甘蔗应答黑穗病菌侵染相关酶活性和内源激素含量的变化
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摘要
【目的】研究不同甘蔗品种与黑穗病菌长期互作过程中相关酶活性及内源激素含量的变化,为甘蔗应答黑穗病菌侵染的抗性机制提供理论依据。【方法】选用抗黑穗病品种桂糖29号和感黑穗病品种崖城71-374,用浸渍法接种黑穗病菌,对照组用无菌水模拟接菌。在接菌后30、60、90和180 d取甘蔗+1位叶为样品,分析几丁质酶、β-1,3-葡聚糖酶和过氧化氢酶(CAT)活性及生长素(IAA)、细胞分裂素(CTK)和乙烯(ETH)含量的变化。【结果】在整个侵染期间,两个甘蔗品种叶片的几丁质酶活性、β-1,3-葡聚糖酶活性及IAA、CTK含量呈逐步降低或波动下降趋势,均在接种后180 d达最低值,而CAT活性整体上呈上升趋势,在接种后180 d达最高值;其中抗性品种桂糖29号的β-1,3-葡聚糖酶活性和ETH含量在整个浸染过程均高于感病品种崖城71-374。【结论】甘蔗在不同时期应答黑穗病菌胁迫的生理生化反应不同,感病品种与抗病品种的防御机制可能存在差异。
【Objective】The changes of related enzyme activities and endogenous hormone contents in different sugarcane varieties during the long term interaction with smut fungus were studied to provide theoretical basis to illustrate the resistance mechanism of sugarcane to smut fungus infection.【Method】The smut-resistant variety GT29 and the smut-susceptible variety Yacheng 71-3744 were selected to be inoculated with smut fungus by dipping method;the control group was treated with sterile water to conduct simulated inoculation with fungus. Sugarcane +1 leaf samples were taken at 30,60,90 and 180 d respectively after the inoculation treatment, the changes of chitinase,β-1, 3-glucanase and catalase(CAT)activities,and auxin(IAA),cytokinin(CTK)and ethylene(ETH)contents were analyzed.【Result】During the entire infection period,activities of chitinase and β-1,3-glucanase and contents of IAA and CTK the two sugarcane varieties leaves showed the trends of gradual decrease or fluctuating decrease,and they reached the lowest level 180 d after inoculation. But the activity of CAT showed an increasing trend and reached the highest level 180 d after inoculation. β-1,3-glucanase activity and ETH content in the smut-resistant variety GT29 were always higher than those in the smut-susceptible variety Yacheng 71-3744 during the whole infection period.【Conclusion】Physiological and biochemical responses of sugarcane to smut fungus stress are different at different growth stages,which may reflect different defense mechanisms between the susceptible variety and the resistant variety.
【Objective】The changes of related enzyme activities and endogenous hormone contents in different sugarcane varieties during the long term interaction with smut fungus were studied to provide theoretical basis to illustrate the resistance mechanism of sugarcane to smut fungus infection.【Method】The smut-resistant variety GT29 and the smut-susceptible variety Yacheng 71-3744 were selected to be inoculated with smut fungus by dipping method;the control group was treated with sterile water to conduct simulated inoculation with fungus. Sugarcane +1 leaf samples were taken at 30,60,90 and 180 d respectively after the inoculation treatment, the changes of chitinase,β-1, 3-glucanase and catalase(CAT)activities,and auxin(IAA),cytokinin(CTK)and ethylene(ETH)contents were analyzed.【Result】During the entire infection period,activities of chitinase and β-1,3-glucanase and contents of IAA and CTK the two sugarcane varieties leaves showed the trends of gradual decrease or fluctuating decrease,and they reached the lowest level 180 d after inoculation. But the activity of CAT showed an increasing trend and reached the highest level 180 d after inoculation. β-1,3-glucanase activity and ETH content in the smut-resistant variety GT29 were always higher than those in the smut-susceptible variety Yacheng 71-3744 during the whole infection period.【Conclusion】Physiological and biochemical responses of sugarcane to smut fungus stress are different at different growth stages,which may reflect different defense mechanisms between the susceptible variety and the resistant variety.
引文
陈爱葵,庄文宋,陈冬梅.2010.植物几丁质酶及其基因工程研究进展[J].广东教育学院学报,30(3):47-53.[Chen A K,Zhuang W S,Chen D M.2010.Advances in the research on genetic engineering of plant chitinase[J].Journal of Guangdong Education Institute,30(3):47-53.]
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顾丽红,张树珍,杨本鹏,蔡文伟,黄东杰,王文治,李娇.2008.几丁质酶和β-1,3-葡聚糖酶基因导入甘蔗[J].分子植物育种,6(2):277-280.[Gu L H,Zhang S Z,Yang B P,Cai W W,Huang D J,Wang W Z,Li J.2008.Introduction of chitin andβ-1,3-glucan into sugarcane[J].Molecular Plant Breeding,6(2):277-280.]
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李欣.2007.植物病原细菌中内源过氧化氢的产生、功能及其在与植物互作中的作用[D].兰州:兰州大学.[Li X.2007.Production and roles of endogenous hydrogen peroxide in phytopathogenic bacteria and its roles in the interaction with plant[D].Lanzhou:Lanzhou University.]
刘守伟,吴凤芝,马艳玲.2009.枯萎病菌对不同抗性黄瓜品种几种酶活性的影响[J].植物保护,35(1):82-85.[Liu S W,Wu F Z,Ma Y L.2009.Effects of fusarium wilt pathogen on the enzyme activity of cucumber cultivars of different resistance[J].Plant Protection,35(1):82-85.]
莫凤连,杨丽涛,潘如科,宋修鹏,李杨瑞.2012.甘蔗黑穗病菌胁迫对甘蔗内源激素含量的影响[J].南方农业学报,43(11):1676-1681.[Mo F L,Yang L T,Pan R K,Song X P,Li Y R.2012.Changes of endogenous hormone content in sugarcane under smut pathogen stress[J].Journal of Southern Agriculture,43(11):1676-1681.]
欧阳石文,赵开军,冯兰香.2002.植物中几丁质酶的作用[J].生物学通报,(6):13-14.[Ouyang S W,Zhao K J,Feng L X.2002.The role of chitinase in plants[J].Bulletin of Biology,(6):13-14.]
苏亚春.2014.甘蔗应答黑穗病菌侵染的转录组与蛋白组研究及抗性相关基因挖掘[D].福州:福建农林大学.[Su Y C.2014.Transcriptomics and proteomics of sugarcane response to Sporisorium scitamineum infection and mining of resistance-related genes[D].Fuzhou:Fujian Agriculture and Forestry University.]
王果萍,王景雪,孙毅,崔贵梅,孟玉平,乔燕祥.2003.几丁质酶基因导入西瓜植株及其抗病性鉴定研究[J].植物遗传资源学报,4(2):104-109.[Wang G P,Wang J X,Sun Y,Cui G M,Meng Y P,Qiao Y X.2003.Transformation of watermelon plants with chintinase gene and evaluation for fusarium wilt resistance[J].Journal of Plant Genetic Resources,4(2):104-109.]
王廷璞,马静静,赵菲佚.2010.β-1,3-葡聚糖酶和几丁质酸在农作物病虫害防治中的研究进展[J].安徽农业科学,38(26):14417-14419.[Wang T P,Ma J J,Zhao F Y.2010.Research advance ofβ-1,3-glucanase and chitinase in controlling insect pests of crops[J].Journal of Anhui A-gricultural Sciences,38(26):14417-14419.]
王晓莉.2008.生长素信号对植物生长与系统性获得抗性的调控作用[D].南京:南京农业大学.[Wang X L.2008.Effects of auxin on the coordinate regulation of plant growth and systemic acquired resistance[D].Nanjing:Nanjing Agricultural University.]
王彦芹,邰红忠,练文明,卢金宝,武刚.2017.基于RNA-Seq的海岛棉“新海17号”黄萎病菌处理的表达谱分析[J].分子植物育种,15(8):2912-2920.[Wang Y Q,Tai H Z,Lian W M,Lu J B,Wu G.2017.Expression profile analysis of gossypium barbadense“Xinhai 17”under verticillium wilt stress by RNA-Seq[J].Molecular Plant Breeding,15(8):2912-2920.]
袁红旭,许新萍,张燕娟,刘晚苟,郭建夫.2009.转抗真菌基因水稻对纹枯病的抗性表达分析[J].植物保护学报,36(6):504-508.[Yuan H X,Xu X P,Zhang Y J,Liu W G,Guo J F.2009.Analysis on resistance to sheath blight of transgenic rice with extra anti-fungual genes[J].Acta Phytophylacica Sinica,36(6):504-508.]
张荣华,何红,张革民,刘海斌,李杨瑞,方锋学,宋焕忠,方位宽,闭少玲.2011.宿根性特强甘蔗新品种桂糖29号的选育[J].中国糖料,(1):1-4.[Zhang R H,He H,Zhang GM,Liu H B,Li Y R,Fang F X,Song H Z,Fang W K,Bi S L.2011.Breeding of new sugarcane variety Guitang29 with high ratoon ability[J].Sugar Crops of China,(1):1-4.]
周思军,李希臣,刘昭军,刘文萍,南相日.2000.通过农杆菌介导将菜豆几丁质酶基因导入马铃薯[J].中国马铃薯,14(2):70-72.[Zhou S J,Li X C,Liu Z J,Liu W P,Nan X R.2000.Introduction of chitinase gene from kidney bean into potato by agrobacterium mediated transformation[J].Chinese Potato,14(2):70-72.]
Adams D J.2004.Fungal cell wall chitinases and glucanases[J].Microbiology,150(7):2029-2035.
Albert H H,Schenck S.1996.PCR amplification from a homolog of the b E mating-type gene as a sensitive assay for the presence of Ustilago scitaminea DNA[J].Plant Disease,80(10):1189-1192.
Broglie K,Chet I,Holliday M,Cressman R,Biddle P,Knowlton S,Mauvais J C,Broglie R.1991.Transgenic plants with enhanced resistance to the fungal pathogen rhizoctonia solani[J].Science,24(5035):1194-1197.
Martinez M,Medina I,Naranjo S,Rodriguez C,Armas R,Pinon D,Vicente C,Legaz M E.2000.Changes of some chemical parameters,involved in sucrose recovery from sugarcane juices,related to the susceptibility or resistance of sugarcane plants to smut(Ustilagoscitaminea)[J].Indian Sugar,50(6):341-346.
Mhamdi A,Queval G,Chaouch S,Vanderauwera S,Van Breusegem F,Noctor G.2010.Catalase function in plants:A focus on Arabidopsis mutants as stress-mimic models[J].Journal of Experimental Botany,61(15):4197-4220.
Peters L P,Carvalho G,Vilhena M B,Creste S,Azevedo R A,Monteiro-Vitorello C B.2017.Functional analysis of oxidative burst in sugarcane smut-resistant and susceptible genotypes[J].Planta,245(4):749-764.
高轶静,张革民,张荣华,宋焕忠,罗霆,段维兴,贤武,廖江雄,周会,游建华.2013.甘蔗优良新品种(系)的黑穗病抗性鉴定[J].中国糖料,(2):25-26.[Gao Y J,Zhang GM,Zhang R H,Song H Z,Luo T,Duan W X,Xian W,Liao J X,Zhou H,You J H.2013.Evaluation of resistance to smut disease in new sugarcane varieties and breeding lines[J].Sugar Crops of China,(2):25-26.]
顾丽红,张树珍,杨本鹏,蔡文伟,黄东杰,王文治,李娇.2008.几丁质酶和β-1,3-葡聚糖酶基因导入甘蔗[J].分子植物育种,6(2):277-280.[Gu L H,Zhang S Z,Yang B P,Cai W W,Huang D J,Wang W Z,Li J.2008.Introduction of chitin andβ-1,3-glucan into sugarcane[J].Molecular Plant Breeding,6(2):277-280.]
蓝海燕,王长海,张丽华,刘桂珍,王岚兰,陈正华,田颖川.2000.导入β-1,3-葡聚糖酶及几丁质酶基因的转基因可育油菜及其抗菌核病的研究[J].生物工程学报,16(2):26-30.[Lan H Y,Wang C H,Zhang L H,Liu G Z,Wang L L,Chen Z H,Tian Y C.2000.Studies on transgenic oilseed rape(Brassica napus)plants transformed with beta-1,3-glucanase and chitinase genes and its resistance to Sclerotinia sclerotiorium[J].Chinese Journal of Biotechnology,16(2):26-30.]
李欣.2007.植物病原细菌中内源过氧化氢的产生、功能及其在与植物互作中的作用[D].兰州:兰州大学.[Li X.2007.Production and roles of endogenous hydrogen peroxide in phytopathogenic bacteria and its roles in the interaction with plant[D].Lanzhou:Lanzhou University.]
刘守伟,吴凤芝,马艳玲.2009.枯萎病菌对不同抗性黄瓜品种几种酶活性的影响[J].植物保护,35(1):82-85.[Liu S W,Wu F Z,Ma Y L.2009.Effects of fusarium wilt pathogen on the enzyme activity of cucumber cultivars of different resistance[J].Plant Protection,35(1):82-85.]
莫凤连,杨丽涛,潘如科,宋修鹏,李杨瑞.2012.甘蔗黑穗病菌胁迫对甘蔗内源激素含量的影响[J].南方农业学报,43(11):1676-1681.[Mo F L,Yang L T,Pan R K,Song X P,Li Y R.2012.Changes of endogenous hormone content in sugarcane under smut pathogen stress[J].Journal of Southern Agriculture,43(11):1676-1681.]
欧阳石文,赵开军,冯兰香.2002.植物中几丁质酶的作用[J].生物学通报,(6):13-14.[Ouyang S W,Zhao K J,Feng L X.2002.The role of chitinase in plants[J].Bulletin of Biology,(6):13-14.]
苏亚春.2014.甘蔗应答黑穗病菌侵染的转录组与蛋白组研究及抗性相关基因挖掘[D].福州:福建农林大学.[Su Y C.2014.Transcriptomics and proteomics of sugarcane response to Sporisorium scitamineum infection and mining of resistance-related genes[D].Fuzhou:Fujian Agriculture and Forestry University.]
王果萍,王景雪,孙毅,崔贵梅,孟玉平,乔燕祥.2003.几丁质酶基因导入西瓜植株及其抗病性鉴定研究[J].植物遗传资源学报,4(2):104-109.[Wang G P,Wang J X,Sun Y,Cui G M,Meng Y P,Qiao Y X.2003.Transformation of watermelon plants with chintinase gene and evaluation for fusarium wilt resistance[J].Journal of Plant Genetic Resources,4(2):104-109.]
王廷璞,马静静,赵菲佚.2010.β-1,3-葡聚糖酶和几丁质酸在农作物病虫害防治中的研究进展[J].安徽农业科学,38(26):14417-14419.[Wang T P,Ma J J,Zhao F Y.2010.Research advance ofβ-1,3-glucanase and chitinase in controlling insect pests of crops[J].Journal of Anhui A-gricultural Sciences,38(26):14417-14419.]
王晓莉.2008.生长素信号对植物生长与系统性获得抗性的调控作用[D].南京:南京农业大学.[Wang X L.2008.Effects of auxin on the coordinate regulation of plant growth and systemic acquired resistance[D].Nanjing:Nanjing Agricultural University.]
王彦芹,邰红忠,练文明,卢金宝,武刚.2017.基于RNA-Seq的海岛棉“新海17号”黄萎病菌处理的表达谱分析[J].分子植物育种,15(8):2912-2920.[Wang Y Q,Tai H Z,Lian W M,Lu J B,Wu G.2017.Expression profile analysis of gossypium barbadense“Xinhai 17”under verticillium wilt stress by RNA-Seq[J].Molecular Plant Breeding,15(8):2912-2920.]
袁红旭,许新萍,张燕娟,刘晚苟,郭建夫.2009.转抗真菌基因水稻对纹枯病的抗性表达分析[J].植物保护学报,36(6):504-508.[Yuan H X,Xu X P,Zhang Y J,Liu W G,Guo J F.2009.Analysis on resistance to sheath blight of transgenic rice with extra anti-fungual genes[J].Acta Phytophylacica Sinica,36(6):504-508.]
张荣华,何红,张革民,刘海斌,李杨瑞,方锋学,宋焕忠,方位宽,闭少玲.2011.宿根性特强甘蔗新品种桂糖29号的选育[J].中国糖料,(1):1-4.[Zhang R H,He H,Zhang GM,Liu H B,Li Y R,Fang F X,Song H Z,Fang W K,Bi S L.2011.Breeding of new sugarcane variety Guitang29 with high ratoon ability[J].Sugar Crops of China,(1):1-4.]
周思军,李希臣,刘昭军,刘文萍,南相日.2000.通过农杆菌介导将菜豆几丁质酶基因导入马铃薯[J].中国马铃薯,14(2):70-72.[Zhou S J,Li X C,Liu Z J,Liu W P,Nan X R.2000.Introduction of chitinase gene from kidney bean into potato by agrobacterium mediated transformation[J].Chinese Potato,14(2):70-72.]
Adams D J.2004.Fungal cell wall chitinases and glucanases[J].Microbiology,150(7):2029-2035.
Albert H H,Schenck S.1996.PCR amplification from a homolog of the b E mating-type gene as a sensitive assay for the presence of Ustilago scitaminea DNA[J].Plant Disease,80(10):1189-1192.
Broglie K,Chet I,Holliday M,Cressman R,Biddle P,Knowlton S,Mauvais J C,Broglie R.1991.Transgenic plants with enhanced resistance to the fungal pathogen rhizoctonia solani[J].Science,24(5035):1194-1197.
Martinez M,Medina I,Naranjo S,Rodriguez C,Armas R,Pinon D,Vicente C,Legaz M E.2000.Changes of some chemical parameters,involved in sucrose recovery from sugarcane juices,related to the susceptibility or resistance of sugarcane plants to smut(Ustilagoscitaminea)[J].Indian Sugar,50(6):341-346.
Mhamdi A,Queval G,Chaouch S,Vanderauwera S,Van Breusegem F,Noctor G.2010.Catalase function in plants:A focus on Arabidopsis mutants as stress-mimic models[J].Journal of Experimental Botany,61(15):4197-4220.
Peters L P,Carvalho G,Vilhena M B,Creste S,Azevedo R A,Monteiro-Vitorello C B.2017.Functional analysis of oxidative burst in sugarcane smut-resistant and susceptible genotypes[J].Planta,245(4):749-764.