转录组解析白三叶根际溶磷菌株RW8的解磷机制

doi:10.11686/cyxb2016399

摘要/Abstract

摘要： 采用转录组测序的方法分析了白三叶根际溶磷菌RW8在含有难溶磷(A组)、可溶磷(B组)与无磷(C组)培养条件下差异基因表达。以可溶磷为对照,在难溶磷条件下,分别检测到4782个基因在RW8中上调表达,447个基因下调表达;在无磷组中,共检测到3630个基因上调表达,209个基因下调表达。GO基因注释发现RW8在A组和C组中的差异基因聚类基本相同。生物学过程主要聚类在代谢过程、细胞过程、单细胞过程、刺激应答、定位以及生物反应调节;细胞组分主要聚类在细胞组分、细胞膜、膜组分与高分子配合体等;分子功能主要聚类在催化活性、结合功能与转运功能。代谢途径分析发现2-α-氧代羧、α-亚麻酸、五碳二元酸、脂肪酸、甘氨酸-丝氨酸-蛋氨酸以及缬氨酸-亮氨酸-异亮氨酸等代谢途径的基因显著被富集。挑选10个差异基因并用荧光定量检测其在A、B、C 3种不同培养条件下的基因表达,发现所有挑选基因的表达变化与转录组结果变化趋势相同。液相色谱检测有机酸组成及含量,发现在A组和C组中乳酸、琥珀酸与柠檬酸显著高于B组,富马酸与苹果酸的含量在C组中显著升高, A组与B组差异不显著;α-酮戊二酸的含量在A组中显著增高, B组和C组差异不显著。

Abstract: We conducted transcriptome sequencing of the white clover (Trifolium repens) rhizosphere phosphorus-solubilizing bacterium RW8 to explore its gene expression profiles. Cells of RW8 were cultured in three different media: one containing insoluble phosphorus (group A), one containing soluble phosphorus (group B), and one lacking phosphorus (group C). Compared with group B cells, group A cells showed 4782 and 447 up- and down-regulated genes, respectively, and group C cells showed 3630 and 209 up- and down-regulated genes, respectively. The up-regulated genes in Groups A and C had similar gene ontology annotations. The main subcategories in the biological process category were metabolic process, cellular process, single-organism process, response to stimulus, localization, and biological regulation. In the cell components category, the main subcategories were cell part, membrane, membrane part, and macromolecular complex. In the molecular function category, the main subcategories were catalytic, binding, and transporter. A metabolic pathway analysis showed that several metabolic pathways were significantly enriched in groups A and C, including 2-oxocarboxylic acid metabolism, alpha-linolenic acid metabolism, C5-branched dibasic acid metabolism, fatty acid metabolism, glycine-serine-threonine metabolism and valine-leucine-isoleucine biosynthesis. Ten differentially expressed genes in three different groups were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The transcriptional patterns detected by qRT-PCR were similar to those detected in the transcriptome sequencing analyses. The composition and concentrations of organic acids were detected by high performance liquid chromatography (HPLC). The lactic acid, succinic acid, and citric acid concentrations were significantly higher in group A and group C than in group B. The fumaric acid and malic acid concentrations were much higher in group C than in groups A and B, and did not differ significantly between groups A and B. The concentration of α-ketoglutaric acid was higher in group A than in group B, and did not differ significantly between groups C and B.

李小冬, 王小利, 陈锡, 蔡璐, 曾庆飞, 舒健虹, 蔡一鸣. 转录组解析白三叶根际溶磷菌株RW8的解磷机制[J]. 草业学报, 2017, 26(8): 168-179.

LI Xiao-Dong, WANG Xiao-Li, CHEN Xi, CAI Lu, ZENG Qing-Fei, SHU Jian-Hong, CAI Yi-Ming. Transcriptome profiling analysis of the phosphate-solubilizing mechanism of the white clover rhizosphere strain RW8[J]. Acta Prataculturae Sinica, 2017, 26(8): 168-179.

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