Institute of Plant Sciences, ARO, the Volcani Center, 50250 Bet Dagan, Israel.
Planta. 2012 Jun;235(6):1341-53. doi: 10.1007/s00425-011-1578-6. Epub 2011 Dec 29.
Potato steroidal glycoalkaloids (SGAs) are toxic secondary metabolites whose total content in tubers must be regulated. SGAs are biosynthesized by the sterol branch of the mevalonic acid/isoprenoid pathway. In a previous study, we showed a correlation between SGA levels and the abundance of transcript coding for HMG-CoA reductase 1 (HMG1) and squalene synthase 1 (SQS1) in potato tissues and potato genotypes varying in SGA content. Here, Solanum tuberosum cv. Desirée (low SGA producer) was transformed with a gene construct containing the coding region of either HMG1 or SQS1 of Solanum chacoense Bitt. clone 8380-1, a high SGA producer. SGA levels in transgenic HMG-plants were either greater than (in eight of 14 plants) or no different from untransformed controls, whereas only four of 12 SQS-transgenics had greater SGA levels than control, as determined by HPLC. Quantitative real-time PCR was used to estimate relative steady-state transcript levels of isoprenoid-, steroid-, and SGA-related genes in leaves of the transgenic plants compared to nontransgenic controls. HMG-transgenic plants exhibited increased transcript accumulation of SQS1, sterol C24-methyltransferase type1 (SMT1), and solanidine glycosyltransferase 2 (SGT2), whereas SQS-transgenic plants, had consistently lower transcript levels of HMG1 and variable SMT1 and SGT2 transcript abundance among different transgenics. HMG-transgenic plants exhibited changes in transcript accumulation for some sterol biosynthetic genes as well. Taken together, the data suggest coordinated regulation of isoprenoid metabolism and SGA secondary metabolism.
马铃薯甾体糖苷生物碱(SGAs)是有毒的次生代谢物,其块茎总含量必须受到调控。SGAs 通过甲羟戊酸/异戊烯途径的甾醇分支生物合成。在之前的研究中,我们表明 SGA 水平与编码 HMG-CoA 还原酶 1(HMG1)和角鲨烯合酶 1(SQS1)的转录物丰度之间存在相关性,这些转录物存在于马铃薯组织和 SGA 含量不同的马铃薯基因型中。在这里,我们用含有 Solanum chacoense Bitt. clone 8380-1 的 HMG1 或 SQS1 编码区的基因构建体转化了马铃薯品种 Desirée(低 SGA 产生者)。S. chacoense Bitt. clone 8380-1 是一种高 SGA 产生者。通过 HPLC 测定,在 14 株 HMG 转化株中,有 8 株的 SGA 水平高于(而非转化对照),而在 12 株 SQS 转化株中,只有 4 株的 SGA 水平高于对照。实时定量 PCR 用于估计转基因植株叶片中异戊烯、甾体和 SGA 相关基因的相对稳态转录水平与非转基因对照相比。HMG 转化株中 SQS1、甾醇 C24-甲基转移酶 1 型(SMT1)和茄碱糖基转移酶 2(SGT2)的转录物积累增加,而 SQS 转化株中 HMG1 的转录物水平持续降低,并且不同转化株中 SMT1 和 SGT2 的转录物丰度也不同。HMG 转化株中一些甾醇生物合成基因的转录物积累也发生了变化。总之,这些数据表明异戊烯代谢和 SGA 次生代谢的协调调控。