Temasek Life Sciences Laboratory, National University of Singapore, Singapore.
Plant Biotechnol J. 2009 Dec;7(9):964-76. doi: 10.1111/j.1467-7652.2009.00457.x.
Jatropha curcas L. is a small, woody tree of the Euphorbiaceae family. This plant can grow on marginal land in the tropical and subtropical regions and produces seeds containing up to 30% oil. Several Asian countries have selected Jatropha for large scale planting as a biodiesel feedstock. Nevertheless, Jatropha also possesses several undesirable traits that may limit its wide adoption. An improved understanding of plant development and the regulation of fatty acid (FA) and triacylglyceride biosynthesis in Jatropha is particularly facilitative for the development of elite crops. Here, we show that a tobacco rattle virus (TRV) vector can trigger virus-induced gene silencing (VIGS) in Jatropha. Our optimized method produced robust and reliable gene silencing in plants agroinoculated with recombinant TRV harbouring Jatropha gene sequences. We used VIGS to investigate possible functions of 13 Jatropha genes of several functional categories, including FA biosynthesis, developmental regulation and toxin biosynthesis, etc. Based on the effects of VIGS on the FA composition of newly emerged leaves, we determined the function of several genes implicated in FA biosynthesis. Moreover, VIGS was able to discriminate independent functions of related gene family members. Our results show that VIGS can be used for high-throughput screening of Jatropha genes whose functions can be assayed in leaves.
麻疯树是大戟科麻疯树属的一种小乔木。这种植物可以生长在热带和亚热带地区的边缘土地上，其种子含油率高达 30%。一些亚洲国家已经选择麻疯树作为生物柴油的原料进行大规模种植。然而，麻疯树也具有一些不良特性，可能限制其广泛应用。因此，深入了解麻疯树的植物发育和脂肪酸（FA）及三酰基甘油生物合成的调控，对于优质作物的开发具有特别重要的意义。在这里，我们展示了烟草脆裂病毒（TRV）载体可以在麻疯树中引发病毒诱导的基因沉默（VIGS）。我们优化的方法在利用含有麻疯树基因序列的重组 TRV 进行农杆菌接种的植物中产生了强大而可靠的基因沉默。我们利用 VIGS 研究了 FA 生物合成、发育调控和毒素生物合成等几个功能类别中的 13 个麻疯树基因的可能功能。基于 VIGS 对新出现叶片中 FA 组成的影响，我们确定了几个参与 FA 生物合成的基因的功能。此外，VIGS 能够区分相关基因家族成员的独立功能。我们的研究结果表明，VIGS 可用于高通量筛选麻疯树基因，其功能可以在叶片中进行检测。
