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反义表型揭示了生长素响应因子OsARF1在转基因水稻中的功能表达。

Antisense phenotypes reveal a functional expression of OsARF1, an auxin response factor, in transgenic rice.

作者信息

Attia Kotb A, Abdelkhalik Amr F, Ammar Megahed H, Wei Chun, Yang Jinshui, Lightfoot David A, El-Sayed Wagih M, El-Shemy Hany A

机构信息

Rice Biotechnology Lab., Rice Research and Training Center, Sakha, Kafr EL-Sheikh, 33717, Egypt.

出版信息

Curr Issues Mol Biol. 2009;11 Suppl 1:i29-34. Epub 2009 Feb 2.

Abstract

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.

摘要

OsARF1是从单子叶植物中克隆出的生长素响应因子(ARF)基因家族的首个全长成员。本研究使用定量逆转录聚合酶链反应发现,OsARF1在胚胎组织中的转录丰度显著高于营养组织。为了研究OsARF1对水稻表型的影响,将OsARF1的一个cDNA片段以反向方向插入载体pBin438中的35S启动子,以产生反义(AS)构建体。通过根癌农杆菌介导的转化,将AS-OsARF1构建体导入水稻(Oryza sativa L. japonica)愈伤组织。对转基因植物的分子分析表明,OsARF1的功能表达在mRNA水平上得到有效抑制。AS-OsARF1植株生长极低、活力差、叶片短且卷曲,分蘖但不育。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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