Changes in Abundance of Enzymes Involved in Organic Acid, Amino Acid and Sugar Metabolism, and Photosynthesis during the Ripening of Blackberry Fruit
Changes in Abundance of Enzymes Involved in Organic Acid, Amino Acid and Sugar Metabolism, and Photosynthesis during the Ripening of Blackberry Fruit
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DOI：
10.21273/jashs.134.2.167
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发表时间：
2009-03
影响因子：
1.9
通讯作者：
F. Famiani;R. P. Walker
中科院分区：
文献类型：
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作者：
F. Famiani;R. P. Walker
虽然关于黑莓(Rubus L.)中糖和有机/氨基酸等各种代谢物的含量已有信息，但人们对其酶组成知之甚少。本研究的目的是研究黑莓成熟过程中各种酶的丰度变化。黑莓是一种聚合果，由花托和与之相连的几个小核果组成，小核果又由果肉(mesocarpplusepicarp)andseedenclosedintheendocarp；组成，因此，这些部分与花梗一起被分开分析。所研究的酶参与有机/氨基酸和糖的代谢以及光合作用，这些过程在水果发育中是重要的。这些酶是：磷酸烯醇式丙酮酸羧激酶(PEPCK(EC：4.1.1.49))、磷酸烯醇式丙酮酸羧基酶(PEPC(EC：4.1.1.31))、丙酮酸正磷酸二激酶(PPDK(EC：2.7.9.1))、胞质天冬氨酸氨基转移酶(Cyt Aspat)(EC：2.6.1.1)、醛缩酶(EC：4.1.2.13)、谷氨酰胺合成酶(GS(EC：6.3.1.2))和核酮糖-1，5-二磷酸羧基/加氧酶(Rubisco(EC：4.1.1.39))。为了避免在测量酶活性时出现问题，采取的方法是使用针对每个酶的特异性抗体，并结合十二烷基硫酸钠聚丙烯酰胺凝胶电泳法进行免疫印迹。在成熟过程中，这些酶中有几种的丰度发生了显著的变化，这些变化取决于所研究的组织。当果肉中有机酸减少时，PEPCK出现，仅在该组织中检测到，而PPDK在任何组织中均未检测到。果肉中Rubisco、胞质GS和胞质醛缩酶的丰度显著降低，而胞质GS、胞质醛缩酶和PEPC的丰度变化不大。种子中所有酶的丰度均降低，而在种皮和花梗中，除了花托中Rubisco的丰度降低外，酶的丰度变化不大。
Although information is available regarding the content of various metabolites such as sugars and organic/ amino acids in blackberry (Rubus L.), little is known about its enzyme composition. The aim of this study was to investigate changes in the abundance of various enzymes during the ripening of blackberry. Blackberry is an aggregate fruit, composed of a receptacle and several drupelets attached to it, which in turn, are composed of the flesh (mesocarpplusepicarp)andseedenclosedintheendocarp; therefore,thesepartswere analyzedseparatelyalongwith the pedicel. The enzymes studied participate in organic/amino acid and sugar metabolism and photosynthesis, processes known to be important in fruit development. These enzymes were phosphoenolpyruvate carboxykinase (PEPCK (EC:4.1.1.49)), phosphoenolpyruvate carboxylase (PEPC (EC:4.1.1.31)), pyruvate, orthophosphate dikinase (PPDK (EC:2.7.9.1)), cytosolic aspartate aminotransferase (cyt AspAT (EC:2.6.1.1)), aldolase (EC:4.1.2.13), glutamine synthetase (GS (EC:6.3.1.2)), and ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO (EC:4.1.1.39)). To avoid problems in measuring enzyme activity, the approach taken was to use antibodies specific for each enzyme in conjunction with immunoblotting of sodium dodecyl sulfate polyacrylamide gel electrophoresis. During ripening, there were marked changes in abundance of several of these enzymes and these changes were dependent on the tissue investigated. PEPCK appeared when organic acids decreased in the flesh and was only detected in this tissue, whereas PPDK was not detected in any tissue. In the flesh, there was a large decrease in abundance of RUBISCO, plastidic GS, and plastidic aldolase, but little change in cytosolic GS, cytosolic aldolase, and PEPC.Inseeds,therewasadecrease intheabundanceofallenzymes.Inthereceptacle andpedicel,apartfromalarge decrease in RUBISCO in the receptacle, there was little change in enzyme abundance.