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Expression Analysis of a Ripening-Specific, Auxin-Repressed Endo-1,4-β-Glucanase Gene in Strawberry

机译:特定于成熟的生长素抑制的表达分析 草莓中的Endo-1,4-β-葡聚糖酶基因

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摘要

A cDNA (Cel1) encoding an endo-1,4-β-glucanase (EGase) was isolated from ripe fruit of strawberry (Fragaria × ananassa). The deduced protein of 496 amino acids contains a presumptive signal sequence, a common feature of cell wall-localized EGases, and one potential N-glycosylation site. Southern- blot analysis of genomic DNA from F. × ananassa, an octoploid species, and that from the diploid species Fragaria vesca indicated that the Cel1 gene is a member of a divergent multigene family. In fruit, Cel1 mRNA was first detected at the white stage of development, and at the onset of ripening, coincident with anthocyanin accumulation, Cel1 mRNA abundance increased dramatically and remained high throughout ripening and subsequent fruit deterioration. In all other tissues examined, Cel1 expression was invariably absent. Antibodies raised to Cel1 protein detected a protein of 62 kD only in ripening fruit. Upon deachenation of young white fruit to remove the source of endogenous auxins, ripening, as visualized by anthocyanin accumulation, and Cel1 mRNA accumulation were both accelerated. Conversely, auxin treatment of white fruit repressed accumulation of both Cel1 mRNA and ripening. These results indicate that strawberry Cel1 is a ripening-specific and auxin-repressed EGase, which is regulated during ripening by a decline in auxin levels originating from the achenes.
机译:从草莓(Fragaria×ananassa)的成熟果实中分离出编码内切1,4-β-葡聚糖酶(EGase)的cDNA(Cel1)。推导的496个氨基酸的蛋白质包含一个假定的信号序列,细胞壁局部EGase的共同特征和一个潜在的N-糖基化位点。来自八倍体F.×ananassa和二倍体草莓Fragaria vesca的基因组DNA的Southern印迹分析表明,Cel1基因是一个有差异的多基因家族的成员。在水果中,首先在发育的白色阶段检测到Cel1 mRNA,并且在成熟开始时,与花色苷的积累同时发生,Cel1 mRNA的丰度急剧增加,并在整个成熟和随后的果实变质中保持较高水平。在检查的所有其他组织中,Cel1表达总是不存在。产生于Cel1蛋白的抗体仅在成熟的果实中检测到62 kD的蛋白。年轻的白色水果脱去卵后, 内源性生长素的来源,成熟,如花色苷所示 积累和Cel1 mRNA积累都是 加速。相反,生长素处理对白色水果的抑制作用 Cel1 mRNA的积累和成熟。这些 结果表明草莓Cel1是 特定于成熟的生长素抑制型EGase 源自瘦果的生长素水平下降而成熟。

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