过氧化氢缓解裸燕麦幼苗低温胁迫的主成分和隶属函数分析
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摘要
为探明信号分子过氧化氢(H_2O_2)提高裸燕麦幼苗耐冷性的作用,以‘定莜6号’沙培幼苗为材料,在3叶期喷施10μmol·L~(-1)H_2O_212 h后于8℃/5℃(昼/夜)条件下低温胁迫,以喷蒸馏水(H_2O)为对照,分别在低温处理的0、1、2、3、4、5 d取幼苗叶片测定超氧阴离子(O_2~·)、H_2O_2、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、抗坏血酸(AsA)、谷胱甘肽(GSH)、可溶性糖(SS)、脯氨酸(Pro)、可溶性蛋白质(SP)和热稳定性蛋白质(HSP) 13项与耐冷性有关的生理指标及低温处理5 d后植株株高和生物量增量,采用主成分和隶属函数分析综合评价H_2O_2对裸燕麦幼苗耐冷性的影响。结果表明,与对照相比,喷施H_2O_2显著提高了低温胁迫下裸燕麦幼苗株高和生物量增量,降低了裸燕麦幼苗叶片O_2~·和MDA及低温胁迫2~5 d的H_2O_2含量,促进低温胁迫期间裸燕麦幼苗叶片SOD、CAT、POD和APX活性提高及AsA、GSH、SS、Pro、SP和HSP积累。主成分分析13项生理指标离差标准化数据,提取的前4个主成分累积方差贡献率达85. 6%;隶属函数综合评价4个主成分得分值显示,喷施H_2O_2显著提高了低温胁迫0~5 d的综合评价值。表明喷施H_2O_2能够通过调控生理生化代谢提高裸燕麦幼苗的耐冷性。
In order to explore whether H_2O_2 could improve the chilling tolerance in naked oat seedlings,threeleaf period ‘Dingyou No. 6'seedlings of sand culture were sprayed with 10 μmol·L~(-1) H_2O_2 for 12 h before8℃/5℃( day/night) low temperature stress,spraying distilled water as the control,and 13 physiological indexes related to cold resistance including superoxide anion( O_2~·), H_2O_2, malondialdehyde( MDA),superoxide dismutase( SOD),catalase( CAT),peroxidase( POD),ascorbic acid peroxidase( APX),ascorbic acid( AsA),glutathione( GSH),soluble sugar( SS),proline( Pro),soluble protein( SP) and heat stable protein( HSP) in the seedling leaves under low temperature stress for 0,1,2,3,4 and 5 d as well as the plant height and biomass increments of the seedlings processed for 5 d were determined,respectively. By principal component and subordinate function analyses,the effect of H_2O_2 on the cold resistance was evaluated in naked oat seedlings. Compared with the control,H_2O_2 treatment could significantly enhance the plant height incrementand biomass increment of naked oat seedlings under low temperature stress,decrease O_2~· and MDA contents in leaves during low temperature stress and H_2O_2 content of low temperature stress for 2-5 d,and increase the activities of SOD,CAT,POD,APX and contents of AsA,GSH,SS,Pro,SP and HSP of naked oat seedling leaves. From 13 physiological indexes converted data matrix and principal component analysis,the cumulative variance contribution of four principal factors extracted was 85. 6%. By subordinate function analysis,spraying H_2O_2 significantly improved the comprehensive evaluation value of the naked oat seedling leaves under low temperature stress for 0-5 d. Therefore,spraying H_2O_2 can improve the cold resistance of naked oat seedlings by regulating the physiological and biochemical metabolism.
In order to explore whether H_2O_2 could improve the chilling tolerance in naked oat seedlings,threeleaf period ‘Dingyou No. 6'seedlings of sand culture were sprayed with 10 μmol·L~(-1) H_2O_2 for 12 h before8℃/5℃( day/night) low temperature stress,spraying distilled water as the control,and 13 physiological indexes related to cold resistance including superoxide anion( O_2~·), H_2O_2, malondialdehyde( MDA),superoxide dismutase( SOD),catalase( CAT),peroxidase( POD),ascorbic acid peroxidase( APX),ascorbic acid( AsA),glutathione( GSH),soluble sugar( SS),proline( Pro),soluble protein( SP) and heat stable protein( HSP) in the seedling leaves under low temperature stress for 0,1,2,3,4 and 5 d as well as the plant height and biomass increments of the seedlings processed for 5 d were determined,respectively. By principal component and subordinate function analyses,the effect of H_2O_2 on the cold resistance was evaluated in naked oat seedlings. Compared with the control,H_2O_2 treatment could significantly enhance the plant height incrementand biomass increment of naked oat seedlings under low temperature stress,decrease O_2~· and MDA contents in leaves during low temperature stress and H_2O_2 content of low temperature stress for 2-5 d,and increase the activities of SOD,CAT,POD,APX and contents of AsA,GSH,SS,Pro,SP and HSP of naked oat seedling leaves. From 13 physiological indexes converted data matrix and principal component analysis,the cumulative variance contribution of four principal factors extracted was 85. 6%. By subordinate function analysis,spraying H_2O_2 significantly improved the comprehensive evaluation value of the naked oat seedling leaves under low temperature stress for 0-5 d. Therefore,spraying H_2O_2 can improve the cold resistance of naked oat seedlings by regulating the physiological and biochemical metabolism.
引文
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3. Jiang J L,Su M,Wang L Y,et al. Exogenous hydrogen peroxide reversibly inhibits root gravitropism and induces horizontal curvature of primary root during grass pea germination[J]. Plant Physiology and Biochemistry,2012,53:84-93.
4. Su L Q,Lan Q Y,Pritchard H W,et al. Reactive oxygen species induced by cold stratification promote germination of Hedysarum scoparium seeds[J]. Plant Physiology and Biochemistry,2016,109:406-415.
5. 庞学群,潘少丽,王海波,等.过氧化氢在采后香蕉果实耐冷性诱导中的作用[J].园艺学报,2007,34(6):1373-1378.Pang X Q,Pan S L,Wang H B,et al. Role of hydrogen peroxide on chilling resistance of postharvest banana fruits[J]. Acta Horticulturae Sinica,2007,34(6):1373-1378.
6. 蒋景龙.外源H2O2对低温胁迫下柑橘叶片抗寒性的影响[J].西北植物学报,2016,36(3):499-505.Jiang J L. Effect of exogenous H2O2treatment on cold resistance of Citrus reticulata leaves under low temperature stress[J]. Acta Botanica Boreali-Occidentalia Sinica,2016,36(3):499-505.
7. 韩娜,冯汉青,李燕鸿,等.外源H2O2对水稻幼苗抗寒性的调节作用[J].西北植物学报,2009,29(6):1214-1219.Han N,Feng H Q,Li Y H,et al. Regulation of exogenous H2O2on rice seedlings chilling tolerance[J]. Acta Botanica Boreali-Occidentalia Sinica,2009,29(6):1214-1219.
8. Yu C W,Murphy T M,Sung W W,et al. H2O2treatment induces glutathione accumulation and chilling tolerance in mung bean[J]. Functional Plant Biology,2002,29(9):1081-1087.
9. 万林,张曼,钟飞燕,等. H2O2浸种处理对晚直播油菜苗期耐寒性的影响[J].中国油料作物学报,2015,37(6):811-819.Wan L,Zhang M,Zhong F Y,et al. Effects of seed soaking with hydrogen peroxide on seedling cold tolerance of late direct-sowing rape[J]. Chinese Journal of Oil Crop Sciences,2015,37(6):811-819.
10. Wang Y,Li J L,Wang J Z,et al. Exogenous H2O2improves the chilling tolerance of manilagrass and mascarenegrass by activating the antioxidative system[J]. Plant Growth Regulation,2010,61(2):195-204.
11. Dong X,Bi H,Wu G,et al. Drought-induced chilling tolerance in cucumber involves membrane stabilisation improved by antioxidant system[J]. International Journal of Plant Production,2013,7(1):67-80.
12. 薛盈文,黄寿光,范博文,等.低温和UV-B复合胁迫对小麦幼苗抗氧化酶和渗透调节物质的影响[J].麦类作物学报,2017,37(6):834-840.Xue Y W,Huang S G,Fan B W,et al. Effect of low temperature and UV-B on the antioxidant enzymes and osmotic substances in wheat seedlings[J]. Journal of Triticeae Crops,2017,37(6):834-840.
13. 张曼,戴蓉,张顺凯,等. H2O2浸种对油菜种子低温萌发的缓解效应[J].南京农业大学学报,2017,40(6):963-970.Zhang M,Dai R,Zhang S K,et al. Alleviation effects of seed soaking with H2O2on seed germination in rape under low temperature stress[J]. Journal of Nanjing Agricultural University,2017,40(6):963-970.
14. 叶朝军,万林,孙继,等.过氧化氢浸种处理对油菜幼苗抗寒生理特性的影响[J].江苏农业科学,2017,45(20):104-106,156.Ye C J,Wan L,Sun J,et al. Effects of H2O2soaking treatment on physiological characteristics of oilseed rape seedlings[J]. Jiangsu Agricultural Sciences,2017,45(20):104-106,156.
15. 康国章,陶均,孙谷畴,等. H2O2和Ca2+对受低温胁迫香蕉幼苗抗冷性的影响[J].园艺学报,2002,29(2):119-122.Kang G Z,Tao J,Sun G C,et al. Physiological effects of H2O2and Ca2+on cold stressed banana seedlings[J]. Acta Horticulturae Sinica,2002,29(2):119-122.
16. 王玲,段晓宇,唐敏,等.镧处理下大花蕙兰耐寒性生理指标的综合评价[J].植物生理学报,2014,50(8):1177-1183.Wang L,Duan X Y,Tang M,et al. Comprehensive evaluation of physiological indices of cold resistance in Cymbidium under lanthanum treatment[J]. Plant Physiology Journal,2014,50(8):1177-1183.
17. 杜庆鑫,刘攀峰,魏艳秀,等.基于主成分与聚类分析的杜仲雄花品质综合评价[J].植物研究,2016,36(6):846-852.Du Q X,Liu P F,Wei Y X,et al. Comprehensive evaluation of Eucommia ulmoides male flowers quality by principal component and cluster analysis[J]. Bulletin of Botanical Research,2016,36(6):846-852.
18. Sui N,Li M,Liu X Y,et al. Response of xanthophyll cycle and chloroplastic antioxidant enzymes to chilling stress in tomato over-expressing glycerol-3-phosphate acyltransferase gene[J]. Photosynthetica,2007,45(3):447-454.
19. Sergiev I,Alexieva V,Karanov E. Effect of spermine,atrazine and combination between them on some endogenous protective systems and stress markers in plants[J].Comptes Rendus de I’Academie Bulgare des Sciences,1997,51(2):121-124.
20. 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:195-197,258-261.Li H S. Principles and techniques of plant physiological biochemical experiment[M]. Beijing:Higher Education Press,2000:195-197; 258-261.
21. 陈建勋,王晓峰.植物生理学实验指导[M].广州:华南理工大学出版社,2002:68-77.Chen J X,Wang X F. Plant physiology experiment instruction[M]. Guangzhou:South China University of Technology Press,2006:68-77.
22. 曾韶西,王以柔,李美如.不同胁迫预处理提高水稻幼苗抗寒性期间蛋白质的变化[J].植物学报,1997,39(2):130-136.Zeng S X,Wang Y R,Li M R. Protein changes in rice seedlings during the enhancement of chilling resistance by different stress pretreatments[J]. Acta Botanica Sinica,1997,39(2):130-136.
23. 何文平,王新霞,张旭强,等.低温胁迫下外源脱落酸对高山离子芥抗氧化酶和渗透调节物质的影响[J].植物研究,2016,36(6):870-877.He W P,Wang X X,Zhang X Q,et al. Exogenous ABA affected on antioxidant enzymes and osmotic adjustment in Choraspora bungeana under low temperature stress[J].Bulletin of Botanical Research,2016,36(6):870-877.
24. Chen L J,Xiang H Z,Miao Y,et al. An overview of cold resistance in plants[J]. Journal of Agronomy and Crop Science,2014,200(4):237-245.
25. Kang G Z,Wang Z X,Sun G C. Rarticipation of H2O2in enhancement of cold chilling by salicylic acid in banana seedlings[J]. Acta Botanica Sinica,2003,45(5):567-573.
26. Morsy M R,Jouve L,Hausman J F,et al. Alteration of oxidative and carbohydrate metabolism under abiotic stress in two rice(Oryza sativa L.)genotypes contrasting in chilling tolerance[J]. Journal of Plant Physiology,2007,164(2):157-167.
27. 刘建新,王金成,王瑞娟,等.混合盐碱胁迫下裸燕麦的种子萌发和幼苗逆境生理特征[J].植物研究,2016,36(2):224-231.Liu J X,Wang J C,Wang R J,et al. Effects of complex saline-alkali stress on seed germination and stress-resistance physiological characteristics of naked oat seedlings[J].Bulletin of Botanical Research,2016,36(2):224-231.
28. 吴顺,赵永进,萧浪涛,等.外施H2O2对黄瓜幼苗抗低温胁迫的影响[J].中南林业科技大学学报,2008,28(5):68-70,76.Wu S,Zhao Y J,Xiao L T,et al. Effects of exogenous H2O2on the chilling stress resistance of cucumber seedlings[J].Journal of Central South University of Forestry&Technology,2008,28(5):68-70,76.
2. 林叶春,曾昭海,任长忠,等.局部根区灌溉对裸燕麦光合特征曲线及叶绿素荧光特性的影响[J].作物学报,2012,38(6):1062-1070.Lin Y C,Zeng Z H,Ren C Z,et al. Effects of partial root zone irrigation on leaf photosynthetic curves and chlorophyll fluorescence parameters in naked oat[J]. Acta Agronomica Sinica,2012,38(6):1062-1070.
3. Jiang J L,Su M,Wang L Y,et al. Exogenous hydrogen peroxide reversibly inhibits root gravitropism and induces horizontal curvature of primary root during grass pea germination[J]. Plant Physiology and Biochemistry,2012,53:84-93.
4. Su L Q,Lan Q Y,Pritchard H W,et al. Reactive oxygen species induced by cold stratification promote germination of Hedysarum scoparium seeds[J]. Plant Physiology and Biochemistry,2016,109:406-415.
5. 庞学群,潘少丽,王海波,等.过氧化氢在采后香蕉果实耐冷性诱导中的作用[J].园艺学报,2007,34(6):1373-1378.Pang X Q,Pan S L,Wang H B,et al. Role of hydrogen peroxide on chilling resistance of postharvest banana fruits[J]. Acta Horticulturae Sinica,2007,34(6):1373-1378.
6. 蒋景龙.外源H2O2对低温胁迫下柑橘叶片抗寒性的影响[J].西北植物学报,2016,36(3):499-505.Jiang J L. Effect of exogenous H2O2treatment on cold resistance of Citrus reticulata leaves under low temperature stress[J]. Acta Botanica Boreali-Occidentalia Sinica,2016,36(3):499-505.
7. 韩娜,冯汉青,李燕鸿,等.外源H2O2对水稻幼苗抗寒性的调节作用[J].西北植物学报,2009,29(6):1214-1219.Han N,Feng H Q,Li Y H,et al. Regulation of exogenous H2O2on rice seedlings chilling tolerance[J]. Acta Botanica Boreali-Occidentalia Sinica,2009,29(6):1214-1219.
8. Yu C W,Murphy T M,Sung W W,et al. H2O2treatment induces glutathione accumulation and chilling tolerance in mung bean[J]. Functional Plant Biology,2002,29(9):1081-1087.
9. 万林,张曼,钟飞燕,等. H2O2浸种处理对晚直播油菜苗期耐寒性的影响[J].中国油料作物学报,2015,37(6):811-819.Wan L,Zhang M,Zhong F Y,et al. Effects of seed soaking with hydrogen peroxide on seedling cold tolerance of late direct-sowing rape[J]. Chinese Journal of Oil Crop Sciences,2015,37(6):811-819.
10. Wang Y,Li J L,Wang J Z,et al. Exogenous H2O2improves the chilling tolerance of manilagrass and mascarenegrass by activating the antioxidative system[J]. Plant Growth Regulation,2010,61(2):195-204.
11. Dong X,Bi H,Wu G,et al. Drought-induced chilling tolerance in cucumber involves membrane stabilisation improved by antioxidant system[J]. International Journal of Plant Production,2013,7(1):67-80.
12. 薛盈文,黄寿光,范博文,等.低温和UV-B复合胁迫对小麦幼苗抗氧化酶和渗透调节物质的影响[J].麦类作物学报,2017,37(6):834-840.Xue Y W,Huang S G,Fan B W,et al. Effect of low temperature and UV-B on the antioxidant enzymes and osmotic substances in wheat seedlings[J]. Journal of Triticeae Crops,2017,37(6):834-840.
13. 张曼,戴蓉,张顺凯,等. H2O2浸种对油菜种子低温萌发的缓解效应[J].南京农业大学学报,2017,40(6):963-970.Zhang M,Dai R,Zhang S K,et al. Alleviation effects of seed soaking with H2O2on seed germination in rape under low temperature stress[J]. Journal of Nanjing Agricultural University,2017,40(6):963-970.
14. 叶朝军,万林,孙继,等.过氧化氢浸种处理对油菜幼苗抗寒生理特性的影响[J].江苏农业科学,2017,45(20):104-106,156.Ye C J,Wan L,Sun J,et al. Effects of H2O2soaking treatment on physiological characteristics of oilseed rape seedlings[J]. Jiangsu Agricultural Sciences,2017,45(20):104-106,156.
15. 康国章,陶均,孙谷畴,等. H2O2和Ca2+对受低温胁迫香蕉幼苗抗冷性的影响[J].园艺学报,2002,29(2):119-122.Kang G Z,Tao J,Sun G C,et al. Physiological effects of H2O2and Ca2+on cold stressed banana seedlings[J]. Acta Horticulturae Sinica,2002,29(2):119-122.
16. 王玲,段晓宇,唐敏,等.镧处理下大花蕙兰耐寒性生理指标的综合评价[J].植物生理学报,2014,50(8):1177-1183.Wang L,Duan X Y,Tang M,et al. Comprehensive evaluation of physiological indices of cold resistance in Cymbidium under lanthanum treatment[J]. Plant Physiology Journal,2014,50(8):1177-1183.
17. 杜庆鑫,刘攀峰,魏艳秀,等.基于主成分与聚类分析的杜仲雄花品质综合评价[J].植物研究,2016,36(6):846-852.Du Q X,Liu P F,Wei Y X,et al. Comprehensive evaluation of Eucommia ulmoides male flowers quality by principal component and cluster analysis[J]. Bulletin of Botanical Research,2016,36(6):846-852.
18. Sui N,Li M,Liu X Y,et al. Response of xanthophyll cycle and chloroplastic antioxidant enzymes to chilling stress in tomato over-expressing glycerol-3-phosphate acyltransferase gene[J]. Photosynthetica,2007,45(3):447-454.
19. Sergiev I,Alexieva V,Karanov E. Effect of spermine,atrazine and combination between them on some endogenous protective systems and stress markers in plants[J].Comptes Rendus de I’Academie Bulgare des Sciences,1997,51(2):121-124.
20. 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:195-197,258-261.Li H S. Principles and techniques of plant physiological biochemical experiment[M]. Beijing:Higher Education Press,2000:195-197; 258-261.
21. 陈建勋,王晓峰.植物生理学实验指导[M].广州:华南理工大学出版社,2002:68-77.Chen J X,Wang X F. Plant physiology experiment instruction[M]. Guangzhou:South China University of Technology Press,2006:68-77.
22. 曾韶西,王以柔,李美如.不同胁迫预处理提高水稻幼苗抗寒性期间蛋白质的变化[J].植物学报,1997,39(2):130-136.Zeng S X,Wang Y R,Li M R. Protein changes in rice seedlings during the enhancement of chilling resistance by different stress pretreatments[J]. Acta Botanica Sinica,1997,39(2):130-136.
23. 何文平,王新霞,张旭强,等.低温胁迫下外源脱落酸对高山离子芥抗氧化酶和渗透调节物质的影响[J].植物研究,2016,36(6):870-877.He W P,Wang X X,Zhang X Q,et al. Exogenous ABA affected on antioxidant enzymes and osmotic adjustment in Choraspora bungeana under low temperature stress[J].Bulletin of Botanical Research,2016,36(6):870-877.
24. Chen L J,Xiang H Z,Miao Y,et al. An overview of cold resistance in plants[J]. Journal of Agronomy and Crop Science,2014,200(4):237-245.
25. Kang G Z,Wang Z X,Sun G C. Rarticipation of H2O2in enhancement of cold chilling by salicylic acid in banana seedlings[J]. Acta Botanica Sinica,2003,45(5):567-573.
26. Morsy M R,Jouve L,Hausman J F,et al. Alteration of oxidative and carbohydrate metabolism under abiotic stress in two rice(Oryza sativa L.)genotypes contrasting in chilling tolerance[J]. Journal of Plant Physiology,2007,164(2):157-167.
27. 刘建新,王金成,王瑞娟,等.混合盐碱胁迫下裸燕麦的种子萌发和幼苗逆境生理特征[J].植物研究,2016,36(2):224-231.Liu J X,Wang J C,Wang R J,et al. Effects of complex saline-alkali stress on seed germination and stress-resistance physiological characteristics of naked oat seedlings[J].Bulletin of Botanical Research,2016,36(2):224-231.
28. 吴顺,赵永进,萧浪涛,等.外施H2O2对黄瓜幼苗抗低温胁迫的影响[J].中南林业科技大学学报,2008,28(5):68-70,76.Wu S,Zhao Y J,Xiao L T,et al. Effects of exogenous H2O2on the chilling stress resistance of cucumber seedlings[J].Journal of Central South University of Forestry&Technology,2008,28(5):68-70,76.