Gene expression of NAD -: Dependent sorbitol dehydrogenase during fruit development of apple (Malus pumila Mill. var. domestica Schneid.)
Gene expression of NAD -: Dependent sorbitol dehydrogenase during fruit development of apple (Malus pumila Mill. var. domestica Schneid.)
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DOI：
10.2503/jjshs.68.1099
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发表时间：
1999-11-01
影响因子：
--
通讯作者：
Yamaki, S
中科院分区：
文献类型：
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作者：
Yamada, K;Mori, H;Yamaki, S
本研究的目的是通过研究NAD-SDH活性以及NAD-SDH蛋白和转录本的水平，建立苹果（Malus pumila Mill. var. Domestica Schneid. cv. Fuji）发育过程中NAD-SDH活性的表达机制。使用 NAD-SDH cDNA 通过 RNA 印迹分析研究基因表达，并使用针对大肠杆菌中表达的 NAD-SDH 蛋白的抗体通过蛋白质印迹测定蛋白质水平。 NAD-SDH 的活性在开花后 80 天 (DAF) 之前非常弱，然后在 104DAF 时暂时增加。此后，随着果实增大而下降；随着果实成熟，它从160 DAF再次迅速上升，并保持在高位直至207 DAF。每克鲜重中 NAD-SDH 的免疫反应蛋白水平较高，分别为 104、181 和 207 DAF，表明 NAD-SDH 的活性与蛋白之间存在密切相关性。此外，RNA印迹分析表明NAD-SDH转录物在整个发育阶段与活性和蛋白质水平成比例。这些结果表明活性的升高可能取决于NAD-SDH基因的表达。
The purpose of this study is to establish the mechanism of expression of NAD-dependent sorbitol dehydrogenase (NAD-SDH) activity during the development of apple (Malus pumila Mill. var. domestica Schneid. cv. Fuji) by investigating the NAD-SDH activity and the levels of NAD-SDH protein and transcript. The gene expression was investigated by RNA blot analysis using NAD-SDH cDNA and the protein level was determined by Western blot using the antibody against NAD-SDH protein expressed in E. coli.The activity of NAD-SDH was very weak up to 80 days after flowering (DAF), then increased temporarily at 104DAF. Thereafter, it declined with fruit enlargement; it rose rapidly again from 160 DAF with fruit maturation and remained high up to 207 DAF. That the levels of the immunoreactive protein of NAD-SDH per g fresh weight were high at 104, 181, and 207 DAF indicates a close correlation between the activity and the protein of NAD-SDH. Furthermore, RNA blot analysis indicated that the NAD-SDH transcript was proportionate to the level of the activity and protein through the developmental stages. These results indicate that the rise in the activity may depend upon the expression of the NAD-SDH gene.