Venkatesan S, Raja J A J, Maruthasalam S, Kumar K K, Ramanathan A, Sudhakar D, Balasubramanian P
Rice Transformation Laboratory, Department of Plant Molecular Biology and Biotechnology, Center for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore, 641003, Tamil Nadu, India.
Virus Genes. 2009 Jun;38(3):445-54. doi: 10.1007/s11262-009-0342-5. Epub 2009 Mar 3.
The nucleocapsid protein (N) gene of a Tospovirus devastating tomato crop in the south Indian state of Tamil Nadu was cloned and characterized. The high identity of the cloned sequence to a Peanut bud necrosis virus (PBNV) tomato isolate (97.8/99.6% nucleotide/amino acid) and a PBNV peanut isolate (94.4/96.3% nucleotide/amino acid) identified the Tospovirus as an isolate of PBNV, designated PBNV Coimbatore tomato (PBNV CT) isolate. Phylogenetic analysis of PBNV CT N gene provided useful insights into the movement and evolution of PBNV within Indian Territory. The characteristic phylogeny of PBNV CT N gene implied its potential to be an efficient transgene to confer effective PBNV resistance on crop plants. The efficacy of PBNV CT N gene in conferring PBNV resistance was studied by generating tobacco (Nicotiana tabacum L. cv Wisconsin) lines transgenic to the sense or antisense version of the gene. Several transgenic lines showed transgenic mRNA and/or protein accumulation, ranging from very high to undetectable levels, accompanied by different degrees of PBNV resistance. The undetectable or very low levels of transgene transcripts in certain PBNV-resistant sense or antisense N gene transgenic lines suggested RNA-mediated resistance by post-transcriptional gene silencing (PTGS) mechanism. However, PBNV resistance of certain transgenic lines with high levels of N gene transcripts was suggestive of possible operation of RNA-mediated non-PTGS mechanism(s) of resistance in those lines. Moreover, the high levels of N protein in certain PBNV-resistant sense N gene transgenic lines suggested protein-mediated resistance. The results predict the potential of PBNV CT N gene to confer effective PBNV resistance on tomato and other economically important crops.
对印度南部泰米尔纳德邦一种严重破坏番茄作物的番茄斑萎病毒的核衣壳蛋白(N)基因进行了克隆和特性分析。克隆序列与花生芽坏死病毒(PBNV)番茄分离株(核苷酸/氨基酸一致性分别为97.8%/99.6%)以及PBNV花生分离株(核苷酸/氨基酸一致性分别为94.4%/96.3%)高度一致,从而确定该番茄斑萎病毒为PBNV的一个分离株,命名为PBNV哥印拜陀番茄(PBNV CT)分离株。对PBNV CT N基因进行系统发育分析,为PBNV在印度境内的传播和进化提供了有用的见解。PBNV CT N基因独特的系统发育关系表明其有可能成为一种有效的转基因,使作物对PBNV产生有效的抗性。通过构建该基因正义或反义版本的转基因烟草(Nicotiana tabacum L. cv Wisconsin)株系,研究了PBNV CT N基因赋予PBNV抗性的效果。几个转基因株系表现出转基因mRNA和/或蛋白质积累,积累水平从非常高到无法检测,同时伴有不同程度的PBNV抗性。某些抗PBNV的正义或反义N基因转基因株系中无法检测到或极低水平的转基因转录本,表明存在通过转录后基因沉默(PTGS)机制介导的RNA抗性。然而,某些N基因转录本水平较高的转基因株系对PBNV的抗性表明,这些株系可能存在RNA介导的非PTGS抗性机制。此外,某些抗PBNV的正义N基因转基因株系中高水平的N蛋白表明存在蛋白质介导的抗性。这些结果预示了PBNV CT N基因在使番茄和其他重要经济作物对PBNV产生有效抗性方面的潜力。