Yang Shu-Jun, Carter Shelly A, Cole Anthony B, Cheng Ning-Hui, Nelson Richard S
Plant Biology Division, Samuel Roberts Noble Foundation, Inc., 2510 Sam Noble Parkway, Ardmore, OK 73401, USA.
Proc Natl Acad Sci U S A. 2004 Apr 20;101(16):6297-302. doi: 10.1073/pnas.0304346101. Epub 2004 Apr 12.
Nicotiana benthamiana often displays more intense symptoms after infection by RNA viruses than do other Nicotiana species. Here, we examined the role of RNA-dependent RNA polymerases (RdRPs) in N. benthamiana antiviral defense. cDNAs representing only two genes encoding RdRPs were identified in N. benthamiana. One RdRP was similar in sequence to SDE1/SGS2 required for maintenance of transgene silencing, whereas the second, named NbRdRP1m, was >90% identical in sequence to the salicylic acid (SA)-inducible RdRP from Nicotiana tabacum required for defense against viruses. NbRdRP1m expression was induced by SA treatment or challenge with Tobacco mosaic virus, but the gene and transcript sequences differed from those of other SA-inducible RdRPs in that they contained a 72-nt insert with tandem in-frame stop codons in the 5' portion of the ORF. N. benthamiana plants transformed with an SA-inducible RdRP gene from Medicago truncatula were more resistant to infection by Tobacco mosaic virus, Turnip vein-clearing virus, and Sunn hemp mosaic virus (members of Tobamovirus genus), but not to Cucumber mosaic virus and Potato virus X (members of different genera than the tobamoviruses). Our results indicate that N. benthamiana lacks an active SA- and virus-inducible RdRP and thus is hypersusceptible to viruses normally limited in their accumulation by this RdRP. These findings are significant for those studying virus-induced gene silencing, the hypersensitive response and systemic acquired resistance.
与其他烟草属物种相比,本氏烟草在受到RNA病毒感染后通常会表现出更强烈的症状。在此,我们研究了RNA依赖性RNA聚合酶(RdRPs)在本氏烟草抗病毒防御中的作用。在本氏烟草中仅鉴定出代表两个编码RdRPs基因的cDNA。一种RdRP在序列上与维持转基因沉默所需的SDE1/SGS2相似,而另一种名为NbRdRP1m的RdRP在序列上与烟草中对抗病毒所需的水杨酸(SA)诱导型RdRP有>90%的同一性。NbRdRP1m的表达通过SA处理或烟草花叶病毒激发诱导,但该基因和转录本序列与其他SA诱导型RdRPs不同,因为它们在开放阅读框(ORF)的5'部分含有一个带有串联读框内终止密码子的72 nt插入片段。用来自蒺藜苜蓿的SA诱导型RdRP基因转化的本氏烟草植株对烟草花叶病毒、芜菁脉明病毒和太阳麻花叶病毒(烟草花叶病毒属成员)的感染更具抗性,但对黄瓜花叶病毒和马铃薯X病毒(与烟草花叶病毒属不同属的成员)不具抗性。我们的结果表明,本氏烟草缺乏一种活跃的SA和病毒诱导型RdRP,因此对通常因这种RdRP而积累受限的病毒高度敏感。这些发现对于研究病毒诱导的基因沉默、过敏反应和系统获得性抗性的人来说具有重要意义。