Gális Ivan, Simek Petr, Narisawa Tomoko, Sasaki Mami, Horiguchi Tatsuya, Fukuda Hiroo, Matsuoka Ken
RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku,Yokohama 230-0045, Japan.
Plant J. 2006 May;46(4):573-92. doi: 10.1111/j.1365-313X.2006.02719.x.
Target metabolic and large-scale transcriptomic analyses of tobacco (Nicotiana tabacum L.) Bright Yellow-2 (BY-2) cells were employed to identify novel gene(s) involved in methyl jasmonate (MJ)-dependent function in plants. At the metabolic level, we describe the specific accumulation of several phenylpropanoid-polyamine conjugates in MJ-treated BY-2 cells. Furthermore, global gene expression analysis of MJ-treated cells using a 16K cDNA microarray containing expressed sequence tags (ESTs) from BY-2 cells revealed 828 genes that were upregulated by MJ treatment within 48 h. Using time-course expression data we identified a novel MJ-inducible R2R3 MYB-type transcription factor (NtMYBJS1) that was co-expressed in a close temporal pattern with the core phenylpropanoid genes phenylalanine ammonia-lyase (PAL) and 4-coumarate:CoA ligase (4CL). Overexpression of NtMYBJS1 in tobacco BY-2 cells caused accumulation of specific phenylpropanoid conjugates in the cells. Subsequent microarray analysis of NtMYBJS1 transgenic lines revealed that a limited number of genes, including PAL and 4CL, were specifically induced in the presence of the NtMYBJS1 transgene. These results, together with results of both antisense expression analysis and of gel mobility shift assays, strongly indicate that the NtMYBJS1 protein functions in tobacco MJ signal transduction, inducing phenylpropanoid biosynthetic genes and the accumulation of phenylpropanoid-polyamine conjugates during stress.
利用烟草(Nicotiana tabacum L.)亮黄-2(BY-2)细胞进行靶向代谢组学和大规模转录组分析,以鉴定参与植物中茉莉酸甲酯(MJ)依赖性功能的新基因。在代谢水平上,我们描述了MJ处理的BY-2细胞中几种苯丙烷类-多胺共轭物的特异性积累。此外,使用包含BY-2细胞表达序列标签(EST)的16K cDNA微阵列对MJ处理的细胞进行全局基因表达分析,结果显示有828个基因在48小时内被MJ处理上调。利用时间进程表达数据,我们鉴定出一个新的MJ诱导型R2R3 MYB类转录因子(NtMYBJS1),它与核心苯丙烷类基因苯丙氨酸解氨酶(PAL)和4-香豆酸:辅酶A连接酶(4CL)以紧密的时间模式共表达。NtMYBJS1在烟草BY-2细胞中的过表达导致细胞中特异性苯丙烷类共轭物的积累。随后对NtMYBJS1转基因系进行微阵列分析,结果显示在存在NtMYBJS1转基因的情况下,包括PAL和4CL在内的有限数量的基因被特异性诱导。这些结果,连同反义表达分析和凝胶迁移率变动分析的结果,有力地表明NtMYBJS1蛋白在烟草MJ信号转导中发挥作用,在胁迫期间诱导苯丙烷类生物合成基因并积累苯丙烷类-多胺共轭物。