蓝光和蛋白酶体抑制剂MG132处理对蛹虫草Cmfrq、Cmwc-1和Cmwc-2转录水平的影响  

Effects of blue light and proteasome inhibitor MG132 treatments on the transcription of Cmfrq,Cmwc-1 and Cmwc-2 in Cordyceps militaris

作　　者：丰磊 李晓然 付鸣佳[1] 黄紫妍 魏伟群 肖世平 FENG Lei;LI Xiaoran;FU Mingjia;HUANG Ziyan;WEI Weiqun;XIAO Shiping(College of Life Sciences,Jiangxi Normal University,Nanchang 330022,Jiangxi,China;Jiangxi Tianjia Bioengineering Co.,Ltd.,Nanchang 330200,Jiangxi,China)

机构地区：[1]江西师范大学生命科学学院,江西南昌330022 [2]江西天佳生物工程股份有限公司,江西南昌330200

出　　处：《菌物学报》2022年第4期587-600,共14页Mycosystema

基　　金：国家自然科学基金(31760601);南昌市重大科技攻关项目(2020)。

摘　　要：生物钟的节律振荡器主要成分之间的关系构成了转录-翻译负反馈环,并以此调控生物体的生理生化反应和生长发育等。以不同时间的蓝光照射和蛋白酶体抑制剂MG132处理蛹虫草菌丝体,通过实时荧光PCR分析其中节律振荡器主要成分的3个基因Cmfrq、Cmwc-1和Cmwc-2转录水平变化,以期确定3个基因在蛹虫草中的相互关系和变化规律。研究表明,持续的黑暗情况下,MG132对Cmfrq转录水平的影响比较大,对Cmwc-1和Cmwc-2转录水平的影响有限。在黑暗培养过程中2 h的蓝光间断条件下,无MG132处理培养后期Cmfrq转录水平急剧下降,Cmwc-1转录水平却急剧上升,Cmfrq和Cmwc-1转录水平呈现生物振荡器负反馈调控特点;Cmfrq转录水平受营养条件的影响,加MG132处理后使得Cmfrq转录对营养的改变不敏感。持续的蓝光照射培养条件下,无MG132处理时Cmfrq转录水平在后期也出现急剧的下降,Cmfrq转录对MG132处理更为敏感;无论有或无MG132处理,这3个基因之间均没有反映出生物振荡器负反馈环的特点。在蓝光12h和黑暗12h的周期交替条件下,前后2个周期这3个基因的转录水平出现不一致的情况,由持续黑暗进入周期交替时,Cmfrq转录水平由高到低;在第2个周期时Cmwc-1和Cmwc-2转录水平均非常低,但MG132导致Cmwc-1和Cmwc-2转录水平升高。与黑暗条件相比,蓝光处理可以导致Cmfrq转录水平升高。Cmwc-1转录水平在多数情况下均非常低,表明该基因是生物振荡器中的关键基因。The relationship between the main components of the circadian oscillator of circadian clock constitutes a transcription-translation negative feedback loop which regulates the physiology,biochemistry,growth and development of the organisms.The mycelium of Cordyceps militaris was treated with different time of blue light irradiation and proteasome inhibitor MG132,and the transcriptional changes of three genes Cmfrq,Cmwc-1 and Cmwc-2,the main components of the circadian oscillator,were analyzed by real-time fluorescent PCR,in order to determine the relationship and variation rule of the three genes in C.militaris.Under the condition of continuous darkness,MG132 has a relatively notable impact on the transcription levels of Cmfrq,but a limited impact on the transcription levels of Cmwc-1 and Cmwc-2.In the process of dark culture,under the condition of 2 h blue light interruption,the Cmfrq transcription level decreased sharply in the later period of culture without MG132 treatment,while the transcription level of Cmwc-1 increased sharply.The Cmfrq and Cmwc-1 transcription levels showed the characteristics of circadian oscillator negative feedback regulation.The transcriptional level of Cmfrq is affected by nutrient conditions,and the addition of MG132 made Cmfrq transcription insensitive to nutrient changes.Under the condition of continuous blue light irradiation,the Cmfrq transcription level also showed a sharp decline in the later period without MG132 treatment,and Cmfrq transcription was more sensitive to MG132 treatment.With or without MG132 treatment,the three genes did not reflect the characteristics of circadian oscillator negative feedback loop.Under the cycle alternation of blue light 12 h and dark 12 h,the transcriptional levels of these three genes were inconsistent between the two cycles.When continuous darkness entered the cycle alternation,Cmfrq transcriptional levels were from high to low.In the second cycle,the transcription levels of Cmwc-1 and Cmwc-2 were very low,but MG132 led to the increase of the

关 键 词：蛹虫草 蓝光 蛋白酶体抑制剂 基因转录水平 

分 类 号：S567.3[农业科学—中草药栽培]