猕猴桃果实采后软化期间淀粉降解关键基因表达分析
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摘要
为探究淀粉降解与果实采后软化的关系,以红阳猕猴桃果实为试验材料,研究外源乙烯和1-甲基环丙烯(1-MCP)处理对猕猴桃淀粉降解的影响。结果表明,猕猴桃果实采后贮藏期间硬度与淀粉含量均呈下降趋势,外源乙烯处理促进果实的软化与淀粉降解,而外源1-MCP处理则能使果实保持较高的硬度与淀粉含量。外源乙烯处理上调了Ac AMY1、Ac AMY3、Ac BAM1和Ac ABAM3的表达;1-MCP处理除了能抑制上述基因表达外,还抑制了Ac LDA1的表达,保持果实贮藏期间较高的Ac PWD和Ac ISA3的基因表达水平。表明,红阳猕猴桃采后淀粉降解与果实软化密切相关,乙烯能抑制Ac PWD表达,促进淀粉颗粒磷酸化进而促进淀粉降解和果实软化;外源1-MCP处理抑制葡聚糖聚合物脱支,延缓淀粉的降解进而减缓果实软化。本研究结果为揭示猕猴桃采后淀粉降解的机制及其与成熟软化进程之间的关系提供了理论依据。
In order to explore the relationship between postharvest fruit softening and starch degradation,the effect of exogenous ethylene and 1-methylcyclopropene( 1-MCP) treatments on starch degradation in ‘Hongyang'kiwifruit were investigated. Results showed that,firmness and starch content in postharvest kiwifruit decreased during storage.Exogenous ethylene treatment promoted softening and starch degradation,while exogenous 1-MCP treatment could maintain higher firmness and starch content. The expression of Ac AMY1,Ac AMY3,Ac BAM1,and Ac ABAM3 were significantly up-regulated by ethylene treatment and inhibited by 1-MCP treatment during storage. Meanwhile,exogenous 1-MCP treatment also inhibited the expression of Ac LDA1,and maintained higher transcription levels of Ac PWD and Ac ISA3 during storage. These results suggest that there is a positive correlation between kiwifruit softening and starch degradation during storage. Ethylene treatment inhibited the expression of Ac PWD,which may accelerate starch phosphorylation and finally promoted in starch degradation and fruit softening. 1-MCP treatment could inhibit the glucan de-branching,which could efficiently delay starch degradation and alleviate kiwifruit softening during storage.These results provided a theoretical basis for revealing relationship between the mechanism of starch degradation and the process of softening in kiwifruit.
In order to explore the relationship between postharvest fruit softening and starch degradation,the effect of exogenous ethylene and 1-methylcyclopropene( 1-MCP) treatments on starch degradation in ‘Hongyang'kiwifruit were investigated. Results showed that,firmness and starch content in postharvest kiwifruit decreased during storage.Exogenous ethylene treatment promoted softening and starch degradation,while exogenous 1-MCP treatment could maintain higher firmness and starch content. The expression of Ac AMY1,Ac AMY3,Ac BAM1,and Ac ABAM3 were significantly up-regulated by ethylene treatment and inhibited by 1-MCP treatment during storage. Meanwhile,exogenous 1-MCP treatment also inhibited the expression of Ac LDA1,and maintained higher transcription levels of Ac PWD and Ac ISA3 during storage. These results suggest that there is a positive correlation between kiwifruit softening and starch degradation during storage. Ethylene treatment inhibited the expression of Ac PWD,which may accelerate starch phosphorylation and finally promoted in starch degradation and fruit softening. 1-MCP treatment could inhibit the glucan de-branching,which could efficiently delay starch degradation and alleviate kiwifruit softening during storage.These results provided a theoretical basis for revealing relationship between the mechanism of starch degradation and the process of softening in kiwifruit.
引文
[1]张慧琴,谢鸣,张琛,杨鲁琼,章镇,肖金平,周利秋.猕猴桃果实发育过程中淀粉积累差异及其糖代谢特性[J].中国农业科学,2014,47(17):3453-3464
[2]Yi J,Kebede B T,Grauwet T,Van Loey A,Hu X,Hendrickx M.A multivariate approach into physicochemical,biochemical and aromatic quality changes of purée based on Hayward kiwifruit during the final phase of ripening[J].Postharvest Biology&Technology,2016,117:206-216
[3]Zeeman S C,Kossmann J,Smith A M.Starch:its metabolism,evolution,and biotechnological modification in plants[J].Annual Review of Plant Biology,2010,61(1):209-234
[4]Richardson A C,Marsh K B,Boldingh H L,Pickering A H,Bulley S M,Frearson N J,Ferguson A R,Thornber S E,Bolitho K M,Macrae E A.High growing temperatures reduce fruit carbohydrate and vitamin C in kiwifruit[J].Plant Cell Environment,2004,27(4):423-435
[5]Nardozza S,Boldingh H L,Richardson A C,Costa G,Marsh H,Macrae E A,Clearwater M J.Variation in carbon content and size in developing fruit of Actinidia deliciosa genotypes[J].Functional Plant Biology,2010,37(6):545-554
[6]Harker F R,Carr B T,Lenjo M,Macrae E A,Wismer W V,Marsh K B,Williamsa M,Whitea A,Lunda C M,Walkera S B,Gunsona F A,Gunson F A,Pereiraa R B.Consumer liking for kiwifruit flavour:a meta-analysis of five studies on fruit quality[J].Food Quality and Preference,2009,20(1):30-41
[7]Nardozza S,Gamble J,Axten L G,Wohlers M W,Clearwater M J,Feng J,Harker F R.Dry matter content and fruit size affect flavour and texture of novel Actinidia deliciosa genotypes[J].Journal of the Science of Food and Agriculture,2011,91(4):742-748
[8]Nardozza S,Boldingh H L,Osorio S,H9hne M,Wohlers M,Gleave A P,MacRae E A,Richardson A C,Atkinson R G,Sulpice R,Fernie A R,Clearwater M J.Metabolic analysis of kiwifruit(Actinidia deliciosa)berries from extreme genotypes reveals hallmarks for fruit starch metabolism[J].Journal of Experimental Botany,2013,64(16):5049-5063
[9]Stevenson D G,Johnson S R,Jane J L,Inglett G E.Chemical and physical properties of kiwifruit(Actinidia deliciosa)starch[J].Starch-Starke,2006,58(7):323-329
[10]Smith A M,Zeeman S C,Smith S M.Starch degradation[J].Annual Review of Plant Biology,2005,56(1):73-98
[11]Baunsgaard L,Lutken H,Mikkelsen R M,Pham T,Blennow A.A novel isoform of glucan,water dikinase phosphorylates prephosphorylatedα-glucans and is involved in starch degradation in Arabidopsis[J].The Plant Journal,2005,41(4):595-605
[12]K9tting O,Pusch K,Tiessen A,Geigenberger P,Steup M,Ritte G.Identification of a novel enzyme required for starch metabolism in Arabidopsis leaves.The phosphoglucan,water dikinase[J].Plant Physiology,2005,137(1):242-252
[13]Edner C,Li J,Albrecht T,Mahlow S,Hejazi M,Hussain H,Kaplan F,Guy C,Smith S M,Steup M,Ritte G.Glucan,water dikinase activity stimulates breakdown of starch granules by plastidialβ-amylases[J].Plant Physiology,2007,145(1):17-28
[14]Silver D M,K9tting O,Moorhead G B G.Phosphoglucan phosphatase function sheds light on starch degradation[J].Trends in Plant Science,2014,19(7):471-478
[15]Asatsuma S,Sawada C,Itoh K,Okito M,Kitajima A,Mitsui T.Involvement ofα-amylase I-1 in starch degradation in rice chloroplasts[J].Plant and Cell Physiology,2005,46(6):858-869
[16]丛庆,张琪,宋丽莉,陈月,蔡洪生,郭长虹.激素在植物冷胁迫应答中的角色[J].核农学报,2016,30(3):614-619
[17]Wei J,Qi X,Guan J,Zhu X.Effect of cold storage and 1-MCP treatment on postharvest changes of fruit quality and cell wall metabolism in sweet cherry[J].Journal of Food,Agriculture&Environment,2011,9(3/4):118-122
[18]Watkins C B.Overview of 1-methylcyclopropene trials and uses for edible horticultural crops[J].Hort Science,2008,43(1):86-94
[19]苗红霞,金志强,刘伟鑫,张建平,孙佩光,徐碧玉.香蕉果实抗性淀粉含量变化及其与其他类型淀粉相关性分析[J].中国农业科学,2013(24):5180-5187
[20]邵佳蓉,宋春波,卞坤,陈伟,杨震峰.桃果实Pp SIZ1基因对低温和外源褪黑素处理的响应[J].园艺学报,2016,43(7):1257-1266
[21]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCTmethod[J].Methods,2001,25(4):402-408
[22]张永平,乔永旭,喻景权,赵智中.园艺植物果实糖积累的研究进展[J].中国农业科学,2008,41(4):1151-1157
[23]Yang X T,Song J,Campbell-Palmer L,Fillmore S,Zhang Z Q.Effect of ethylene and 1-MCP on expression of genes involved in ethylene biosynthesis and perception during ripening of apple fruit[J].Postharvest Biology Technology,2013,78(4):55-66
[24]于年文,李俊才,王家珍,蔡忠民,沙守峰,李宏军,郭修武.不同时期采收的南果梨后熟过程中糖分的变化规律[J].中国农学通报,2011,27(20):123-126
[25]林河通,席玙芳,陈绍军.黄花梨果实采后软化生理基础[J].中国农业科学,2003,36(3):349-352
[26]齐秀东,魏建梅,李永红.苹果果实质地软化过程中碳水化合物代谢及其关键酶基因表达的变化[J].园艺学报,2015,42(3):409-417
[27]李映志,董黎梨,汪永保,李洪波,张佳琦,叶春海.干、湿苞木菠萝果实成熟过程中糖代谢特性[J].果树学报,2014,31(1):85-90
[28]齐秀东,魏建梅,赵美微,彭红丽,张海娥.‘京白梨’果实后熟软化与糖、淀粉代谢及其基因表达的关系[J].中国农业科学,2015,48(13):2591-2599
[29]K9tting O,Kossmann J,Zeeman S C,Lloyd J R.Regulation of starch metabolism:the age of enlightenment?[J].Current Opinion in Plant Biology,2010,13(3):320-328
[30]Ritte G,Lloyd J R,Eckermann N,Rottmann A,Kossmann J,Steup M.The starch-related R1 protein is anα-glucan,water dikinase[J].Proceedings of the National Academy of Sciences,2002,99(10):7166-7171
[31]Peroni F H G,Koike C,Louro R P,Purgatto E,do Nascimento J R O,Lajolo F M,Cordenunsi B R.Mango starch degradation.II.The binding ofα-amylase andβ-amylase to the starch granule[J].Journal of Agricultural and Food Chemistry,2008,56(16):7416-7421
[32]Richardson A C,Boldingh H L,Mc Atee P A,Gunaseelan K,Luo Z,Atkinson R G,Schaffer R J.Fruit development of the diploid kiwifruit,Actinidia chinensis‘Hort16A’[J].BMC Plant Biology,2011,11(1):182
[33]Maria T,Tsaniklidis G,Delis C,Nikolopoulou A E,Nikoloudakis N,Karapanos I,Aivalakis G.Gene transcript accumulation and enzyme activity ofβ-amylases suggest involvement in the starch depletion during the ripening of cherry tomatoes[J].Plant Gene,2016,5:8-12
[34]do Nascimento J R O,Júnior A V,Bassinello P Z,Cordenunsi B R,Mainardi J A,Purgatto E,Lajolo F M.Beta-amylase expression and starch degradation during banana ripening[J].Postharvest Biology and Technology,2006,40(1):41-47
[35]Sorndech W,Sagnelli D,Meier S,Jansson A M,Lee B H,Hamaker B R,Rolland-SabatéA,Hebelstrup K H,Tongta S,Blennow A.Structure of branching enzyme-and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates[J].Carbohydrate Polymers,2016,152:51-61
[36]Seung D,Thalmann M,Sparla F,Hachem M A,Lee S K,Issakidis-Bourguet E,Svensson B,Zeeman S C,Santelia D.Arabidopsis thaliana AMY3 is a unique redox-regulated chloroplasticα-amylase[J].Journal of Biological Chemistry,2013,288(47):33620-33633
[37]Gao H,Huang S,Dong T,Yang Q,Yi G.Analysis of resistant starch degradation in postharvest ripening of two banana cultivars:Focus on starch structure and amylases[J].Postharvest Biology and Technology,2016,119:1-8
[38]Soares C A,Peroni-Okita F H G,Cardoso M B,Shitakubo R,Lajolo F M,Cordenunsi B R.Plantain and banana starches:granule structural characteristics explain the differences in their starch degradation patterns[J].Journal of Agricultural and Food Chemistry,2011,59(12):6672-6681
[39]Delatte T,Umhang M,Trevisan M,Eicke S,Thorneycroft D,Smith S M,Zeeman S C.Evidence for distinct mechanisms of starch granule breakdown in plants[J].Journal of Biological Chemistry,2006,281(17):12050-12059
[40]Wattebled F,Dong Y,Dumez S,DelvalléD,Planchot V,Berbezy P,Vyas D,Colonna P,Chatterjee M,Ball S,D'Hulst C.Mutants of Arabidopsis lacking a chloroplastic isoamylase accumulate phytoglycogen and an abnormal form of amylopectin[J].Plant Physiology,2005,138(1):184-195
[41]Shangguan L,Song C,Leng X,Kayesh E,Sun X,Fang J.Mining and comparison of the genes encoding the key enzymes involved in sugar biosynthesis in apple,grape,and sweet orange[J].Scientia Horticulturae,2014,165(3):311-318
[2]Yi J,Kebede B T,Grauwet T,Van Loey A,Hu X,Hendrickx M.A multivariate approach into physicochemical,biochemical and aromatic quality changes of purée based on Hayward kiwifruit during the final phase of ripening[J].Postharvest Biology&Technology,2016,117:206-216
[3]Zeeman S C,Kossmann J,Smith A M.Starch:its metabolism,evolution,and biotechnological modification in plants[J].Annual Review of Plant Biology,2010,61(1):209-234
[4]Richardson A C,Marsh K B,Boldingh H L,Pickering A H,Bulley S M,Frearson N J,Ferguson A R,Thornber S E,Bolitho K M,Macrae E A.High growing temperatures reduce fruit carbohydrate and vitamin C in kiwifruit[J].Plant Cell Environment,2004,27(4):423-435
[5]Nardozza S,Boldingh H L,Richardson A C,Costa G,Marsh H,Macrae E A,Clearwater M J.Variation in carbon content and size in developing fruit of Actinidia deliciosa genotypes[J].Functional Plant Biology,2010,37(6):545-554
[6]Harker F R,Carr B T,Lenjo M,Macrae E A,Wismer W V,Marsh K B,Williamsa M,Whitea A,Lunda C M,Walkera S B,Gunsona F A,Gunson F A,Pereiraa R B.Consumer liking for kiwifruit flavour:a meta-analysis of five studies on fruit quality[J].Food Quality and Preference,2009,20(1):30-41
[7]Nardozza S,Gamble J,Axten L G,Wohlers M W,Clearwater M J,Feng J,Harker F R.Dry matter content and fruit size affect flavour and texture of novel Actinidia deliciosa genotypes[J].Journal of the Science of Food and Agriculture,2011,91(4):742-748
[8]Nardozza S,Boldingh H L,Osorio S,H9hne M,Wohlers M,Gleave A P,MacRae E A,Richardson A C,Atkinson R G,Sulpice R,Fernie A R,Clearwater M J.Metabolic analysis of kiwifruit(Actinidia deliciosa)berries from extreme genotypes reveals hallmarks for fruit starch metabolism[J].Journal of Experimental Botany,2013,64(16):5049-5063
[9]Stevenson D G,Johnson S R,Jane J L,Inglett G E.Chemical and physical properties of kiwifruit(Actinidia deliciosa)starch[J].Starch-Starke,2006,58(7):323-329
[10]Smith A M,Zeeman S C,Smith S M.Starch degradation[J].Annual Review of Plant Biology,2005,56(1):73-98
[11]Baunsgaard L,Lutken H,Mikkelsen R M,Pham T,Blennow A.A novel isoform of glucan,water dikinase phosphorylates prephosphorylatedα-glucans and is involved in starch degradation in Arabidopsis[J].The Plant Journal,2005,41(4):595-605
[12]K9tting O,Pusch K,Tiessen A,Geigenberger P,Steup M,Ritte G.Identification of a novel enzyme required for starch metabolism in Arabidopsis leaves.The phosphoglucan,water dikinase[J].Plant Physiology,2005,137(1):242-252
[13]Edner C,Li J,Albrecht T,Mahlow S,Hejazi M,Hussain H,Kaplan F,Guy C,Smith S M,Steup M,Ritte G.Glucan,water dikinase activity stimulates breakdown of starch granules by plastidialβ-amylases[J].Plant Physiology,2007,145(1):17-28
[14]Silver D M,K9tting O,Moorhead G B G.Phosphoglucan phosphatase function sheds light on starch degradation[J].Trends in Plant Science,2014,19(7):471-478
[15]Asatsuma S,Sawada C,Itoh K,Okito M,Kitajima A,Mitsui T.Involvement ofα-amylase I-1 in starch degradation in rice chloroplasts[J].Plant and Cell Physiology,2005,46(6):858-869
[16]丛庆,张琪,宋丽莉,陈月,蔡洪生,郭长虹.激素在植物冷胁迫应答中的角色[J].核农学报,2016,30(3):614-619
[17]Wei J,Qi X,Guan J,Zhu X.Effect of cold storage and 1-MCP treatment on postharvest changes of fruit quality and cell wall metabolism in sweet cherry[J].Journal of Food,Agriculture&Environment,2011,9(3/4):118-122
[18]Watkins C B.Overview of 1-methylcyclopropene trials and uses for edible horticultural crops[J].Hort Science,2008,43(1):86-94
[19]苗红霞,金志强,刘伟鑫,张建平,孙佩光,徐碧玉.香蕉果实抗性淀粉含量变化及其与其他类型淀粉相关性分析[J].中国农业科学,2013(24):5180-5187
[20]邵佳蓉,宋春波,卞坤,陈伟,杨震峰.桃果实Pp SIZ1基因对低温和外源褪黑素处理的响应[J].园艺学报,2016,43(7):1257-1266
[21]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCTmethod[J].Methods,2001,25(4):402-408
[22]张永平,乔永旭,喻景权,赵智中.园艺植物果实糖积累的研究进展[J].中国农业科学,2008,41(4):1151-1157
[23]Yang X T,Song J,Campbell-Palmer L,Fillmore S,Zhang Z Q.Effect of ethylene and 1-MCP on expression of genes involved in ethylene biosynthesis and perception during ripening of apple fruit[J].Postharvest Biology Technology,2013,78(4):55-66
[24]于年文,李俊才,王家珍,蔡忠民,沙守峰,李宏军,郭修武.不同时期采收的南果梨后熟过程中糖分的变化规律[J].中国农学通报,2011,27(20):123-126
[25]林河通,席玙芳,陈绍军.黄花梨果实采后软化生理基础[J].中国农业科学,2003,36(3):349-352
[26]齐秀东,魏建梅,李永红.苹果果实质地软化过程中碳水化合物代谢及其关键酶基因表达的变化[J].园艺学报,2015,42(3):409-417
[27]李映志,董黎梨,汪永保,李洪波,张佳琦,叶春海.干、湿苞木菠萝果实成熟过程中糖代谢特性[J].果树学报,2014,31(1):85-90
[28]齐秀东,魏建梅,赵美微,彭红丽,张海娥.‘京白梨’果实后熟软化与糖、淀粉代谢及其基因表达的关系[J].中国农业科学,2015,48(13):2591-2599
[29]K9tting O,Kossmann J,Zeeman S C,Lloyd J R.Regulation of starch metabolism:the age of enlightenment?[J].Current Opinion in Plant Biology,2010,13(3):320-328
[30]Ritte G,Lloyd J R,Eckermann N,Rottmann A,Kossmann J,Steup M.The starch-related R1 protein is anα-glucan,water dikinase[J].Proceedings of the National Academy of Sciences,2002,99(10):7166-7171
[31]Peroni F H G,Koike C,Louro R P,Purgatto E,do Nascimento J R O,Lajolo F M,Cordenunsi B R.Mango starch degradation.II.The binding ofα-amylase andβ-amylase to the starch granule[J].Journal of Agricultural and Food Chemistry,2008,56(16):7416-7421
[32]Richardson A C,Boldingh H L,Mc Atee P A,Gunaseelan K,Luo Z,Atkinson R G,Schaffer R J.Fruit development of the diploid kiwifruit,Actinidia chinensis‘Hort16A’[J].BMC Plant Biology,2011,11(1):182
[33]Maria T,Tsaniklidis G,Delis C,Nikolopoulou A E,Nikoloudakis N,Karapanos I,Aivalakis G.Gene transcript accumulation and enzyme activity ofβ-amylases suggest involvement in the starch depletion during the ripening of cherry tomatoes[J].Plant Gene,2016,5:8-12
[34]do Nascimento J R O,Júnior A V,Bassinello P Z,Cordenunsi B R,Mainardi J A,Purgatto E,Lajolo F M.Beta-amylase expression and starch degradation during banana ripening[J].Postharvest Biology and Technology,2006,40(1):41-47
[35]Sorndech W,Sagnelli D,Meier S,Jansson A M,Lee B H,Hamaker B R,Rolland-SabatéA,Hebelstrup K H,Tongta S,Blennow A.Structure of branching enzyme-and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates[J].Carbohydrate Polymers,2016,152:51-61
[36]Seung D,Thalmann M,Sparla F,Hachem M A,Lee S K,Issakidis-Bourguet E,Svensson B,Zeeman S C,Santelia D.Arabidopsis thaliana AMY3 is a unique redox-regulated chloroplasticα-amylase[J].Journal of Biological Chemistry,2013,288(47):33620-33633
[37]Gao H,Huang S,Dong T,Yang Q,Yi G.Analysis of resistant starch degradation in postharvest ripening of two banana cultivars:Focus on starch structure and amylases[J].Postharvest Biology and Technology,2016,119:1-8
[38]Soares C A,Peroni-Okita F H G,Cardoso M B,Shitakubo R,Lajolo F M,Cordenunsi B R.Plantain and banana starches:granule structural characteristics explain the differences in their starch degradation patterns[J].Journal of Agricultural and Food Chemistry,2011,59(12):6672-6681
[39]Delatte T,Umhang M,Trevisan M,Eicke S,Thorneycroft D,Smith S M,Zeeman S C.Evidence for distinct mechanisms of starch granule breakdown in plants[J].Journal of Biological Chemistry,2006,281(17):12050-12059
[40]Wattebled F,Dong Y,Dumez S,DelvalléD,Planchot V,Berbezy P,Vyas D,Colonna P,Chatterjee M,Ball S,D'Hulst C.Mutants of Arabidopsis lacking a chloroplastic isoamylase accumulate phytoglycogen and an abnormal form of amylopectin[J].Plant Physiology,2005,138(1):184-195
[41]Shangguan L,Song C,Leng X,Kayesh E,Sun X,Fang J.Mining and comparison of the genes encoding the key enzymes involved in sugar biosynthesis in apple,grape,and sweet orange[J].Scientia Horticulturae,2014,165(3):311-318