烯效唑对干旱胁迫下薏苡抗氧化酶活性及DNA甲基化水平的影响
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摘要
目的探讨烯效唑(S3307)对干旱胁迫下薏苡抗氧化酶活性及DNA甲基化水平的影响。方法将薏辽5号种子培养至幼苗的两叶一心期,实验分为3组:对照组(control):用Hoagland培养液处理;干旱胁迫组(PEG):用25%PEG的营养液处理;S3307缓解组(PEG+S3307组):经S3307预处理后,再用25%PEG处理。检测各组幼苗的形态指标(苗高、茎粗、根长、根数)及过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)的活性,同时采用高效液相色谱(HPLC)法测定各组幼苗的DNA甲基化水平。结果经S3307预处理后,幼苗的株高和根长显著降低(P<0.05),茎粗及根冠比显著增加(P<0.05);SOD、CAT和POD抗氧化酶的活性显著升高(P<0.05);幼苗叶片中5-甲基胞嘧啶(5-m C)比例比control组增加了34.29%(P<0.001),增加幅度比PEG组降低了12.99%。结论 S3307可缓解干旱胁迫下薏苡的膜脂过氧化损伤及DNA甲基化水平的上升,具有良好的壮苗作用,为薏苡的抗旱性机理研究奠定了基础。
Objective To investigate the effect of S3307 on anti-oxidant enzyme activity and DNA methylation level of Coix seedlings under drought stress.Methods The coix cultivar,"No 5 Yiliao",was cultured to the"two leaves and one core"stage,and divided into three groups.The coix seedlings in control group were treated with Hoagland medium,while those in PEG group with the nutrient medium containing 25% PEG,and those in PEG + S3307 group were pre-treated with S3307 then treated with 25% PEG.The morphological indexes of seedlings,such as height of seedling,diameter of stem,length and number of root in various groups were observed,while the activities of peroxidase(POD),superoxide dismutase(SOD)and catalase(CAT)were determined,and the DNA metylation levels were measured by HPLC.Results After pre-treatment with S3307,the height of seedlings and length of root decreased significantly(P < 0.05),while the diameter of stem and root cap ratio as well as the activities of SOD,CAT and POD increased significantly(P < 0.05).The percentage of 5-m C in leaves of seedlings increased by 34.29% as compared with that in control group,of which the increased extent decreased by 12.99% as compared with that in PEG group.Conclusion S3307 relieved the peroxide damage of membrane lipid and increase of DNA metylation level in coix seedlings under drought stress,which promoted the growth of seedlings.It laid a foundation of study on drought-relief mechanism of coix.
Objective To investigate the effect of S3307 on anti-oxidant enzyme activity and DNA methylation level of Coix seedlings under drought stress.Methods The coix cultivar,"No 5 Yiliao",was cultured to the"two leaves and one core"stage,and divided into three groups.The coix seedlings in control group were treated with Hoagland medium,while those in PEG group with the nutrient medium containing 25% PEG,and those in PEG + S3307 group were pre-treated with S3307 then treated with 25% PEG.The morphological indexes of seedlings,such as height of seedling,diameter of stem,length and number of root in various groups were observed,while the activities of peroxidase(POD),superoxide dismutase(SOD)and catalase(CAT)were determined,and the DNA metylation levels were measured by HPLC.Results After pre-treatment with S3307,the height of seedlings and length of root decreased significantly(P < 0.05),while the diameter of stem and root cap ratio as well as the activities of SOD,CAT and POD increased significantly(P < 0.05).The percentage of 5-m C in leaves of seedlings increased by 34.29% as compared with that in control group,of which the increased extent decreased by 12.99% as compared with that in PEG group.Conclusion S3307 relieved the peroxide damage of membrane lipid and increase of DNA metylation level in coix seedlings under drought stress,which promoted the growth of seedlings.It laid a foundation of study on drought-relief mechanism of coix.
引文
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[7]闫艳红,李波,杨文钰.烯效唑浸种对大豆苗期抗旱性的影响[J].中国油料作物学报,2009,31(4):480-485.YAN Y H,LI B,YANG W Y.Effects of uniconazole soaking on drought tolerance of soybean seedling[J].Chin J Oil Crop Sci,2009,31(4):480-485.(in Chinese)
[8]LI N Y,WANG J Z,HAN X F.Effects of Uniconazole on leaf anatomical structure and main osmotic regulatory metabolites contents in Petunia hybrida under salt stress[C].The 3rd Conference on Key Technology of Horticulture,Shenyang,2011:403-406.
[9]MA Y H,SONG Y,ZHANG H R.Effects of uniconazole on the drought resistance of perennial ryegrass[J].Pratacult Sci,2009,26(5):169-173.(in Chinese)马艳华,宋瑜,张洪荣.烯效唑对黑麦草抗旱生理特性的影响[J].草业科学,2009,26(5):169-173.
[10]WU C T,MORRIS J R.Genes,genetics and epigenetics:a correspondence[J].Science,2001,293(5532):1103-1105.
[11]DOWEN R H,PELIZZOLA M,SCHMITZ R J,et al.Widespread dynamic DNA methylation in response to biotic stress[J].Proc Natl Acad Sci USA,2012,109(32):E2183-E2191.doi:10.1073/pnas.1209329109.
[12]ZHAO Y L,YE W W,WANG J J,et al.Review of DNAmethylation and plant stress-tolerance[J].Acta Bot BorealiOccident Sin,2009,29(7):1479-1489.(in Chinese)赵云雷,叶武威,王俊娟,等.DNA甲基化与植物抗逆性研究进展[J].西北植物学报,2009,29(7):1479-1489.
[13]FRAGA M F,ESTELLER M.DNA methylation:a profile of methods and applications[J].Biotechniques,2002,33(3):632,634,636-649.
[14]HUANG X.Study on physiological and molecular mechanism of cold resistance enhanced by exogenous ABA application in sugarcane[D].Nanning:Guangxi University,2012.(in Chinese).黄杏.外源ABA提高甘蔗抗寒性的生理及分子机制研究[D].广西南宁:广西大学,2012.
[15]PAN Y J,FU B Y,WANG D,et al.Spatial and temporal profiling of DNA methylation induced by drought stress in rice[J].Scient Agr Sin,2009,42(9):3009-3018.(in Chinese)潘雅姣,傅彬英,王迪,等.水稻干旱胁迫诱导DNA甲基化时空变化特征分析[J].中国农业科学,2009,42(9):3009-3018.
[16]HUANG X,LIANG Y S,YANG L T,et al.Effects of abscisic acid and its biosynthesis inhibitor on the activities of antioxidant system of sugarcanes treated by cold stress[J].J South China Agr Univ,2013,34(3):356-361.(in Chinese)黄杏,梁勇生,杨丽涛,等.低温胁迫下脱落酸及合成抑制剂对甘蔗幼苗抗氧化系统的影响[J].华南农业大学学报,2013,34(3):356-361.
[17]WANG H Y,WANG Y F,YANG K J,et al.Effect of exohenous nitric oxide on lipid peroxidation and antioxidant enzymes activity in maize seedings under salt stress[J].J Maize Sci,2014,22(4):84-87.(in Chinese)王洪义,王玉凤,杨克军,等.外源NO对Na Cl胁迫下玉米幼苗脂质过氧化和抗氧化酶活性的影响[J].玉米科学,2014,22(4):84-87.
[18]YAN H,TIAN Z H,XU Y G.Effects of uniconazole on seed germination and physiological characters of sweet corn[J].JChangjiang Vegetables,2009,26(15):40-42.(in Chinese)严寒,田志宏,徐勇刚.烯效唑对甜玉米种子萌发及幼苗生理特性的影响[J].长江蔬菜,2009,26(15):40-42.
[19]CHENG S M.Effects of uniconazole on the stage bulbil of drought-tolerant characteristics of pinellia ternate(Thunb.)Breit[D].Chengdu:Sichuan Agicultural University,2010.(in Chinese)程仕明.烯效唑对半夏珠芽期耐干旱特性的影响[D].成都:四川农业大学,2010.
[20]HAN R L.Research progress of ascorbate peroxidase in tomato[J].J Shanxi Agr Sci,2013,41(6):650-652.(in Chinese)韩瑞丽.番茄抗坏血酸过氧化物酶研究进展[J].山西农业科学,2013,41(6):650-652.
[21]YANG S Q,ZHOU R L,LIANG H M,et al.Relationship of antioxidant enzyme and osmoregulation substance to photosynthese in several desert plants[J].J Desert Res,2015,35(6):1557-1564.(in Chinese)杨淑琴,周瑞莲,梁慧敏,等.沙漠植物抗氧化酶活性及渗透调节物质含量与光合作用的关系[J].中国沙漠,2015,35(6):1557-1564.
[22]SHI Y H,ZHANG Y Q.Effects of soaking seed with uniconazole on the growth of tartary buckwheat under different water conditions[J].Hubei Agr Sci,2015,54(6):1289-1295.(in Chinese)石艳华,张永清.烯效唑浸种对不同水分条件下苦荞生长的影响[J].湖北农业科学,2015,54(6):1289-1295.
[23]YANG W Y,FAN G Q,REN W J,et al.Physiological effect of uniconazole waterless-dressed seeds on root and leaf of wheat[J].Scient Agr Sin,2005,38(7):1339-1345.(in Chinese)杨文钰,樊高琼,任万军,等.烯效唑干拌种对小麦根叶生理功能的影响[J].中国农业科学,2005,38(7):1339-1345.
[24]CHEN Q.The effects of ABA or SA pretreatment on the total level of DNA methylation under water and salt stress[D].Shenyang:Shenyang Normal University,2012.(in Chinese)陈强.ABA、SA预处理对水分和盐胁迫下水稻DNA甲基化水平的影响[D].辽宁沈阳:沈阳师范大学,2012.
[25]DU Z H,WANG B Q,XIE B T,et al.Effects of plant growth regulators on physiological and biochemical characteristics of roots in summer sweetpotato seeding under simulated drought stress[J].Acta Agr Boreali-Occident Sin,2014,23(10):97-104.(in Chinese)杜召海,汪宝卿,解备涛,等.模拟干旱条件下植物生长调节剂对夏薯苗期根系生理生化特性的影响[J].西北农业学报,2014,23(10):97-104.
[26]WANG J W.Response of speckled kidney bean to drought stress and regulation of uniconazole[D].Shenyang:Shenyang Agricultural University,2014.(in Chinese)王景伟.奶花芸豆对干旱胁迫及烯效唑调控的响应[D].辽宁沈阳:沈阳农业大学,2014.
[27]YUAN Z,WANG B Q,JIANG Y,et al.Effects of Uniconazole on physiological and biochemical properties of roots of different Sweetpotato cultivars at seedlings stage[J].Agr Sci Technol,2015,16(4):629-638.
[28]LI N Y,YU Y,FU Y D,et al.Effects of bulb soaking with S3307 on growth and endogenous hormone contents of potting lily(Lilium brownie F.E.brown asiatic hybrids)[J].Plant Physiol Commun,2007,43(1):104-106.(in Chinese)李宁毅,于洋,付印东,等.S3307浸球对盆栽百合生长和内源激素含量的影响[J].植物生理学通讯,2007,43(1):104-106.
[2]李小雁.干旱地区土壤-植被-水文耦合、响应与适应机制[J].中国科学:地球科学,2011,41(12):1271-1230.
[3]麻继仙,杨长楷,但忠,等.50%矮壮素水剂对不同苗龄番茄幼苗的影响[J].现代农业科技,2012,41(4):211,213.
[4]甘小虎,何从亮,阎庆久,等.矮壮素、多效唑对高温季节黄瓜育苗的影响[J].蔬菜,2012,31(7):64-66.
[5]ZHEN H L,YAN Z H,FENG L J,et al.Effects of chlormequat chloride on the growth and endogenous hormones contents of Dahlia pinnata and their correlation analysis[J].Pratacult Sci,2012,29(1):76-82.(in Chinese)甄红丽,苑兆和,冯立娟,等.矮壮素对大丽花生长发育和内源激素含量的影响及相关性分析[J].草业科学,2012,29(1):76-82.
[6]SUN F D,FENG N J,ZHENG D F,et al.Effects of plant growth regulators S3307 and DTA-6 on physiological metabolism and Gm AC gene expression in soybean[J].Scient Agr Sin,2016,49(7):1267-1276.(in Chinese)孙福东,冯乃杰,郑殿峰,等.植物生长调节剂S3307和DTA-6对大豆荚的生理代谢及Gm AC的影响[J].中国农业科学,2016,49(7):1267-1276.
[7]闫艳红,李波,杨文钰.烯效唑浸种对大豆苗期抗旱性的影响[J].中国油料作物学报,2009,31(4):480-485.YAN Y H,LI B,YANG W Y.Effects of uniconazole soaking on drought tolerance of soybean seedling[J].Chin J Oil Crop Sci,2009,31(4):480-485.(in Chinese)
[8]LI N Y,WANG J Z,HAN X F.Effects of Uniconazole on leaf anatomical structure and main osmotic regulatory metabolites contents in Petunia hybrida under salt stress[C].The 3rd Conference on Key Technology of Horticulture,Shenyang,2011:403-406.
[9]MA Y H,SONG Y,ZHANG H R.Effects of uniconazole on the drought resistance of perennial ryegrass[J].Pratacult Sci,2009,26(5):169-173.(in Chinese)马艳华,宋瑜,张洪荣.烯效唑对黑麦草抗旱生理特性的影响[J].草业科学,2009,26(5):169-173.
[10]WU C T,MORRIS J R.Genes,genetics and epigenetics:a correspondence[J].Science,2001,293(5532):1103-1105.
[11]DOWEN R H,PELIZZOLA M,SCHMITZ R J,et al.Widespread dynamic DNA methylation in response to biotic stress[J].Proc Natl Acad Sci USA,2012,109(32):E2183-E2191.doi:10.1073/pnas.1209329109.
[12]ZHAO Y L,YE W W,WANG J J,et al.Review of DNAmethylation and plant stress-tolerance[J].Acta Bot BorealiOccident Sin,2009,29(7):1479-1489.(in Chinese)赵云雷,叶武威,王俊娟,等.DNA甲基化与植物抗逆性研究进展[J].西北植物学报,2009,29(7):1479-1489.
[13]FRAGA M F,ESTELLER M.DNA methylation:a profile of methods and applications[J].Biotechniques,2002,33(3):632,634,636-649.
[14]HUANG X.Study on physiological and molecular mechanism of cold resistance enhanced by exogenous ABA application in sugarcane[D].Nanning:Guangxi University,2012.(in Chinese).黄杏.外源ABA提高甘蔗抗寒性的生理及分子机制研究[D].广西南宁:广西大学,2012.
[15]PAN Y J,FU B Y,WANG D,et al.Spatial and temporal profiling of DNA methylation induced by drought stress in rice[J].Scient Agr Sin,2009,42(9):3009-3018.(in Chinese)潘雅姣,傅彬英,王迪,等.水稻干旱胁迫诱导DNA甲基化时空变化特征分析[J].中国农业科学,2009,42(9):3009-3018.
[16]HUANG X,LIANG Y S,YANG L T,et al.Effects of abscisic acid and its biosynthesis inhibitor on the activities of antioxidant system of sugarcanes treated by cold stress[J].J South China Agr Univ,2013,34(3):356-361.(in Chinese)黄杏,梁勇生,杨丽涛,等.低温胁迫下脱落酸及合成抑制剂对甘蔗幼苗抗氧化系统的影响[J].华南农业大学学报,2013,34(3):356-361.
[17]WANG H Y,WANG Y F,YANG K J,et al.Effect of exohenous nitric oxide on lipid peroxidation and antioxidant enzymes activity in maize seedings under salt stress[J].J Maize Sci,2014,22(4):84-87.(in Chinese)王洪义,王玉凤,杨克军,等.外源NO对Na Cl胁迫下玉米幼苗脂质过氧化和抗氧化酶活性的影响[J].玉米科学,2014,22(4):84-87.
[18]YAN H,TIAN Z H,XU Y G.Effects of uniconazole on seed germination and physiological characters of sweet corn[J].JChangjiang Vegetables,2009,26(15):40-42.(in Chinese)严寒,田志宏,徐勇刚.烯效唑对甜玉米种子萌发及幼苗生理特性的影响[J].长江蔬菜,2009,26(15):40-42.
[19]CHENG S M.Effects of uniconazole on the stage bulbil of drought-tolerant characteristics of pinellia ternate(Thunb.)Breit[D].Chengdu:Sichuan Agicultural University,2010.(in Chinese)程仕明.烯效唑对半夏珠芽期耐干旱特性的影响[D].成都:四川农业大学,2010.
[20]HAN R L.Research progress of ascorbate peroxidase in tomato[J].J Shanxi Agr Sci,2013,41(6):650-652.(in Chinese)韩瑞丽.番茄抗坏血酸过氧化物酶研究进展[J].山西农业科学,2013,41(6):650-652.
[21]YANG S Q,ZHOU R L,LIANG H M,et al.Relationship of antioxidant enzyme and osmoregulation substance to photosynthese in several desert plants[J].J Desert Res,2015,35(6):1557-1564.(in Chinese)杨淑琴,周瑞莲,梁慧敏,等.沙漠植物抗氧化酶活性及渗透调节物质含量与光合作用的关系[J].中国沙漠,2015,35(6):1557-1564.
[22]SHI Y H,ZHANG Y Q.Effects of soaking seed with uniconazole on the growth of tartary buckwheat under different water conditions[J].Hubei Agr Sci,2015,54(6):1289-1295.(in Chinese)石艳华,张永清.烯效唑浸种对不同水分条件下苦荞生长的影响[J].湖北农业科学,2015,54(6):1289-1295.
[23]YANG W Y,FAN G Q,REN W J,et al.Physiological effect of uniconazole waterless-dressed seeds on root and leaf of wheat[J].Scient Agr Sin,2005,38(7):1339-1345.(in Chinese)杨文钰,樊高琼,任万军,等.烯效唑干拌种对小麦根叶生理功能的影响[J].中国农业科学,2005,38(7):1339-1345.
[24]CHEN Q.The effects of ABA or SA pretreatment on the total level of DNA methylation under water and salt stress[D].Shenyang:Shenyang Normal University,2012.(in Chinese)陈强.ABA、SA预处理对水分和盐胁迫下水稻DNA甲基化水平的影响[D].辽宁沈阳:沈阳师范大学,2012.
[25]DU Z H,WANG B Q,XIE B T,et al.Effects of plant growth regulators on physiological and biochemical characteristics of roots in summer sweetpotato seeding under simulated drought stress[J].Acta Agr Boreali-Occident Sin,2014,23(10):97-104.(in Chinese)杜召海,汪宝卿,解备涛,等.模拟干旱条件下植物生长调节剂对夏薯苗期根系生理生化特性的影响[J].西北农业学报,2014,23(10):97-104.
[26]WANG J W.Response of speckled kidney bean to drought stress and regulation of uniconazole[D].Shenyang:Shenyang Agricultural University,2014.(in Chinese)王景伟.奶花芸豆对干旱胁迫及烯效唑调控的响应[D].辽宁沈阳:沈阳农业大学,2014.
[27]YUAN Z,WANG B Q,JIANG Y,et al.Effects of Uniconazole on physiological and biochemical properties of roots of different Sweetpotato cultivars at seedlings stage[J].Agr Sci Technol,2015,16(4):629-638.
[28]LI N Y,YU Y,FU Y D,et al.Effects of bulb soaking with S3307 on growth and endogenous hormone contents of potting lily(Lilium brownie F.E.brown asiatic hybrids)[J].Plant Physiol Commun,2007,43(1):104-106.(in Chinese)李宁毅,于洋,付印东,等.S3307浸球对盆栽百合生长和内源激素含量的影响[J].植物生理学通讯,2007,43(1):104-106.