1 Agriculture and Agri-Food Canada, Fredericton Research and Development Centre, Fredericton, NB, Canada.
2 Agriculture and Agri-Food Canada, Kentville Research and Development Centre, Kentville, NS, Canada.
Mol Plant Microbe Interact. 2017 Nov;30(11):876-885. doi: 10.1094/MPMI-02-17-0033-R. Epub 2017 Sep 6.
Steroidal glycoalkaloids (SGAs) are major secondary metabolites constitutively produced in cultivated potato Solanum tuberosum, and α-solanine and α-chaconine are the most abundant SGAs. SGAs are toxic to humans at high levels but their role in plant protection against pests and pathogens is yet to be established. In this study, levels of SGAs in potato were reduced by RNA interference (RNAi)-mediated silencing of GLYCOALKALOID METABOLISM 4 (GAME4)-a gene encoding cytochrome P450, involved in an oxidation step in the conversion of cholesterol to SGA aglycones. Two GAME4 RNAi lines, T8 and T9, were used to investigate the effects of manipulation of the SGA biosynthetic pathway in potato. Growth and development of an insect pest, Colorado potato beetle (CPB), were affected in these lines. While no effect on CPB leaf consumption or weight gain was observed, early instar larval death and accelerated development of the insect was found while feeding on leaves of GAME4 RNAi lines. Modulation of SGA biosynthetic pathway in GAME4 RNAi plants was associated with a larger alteration to the metabolite profile, including increased levels of one or both the steroidal saponins or phytoecdysteroids, which could affect insect mortality as well as development time. Colonization by Verticillium dahliae on GAME4 RNAi plants was also tested. There were increased pathogen levels in the T8 GAME4 RNAi line but not in the T9. Metabolite differences between T8 and T9 were found and may have contributed to differences in V. dahliae infection. Drought responses created by osmotic stress were not affected by modulation of SGA biosynthetic pathway in potato.
甾体糖苷生物碱(SGAs)是在栽培马铃薯(Solanum tuberosum)中产生的主要次生代谢产物,α-茄碱和α-卡茄碱是最丰富的 SGAs。在高浓度下,SGAs 对人类有毒,但它们在植物抵御害虫和病原体方面的作用尚未确定。在这项研究中,通过 RNA 干扰(RNAi)介导的糖基生物碱代谢 4(GAME4)基因沉默来降低马铃薯中的 SGAs 水平,GAME4 基因编码细胞色素 P450,参与胆固醇转化为 SGA 糖苷配基的氧化步骤。使用 GAME4 RNAi 线 T8 和 T9 来研究操纵马铃薯 SGA 生物合成途径的影响。这些线中昆虫害虫科罗拉多马铃薯甲虫(CPB)的生长和发育受到影响。虽然在这些线中观察到对 CPB 叶片消耗或体重增加没有影响,但发现当幼虫在 GAME4 RNAi 线的叶片上取食时,早期幼虫死亡和昆虫发育加速。在 GAME4 RNAi 植物中调节 SGA 生物合成途径与代谢物谱的更大改变相关联,包括一个或两个甾体皂苷或植物固醇水平的增加,这可能会影响昆虫死亡率以及发育时间。还测试了 GAME4 RNAi 植物上的Verticillium dahliae 的定殖。在 T8 GAME4 RNAi 线中病原体水平增加,但在 T9 中没有增加。在 T8 和 T9 之间发现了代谢物差异,这可能导致 V. dahliae 感染的差异。通过渗透胁迫引起的干旱反应不受马铃薯中 SGA 生物合成途径调节的影响。