番茄果实中甘露聚糖酶与其它细胞壁降解酶的协同关系研究
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摘要
番茄(Lycopersicon esculentum Mill)是一种世界性蔬菜,其果实成熟的提前和延后都会带来重大的社会效益与经济效益。果实软化是果实成熟的一个明显特征,也是导致果实成熟和可食用性的直接原因。果实软化的主要原因是细胞壁结构的变化和胞壁物质的降解,细胞壁物质的降解主要是由细胞壁降解酶引起的。前人通过反义技术和RNA干扰技术将甘露聚糖酶基因转入到番茄果实中,表明甘露聚糖酶并非是果实软化的主要调控因子,本实验主要研究甘露聚糖酶与其它细胞壁降解酶协同作用以及植物激素对这些细胞壁降解酶的调控的作用,这样有助于我们更全面地了解番茄果实成熟软化机制。本实验对ABA突变体、GA突变体、microtom、rin和nor果实的果皮和果肉中甘露聚糖酶及其它细胞壁降解酶的活性进行了测定,试验结果表明如下。
1.果皮中甘露聚糖酶活性与硬度达到极显著的负相关(R=-0.50**);同多聚半乳糖醛酸酶活性达到显著正相关(R=0.45*);同可溶性固形物达到极显著正相关(R=0.53**)。同时也表明:番茄果肉中甘露聚糖酶的活性同果实的硬度活性达到极显著的负相关(R=-0.55**);同多聚半乳糖醛酸酶的活性达到极显著的正相关(R=0.48**);同果胶甲酯酶活性达到显著正相关(R=0.46*);同纤维素酶达到极显著正相关(R=0.52**);同甘露糖苷酶达到显著正相关(R=0.38*);同可溶性固形物达到极显著正相关(R=0.59**)。甘露聚糖酶与PME,PG和纤维素酶之间存在协同关系,并影响着果实的软化。
2.通过GA和ABA处理GA突变体,rin和nor果皮中细胞壁降解酶,表明GA显著的提高了半乳糖苷酶、PG、甘露聚糖酶、甘露糖苷酶和PME的活性而抑制了木糖苷酶的活性,部分提高和部分降低了阿拉伯糖苷酶,GA提高了rin和nor中甘露聚糖酶的活性,ABA协同GA提高甘露糖苷酶的活性。GA正调控半乳糖苷酶、PG、甘露聚糖酶、甘露糖苷酶和PME的活性而负调控木糖苷酶的活性,部分正调控和部分负调控阿拉伯糖苷酶。GA,rin和甘露聚糖酶在同一信号传导通路上;GA,nor和甘露聚糖酶在同一信号传导通路上。ABA协同GA正调控甘露糖苷酶。
3.在GA提高了半乳糖苷酶、木糖苷酶和PME的活性,降低了甘露糖苷酶,部分调控和部分抑制阿拉伯糖苷酶、PG和甘露聚糖酶。而GA提高了rin和nor突变体果肉中木糖苷酶的活性。ABA协同GA提高了半乳糖苷酶的活性。在GA正调控半乳糖苷酶、木糖苷酶和PME;而负调控甘露糖苷酶,部分正调控和部分负调控阿拉伯糖苷酶、PG和甘露聚糖酶。而GA,rin和nor和木糖苷酶在同一信号传导通路上;GA,nor和木糖苷酶在同一信号传导通路。ABA协同GA正调控半乳糖苷酶。
4.在果皮中ABA,rin,甘露聚糖酶,阿拉伯糖苷酶,PME和PG在同一信号通路中;ABA,nor,半乳糖苷酶,PME,甘露聚糖酶,PG和木糖苷酶在同一信号通路中;GA,rin,木糖苷酶,PME,PG和阿拉伯糖苷酶在同一信号通路中;GA,nor,半乳糖苷酶和木糖苷酶在同一信号通路中。
5.在果肉中ABA,rin,PG和甘露聚糖酶在同一信号通路中;ABA,nor,木糖苷酶,甘露糖苷酶,PG,PME和甘露聚糖酶在同一信号通路中;GA,rin,PG,甘露聚糖酶和半乳糖苷酶在同一信号通路中;GA,nor,PG和甘露聚糖酶在同一信号通路中。
Tomato(Lycopersicon esculentum Mill)is a worldwise-grown vegetable. That tomato fruits ripen in advance or postponed will produce social and economic efficiency. Softening is an obvious character and also a direct factor which is related to ediblity.The change of cell wall and degradation of cell wall materials were considered as the main factor involved in change of fruit texture, cell wall degrading enzyme degraded the cell wall material. Transferred LeMan4 in tomato through antisense RNA and RNAi showed that endo-β-mannanase was not the key factor in fruit softening.The experiment has studyed the coordination between the endo-β-mannanase and other cell wall degrading enzyme . How the plant hormone regualted the cell wal degrading enzyme .So we can facilitate the overall understanding of tomato fruit softening,the experiments results showed below.
1.The experiments were measured endo-β-mannanase and other cell wall degrading enzyme of skin and pericarp of ABA-deficent, GA-deficient, microtom, ripening -inhibitor and non-ripening . In the skin of tomato , a negative correlation existed between endo-β-mannanase activity and fruit firmness with a correlation coefficient of -0.50, significant at 0.01 level;a positive correlation existed between endo-β-mannanase and PG activity with a correlation coefficient of 0.45,significantat 0.05 level:a positive correlation existedbetweenendo-β-mannanase activity and soluble solid content with a correlation coefficient of 0.53,significant at 0.01 level. In the pericarp of tomato ,the relation among endo-β-mannanase activity, cell wall degrading engzyme,soluble solid content and fruit firmness, endo-β-mannanase activity were different in differernt varity, the higher the endo-β-mannanase activity,the smaller the fruit firmness . A negative correlation existed between endo-β-mannanase activity and fruit firmness with a correlation coefficient of -0.55, significant at 0.01 level;a positive correlation existed between endo-β-mannanase activity and PG activity with a correlation coefficient of 0.48,significant at 0.01 level:a positive correlation existed between endo-β-mannanase activity and PME activity with a correlation coefficient of 0.46,significant at 0.05level: a positive correlation existed between endo-β-mannanase activity and cellulaus activity with a correlation coefficient of 0.55,significant at 0.01level: a positive correlation existed between endo-β-mannanase activity andβ-mannosidase activity with a correlation coefficient of 0.38,significant at 0.05 level: a positive correlation existed between endo-β-mannanase activity and soluble solid content with a correlation coefficient of 0.59,significant at 0.01 level. There were correlation existed among endo-β-mannanase ,PME,PG and cellulaus and effeted the fruit softening.
2.We deteminated the GA-deficient,ABA-deficient ,ripenging-inhibitor and non-ripening enzymeof cell wall degrading.GA significantly increased the activity of galactosidase, PG, mannanase, mannosidase, PME and decreasedβ-D-xylosidase activity the in the GA-deficient tomato skin and part increased and part decreased the a-L-arabinofuranosidase activity. GA increased endo-β-mannanase activity of ripening–inhibitor and non-ripening tomato skin.So GA positive regulated the galactosidase, PG, mannanase, mannosidase, PME. GA negative regulatatedβ-D-xylosidase, and negative and positive regulatied a-L-arabinofuranosidase and we speculated that the rin,nor,leman4 in the GA signal transduction . ABA coordinated with GA regulated the galactosidase activity .
2. Through determinated the cell wall degrading enzyme of GA-deficient,ABA-deficient of tomato pericarp, GA significantly increased the activity of galactosidase,β-D-xylosidase ,PME and decreased the mannosidase activity and part increased and part decreased PG, endo-β-mannanase and a-L-arabinofuranosidase activity.GA decreased the acitivity ofβ-D-xylosidase of ripeing-inhibitor and non-ripening tomato pericarp. ABA coordinated with GA increased galactosidase activity .So GA positive regulated galactosidase,β-D-xylosidase ,PME and negative regulated the mannosidase and part increased and part negative regulated PG, endo-β-mannanase and a-L-arabinofuranosidase activity.GA,rin andβ-D-xylosidase in same signal transduction.GA,nor andβ-D-xylosidase in same signal transduction. ABA coordinated with GA positive regulated galactosidase .
2.In tomato skin, ABA,rin,endo-β-mannanase,a-L-arabinofuranosidase,PME,PG insame signal transduction. ABA,nor ,endo-β-mannanase ,a-L-arabinofuranosidase,PME,PG insame signal transduction.GA,rin,β-D-xylosidase,PME,PG and a-L-arabinofuranosidase in same signal transduction. GA,nor, galactosidase ,β-D-xylosidase in same signal transduction.
5. In tomato pericarp , ABA,rin,PG, endo-β-mannanase in same signal transduction.ABA,nor ,β-D-xylosidase,mannosidase,PG,PMEand endo-β-mannanase in same signal transduction.GA,nor, PG, endo-β-mannanase, galactosidase in same signal transduction .GA,nor,PG, endo-β-mannanase in same signal transduction.
1.果皮中甘露聚糖酶活性与硬度达到极显著的负相关(R=-0.50**);同多聚半乳糖醛酸酶活性达到显著正相关(R=0.45*);同可溶性固形物达到极显著正相关(R=0.53**)。同时也表明:番茄果肉中甘露聚糖酶的活性同果实的硬度活性达到极显著的负相关(R=-0.55**);同多聚半乳糖醛酸酶的活性达到极显著的正相关(R=0.48**);同果胶甲酯酶活性达到显著正相关(R=0.46*);同纤维素酶达到极显著正相关(R=0.52**);同甘露糖苷酶达到显著正相关(R=0.38*);同可溶性固形物达到极显著正相关(R=0.59**)。甘露聚糖酶与PME,PG和纤维素酶之间存在协同关系,并影响着果实的软化。
2.通过GA和ABA处理GA突变体,rin和nor果皮中细胞壁降解酶,表明GA显著的提高了半乳糖苷酶、PG、甘露聚糖酶、甘露糖苷酶和PME的活性而抑制了木糖苷酶的活性,部分提高和部分降低了阿拉伯糖苷酶,GA提高了rin和nor中甘露聚糖酶的活性,ABA协同GA提高甘露糖苷酶的活性。GA正调控半乳糖苷酶、PG、甘露聚糖酶、甘露糖苷酶和PME的活性而负调控木糖苷酶的活性,部分正调控和部分负调控阿拉伯糖苷酶。GA,rin和甘露聚糖酶在同一信号传导通路上;GA,nor和甘露聚糖酶在同一信号传导通路上。ABA协同GA正调控甘露糖苷酶。
3.在GA提高了半乳糖苷酶、木糖苷酶和PME的活性,降低了甘露糖苷酶,部分调控和部分抑制阿拉伯糖苷酶、PG和甘露聚糖酶。而GA提高了rin和nor突变体果肉中木糖苷酶的活性。ABA协同GA提高了半乳糖苷酶的活性。在GA正调控半乳糖苷酶、木糖苷酶和PME;而负调控甘露糖苷酶,部分正调控和部分负调控阿拉伯糖苷酶、PG和甘露聚糖酶。而GA,rin和nor和木糖苷酶在同一信号传导通路上;GA,nor和木糖苷酶在同一信号传导通路。ABA协同GA正调控半乳糖苷酶。
4.在果皮中ABA,rin,甘露聚糖酶,阿拉伯糖苷酶,PME和PG在同一信号通路中;ABA,nor,半乳糖苷酶,PME,甘露聚糖酶,PG和木糖苷酶在同一信号通路中;GA,rin,木糖苷酶,PME,PG和阿拉伯糖苷酶在同一信号通路中;GA,nor,半乳糖苷酶和木糖苷酶在同一信号通路中。
5.在果肉中ABA,rin,PG和甘露聚糖酶在同一信号通路中;ABA,nor,木糖苷酶,甘露糖苷酶,PG,PME和甘露聚糖酶在同一信号通路中;GA,rin,PG,甘露聚糖酶和半乳糖苷酶在同一信号通路中;GA,nor,PG和甘露聚糖酶在同一信号通路中。
Tomato(Lycopersicon esculentum Mill)is a worldwise-grown vegetable. That tomato fruits ripen in advance or postponed will produce social and economic efficiency. Softening is an obvious character and also a direct factor which is related to ediblity.The change of cell wall and degradation of cell wall materials were considered as the main factor involved in change of fruit texture, cell wall degrading enzyme degraded the cell wall material. Transferred LeMan4 in tomato through antisense RNA and RNAi showed that endo-β-mannanase was not the key factor in fruit softening.The experiment has studyed the coordination between the endo-β-mannanase and other cell wall degrading enzyme . How the plant hormone regualted the cell wal degrading enzyme .So we can facilitate the overall understanding of tomato fruit softening,the experiments results showed below.
1.The experiments were measured endo-β-mannanase and other cell wall degrading enzyme of skin and pericarp of ABA-deficent, GA-deficient, microtom, ripening -inhibitor and non-ripening . In the skin of tomato , a negative correlation existed between endo-β-mannanase activity and fruit firmness with a correlation coefficient of -0.50, significant at 0.01 level;a positive correlation existed between endo-β-mannanase and PG activity with a correlation coefficient of 0.45,significantat 0.05 level:a positive correlation existedbetweenendo-β-mannanase activity and soluble solid content with a correlation coefficient of 0.53,significant at 0.01 level. In the pericarp of tomato ,the relation among endo-β-mannanase activity, cell wall degrading engzyme,soluble solid content and fruit firmness, endo-β-mannanase activity were different in differernt varity, the higher the endo-β-mannanase activity,the smaller the fruit firmness . A negative correlation existed between endo-β-mannanase activity and fruit firmness with a correlation coefficient of -0.55, significant at 0.01 level;a positive correlation existed between endo-β-mannanase activity and PG activity with a correlation coefficient of 0.48,significant at 0.01 level:a positive correlation existed between endo-β-mannanase activity and PME activity with a correlation coefficient of 0.46,significant at 0.05level: a positive correlation existed between endo-β-mannanase activity and cellulaus activity with a correlation coefficient of 0.55,significant at 0.01level: a positive correlation existed between endo-β-mannanase activity andβ-mannosidase activity with a correlation coefficient of 0.38,significant at 0.05 level: a positive correlation existed between endo-β-mannanase activity and soluble solid content with a correlation coefficient of 0.59,significant at 0.01 level. There were correlation existed among endo-β-mannanase ,PME,PG and cellulaus and effeted the fruit softening.
2.We deteminated the GA-deficient,ABA-deficient ,ripenging-inhibitor and non-ripening enzymeof cell wall degrading.GA significantly increased the activity of galactosidase, PG, mannanase, mannosidase, PME and decreasedβ-D-xylosidase activity the in the GA-deficient tomato skin and part increased and part decreased the a-L-arabinofuranosidase activity. GA increased endo-β-mannanase activity of ripening–inhibitor and non-ripening tomato skin.So GA positive regulated the galactosidase, PG, mannanase, mannosidase, PME. GA negative regulatatedβ-D-xylosidase, and negative and positive regulatied a-L-arabinofuranosidase and we speculated that the rin,nor,leman4 in the GA signal transduction . ABA coordinated with GA regulated the galactosidase activity .
2. Through determinated the cell wall degrading enzyme of GA-deficient,ABA-deficient of tomato pericarp, GA significantly increased the activity of galactosidase,β-D-xylosidase ,PME and decreased the mannosidase activity and part increased and part decreased PG, endo-β-mannanase and a-L-arabinofuranosidase activity.GA decreased the acitivity ofβ-D-xylosidase of ripeing-inhibitor and non-ripening tomato pericarp. ABA coordinated with GA increased galactosidase activity .So GA positive regulated galactosidase,β-D-xylosidase ,PME and negative regulated the mannosidase and part increased and part negative regulated PG, endo-β-mannanase and a-L-arabinofuranosidase activity.GA,rin andβ-D-xylosidase in same signal transduction.GA,nor andβ-D-xylosidase in same signal transduction. ABA coordinated with GA positive regulated galactosidase .
2.In tomato skin, ABA,rin,endo-β-mannanase,a-L-arabinofuranosidase,PME,PG insame signal transduction. ABA,nor ,endo-β-mannanase ,a-L-arabinofuranosidase,PME,PG insame signal transduction.GA,rin,β-D-xylosidase,PME,PG and a-L-arabinofuranosidase in same signal transduction. GA,nor, galactosidase ,β-D-xylosidase in same signal transduction.
5. In tomato pericarp , ABA,rin,PG, endo-β-mannanase in same signal transduction.ABA,nor ,β-D-xylosidase,mannosidase,PG,PMEand endo-β-mannanase in same signal transduction.GA,nor, PG, endo-β-mannanase, galactosidase in same signal transduction .GA,nor,PG, endo-β-mannanase in same signal transduction.
引文
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