辐照降解壳聚糖对小麦幼苗抗旱的生理生化特性影响的研究
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摘要
小麦是世界上广为种植的一种主要的粮食作物,干旱对小麦的影响广泛而严重。壳聚糖是几丁质脱乙酰基后得到的一种氨基多糖,作为高效绿色农药及植物生长调节剂近几年在农业领域得到了广泛的研究与应用。本文以苗期的小麦为实验对象,采用辐照降解后的壳聚糖醋酸溶液浸种并结合叶面喷洒的方法,观测低聚壳聚糖对小麦幼苗抗旱的生理作用。结果表明:
1.选用两种苗期抗旱性不同的小麦,皖46号小麦(抗旱性较弱)和旱选10号(抗旱性较强),利用三种辐照剂量(50kGy、200kGy、500kGy)处理壳聚糖,发现在干旱胁迫下这三种处理的壳聚糖溶液均能明显的提高两种小麦的出苗率、含水量、根干重和根冠比,只是提高的程度不同。其中200kGy处理的壳聚糖溶液对于皖46号小麦幼苗形态性状的影响最显著,50kGy处理的壳聚糖溶液对于旱选10号小麦幼苗形态性状的影响最显著。
2.通过对幼苗生理生化指标的测定,得出:在干旱胁迫期间(7天),200 kGy处理壳聚糖的醋酸溶液作用的皖46号小麦幼苗的可溶性蛋白、叶绿素的含量较对照均有显著的提高(P＜0.05),MDA含量较对照有显著的降低(P＜0.05),抗氧化酶(SOD、POD、CAT)活力在干旱胁迫的第7天与对照达到极显著水平(P＜0.01);50kGy处理壳聚糖的醋酸溶液作用的旱选10号小麦幼苗的可溶性蛋白、叶绿素的含量较对照均有显著的提高(P＜0.05),MDA含量较对照有极显著的降低(P＜0.01),抗氧化酶(SOD、POD、CAT)活力在干旱胁迫的第7天与对照达到极显著水平(P＜0.01)。表明辐照降解后的壳聚糖具有增强活性氧清除能力、保护生物膜功能、提高小麦幼苗抗旱的协同生理作用。
As a major crop,wheat is planted abroad in the world.The effect of drought to wheat is seriously.Chitosan is a kind of aminoploysaccharides,derived from chitin by deacetylating.In recent years,chitosan has been investigated and used abroad as non-contamination pesticide and growth regulator in agricultural area.
This experiment was conducted with drought light-resistant wheat cultivar Wan NO.46 and drought resistant wheat cultivar Hanxuan NO.10,irradiated chitosan with three kinds of dose(50kGy、200kGy、500kGy) acetum was dealt with the wheat seeds and was prayed on the leaves of wheat seedlings respectively,then observed the physiological and biochemical effect of irradiated chitosan on drought resistance of wheat.The results showed that:
The three kinds of chitosan solution could improve the germination percentage of seed,moisture percentage of seedling,the dry weight of root,ratio of root to shoot of two kinds of wheat compared with the control,but increased with different degree.Chitosan with 200kGy dose treatment has great effect on the seedling shape character of wheat Wan NO.46 than another two treatments.In addition,chitosan with 50kGy dose treatment has great effect on the seedling shape character of wheat Hanxuan NO.10.
The physiological and biochemical indexs of wheat seedlings were measured,the results indicated that:During the whole stage of drought stress,for the wheat Wan NO.46 seedlings treated by chitosan with 200kGy dose and the wheat Hanxuan NO.10 seedlings treated by chitosan with 50kGy,the soluble protein contents and the contents ofchlorophyll were increased remarkably compared with the control(P＜0.05),the contents of malondialdehyde (MDA)were decreased obviously(P＜0.05).The activities of antioxidases such as superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)were increased significantly compared with the control(P＜0.01),which could make the performance period last more than 7 days.This study indicated that irradiated chitosan was capable of improving the ability to eliminate reactive oxygen species,protecting the functions of bio-membrane, raising the physiological synergisms in drought resistance of wheat seedlings.
1.选用两种苗期抗旱性不同的小麦,皖46号小麦(抗旱性较弱)和旱选10号(抗旱性较强),利用三种辐照剂量(50kGy、200kGy、500kGy)处理壳聚糖,发现在干旱胁迫下这三种处理的壳聚糖溶液均能明显的提高两种小麦的出苗率、含水量、根干重和根冠比,只是提高的程度不同。其中200kGy处理的壳聚糖溶液对于皖46号小麦幼苗形态性状的影响最显著,50kGy处理的壳聚糖溶液对于旱选10号小麦幼苗形态性状的影响最显著。
2.通过对幼苗生理生化指标的测定,得出:在干旱胁迫期间(7天),200 kGy处理壳聚糖的醋酸溶液作用的皖46号小麦幼苗的可溶性蛋白、叶绿素的含量较对照均有显著的提高(P＜0.05),MDA含量较对照有显著的降低(P＜0.05),抗氧化酶(SOD、POD、CAT)活力在干旱胁迫的第7天与对照达到极显著水平(P＜0.01);50kGy处理壳聚糖的醋酸溶液作用的旱选10号小麦幼苗的可溶性蛋白、叶绿素的含量较对照均有显著的提高(P＜0.05),MDA含量较对照有极显著的降低(P＜0.01),抗氧化酶(SOD、POD、CAT)活力在干旱胁迫的第7天与对照达到极显著水平(P＜0.01)。表明辐照降解后的壳聚糖具有增强活性氧清除能力、保护生物膜功能、提高小麦幼苗抗旱的协同生理作用。
As a major crop,wheat is planted abroad in the world.The effect of drought to wheat is seriously.Chitosan is a kind of aminoploysaccharides,derived from chitin by deacetylating.In recent years,chitosan has been investigated and used abroad as non-contamination pesticide and growth regulator in agricultural area.
This experiment was conducted with drought light-resistant wheat cultivar Wan NO.46 and drought resistant wheat cultivar Hanxuan NO.10,irradiated chitosan with three kinds of dose(50kGy、200kGy、500kGy) acetum was dealt with the wheat seeds and was prayed on the leaves of wheat seedlings respectively,then observed the physiological and biochemical effect of irradiated chitosan on drought resistance of wheat.The results showed that:
The three kinds of chitosan solution could improve the germination percentage of seed,moisture percentage of seedling,the dry weight of root,ratio of root to shoot of two kinds of wheat compared with the control,but increased with different degree.Chitosan with 200kGy dose treatment has great effect on the seedling shape character of wheat Wan NO.46 than another two treatments.In addition,chitosan with 50kGy dose treatment has great effect on the seedling shape character of wheat Hanxuan NO.10.
The physiological and biochemical indexs of wheat seedlings were measured,the results indicated that:During the whole stage of drought stress,for the wheat Wan NO.46 seedlings treated by chitosan with 200kGy dose and the wheat Hanxuan NO.10 seedlings treated by chitosan with 50kGy,the soluble protein contents and the contents ofchlorophyll were increased remarkably compared with the control(P＜0.05),the contents of malondialdehyde (MDA)were decreased obviously(P＜0.05).The activities of antioxidases such as superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)were increased significantly compared with the control(P＜0.01),which could make the performance period last more than 7 days.This study indicated that irradiated chitosan was capable of improving the ability to eliminate reactive oxygen species,protecting the functions of bio-membrane, raising the physiological synergisms in drought resistance of wheat seedlings.
引文
[1]马鹏鹏.甲壳素及其衍生物在农业生产中的应用.植物生理学通讯,2001,37(5):475-478
[2]陈薄智.[c].中国第一届甲壳质化学学术会议论文摘要集,1996,1
[3]刘万顺,等.[M].中国甲壳资源研讨会论文集.1997上册:5
[4]何福相.以壳聚糖为原料的种子处理剂制造方法[P].CN:1044 748,1990-08
[5]严瑞宣.水溶性高分子[M].北京:化学工业出版社,1989.562
[6]蒋挺大.甲壳素[M].北京:中国环境科学出版社,1996
[7]铃木隆司.JP(Japen Patent,日本专利)昭64-13946
[8]陈安和,孙敏,李坤培.几丁质对草莓的保鲜作用研究[J].西南农业大学学报,1994,16(4):333-335
[9]杨铭铎,曲志华,李元瑞.几丁质/几丁聚糖及其衍生物在食品中的作用[J].食品科学,2001,22(5):97-99.
[10]Benhamou N,Lafontainc P J,Nicolc M.Induction of system resistance to fusarium crown and rot in tomato plants by seed treatment with chitosan.Phytopathology[J].1994,84(12):1432-1444
[11]E.haouth A,A rul J,Grenicr J,et al.Effect of chitosan on cucumber plants suppression of Pythiumap hanidermatum and induction of defense reactions[J].Phytopathology.1994,84(3):313-320
[12]Hirano S,HayashiM,N agao N,et al.Chitinase activity of some seeds during their germination process and its induction by treating with chitosan and derivatives.In:Chitin and Chitosan:Sources,Chemistry,Biochcrnistry,Physical,Properties,and Appications.Edited by Skjak B,Gudmund,London U K.1998:743-747
[13]于汉寿,吴汉章,张益明,等.水溶性壳聚糖对水稻恶苗病和油菜菌核病的作用[J].江苏农业科学,1998,(5):38-401
[14]E.haouth A,A rul J,Grenier J,et al.Antifungal activity of chitosan on two postharvest pathogens of strawberry Fruits[J].Phytopathology.1992,82:398-402.
[15]刘鹏飞,刘西莉.壳聚糖作为种衣剂成膜剂应用效果研究[J].农药,2004,43(7):312-314
[16]刘环淑[J].食品工业(台湾).1994,26(1):26-36
[17]孙多先[M]中国第一届甲壳质化学学术会议论文摘要集.1996,14
[18]夏文水,吴焱楠.甲壳低聚糖功能性质[J]无锡轻工业大学学报,1996,15(4):297
[19]Tamo Fukamizo.[J].Agric Biol Chem.1991,(55):2653
[20]A Domard,et al.[J].Advances in chitin science.1997,2:156
[21]福田猛他.[J].纤维学会志,1991.47:452
[22]Puhakainen T,Hess M W,Makela P et al.Overexpression of multiple dehydfin genes enhances tolerance to freezing stress in Arabidopsis.Plant Mol.Biol.,2004,51:1-11
[23]Ashraf M,Foolad M R.Roles of glycine betaine and praline in improving plant abiotic stress resistance.Environ.Exp.Bot.,2006,59:206-216
[24]李原园,李英能,苏人琼等编.中国农业水危机及其对策[R].北京:中国国家科学技术委员会农村科技司,1997:52-54
[25]薛国希,高辉远,李鹏民,邹琦,低温下壳聚糖处理对黄瓜幼苗生理生化特性的影响,植物生理与分子生物学学报 2004,30(4):441-448
[26]舒英杰,时侠清,张子学,隋益虎,壳聚糖对黄瓜种子萌发及幼苗抗冷性的效应,种子,2007,26(1):22-25
[27]李茂富,李绍鹏,赵维峰,壳聚糖提高香蕉幼苗抗冷性的效应,植物生理学通讯,2005,41(4):464-466
[28]宋士清,尚庆茂,郭世荣,张志刚,壳聚糖对黄瓜幼苗抗盐的协同生理作用研究,西北植物学报,2006,26(3):435-441
[29]阮松林,薛庆中,壳聚糖包衣对杂交水稻种子发芽和幼苗耐盐性的影响,作物学报,2002,28(6):803-808
[30]刘晚苟,黄小明,杨秀坚。壳聚糖对芒果离体叶片渗透胁迫的缓解效应,干旱地区农业研究,2004,22(4):195-197
[31]刘晚苟,廖锡佳。壳聚糖对沿阶草叶片抗渗透胁迫的影响。湛江师范学院学报,2004,25(3):55-57
[32]Jaroslaw M,Wasikiewic z,Fumio Yoshii,Naotsugu Nagasawa,Degradation of chitosan and sodium alginate by gamma radiation,sonochemical and ultraviolet methods,Radiation Physics and Chemistry,2005,287-295
[33]Maolin Zhai,Long Zhao,Fumio Yoshii,Tamikazu Kume,Study on antibacterial starch/chitosan blend film formed under the action of irradiation,Carbohydrate Polymers,2004,83-88
[34]张志良.植物生理学实验指导[M].北京:高等教育出版社,1999
[35]梁新华,徐兆桢,等.小麦抗旱生理研究现状与思考,甘肃农业科技,2001(2):24-27
[36]Mc CORD J M.FRIDOVICH I.Super Oxide Dismutasean Enzymatic Function for Erythrocuprein(hemocuprein)[J].J.Biol.Chem.1969(244):6049-6055
[37]王利军,黄卫东,李家永。SA对葡萄幼苗叶片膜脂过氧化的影响[J]中国农业科学,2003,36(9):1076-1080
[38]Huff A.Peroxides—catalysed oxidation of chlorophyll by hydrogen peroxide[J].Phytochem.,1982,21:261-265
[39]石瑛,杜昱光,白雪芳.寡聚糖诱导的植物抗性信号转导.生命科学研究,2001,5:29-32
[40]Zhao X M,She X P,Du Y G et al.Induction of antiviral resistance and stimulary efect by oligochitosan in tobacco.Pesticide Biochemistry and Physiology,2007,87:78-84
[41]She X P,Song X G,He J M.Role and Relationship of Nitric Oxide and Hydrogen Peroxide in"Light/Dak—regulated Stomatal Movement in Vi-cia faba.Acta Botanica Sinica,2004,46:1292-1300
[42]张学昆,唐章林,谌利等.脱乙酰化几丁质的乙酰化程度对诱导油菜抗性的影响.中国农业科学,2002,35(3):287-290
[43]Bhaskara R M V,Arul J,Angers P et al.Chitosan treatment of wheat seeds induces resistance to Fusarium graminearum and improves seed quality.J AgIic.Food.Chem.,1999,47:1208-1216)
[44]CONRA TH U DOMARD A,KAU SS H.Chitosan-elicited synthesis of callose and of coumarin derivatives in parsley cell suspension cultures[J].Plant Cell R ep ort,1989,8:152-155
[45]KOHLE H,JEBL ICK W,POTEN F,BLASCHEK W,KAUSS H.Chitosan-elicited callose synthesis in soybean cells as a Ca~(2+)-dependent process[J].P lant P hysiol.,1985,77(3):544-551
[46]WALKER-SMMON SM,RYAN C A.P roteinase inhibitor synthesis in tomato leaves[J].P lant Physiol.,1984,76(3):787-790
[47]Lee S,Choi H,Suh S et al.Oligogalaturonic acid and chitosan reduce stomatal aperture by inducing the evolution of reactive oxygen species from guard cells of tomato and Commelina communis[J].Plant Physiol.,1999,121:147-152
[48]Marco B,Markus F,Gaylon C S et al.Reduction of transpiration through foliar application of Chitosan.AgIic For.Meteorol.,2001,107:167-175
[2]陈薄智.[c].中国第一届甲壳质化学学术会议论文摘要集,1996,1
[3]刘万顺,等.[M].中国甲壳资源研讨会论文集.1997上册:5
[4]何福相.以壳聚糖为原料的种子处理剂制造方法[P].CN:1044 748,1990-08
[5]严瑞宣.水溶性高分子[M].北京:化学工业出版社,1989.562
[6]蒋挺大.甲壳素[M].北京:中国环境科学出版社,1996
[7]铃木隆司.JP(Japen Patent,日本专利)昭64-13946
[8]陈安和,孙敏,李坤培.几丁质对草莓的保鲜作用研究[J].西南农业大学学报,1994,16(4):333-335
[9]杨铭铎,曲志华,李元瑞.几丁质/几丁聚糖及其衍生物在食品中的作用[J].食品科学,2001,22(5):97-99.
[10]Benhamou N,Lafontainc P J,Nicolc M.Induction of system resistance to fusarium crown and rot in tomato plants by seed treatment with chitosan.Phytopathology[J].1994,84(12):1432-1444
[11]E.haouth A,A rul J,Grenicr J,et al.Effect of chitosan on cucumber plants suppression of Pythiumap hanidermatum and induction of defense reactions[J].Phytopathology.1994,84(3):313-320
[12]Hirano S,HayashiM,N agao N,et al.Chitinase activity of some seeds during their germination process and its induction by treating with chitosan and derivatives.In:Chitin and Chitosan:Sources,Chemistry,Biochcrnistry,Physical,Properties,and Appications.Edited by Skjak B,Gudmund,London U K.1998:743-747
[13]于汉寿,吴汉章,张益明,等.水溶性壳聚糖对水稻恶苗病和油菜菌核病的作用[J].江苏农业科学,1998,(5):38-401
[14]E.haouth A,A rul J,Grenier J,et al.Antifungal activity of chitosan on two postharvest pathogens of strawberry Fruits[J].Phytopathology.1992,82:398-402.
[15]刘鹏飞,刘西莉.壳聚糖作为种衣剂成膜剂应用效果研究[J].农药,2004,43(7):312-314
[16]刘环淑[J].食品工业(台湾).1994,26(1):26-36
[17]孙多先[M]中国第一届甲壳质化学学术会议论文摘要集.1996,14
[18]夏文水,吴焱楠.甲壳低聚糖功能性质[J]无锡轻工业大学学报,1996,15(4):297
[19]Tamo Fukamizo.[J].Agric Biol Chem.1991,(55):2653
[20]A Domard,et al.[J].Advances in chitin science.1997,2:156
[21]福田猛他.[J].纤维学会志,1991.47:452
[22]Puhakainen T,Hess M W,Makela P et al.Overexpression of multiple dehydfin genes enhances tolerance to freezing stress in Arabidopsis.Plant Mol.Biol.,2004,51:1-11
[23]Ashraf M,Foolad M R.Roles of glycine betaine and praline in improving plant abiotic stress resistance.Environ.Exp.Bot.,2006,59:206-216
[24]李原园,李英能,苏人琼等编.中国农业水危机及其对策[R].北京:中国国家科学技术委员会农村科技司,1997:52-54
[25]薛国希,高辉远,李鹏民,邹琦,低温下壳聚糖处理对黄瓜幼苗生理生化特性的影响,植物生理与分子生物学学报 2004,30(4):441-448
[26]舒英杰,时侠清,张子学,隋益虎,壳聚糖对黄瓜种子萌发及幼苗抗冷性的效应,种子,2007,26(1):22-25
[27]李茂富,李绍鹏,赵维峰,壳聚糖提高香蕉幼苗抗冷性的效应,植物生理学通讯,2005,41(4):464-466
[28]宋士清,尚庆茂,郭世荣,张志刚,壳聚糖对黄瓜幼苗抗盐的协同生理作用研究,西北植物学报,2006,26(3):435-441
[29]阮松林,薛庆中,壳聚糖包衣对杂交水稻种子发芽和幼苗耐盐性的影响,作物学报,2002,28(6):803-808
[30]刘晚苟,黄小明,杨秀坚。壳聚糖对芒果离体叶片渗透胁迫的缓解效应,干旱地区农业研究,2004,22(4):195-197
[31]刘晚苟,廖锡佳。壳聚糖对沿阶草叶片抗渗透胁迫的影响。湛江师范学院学报,2004,25(3):55-57
[32]Jaroslaw M,Wasikiewic z,Fumio Yoshii,Naotsugu Nagasawa,Degradation of chitosan and sodium alginate by gamma radiation,sonochemical and ultraviolet methods,Radiation Physics and Chemistry,2005,287-295
[33]Maolin Zhai,Long Zhao,Fumio Yoshii,Tamikazu Kume,Study on antibacterial starch/chitosan blend film formed under the action of irradiation,Carbohydrate Polymers,2004,83-88
[34]张志良.植物生理学实验指导[M].北京:高等教育出版社,1999
[35]梁新华,徐兆桢,等.小麦抗旱生理研究现状与思考,甘肃农业科技,2001(2):24-27
[36]Mc CORD J M.FRIDOVICH I.Super Oxide Dismutasean Enzymatic Function for Erythrocuprein(hemocuprein)[J].J.Biol.Chem.1969(244):6049-6055
[37]王利军,黄卫东,李家永。SA对葡萄幼苗叶片膜脂过氧化的影响[J]中国农业科学,2003,36(9):1076-1080
[38]Huff A.Peroxides—catalysed oxidation of chlorophyll by hydrogen peroxide[J].Phytochem.,1982,21:261-265
[39]石瑛,杜昱光,白雪芳.寡聚糖诱导的植物抗性信号转导.生命科学研究,2001,5:29-32
[40]Zhao X M,She X P,Du Y G et al.Induction of antiviral resistance and stimulary efect by oligochitosan in tobacco.Pesticide Biochemistry and Physiology,2007,87:78-84
[41]She X P,Song X G,He J M.Role and Relationship of Nitric Oxide and Hydrogen Peroxide in"Light/Dak—regulated Stomatal Movement in Vi-cia faba.Acta Botanica Sinica,2004,46:1292-1300
[42]张学昆,唐章林,谌利等.脱乙酰化几丁质的乙酰化程度对诱导油菜抗性的影响.中国农业科学,2002,35(3):287-290
[43]Bhaskara R M V,Arul J,Angers P et al.Chitosan treatment of wheat seeds induces resistance to Fusarium graminearum and improves seed quality.J AgIic.Food.Chem.,1999,47:1208-1216)
[44]CONRA TH U DOMARD A,KAU SS H.Chitosan-elicited synthesis of callose and of coumarin derivatives in parsley cell suspension cultures[J].Plant Cell R ep ort,1989,8:152-155
[45]KOHLE H,JEBL ICK W,POTEN F,BLASCHEK W,KAUSS H.Chitosan-elicited callose synthesis in soybean cells as a Ca~(2+)-dependent process[J].P lant P hysiol.,1985,77(3):544-551
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