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一个在黑暗中表达且能被赤霉素迅速诱导的豌豆cDNA的分子克隆与分析。

Molecular cloning and analysis of a pea cDNA that is expressed in darkness and very rapidly induced by gibberellic acid.

作者信息

Li H Y, Guo Z F, Zhu Y X

机构信息

The National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing, People's Republic of China.

出版信息

Mol Gen Genet. 1998 Sep;259(4):393-7. doi: 10.1007/s004380050828.

Abstract

Using cDNA representational difference analysis (cDNA RDA), we isolated a cDNA named GDA-1 from a cDNA library constructed with mRNA from short-day (SD) grown G2 pea apical tissue. The amino acid sequence deduced from GDA-1 shares partial identity with the B2 protein which is expressed during embryogenesis of carrot cells. Northern analysis showed that GDA-1 mRNA is abundant in SD-grown G2 pea apical buds. In long-day (LD) conditions, there was almost no detectable GDA-1 mRNA. When LD-grown G2 peas were kept in continuous darkness for 24 h, the GDA-1 mRNA content reached a level equivalent to about 50% of that in the SD samples. On the other hand, when SD-grown peas were transferred into the light for 24 h, the amount of hybridizable GDA-1 mRNA dropped to the same as that of LD-grown plants. GDA-1 expression was found to be independent of flower initiation time. GA3 application in vitro resulted in rapid accumulation of GDA-1 mRNA in LD-grown G2 pea apical buds, which is compatible with its delaying effect on apical senescence. Time-course experiments revealed that GDA-1 is induced within 15 min of GA3 application. Exogenous GA3 did not influence the expression of GDA-1 in SD-grown G2 peas. Since both photoperiod and GA induce the expression of GDA-1, we speculate that they may activate similar signal transduction pathways in G2 peas. Our work also shows that photoperiod may change the efficiency of gibberellin perception by plants.

摘要

利用cDNA代表性差异分析(cDNA RDA),我们从用短日(SD)生长的G2豌豆顶端组织的mRNA构建的cDNA文库中分离出一个名为GDA-1的cDNA。从GDA-1推导的氨基酸序列与胡萝卜细胞胚胎发生过程中表达的B2蛋白具有部分同一性。Northern分析表明,GDA-1 mRNA在SD生长的G2豌豆顶端芽中丰富。在长日(LD)条件下,几乎检测不到GDA-1 mRNA。当LD生长的G2豌豆在连续黑暗中保持24小时时,GDA-1 mRNA含量达到相当于SD样品中约50%的水平。另一方面,当SD生长的豌豆转移到光照下24小时时,可杂交的GDA-1 mRNA量下降到与LD生长的植物相同的水平。发现GDA-1的表达与花起始时间无关。体外施用GA3导致LD生长的G2豌豆顶端芽中GDA-1 mRNA迅速积累,这与其对顶端衰老的延迟作用一致。时间进程实验表明,在施用GA3后15分钟内诱导了GDA-1。外源GA3不影响SD生长的G2豌豆中GDA-1的表达。由于光周期和GA都诱导GDA-1的表达,我们推测它们可能在G2豌豆中激活相似的信号转导途径。我们的工作还表明,光周期可能改变植物对赤霉素的感知效率。

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