Bravo Juan Manuel, Campo Sonia, Murillo Isabel, Coca Mária, San Segundo Blanca
Departamento de Genética Molecular, Instituto de Biología Molecular de Barcelona. Centro de Investigación y Desarrollo, CSIC, Jordi Girona 18, 08034 Barcelona, Spain.
Plant Mol Biol. 2003 Jul;52(4):745-59. doi: 10.1023/a:1025016416951.
Pathogenesis-related (PR) proteins are plant proteins that are induced in response to pathogen attack. PR proteins are grouped into independent families based on their sequences and properties. The PR-4 family comprises class I and class II chitinases. We have isolated a full-length cDNA encoding a chitinase from maize which shares a high degree of nucleotide and amino acid sequence homology with the class II chitinases of the PR-4 family of PR proteins. Our results indicate that fungal infection, and treatment either with fungal elicitors or with moniliformin, a mycotoxin produced by the fungus Fusarium moniliforme, increase the level of ZmPR4 mRNA. In situ mRNA hybridization analysis in sections obtained from fungus-infected germinating embryos revealed that ZmPR4 mRNA accumulation occurs in those cell types that first establish contact with the pathogen. ZmPR4 mRNA accumulation is also stimulated by treatment with silver nitrate whereas the application of the hormones gibberellic acid or acetylsalicylic acid has no effect. Wounding, or treatment with abscisic acid or methyl jasmonate, results in accumulation of ZmPR4 mRNA in maize leaves. Furthermore, the ZmPR4 protein was expressed in Escherichia coli, purified and used to obtain polyclonal antibodies that specifically recognized ZmPR4 in protein extracts from fungus-infected embryos. Accumulation of ZmPR4 mRNA in fungus-infected maize tissues was accompanied by a significant accumulation of the corresponding protein. The possible implications of these findings as part of the general defence response of maize plants against pathogens are discussed.
病程相关(PR)蛋白是植物在受到病原体攻击时诱导产生的蛋白质。PR蛋白根据其序列和特性被分为不同的独立家族。PR - 4家族包括I类和II类几丁质酶。我们从玉米中分离出了一个编码几丁质酶的全长cDNA,它与PR蛋白的PR - 4家族的II类几丁质酶具有高度的核苷酸和氨基酸序列同源性。我们的结果表明,真菌感染以及用真菌激发子或用串珠镰刀菌产生的霉菌毒素串珠镰孢菌素处理,都会增加ZmPR4 mRNA的水平。对真菌感染的萌发胚切片进行原位mRNA杂交分析表明,ZmPR4 mRNA积累发生在那些首先与病原体建立接触的细胞类型中。用硝酸银处理也会刺激ZmPR4 mRNA的积累,而施用赤霉素或乙酰水杨酸则没有效果。创伤,或用脱落酸或茉莉酸甲酯处理,会导致玉米叶片中ZmPR4 mRNA的积累。此外,ZmPR4蛋白在大肠杆菌中表达、纯化,并用于制备能特异性识别来自真菌感染胚的蛋白提取物中ZmPR4的多克隆抗体。真菌感染的玉米组织中ZmPR4 mRNA的积累伴随着相应蛋白的显著积累。本文讨论了这些发现作为玉米植株对病原体的一般防御反应一部分的可能意义。