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豇豆种子子叶中储存的mRNA:一种储存mRNA的克隆cDNA的核苷酸序列及其通过早熟萌发诱导合成

Stored mRNA in cotyledons of Vigna unguiculata seeds: nucleotide sequence of cloned cDNA for a stored mRNA and induction of its synthesis by precocious germination.

作者信息

Ishibashi N, Yamauchi D, Minamikawa T

机构信息

Department of Biology, Tokyo Metropolitan University, Japan.

出版信息

Plant Mol Biol. 1990 Jul;15(1):59-64. doi: 10.1007/BF00017724.

Abstract

By differential hybridization screening, we previously selected a class of cDNA clones from a lambda gt10 cDNA library that was constructed from the total poly(A)+ RNA of mature cowpea cotyledons (Plant Cell Physiol 31: 39-44, 1990). pSAS10, a clone of this class, hybridized with a cDNA probe complementary to poly(A)+ RNA from cotyledons collected 1 day after the onset of imbibition (DAI), but not with the cDNA probe from cotyledons at developmental stage II (13 to 15 days after flowering, DAF). pSAS10 mRNA was detectable only in cotyledons at developmental stage III (17 to 19 DAF) or later, and its level began to decline when seeds germinated. We have suggested that pSAS10 mRNA is likely to belong to the class of 'stored' mRNA or the mRNA that is formed at the late stage of seed maturation, is conserved in quiescent seeds and becomes functional at the early stage of germination. We determined the nucleotide sequence of pSAS10 cDNA consisting of 459 bp and an approximately 36 bp poly(A) tract, and deduced the amino acid sequence of its product, a 10-kDa cysteine-rich polypeptide. Synthesis of pSAS10 mRNA was induced just before germination began, not only in mature seeds but also in immature seeds even at stages I (9 to 11 DAF) and II (13 to 15 DAF) if they were placed under conditions suitable for germination.

摘要

通过差异杂交筛选,我们先前从一个λgt10 cDNA文库中挑选出一类cDNA克隆,该文库是用成熟豇豆子叶的总poly(A)+ RNA构建的(《植物细胞生理学》31: 39 - 44, 1990)。这类克隆中的pSAS10与一个cDNA探针杂交,该探针与吸胀开始后1天(DAI)收集的子叶中的poly(A)+ RNA互补,但不与发育阶段II(开花后13至15天,DAF)的子叶中的cDNA探针杂交。pSAS10 mRNA仅在发育阶段III(17至19 DAF)及以后的子叶中可检测到,并且在种子萌发时其水平开始下降。我们推测pSAS10 mRNA可能属于“储存”mRNA类别,或者是在种子成熟后期形成、在静止种子中保守并在萌发早期发挥功能的mRNA。我们确定了由459 bp和大约36 bp poly(A)尾组成的pSAS10 cDNA的核苷酸序列,并推导了其产物(一种10 kDa富含半胱氨酸的多肽)的氨基酸序列。pSAS10 mRNA的合成在萌发开始前被诱导,不仅在成熟种子中如此,在未成熟种子中,即使处于阶段I(9至11 DAF)和II(13至15 DAF),如果将它们置于适合萌发的条件下也会如此。

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