State Key Laboratory of Agrobiotecnology, College of Biology Science, China Agricultural University, 100193, Beijing, China.
Mol Biol Rep. 2010 Feb;37(2):865-74. doi: 10.1007/s11033-009-9692-2. Epub 2009 Aug 15.
RNA-dependent RNA polymerases (RdRPs) in plants have been reported to be involved in post-transcriptional gene silencing (PTGS) and antiviral defense. In this report, an RdRP gene from maize (ZmRdRP1) was obtained by rapid amplification of cDNA ends (RACE) and RT-PCR. The mRNA of ZmRdRP1 was composed of 3785 nucleotides, including a 167 nt 5' untranslated region (UTR), a 291 nt 3'UTR and a 3327 nt open reading frame (ORF), which encodes a putative protein of 1108 amino acids with an estimated molecular mass of 126.9 kDa and a predicated isoelectric point (pI) of 8.37. Real-time quantitative RT-PCR analysis showed that ZmRdRP1 was elicited by salicylic acid (SA) treatment, methyl jasmonate (MeJA) treatment and sugarcane mosaic virus (SCMV) infection. We silenced ZmRdRP1 by constitutively expressing an inverted-repeat fragment of ZmRdRP1 (ir-RdRP1) in transgenic maize plants. Further studies revealed that the ir-RdRP1 transgenic plants were more susceptible to SCMV infection than wild type plants. Virus-infected transgenic maize plants developed more serious disease symptoms and accumulated more virus than wild type plants. These findings suggested that ZmRdRP1 was involved in antiviral defense in maize.
植物中的 RNA 依赖性 RNA 聚合酶 (RdRPs) 已被报道参与转录后基因沉默 (PTGS) 和抗病毒防御。在本报告中,通过快速扩增 cDNA 末端 (RACE) 和 RT-PCR 获得了来自玉米的 RdRP 基因 (ZmRdRP1)。ZmRdRP1 的 mRNA 由 3785 个核苷酸组成,包括 167 nt 的 5'非翻译区 (UTR)、291 nt 的 3'UTR 和 3327 nt 的开放阅读框 (ORF),编码一个推定的 1108 个氨基酸的蛋白质,估计分子量为 126.9 kDa,预测等电点 (pI) 为 8.37。实时定量 RT-PCR 分析表明,ZmRdRP1 被水杨酸 (SA) 处理、茉莉酸甲酯 (MeJA) 处理和甘蔗花叶病毒 (SCMV) 感染所诱导。我们通过在转基因玉米植株中组成型表达 ZmRdRP1 的反向重复片段 (ir-RdRP1) 来沉默 ZmRdRP1。进一步的研究表明,ir-RdRP1 转基因植物比野生型植物更容易感染 SCMV。感染病毒的转基因玉米植物比野生型植物表现出更严重的病症和更多的病毒积累。这些发现表明 ZmRdRP1 参与了玉米的抗病毒防御。