钙离子调控樟芝深层发酵无性产孢及其分子机制  

Effect of calcium on sporulation of Taiwanofungus camphoratus in submerged fermentation

作　　者：李华祥[1] 陆震鸣[1] 朱青[1] 耿燕[1] 史劲松[1] 许正宏[1,2] 马延和[2] 

机构地区：[1]江南大学药学院工业生物技术教育部重点实验室粮食发酵工艺与技术国家工程实验室,江苏无锡214122 [2]中国科学院天津工业生物技术研究所天津市工业生物系统与过程工程重点实验室,天津300308

出　　处：《生物工程学报》2017年第7期1124-1135,共12页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.31401931)资助~~

摘　　要：樟芝是原产于台湾的珍稀药用真菌,具有保肝、抗癌等活性。樟芝在深层发酵过程中能够产生大量无性孢子,但是目前对樟芝无性产孢的影响因素及其分子机制尚不明确。本研究报道了樟芝发酵培养基中Ca^(2+)浓度能够有效调控樟芝无性产孢的现象;采用双向电泳(2-DE)技术,对不添加Ca^(2+)培养(对照)的菌丝体、添加1 mmol/L Ca^(2+)培养(促进产孢)的菌丝体及添加200 mmol/L Ca^(2+)培养(抑制产孢)的菌丝体进行差异蛋白质组学分析,鉴定了参与Ca^(2+)/钙调素信号通路的CaM蛋白和HSP90蛋白,以及参与FluG调控产孢信号通路的AbaA蛋白;进一步通过生物信息学分析,预测了Ca^(2+)/钙调素和FluG介导的樟芝无性产孢信号通路模型图;采用实时定量PCR(RT-qPCR)技术,对该通路上23个功能基因的转录水平进行了分析,发现了受Ca^(2+)调控最为灵敏的7个产孢相关功能基因:crz1、hsp90、flbB、brlA、abaA、wetA及fadA。本研究结果为解析樟芝无性产孢机制提供了实验依据。Taiwanofungus camphoratus is a valuable and rare medicinal mushroom with various bioactivities, such as liver protection and anti-cancer. T. camphoratus can produce many arthroconidia at the end of submerged fermentation, but molecular mechanism underlying this submerged conidiation remains unknown. In this study, we found that Ca2＋ concentration in culture medium significantly affected the arthroconidium production of T. camphoratus. Then, we identified two proteins （CaM and HSP90） involved in Ca2＋/calmodulin signaling pathway and one protein （AbaA） involved in FluG-mediated conidiation pathway by two-dimensional electrophoresis analyses. Furthermore, we proposed a Ca2＋/calmodulin- and FluG-mediated signaling pathway by bioinformatics analysis. By real-time quantitative PCR analyses of 23 key genes in the Ca24/calmodulin- and FluG-mediated conidiation pathway, we found that expression levels of 7 genes （crzl, hsp90,flbB, brlA, abaA, wetA andfadA） showed significant responses to Ca2＋ concentration in fermentation medium. Our research is beneficial for elucidating the underlying mechanism of submerged fermentation conidiation for T. camphoratus.

关 键 词：樟芝 无性产孢 分子机制 钙离子 钙调蛋白 

分 类 号：S567.3[农业科学—中草药栽培]