Trocha P, Daly J M, Langenbach R J
Department of Biochemistry and Nutrition, University of Nebraska, Lincoln, Nebraska 68503.
Plant Physiol. 1974 Apr;53(4):519-26. doi: 10.1104/pp.53.4.519.
Synthesis of germ tube wall is a major quantitative event during germination penetration of fungi on host plants, but little is known of germ tube composition or metabolic regulation. Sonic oscillation was used to separate germ tubes from germinating uredospores of Uromyces phaseoli var. typica. Uniformly (14)C-labeled wall fractions from both structures were prepared by repeated low speed centrifugation and extraction with polar and nonpolar solvents. Based on amino acid analysis, approximately 6 and 16% of the carbon from uredospore and germ tube walls, respectively, was present in amino acids readily accessible to protease. Covalent linkages between amino acid and carbohydrate of walls was indicated by analysis of fragments prepared by mild hydrolytic procedures and separated by column chromatography and paper electrophoresis. The existence of protein in wall structures may resolve some previous uncertainty about the occurrence of protein biosynthesis during germination of rust fungi. Glucose, mannose, and glucosamine were the only carbohydrate components identified in both germ tubes and uredospore walls but different percentages were observed (germ tubes 28: 16: 16; uredospore 6: 36: 6). In germ tubes, most of the glucosamine was present in linkages hydrolyzed only by strong acid treatment, suggesting chitin-like polymers. In uredospore walls, glucosamine appears to be associated with red uredospore pigment which has properties similar to those of a melanin. Approximately 20% of the carbon in walls could not be identified with known compounds, partially because of degradation during the analytical procedures.
芽管细胞壁的合成是真菌在寄主植物上萌发侵入过程中的一个主要定量事件,但对于芽管的组成或代谢调控却知之甚少。采用声波振荡法从菜豆单胞锈菌典型变种的萌发夏孢子中分离出芽管。通过反复低速离心并用极性和非极性溶剂萃取,制备了来自这两种结构的均匀(14)C标记的细胞壁组分。基于氨基酸分析,夏孢子壁和芽管细胞壁中分别约有6%和16%的碳存在于蛋白酶易于作用的氨基酸中。通过对温和水解程序制备的片段进行分析,并通过柱色谱和纸电泳进行分离,表明了细胞壁中氨基酸与碳水化合物之间的共价键。细胞壁结构中蛋白质的存在可能解决了先前关于锈菌萌发过程中蛋白质生物合成发生情况的一些不确定性。葡萄糖、甘露糖和氨基葡萄糖是在芽管和夏孢子壁中鉴定出的仅有的碳水化合物成分,但观察到的百分比不同(芽管为28:16:16;夏孢子为6:36:6)。在芽管中,大部分氨基葡萄糖以仅能通过强酸处理水解的键存在,表明存在几丁质样聚合物。在夏孢子壁中,氨基葡萄糖似乎与红色夏孢子色素有关,该色素具有类似于黑色素的性质。细胞壁中约20%的碳无法用已知化合物鉴定,部分原因是在分析过程中发生了降解。