Flott Berenike E, Moerschbacher Bruno M, Reisener Hans-Joachim
Institut für Biologie III (Pflanzenphysiologie) RWTH Aachen, Worringerweg, H-5100 Aachen, West Germany.
New Phytol. 1989 Mar;111(3):413-421. doi: 10.1111/j.1469-8137.1989.tb00704.x.
Near-isogenic lines of the wheat (Triticum aestivum L.) cultivar Prelude, currying either the sr5 gene for susceptibility or the Sr5 allele for resistance to stem rust (Puccinia graminis Pers. f. sp. tritici Erics. & E. Henn.) were inoculated with race 32 of the fungus which possesses the P5 gene for avirulence. Enzyme extracts prepared at different times after inoculation were separated by non-denaturing gel electrophoresis and the gels were stained for peroxidase activity using different substrates. Both anodic and cathodic gels were run. In anodic separations, resistant and susceptible plants initially both exhibited decreases in the activity of two isoenzymes of low molecular mass, increases in two enzymes of intermediate molecular mass, and induction of two low high molecular mass isoperoxidases. In susceptible plants, these changes proceeded until 3 days after inoculation, when a general decrease in all isoenzymes was observed until 8 days after inoculation. In contrast, in resistant plants, the increases continued up to 7 days after inoculation. In addition, one isoperoxidase appeared about 2-3 days after inoculation only in the incompatible interaction. Cathodic separations revealed that basic isoperoxidases increased after inoculation as well. In the group of small basic isoenzymes, two new bands appeared following inoculation. Only slight differences were seen between the two isolines. Injecting an elicitor isolated from germinated urediniospores into the intercellular spaces of wheat leaves induces typical symptoms of the resistant reaction in both isolines. Correspondingly, the isoperoxidase pattern of the incompatible interaction, including the resistance related isoenzyme, was induced in both isolines. The results are compared with previously reported changes in enzyme activities of the lignin biosynthetic Pathway, including the total activity of peroxidase. The relationship of different isoenzymes to the lignification response as a decisive mechanism of the hypersensitive resistant reaction is discussed.
将携带感病基因sr5或抗秆锈病(Puccinia graminis Pers. f. sp. tritici Erics. & E. Henn.)的Sr5等位基因的小麦(Triticum aestivum L.)品种Prelude的近等基因系,用具有无毒基因P5的该真菌小种32进行接种。对接种后不同时间制备的酶提取物进行非变性凝胶电泳分离,并用不同底物对凝胶进行过氧化物酶活性染色。同时进行阳极和阴极凝胶电泳。在阳极分离中,抗病和感病植株最初均表现出两种低分子量同工酶活性降低、两种中等分子量酶活性增加以及两种高分子量过氧化物同工酶的诱导。在感病植株中,这些变化持续到接种后3天,此时观察到所有同工酶普遍下降,直至接种后8天。相反,在抗病植株中,这些增加持续到接种后7天。此外,一种过氧化物同工酶仅在接种后2 - 3天出现在不亲和互作中。阴极分离表明,碱性过氧化物同工酶在接种后也增加。在小的碱性同工酶组中,接种后出现了两条新带。两个近等基因系之间仅观察到细微差异。将从发芽夏孢子中分离的激发子注射到小麦叶片的细胞间隙中,在两个近等基因系中均诱导出典型的抗病反应症状。相应地,在两个近等基因系中均诱导出不亲和互作的过氧化物同工酶模式,包括与抗性相关的同工酶。将这些结果与先前报道的木质素生物合成途径中酶活性的变化,包括过氧化物酶的总活性进行了比较。讨论了不同同工酶与作为过敏反应抗性反应决定性机制的木质化反应之间的关系。