Analysis of Saprolegnia parasitica transcriptome following treatment with copper sulfate

Kun Hu a,#, Rong-Rong Ma a,b,#, Jun-Ming Cheng a, #, Xin Ye a, Qi Sun a, Hai-Lan Yuan a, Nan Liang a, Hao-Ran Li a and Xian-Le Yang a *
a National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, 999 Hucheng Huan Road, Shanghai 201306, China;
b East China Sea Fisheries Research Institute Chinese Academy of Fishery Sciences , Shanghai 201306, PR China
 Xian-Le Yang, Ph.D
National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, 999 Hucheng Huan Road, Lingang New City Shanghai 201306, P. R. China.
Tel: 86-21-61900453, Fax: 86-21-61900453;
E-mail: xlyang@shou.edu.cn

Background: Massive infection caused by oomycete fungus Saprolegnia parasitica is detrimental to freshwater fish. Recently, we showed that copper sulfate demonstrated good efficacy for controlling S. parasitica infection in grass carp. In this study, we investigated the mechanism of inhibition of S. parasitica growth by copper sulfate by analyzing the transcriptome of copper sulfate-treated S. parasitica. To examine the mechanism of copper sulfate inhibiting S. parasitica, we utilized RNA-seq technology to compare differential gene expression in S. parasitica treated with or without copper sulfate.  

Results: The total mapped rates of the reads with the reference genome were 90.50% in the control group and 73.50% in the experimental group. In the control group, annotated splice junctions, partial novel splice junctions and complete novel splice junctions were about 83%, 3% and 14%, respectively. In the treatment group, the corresponding values were about 75%, 6% and 19%. Following copper sulfate treatment, a total 310 genes were markedly upregulated and 556 genes were markedly downregulated in S. parasitica. Material metabolism related GO terms including cofactor binding (33 genes), 1,3-beta-D-glucan synthase complex (4 genes), carboxylic acid metabolic process (40 genes) were the most significantly enriched. KEGG pathway analysis also determined that the metabolism-related biological pathways were significantly enriched, including the metabolic pathways (98 genes), biosynthesis of secondary metabolites pathways (42 genes), fatty acid metabolism (13 genes), phenylalanine metabolism (7 genes), starch and sucrose metabolism pathway (12 genes). The qRT-PCR results were largely consistent with the RNA-Seq results.

Conclusion: Our results indicate that copper sulfate inhibits S. parasitica growth by affecting multiple biological functions, including protein synthesis, energy biogenesis, and metabolism.