Cloning of UePkc1 in Ustilago esculenta and Its Expression Analysis in Hypha Growth and the Infected Zizania latifolia
QI Xiao-Qing, ZHOU Ya-Ping, LIU Jing-Xi, ZHENG Yu-Jie, SHANG Yuan-Yuan, TANG Jin-Tian, XIA Wen-Qiang, ZHANG Ya-Fen, YE Zi-Hong, CUI Hai-Feng*
College of Life Sciences/Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, China
Abstract：The development of swollen stem was induced by the infection of Ustilago esculenta in the stem of Zizania latifolia. The mycelium growth of U. esculenta and its response to environmental factors played an important role in regulating the development of swollen stem in Z. latifolia. Protein kinase C1 (Pkc1) was an important regulator of fungi which regulated the cell polar growth and responsed to stimulus feedback. In this research, the UePkc1 gene (GenBank No. OQ572333) was cloned with the cDNA length of 3 573 bp, which encoded 1 190 amino acids and had the STKc_PKC conserved domain of PKC protein. After treatment with 3 types cell wall interferers (Congo red, fluorescent brightener and sodium dodecyl sulfate) and 2 different temperatures (15 ℃ and 35 ℃) during culture of U. esculenta in vitro, the swelling and shortening of basidiospore cells were observed by treatments with cell wall interfering agents and high temperature. The fusion mycelium of U. esculenta was shortened in the treatments with cell wall interfering agents, while no fusion mycelium was found in the 2 treatments with different temperatures. qPCR analysis showed that the expression of UePkc1 in U. esculenta was significantly increased after treatment with cell wall interferers for 48 h, and the expression of UePkc1 in U. esculenta treated with low temperature for 72 h was also significantly increased, while it was significantly decreased after treatment with high temperature. These results suggested that UePkc1 gene was involved in the maintenance of cell wall integrity and responsed to ambient temperature regulation in U. esculenta. Besides, there were significant differences in the expression of UePkc1 among the development stems in Z. latifolia between different swelling phenotypes. The expression of UePkc1 was the highest in the development stem of 10 cm in Grey-Jiaobai, while it was the highest at the early stage of stem in Normal-Jiaobai. Combined with the changes of growth and distribution of U. esculenta during stem development of Grey-Jiaobai and normal Jiaobai, It was preliminarily clarified that the mycelial growth of U.esculanta in the stem of plants was regulated by the expression of UePkc1, which might be positively correlated with the increasing of mycelial quantity related to the swollen of the stem. This study provides technical reference and theoretical basis for the research on the interaction mechanism of inducing the swollen and development of the stem in Z. latifolia infected by U. esculenta.
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