López-Meyer M, Nessler C L
Department of Biology, Texas A&M University, College Station 77843-3258, USA.
Plant J. 1997 Jun;11(6):1167-75. doi: 10.1046/j.1365-313x.1997.11061167.x.
Camptothecin (CPT) is a valuable anti-cancer monoterpene alkaloid produced by the Chinese tree Camptotheca acuminata. Tryptophan decarboxylase (TDC) supplies tryptamine for the indole moiety of CPT and its derivatives, and is considered a key step in monoterpene indole alkaloid biosynthesis as it links primary and secondary metabolism. This report describes the isolation and characterization of tdc1 and tdc2, two autonomously regulated TDC genes from Camptotheca. When expressed in Escherichia coli, the products of each gene could decarboxylate tryptophan, but were inactive against tyrosine, phenylalanine and 3,4-dihydroxyphenylalanine (dopa), tdc1 was developmentally regulated, having its highest expression level in the apex, young stem and bark, tissues which also contain the highest levels of CPT. Expression of tdc1 also increased during seedling development and was correlated with alkaloid accumulation during germination. tdc2 expression was induced in Camptotheca leaf discs and cell suspension cultures treated with fungal elicitor or methyl jasmonate, treatments which did not affect tdc1 expression. Unlike tdc1, tdc2 expression was not detected in any unstressed Camptotheca tissues nor in developing seedlings. These data suggest that tdc1 may be part of a developmentally regulated chemical defense system in Camptotheca, while tdc2 serves as part of a defense system induced during pathogen challenge.
喜树碱(CPT)是一种珍贵的抗癌单萜生物碱,由中国的喜树(Camptotheca acuminata)产生。色氨酸脱羧酶(TDC)为CPT及其衍生物的吲哚部分提供色胺,被认为是单萜吲哚生物碱生物合成中的关键步骤,因为它连接了初级和次级代谢。本报告描述了tdc1和tdc2的分离和特性,这两个是来自喜树的自主调节的TDC基因。当在大肠杆菌中表达时,每个基因的产物都能使色氨酸脱羧,但对酪氨酸、苯丙氨酸和3,4-二羟基苯丙氨酸(多巴)无活性。tdc1受发育调控,在顶端、幼茎和树皮中表达水平最高,这些组织中CPT的含量也最高。tdc1的表达在幼苗发育过程中也增加,并且与发芽过程中的生物碱积累相关。tdc2的表达在经真菌激发子或茉莉酸甲酯处理的喜树叶片圆盘和细胞悬浮培养物中被诱导,这些处理不影响tdc1的表达。与tdc1不同,在任何未受胁迫的喜树组织或发育中的幼苗中均未检测到tdc2的表达。这些数据表明,tdc1可能是喜树中受发育调控的化学防御系统的一部分,而tdc2则是病原体攻击期间诱导的防御系统的一部分。