Departament of Microbiology/BIOAGRO, Federal University of Viçosa, Viçosa, 36570-000, Minas Gerais, Brazil.
Mycorrhiza. 2010 Nov;20(8):531-40. doi: 10.1007/s00572-010-0301-y. Epub 2010 Feb 23.
The pre-symbiotic phase that precedes physical contact between symbionts is a crucial phase in determining their compatibility, allowing the formation of the ectomycorrhiza. A subtractive cDNA library representing the differentially expressed genes of the fungus Hydnangium sp. in the pre-symbiotic phase was constructed using fungal mycelia obtained through the in vitro mycorrhization technique. The fungus was cultured in the presence of Eucalyptus grandis roots, but with no contact between the hyphae and the root system of the host plant. Genes that code for proteins related to carbohydrate, amino acid, and energy metabolisms, transcription, and protein synthesis, cellular communication, signal transduction, stress response, transposons, and proteins related to the biogenesis of cell components were identified among the 131 expressed sequence tags. Expression of the genes that code for acetyl-CoA acetyltransferase, pyruvate dehydrogenase, ATP synthase, a voltage-dependent protein from the selective ion channel, and hydrophobin was evaluated by the RT-qPCR technique, confirming the activation of these genes in this phase of the association.
共生前阶段是共生体发生物理接触之前的关键阶段,决定着共生体之间的兼容性,从而形成外生菌根。本研究采用体外共生技术获得真菌菌丝体,构建了代表共生前阶段Hydnangium sp.差异表达基因的消减 cDNA 文库。在真菌培养过程中,我们向体系中添加了巨桉根系,但并未使菌丝体与宿主植物根系发生接触。在 131 条表达序列标签中,鉴定到了与碳水化合物、氨基酸和能量代谢、转录和蛋白质合成、细胞通讯、信号转导、应激反应、转座子以及与细胞成分生物发生相关的蛋白质编码基因。通过 RT-qPCR 技术评估了编码乙酰辅酶 A 乙酰转移酶、丙酮酸脱氢酶、ATP 合酶、选择性离子通道的电压依赖性蛋白和疏水性蛋白的基因的表达情况,证实了这些基因在共生体的这一阶段被激活。