Cloning and Functional Study of UePkaC Gene in Ustilago esculenta
WANG Pei-You*, ZHANG Ya-Fen*, GE Qian-Wen, XIA Wen-Qiang, CUI Hai-Feng, YU Xiao-Ping, YE Zi-Hong**
Zhejiang Provincial Key Laboratory of Biometrology and Inspection Quarantine/College of Life Sciences, China Jiliang University, Hangzhou 310018, China
AbstractUstilago esculenta is a dimorpic fungi with the ability to infect Zizania latifolia and causes stem enlargement. The fungal dimorphism is vital for the infection ability and pathogenicity of U. esculenta.The mitogen-activated protein kinase (MAPK) cascades and cyclic adenosine monophosphate-protein kinase A(cAMP-PKA) pathway are the core of signaling pathways in regulation of the fungal dimorphism. In this study, the phenomenon of multi-budding growth of cells after the addition of cAMP was found, and the hyphal growth was affected during the mating process. Furthermore, a gene encoding PKA catalytic submit (GenBank No. KR870334.1) was cloned. Sequence analysis results showed that the gene had a total length of 1 346 bp, with 1 intron and 2 exons forming 1 233 bp open reading frame, encoding 410 amino acids. This amino acid sequence was homologous to the amino acid sequence of PKA catalytic subunit in other species, such as U. hordei and U. maydis. The expression pattern analysis showed that UePkaC was up-regulated in the process of mating. The phenotype analysis of the UePkaC deletion mutants showed that they were longer and had no mating ability and pathogenicity. Furthermore, the ability of conjunction tube was lost in UePkaC mutant during mating and the expression of pheromone synthesis gene was significantly inhibited, indicating that UePkaC affects the conjugation tube formation and mating process by regulating the expression of a genes.The above results showed that UePkaC gene played a key role in the haploid growth and dimorphism of U. esculenta, which provided basic data for studying the pathogenic mechanism of U. esculenta.
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