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Antisense Phenotypes Reveal a Functional Expression of OsARF1, an Auxin Response Factor, in Transgenic Rice

机译:反义表型揭示转基因水稻中OsARF1,一种生长素反应因子的功能性表达。

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OsARF1 is the first full-length member of auxin response factor (ARF) gene family to be cloned from monocot plant. Using quantitative RT-PCR this study found that, the transcript abundance of OsARF1 was significantly higher in embryonic tissues than in vegetative tissues. To investigate the effect of OsARF1 on the phenotype of rice, a cDNA fragment of OsARF1 was inserted in inverse orientation to the 35S promoter in vector pBin438 to produce an antisense (AS) construction. The AS-OsARF1 construct was transferred into rice (Oryza sativa L. japonica ) calli via Agrobacterium tumefaciens -mediated transformation. Molecular analysis of transgenic plants showed that the functional expression of OsARF1 was inhibited at mRNA level efficiently. The AS-OsARF1 plants showed extremely low growth, poor vigor, short curled leaves and tillered but were sterile. Therefore, the OsARF1 was shown to be essential for growth in vegetative organs and seed development.
机译:OsARF1是从单子叶植物中克隆的生长素反应因子(ARF)基因家族的第一个全长成员。使用定量RT-PCR的这项研究发现,OsARF1的转录本丰度在胚胎组织中显着高于营养组织。为了研究OsARF1对水稻表型的影响,将OsARF1的cDNA片段以反方向插入载体pBin438中的35S启动子,以产生反义(AS)结构。通过根癌农杆菌介导的转化将AS-OsARF1构建体转移到水稻(Oryza sativa L.japonica)愈伤组织中。对转基因植物的分子分析表明,OsARF1的功能性表达在mRNA水平受到有效抑制。 AS-OsARF1植物显示出极低的生长,活力差,叶片短而卷曲和分till,但不育。因此,OsARF1被证明对营养器官的生长和种子发育至关重要。

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