Bell J N, Ryder T B, Wingate V P, Bailey J A, Lamb C J
Mol Cell Biol. 1986 May;6(5):1615-23. doi: 10.1128/mcb.6.5.1615-1623.1986.
Phenylalanine ammonia-lyase and chalcone synthase catalyze the first reaction of phenylpropanoid biosynthesis and the first reaction of a branch pathway specific for flavonoid-isoflavonoid biosynthesis, respectively. These enzymes are key control elements in the synthesis of kievitone, phaseollin, and related isoflavonoid-derived phytoalexins. RNA blot hybridization with 32P-labeled cDNA sequences was used to demonstrate marked accumulation of phenylalanine ammonia-lyase and chalcone synthase mRNAs in excision-wounded hypocotyls of Phaseolus vulgaris L. (dwarf French bean) and during race-cultivar-specific interactions between hypocotyls of P. vulgaris and the partially biotrophic fungus Colletotrichum lindemuthianum, the causal agent of anthracnose. In an incompatible interaction (host resistant), early concomitant accumulation of phenylalanine ammonia-lyase and chalcone synthase mRNAs, localized mainly but not entirely in tissue adjacent to the site of infection, was observed prior to the onset of phytoalexin accumulation and expression of localized, hypersensitive resistance. In contrast, in a compatible interaction (host susceptible) there was no early accumulation of these transcripts; instead, there was a delayed widespread response associated with phytoalexin accumulation during attempted lesion limitation. Two-dimensional gel electrophoresis of [35S]methionine-labeled polypeptides synthesized in vitro by translation of isolated polysomal RNA demonstrated stimulation of the synthesis of characteristic sets of phenylalanine ammonia-lyase and chalcone synthase isopolypeptides in directly infected tissue and distant, hitherto uninfected tissue in both compatible and incompatible interactions. Our data show that specific accumulation of plant defense gene transcripts is a key early component in the sequence of events leading to expression of defense responses in wounded tissue and in infected tissue during race-cultivar-specific interactions and that an elicitation signal is transmitted intercellularly in response to infection.
苯丙氨酸解氨酶和查尔酮合酶分别催化苯丙烷类生物合成的第一步反应以及类黄酮 - 异黄酮生物合成特定分支途径的第一步反应。这些酶是合成基维酮、菜豆素以及相关异黄酮衍生植保素的关键调控元件。利用与32P标记的cDNA序列进行RNA印迹杂交,证明了在菜豆(矮生菜豆)切除创伤的下胚轴中,以及在菜豆下胚轴与炭疽病病原体、部分活体营养型真菌林氏炭疽菌的小种 - 品种特异性相互作用过程中,苯丙氨酸解氨酶和查尔酮合酶mRNA显著积累。在不亲和相互作用(寄主抗性)中,在植保素积累和局部过敏反应抗性表达之前,观察到苯丙氨酸解氨酶和查尔酮合酶mRNA早期同时积累,主要但并非完全定位于感染部位附近的组织中。相反,在亲和相互作用(寄主感病)中,这些转录本没有早期积累;取而代之的是,在试图限制病斑形成期间,与植保素积累相关的延迟广泛反应。通过对分离的多聚核糖体RNA进行体外翻译合成的[35S]甲硫氨酸标记多肽进行二维凝胶电泳,结果表明,在亲和与不亲和相互作用中,直接感染的组织以及远处尚未感染的组织中,苯丙氨酸解氨酶和查尔酮合酶同工多肽的特征性合成集均受到刺激。我们的数据表明,植物防御基因转录本的特异性积累是导致在小种 - 品种特异性相互作用期间受伤组织和感染组织中防御反应表达的一系列事件中的关键早期组成部分,并且响应感染会在细胞间传递诱导信号。