菰黑粉菌UePkc1克隆及其在菌丝生长与被侵染菰中的表达分析
Cloning of UePkc1 in Ustilago esculenta and Its Expression Analysis in Hypha Growth and the Infected Zizania latifolia
戚晓清 1周雅萍 1柳菁溪 1郑宇捷 1尚袁媛 1汤近天 1夏文强 1张雅芬 1叶子弘 1崔海峰1
作者信息
- 1. 中国计量大学生命科学学院/浙江省生物计量与检验检疫技术重点实验室,杭州 310018
- 折叠
摘要
菰黑粉菌(Ustilago esculenta)侵染菰(Zizania latifolia)植株后能够诱导其茎部膨大发育形成可食用肉质茎,菰黑粉菌菌丝生长及其对环境因子的响应对菰植株茎部膨大发育具有重要调控作用.蛋白激酶C1(protein kinase C1,Pkc1)是真菌调控细胞极性生长和应答刺激反馈的重要调节因子.本研究克隆获得菰黑粉菌UePkc1基因(GenBank No.OQ572333),cDNA全长3 573 bp,可编码1 190个氨基酸,具有PKC蛋白的STKc_PKC保守结构域;分别采用3种细胞壁干扰剂(刚果红,荧光增白剂和十二烷基硫酸钠)及2个温度(15℃及35℃)处理体外培养的菰黑粉菌,显微观察发现,细胞壁干扰剂及高温处理均导致菰黑粉菌担孢子细胞肿胀缩短,而低温处理的菰黑粉菌担孢子细胞较为细长;细胞壁干扰剂处理后菰黑粉菌融合菌丝长度缩短,而2个温度处理组均未见融合菌丝生成.qPCR分析表明,细胞壁干扰剂处理48h菰黑粉菌中UePkc1表达显著升高(P＜0.05),低温处理72h菰黑粉菌UePkc1表达显著升高(P＜0.05),而高温处理后菰黑粉菌UePkc1表达显著下调(P＜0.05).上述结果初步表明UePkc1基因参与菰黑粉菌细胞壁完整性维持并响应环境温度的调节作用.此外,不同膨大表型植株茎部的菰黑粉菌UePkc1表达存在显著差异,UePkc1在灰茭膨大发育为10 cm时表达量最高,而在正常茭白膨大发育初期时UePkc1表达最高,结合正常茭白与灰茭植株2种膨大表型茎部发育期间菰黑粉菌的生长分布变化,初步阐明菰植株茎部的菰黑粉菌菌丝生长受UePkc1表达调控,其表达可能与植株茎部膨大相关的菌丝量增长呈正相关.本研究为深入研究菰黑粉菌侵染诱导菰植株茎部膨大发育的互作机制提供技术参考及理论依据.
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.
关键词
菰黑粉菌/UePkc1/菌丝生长/菰/茎部膨大发育Key words
Ustilago esculenta/UePkc1/Mycelial growth/Zizania latifolia/Swelling and development of stem引用本文复制引用
基金项目
浙江省重点研发计划项目(2021C02058)
浙江省重点研发计划项目(2020C02053)
国家自然科学基金(U20A2043)
浙江省基础公益研究计划项目(LGN21C150012)
浙江省"万人计划"科技创新领军人才项目(2019R52018)
出版年
2024
国家科技期刊平台
NSTL
万方数据