Phytopathology. 1997 Nov;87(11):1174-8. doi: 10.1094/PHYTO.1997.87.11.1174.
ABSTRACT This study examined protein induction and accumulation during imbibition and germination of corn kernels, as well as antifungal activities of extracts from germinating kernels against Aspergillus flavus and Fusarium moniliforme. Genotypes studied included GT-MAS:gk and Mp420, which are resistant to A. flavus infection and aflatoxin accumulation, and Pioneer 3154 and Deltapine G-4666, which are susceptible to A. flavus infection and aflatoxin accumulation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved five protein bands that were present at higher concentrations in germinated kernels than in nongerminated kernels. Western blot analyses revealed that one of these proteins reacted with the 22-kDa zeamatin antiserum, and a zeamatin-like protein accumulated to a higher concentration in germinated kernels. Two protein bands from dry kernels that reacted with ribosome-inactivating protein (RIP) antiserum were identified as the 32-kDa proRIP-like form and an 18-kDa peptide of the two peptides that form active RIP. However, in germinated kernels, two protein bands that reacted with RIP antiserum were identified as two RIP-like peptides with a molecular mass of approximately 18 and 9 kDa. Purified RIP and zeamatin from corn inhibited growth of A. flavus. Bioassays of germinated kernel extracts from all four genotypes exhibited antifungal activity against A. flavus and F. moniliforme, with extracts from the susceptible genotypes showing greater inhibition zones. This study provides evidence of protein induction in corn kernels during imbibition or the early stages of germination, and the induced proteins may be related to our previous findings of germination-associated resistance in the corn kernel, especially in the susceptible kernels.
摘要 本研究考察了玉米种子吸水和萌发过程中的蛋白诱导和积累,以及发芽种子提取物对黄曲霉和串珠镰刀菌的抗真菌活性。研究的基因型包括 GT-MAS:gk 和 Mp420,它们对黄曲霉感染和黄曲霉毒素积累具有抗性,以及 Pioneer 3154 和 Deltapine G-4666,它们对黄曲霉感染和黄曲霉毒素积累敏感。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离出在发芽种子中浓度高于未发芽种子的 5 条蛋白带。Western blot 分析表明,其中一种蛋白与 22 kDa 的 zeamatin 抗血清反应,且发芽种子中 zeamatin 样蛋白积累浓度更高。与核糖体失活蛋白(RIP)抗血清反应的干种子中的两条蛋白带被鉴定为 32 kDa 的 proRIP 样形式和形成活性 RIP 的两个肽之一的 18 kDa 肽。然而,在发芽种子中,与 RIP 抗血清反应的两条蛋白带被鉴定为两种 RIP 样肽,分子量约为 18 和 9 kDa。从玉米中纯化的 RIP 和 zeamatin 抑制了黄曲霉的生长。来自四个基因型的发芽种子提取物的生物测定均表现出对黄曲霉和串珠镰刀菌的抗真菌活性,敏感基因型的提取物显示出更大的抑制区。本研究提供了在玉米种子吸水或萌发早期阶段诱导蛋白的证据,诱导的蛋白可能与我们之前在玉米种子中发现的与萌发相关的抗性有关,尤其是在敏感的种子中。