Ye Jian, Qu Jing, Mao Hui-Zhu, Ma Zhi-Gang, Rahman Nur Estya Binte, Bai Chao, Chen Wen, Jiang Shu-Ye, Ramachandran Srinivasan, Chua Nam-Hai
Temasek Life Sciences Laboratory, NO.1 Research Link, National University of Singapore, Singapore, 117604 Singapore ; State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, NO.1 Beichen West Road, Beijing, 100101 China.
Temasek Life Sciences Laboratory, NO.1 Research Link, National University of Singapore, Singapore, 117604 Singapore.
Biotechnol Biofuels. 2014 Oct 21;7(1):149. doi: 10.1186/s13068-014-0149-z. eCollection 2014.
Jatropha curcus is a good candidate plant for biodiesel production in tropical and subtropical regions. However, J. curcus is susceptible to the geminivirus Indian cassava mosaic virus (ICMV), and frequent viral disease outbreaks severely limit productivity. Therefore the development of J. curcus to carry on durable virus resistance remains crucial and poses a major biotechnological challenge.
We generated transgenic J. curcus plants expressing a hairpin, double-stranded (ds) RNA with sequences homologous to five key genes of ICMV-Dha strain DNA-A, which silences sequence-related viral genes thereby conferring ICMV resistance. Two rounds of virus inoculation were conducted via vacuum infiltration of ICMV-Dha. The durability and heritability of resistance conferred by the dsRNA was further tested to ascertain that T1 progeny transgenic plants were resistant to the ICMV-SG strain, which shared 94.5% nucleotides identity with the ICMV-Dha strain. Quantitative PCR analysis showed that resistant transgenic lines had no detectable virus.
In this study we developed transgenic J. curcus plants to include a resistance to prevailing geminiviruses in Asia. These virus-resistant transgenic J. curcus plants can be used in various Jatropha breeding programs.
麻疯树是热带和亚热带地区生物柴油生产的优良候选植物。然而,麻疯树易受双生病毒印度木薯花叶病毒(ICMV)感染,频繁的病毒病爆发严重限制了其生产力。因此,培育具有持久病毒抗性的麻疯树仍然至关重要,并且是一项重大的生物技术挑战。
我们培育了表达发夹状双链(ds)RNA的转基因麻疯树植株,该dsRNA的序列与ICMV-Dha株系DNA-A的五个关键基因同源,可使序列相关的病毒基因沉默,从而赋予对ICMV的抗性。通过ICMV-Dha的真空渗透进行了两轮病毒接种。进一步测试了dsRNA赋予的抗性的持久性和遗传性,以确定T1代转基因植株对ICMV-SG株系具有抗性,该株系与ICMV-Dha株系具有94.5%的核苷酸同一性。定量PCR分析表明,抗性转基因株系未检测到病毒。
在本研究中,我们培育了对亚洲流行双生病毒具有抗性的转基因麻疯树植株。这些抗病毒转基因麻疯树植株可用于各种麻疯树育种计划。