M.Root exudation and rhizosphere biology.Plant Physiology,2003,132(1):44-51.
[2]Xie X N,Kusumoto D,Takeuchi Y,Yoneyama K,Yamada Y,Yoneyama K.2'-epi-orobanchol and solanacol,two unique strigolactones,germination stimulants for root parasitic weeds,produced by tobacco.Journal of Agricultural and Food Chemistry,2007,55(20):8067-8072.
[3]晋艳,杨宇虹,段玉琪,龙玉华,叶成碧.烤烟连作对烟叶产量和质量的影响研究初报.烟草科技,2002,(1):41-45.
[4]邓阳春,黄建国.长期连作对烤烟产量和土壤养分的影响.植物营养与肥料学报,2010,16(4):840-845.
[5]Weir T L,Park S W,Vivanco J M.Biochemical and physiological mechanisms mediated by allelochemicals.Current Opinion in Plant Biology,2004,7(4):472-479.
[6]Jia Z H,Yi J H,Su Y R,Shen H.Autotoxic substances in the root exudates from continuous tobacco cropping.Allelopathy Journal,2011,27(1):87-96.
[7]Sun Y Y,Jiang G Y,Wei X C,Liu J G.Autotoxicity effects of soils continuously cropped with tomato.Allelopathy Journal,2011,28(2):135-144.
[8]刘苹,赵海军,仲子文,孙明,庞亚群,马征,万书波.三种根系分泌脂肪酸对花生生长和土壤酶活性的影响.生态学报,2013,33(11):3332-3339.
[9]郭亚利,李明海,吴洪田,袁玲,黄建国.烤烟根系分泌物对烤烟幼苗生长和养分吸收的影响.植物营养与肥料学报,2007,13(3):458-463.
[10]Yeasmin R,Nakamatsu K,Matsumoto H,Motoki S,Nishihara E,Yamamoto S.Inference of allelopathy and autotoxicity to varietal resistance of asparagus(Asparagus officinalis L.),Australian Journal of Crop Science,2014,8(2):251-256.
[11]Liu P,Wan S B,Jiang L H,Wang C B,Liu Z H,Zhao H J,Yu S F,Yang L.Autotoxic potential of root exudates of peanut(Arachis hypogaea L.).Allelopathy Journal,2010,26(2):197-205.
[12]Mazzoleni S,Bonanomi G,Incerti G,Chiusano M L,Termolino P,Mingo A,Senatore M,Giannino F,CartenìF,Rietkerk M,Lanzotti V.Inhibitory and toxic effects of extracellular self-DNA in litter:a mechanism for negative plant-soil feedbacks?.New Phytologist,2015,205(3):1195-1210.
[13]Murashige T,Skoog F.A revised medium for rapid growth and bio-assays with tobacco tissue cultures.Physiologia Plantarum,1962,15(3):473-497.
[14]Walch-Liu P,Neumann G,Bangerth F,Engels C.Rapid eects of nitrogen form on leaf morphogenesis in tobacco.Journal of Experimental Botany,2000,51(343):227-237.
[15]Yi J H,Jia Z H,Lin Q,Lv H Z,Shen H.Allelopathic effects of decaying tobacco leaves on tobacco seedlings.Allelopathy Journal,2012,29(1):51-61.
[16]Dhindsa R S,Matowe W.Drought tolerance in two mosses:correlated with enzymatic defence against lipid peroxidation.Journal of Experimental Botany,1981,32(1):79-91.
[17]Beers R F Jr,Sizer I W.A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase.The Journal of Biological Chemistry,1952,195(1):133-140.
[18]Heath R L,Packer L.Photoperoxidation in isolated chloroplast.I.Kinetics and stochiometry of fatty acid peroxidation.Archives of Biochemistry and Biophysics,1968,125(1):189-198.
[19]Elstner E F,Heupel A.Formation of hydrogen peroxide by isolated cell walls from horseradish(Armoracia lapathifolia Gilib.).Planta,1976,130(2):175-180.
[20]Tang C S,Takenaka T.Quantitation of a bioactive metabolite in undisturbed rhizosphere-benzyl isothiocyanate From Carica Papaya L.Journal of Chemical Ecology,1983,9(8):1247-1253.
[21]Rice E L.Allelopathy.2nd ed.Orlando,Florida:Academic Press,1984.
[22]Inderjit.Plant phenolics in allelopathy.The Botanical Review,1996,62(2):186-202.
[23]Weller D E.The interspecific size-density relationship among crowded plant stands and its implications for the-3/2 power rule of self-thinning.The American Naturalist.1989,133(1):20-41.
[24]Singh H P,Batish D R,Kohli R K.Autotoxicity:concept,organisms,and ecological significance.Critical Reviews in Plant Sciences,1999,18(6):757-772.
[25]Inderjit,Callaway R M,Vivanco J M.Can plant biochemistry contribute to understanding of invasion ecology?Trends in Plant Science,2006,11(12):574-580.
[26]Kato-Noguchi H.Allelopathic substance in rice root exudates:Rediscovery of momilactone B as an allelochemical.Journal of Plant Physiology,2004,161(3):271-276.
[27]El-Halmouch Y,Benharrat H,Thalouarn P.Effect of root exudates from different tomato genotypes on broomrape(O.aegyptiaca)seed germination and tubercle development.Crop Protection,2006,25(5):501-507.
[28]Neumann G,Massonneau A,Martinoia E,Rmheld V.Physiological adaptations to phosphorus deficiency during proteoid root development in white lupin.Planta,1999,208(3):373-382.
[29]Tyler G,Strm L.Differing organic acid exudation pattern explains calcifuge and acidifuge behaviour of plants.Annals of Botany,1995,75(1):75-78.
[30]Schumacher W J,Thill D C,Lee G A.Allelopathic potential of wild oat(Avena fatua)on spring wheat(Triticum aestivum)growth.Journal of Chemical Ecology,1983,9(8):1235-1245.
[31]Cies'liński G,van Rees K C J,Szmigielska A M,Krishnamurti G S R,Huang P M.Low-molecular-weight organic acids in rhizosphere soils of durum wheat and their effect on cadmium bioaccumulation.Plant and Soil,1998,203(1):109-117
[32]屠振力,钟儒杰.环境激素邻苯二甲酸丁基苄酯对家蚕生殖的影响.生态学报,2014,34(19):5470-5476.
[33]Alías J C,Sosa T,Escudero J C,Chaves N.Autotoxicity against germination and seedling emergence in Cistus ladanifer L.Plant and Soil,2006,282(1/2):327-332.
[34]Ma Y Q,Jia J N,An Y,Wang Z,Mao J C.Potential of some hybrid maize lines to induce germination of Sunflower Broomrape.Crop Science,2013,53(1):260-270.
[35]Foyer C H,Descourvières P,Kunert K J.Protection against oxygen radicals:an important defence mechanism studied in transgenic plants.Plant,Cell&Environment,1994,17(5):507-523.
[36]Halliwell B,Gutteridge J M C.Free radicals in biology and medicine.4th ed.Oxford:Glarendon Press,1985.
[37]Papadakis A K,Roubelakis-Angelakis K A.The generation of active oxygen species differs in tobacco and grapevine mesophyll protoplasts.Plant Physiology,1999,121(1):197-206.
[38]邓家军,胡继伟,李继新,苏贤坤,黄先飞,刘峰.重金属离子对烤烟叶片中铜锌超氧化物歧化酶活性的影响.中国烟草学报,2010,16(3):1-6.
[39]Mobin M,Khan N A.Photosynthetic activity,pigment composition and antioxidative response of two mustard(Brassica juncea)cultivars differing in photosynthetic capacity subjected to cadmium stress.Journal of Plant Physiology,2007,164(5):601-610.
[40]Van Camp W,INZD,Van Montagu M.The regulation and function of tobacco superoxide dismutases.Free Radical Biology and Medicine,1997,23(3):515-520.
[41]Willekens H,Chamnongpol S,Davey M,Schraudner M,Langebartels C,Van Montagu M,INZD,Van Camp W.Catalase is a sink for H2O2and is indispensable for stress defense in C3plants.the EMBO Journal,1997,16(16):4806-4816.
[42]Bais H P,Vepachedu R,Gilroy S,Callaway R M,Vivanco J M.Allelopathy and exotic plant invasion:from molecules and genes tospecies interactions.Science,2003,301(5638):1377-1380.
[43]李志萍,张文辉,崔豫川.Na Cl和Na2CO3胁迫对栓皮栎种子萌发及幼苗生长的影响.生态学报,2015,35(3):742-751.
[44]Foyer C H,Lopez-Delgado H,Dat J F,Scott I M.Hydrogen peroxide-and glutathione-associated mechanisms of acclimatory stress tolerance and signalling.Physiologia Plantarum,1997,100(2):241-254.
[45]Baziramakenga R,Leroux G D,Simard R R.Effects of benzoic and cinnamic acids on membrane permeability of soybean roots.Journal of Chemical Ecology,1995,21(9):1271-1285.
[46]Ye S F,Zhou Y H,Sun Y,Zou L Y,Yu J Q.Cinnamic acid causes oxidative stress in cucumber roots,and promotes incidence of Fusarium wilt.Environmental and Experimental Botany,2006,56(3):255-262.
[47]Yu J Q,Matsui Y.Effects of root exudates of cucumber(Cucumis sativus)and allelochemicals on ion uptake by cucumber seedlings.Journal of Chemical Ecology,1997,23(3):817-827.
[2]Xie X N,Kusumoto D,Takeuchi Y,Yoneyama K,Yamada Y,Yoneyama K.2'-epi-orobanchol and solanacol,two unique strigolactones,germination stimulants for root parasitic weeds,produced by tobacco.Journal of Agricultural and Food Chemistry,2007,55(20):8067-8072.
[3]晋艳,杨宇虹,段玉琪,龙玉华,叶成碧.烤烟连作对烟叶产量和质量的影响研究初报.烟草科技,2002,(1):41-45.
[4]邓阳春,黄建国.长期连作对烤烟产量和土壤养分的影响.植物营养与肥料学报,2010,16(4):840-845.
[5]Weir T L,Park S W,Vivanco J M.Biochemical and physiological mechanisms mediated by allelochemicals.Current Opinion in Plant Biology,2004,7(4):472-479.
[6]Jia Z H,Yi J H,Su Y R,Shen H.Autotoxic substances in the root exudates from continuous tobacco cropping.Allelopathy Journal,2011,27(1):87-96.
[7]Sun Y Y,Jiang G Y,Wei X C,Liu J G.Autotoxicity effects of soils continuously cropped with tomato.Allelopathy Journal,2011,28(2):135-144.
[8]刘苹,赵海军,仲子文,孙明,庞亚群,马征,万书波.三种根系分泌脂肪酸对花生生长和土壤酶活性的影响.生态学报,2013,33(11):3332-3339.
[9]郭亚利,李明海,吴洪田,袁玲,黄建国.烤烟根系分泌物对烤烟幼苗生长和养分吸收的影响.植物营养与肥料学报,2007,13(3):458-463.
[10]Yeasmin R,Nakamatsu K,Matsumoto H,Motoki S,Nishihara E,Yamamoto S.Inference of allelopathy and autotoxicity to varietal resistance of asparagus(Asparagus officinalis L.),Australian Journal of Crop Science,2014,8(2):251-256.
[11]Liu P,Wan S B,Jiang L H,Wang C B,Liu Z H,Zhao H J,Yu S F,Yang L.Autotoxic potential of root exudates of peanut(Arachis hypogaea L.).Allelopathy Journal,2010,26(2):197-205.
[12]Mazzoleni S,Bonanomi G,Incerti G,Chiusano M L,Termolino P,Mingo A,Senatore M,Giannino F,CartenìF,Rietkerk M,Lanzotti V.Inhibitory and toxic effects of extracellular self-DNA in litter:a mechanism for negative plant-soil feedbacks?.New Phytologist,2015,205(3):1195-1210.
[13]Murashige T,Skoog F.A revised medium for rapid growth and bio-assays with tobacco tissue cultures.Physiologia Plantarum,1962,15(3):473-497.
[14]Walch-Liu P,Neumann G,Bangerth F,Engels C.Rapid eects of nitrogen form on leaf morphogenesis in tobacco.Journal of Experimental Botany,2000,51(343):227-237.
[15]Yi J H,Jia Z H,Lin Q,Lv H Z,Shen H.Allelopathic effects of decaying tobacco leaves on tobacco seedlings.Allelopathy Journal,2012,29(1):51-61.
[16]Dhindsa R S,Matowe W.Drought tolerance in two mosses:correlated with enzymatic defence against lipid peroxidation.Journal of Experimental Botany,1981,32(1):79-91.
[17]Beers R F Jr,Sizer I W.A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase.The Journal of Biological Chemistry,1952,195(1):133-140.
[18]Heath R L,Packer L.Photoperoxidation in isolated chloroplast.I.Kinetics and stochiometry of fatty acid peroxidation.Archives of Biochemistry and Biophysics,1968,125(1):189-198.
[19]Elstner E F,Heupel A.Formation of hydrogen peroxide by isolated cell walls from horseradish(Armoracia lapathifolia Gilib.).Planta,1976,130(2):175-180.
[20]Tang C S,Takenaka T.Quantitation of a bioactive metabolite in undisturbed rhizosphere-benzyl isothiocyanate From Carica Papaya L.Journal of Chemical Ecology,1983,9(8):1247-1253.
[21]Rice E L.Allelopathy.2nd ed.Orlando,Florida:Academic Press,1984.
[22]Inderjit.Plant phenolics in allelopathy.The Botanical Review,1996,62(2):186-202.
[23]Weller D E.The interspecific size-density relationship among crowded plant stands and its implications for the-3/2 power rule of self-thinning.The American Naturalist.1989,133(1):20-41.
[24]Singh H P,Batish D R,Kohli R K.Autotoxicity:concept,organisms,and ecological significance.Critical Reviews in Plant Sciences,1999,18(6):757-772.
[25]Inderjit,Callaway R M,Vivanco J M.Can plant biochemistry contribute to understanding of invasion ecology?Trends in Plant Science,2006,11(12):574-580.
[26]Kato-Noguchi H.Allelopathic substance in rice root exudates:Rediscovery of momilactone B as an allelochemical.Journal of Plant Physiology,2004,161(3):271-276.
[27]El-Halmouch Y,Benharrat H,Thalouarn P.Effect of root exudates from different tomato genotypes on broomrape(O.aegyptiaca)seed germination and tubercle development.Crop Protection,2006,25(5):501-507.
[28]Neumann G,Massonneau A,Martinoia E,Rmheld V.Physiological adaptations to phosphorus deficiency during proteoid root development in white lupin.Planta,1999,208(3):373-382.
[29]Tyler G,Strm L.Differing organic acid exudation pattern explains calcifuge and acidifuge behaviour of plants.Annals of Botany,1995,75(1):75-78.
[30]Schumacher W J,Thill D C,Lee G A.Allelopathic potential of wild oat(Avena fatua)on spring wheat(Triticum aestivum)growth.Journal of Chemical Ecology,1983,9(8):1235-1245.
[31]Cies'liński G,van Rees K C J,Szmigielska A M,Krishnamurti G S R,Huang P M.Low-molecular-weight organic acids in rhizosphere soils of durum wheat and their effect on cadmium bioaccumulation.Plant and Soil,1998,203(1):109-117
[32]屠振力,钟儒杰.环境激素邻苯二甲酸丁基苄酯对家蚕生殖的影响.生态学报,2014,34(19):5470-5476.
[33]Alías J C,Sosa T,Escudero J C,Chaves N.Autotoxicity against germination and seedling emergence in Cistus ladanifer L.Plant and Soil,2006,282(1/2):327-332.
[34]Ma Y Q,Jia J N,An Y,Wang Z,Mao J C.Potential of some hybrid maize lines to induce germination of Sunflower Broomrape.Crop Science,2013,53(1):260-270.
[35]Foyer C H,Descourvières P,Kunert K J.Protection against oxygen radicals:an important defence mechanism studied in transgenic plants.Plant,Cell&Environment,1994,17(5):507-523.
[36]Halliwell B,Gutteridge J M C.Free radicals in biology and medicine.4th ed.Oxford:Glarendon Press,1985.
[37]Papadakis A K,Roubelakis-Angelakis K A.The generation of active oxygen species differs in tobacco and grapevine mesophyll protoplasts.Plant Physiology,1999,121(1):197-206.
[38]邓家军,胡继伟,李继新,苏贤坤,黄先飞,刘峰.重金属离子对烤烟叶片中铜锌超氧化物歧化酶活性的影响.中国烟草学报,2010,16(3):1-6.
[39]Mobin M,Khan N A.Photosynthetic activity,pigment composition and antioxidative response of two mustard(Brassica juncea)cultivars differing in photosynthetic capacity subjected to cadmium stress.Journal of Plant Physiology,2007,164(5):601-610.
[40]Van Camp W,INZD,Van Montagu M.The regulation and function of tobacco superoxide dismutases.Free Radical Biology and Medicine,1997,23(3):515-520.
[41]Willekens H,Chamnongpol S,Davey M,Schraudner M,Langebartels C,Van Montagu M,INZD,Van Camp W.Catalase is a sink for H2O2and is indispensable for stress defense in C3plants.the EMBO Journal,1997,16(16):4806-4816.
[42]Bais H P,Vepachedu R,Gilroy S,Callaway R M,Vivanco J M.Allelopathy and exotic plant invasion:from molecules and genes tospecies interactions.Science,2003,301(5638):1377-1380.
[43]李志萍,张文辉,崔豫川.Na Cl和Na2CO3胁迫对栓皮栎种子萌发及幼苗生长的影响.生态学报,2015,35(3):742-751.
[44]Foyer C H,Lopez-Delgado H,Dat J F,Scott I M.Hydrogen peroxide-and glutathione-associated mechanisms of acclimatory stress tolerance and signalling.Physiologia Plantarum,1997,100(2):241-254.
[45]Baziramakenga R,Leroux G D,Simard R R.Effects of benzoic and cinnamic acids on membrane permeability of soybean roots.Journal of Chemical Ecology,1995,21(9):1271-1285.
[46]Ye S F,Zhou Y H,Sun Y,Zou L Y,Yu J Q.Cinnamic acid causes oxidative stress in cucumber roots,and promotes incidence of Fusarium wilt.Environmental and Experimental Botany,2006,56(3):255-262.
[47]Yu J Q,Matsui Y.Effects of root exudates of cucumber(Cucumis sativus)and allelochemicals on ion uptake by cucumber seedlings.Journal of Chemical Ecology,1997,23(3):817-827.