蚁巢伞属真菌Termitomyces clypeatus实时荧光定量PCR内参基因的筛选  

Screening of the reference genes for RT-qPCR analysis of gene expressions in Termitomyces clypeatus

作　　者：周琳琳 赵玉 李夏雨 桂昊 王雨婷 龙雁华[1] ZHOU Linlin;ZHAO Yu;LI Xiayu;GUI Hao;WANG Yuting;LONG Yanhua(School of Life Sciences,Anhui Agricultural University,Hefei 230036,Anhui,China)

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036

出　　处：《菌物学报》2022年第10期1597-1606,共10页Mycosystema

基　　金：国家自然科学基金(31300426,32072421);安徽农业大学“大学生创新创业训练计划”创新训练项目(S202110364232)。

摘　　要：蚁巢伞属真菌尖盾蚁巢伞Termitomyces clypeatus是一类与大白蚁亚科昆虫共生的野生食用真菌,因味道鲜美备受消费者喜爱。为进一步对蚁巢伞属真菌开展相关生物学和遗传学研究,本试验选取8个候选内参基因[3-磷酸甘油醛脱氢酶(GAPDH)、磷酸葡萄糖变异酶(PGM)、β微管蛋白(TUB)、β肌动蛋白(ACT)、翻译延长因子1-α(EF1)、蛋白磷酸酶2A(PP2A)、聚泛素(UBQ)和翻译延伸因子2(EF2)],对其在T.clypeatus菌株不同生长发育时期(菌丝体、巢内萌发期及成熟子实体)的表达稳定性进行评估,通过4种软件(geNorm、NormFinder、BestKeeper以及RefFinder)进行数据分析,结果表明:在供试条件下,ACT、EF1和TUB的相对表达量处于较稳定的状态,可作为蚁巢伞属真菌功能基因转录水平分析的内参基因。Termitomyces clypeatus is a wild edible fungus that forms a symbiotic relationship with the ants belonging to Macrotermitinae family,and it is famous for its delicious taste.In order to carry out relevant biological and genetic research of T.clypeatus,eight candidate internal reference genes,glyceraldehyde-3-phosphate dehydrogenase(GAPDH),glucose phosphate mutant enzyme(PGM),β-tubulin(TUB),β-actin(ACT),translation prolonging factor 1-α(EF1),protein phosphatase 2A(PP2A),polyubiquitin(UBQ),and translation elongation factor 2(EF2),were used for evaluating their expression stabilities in different development stages(mycelium,germination stage in ant nest and mature fruiting body).Four algorithms(geNorm,NormFinder,BestKeeper and RefFinder)were used for data analysis.The results showed that the relative expression level of ACT,EF1,and TUB were relatively stable under the experimental conditions,and they can be used as internal reference genes for the further biological and genetic studies of gene expressions in T.clypeatus.

关 键 词：尖盾蚁巢伞 实时荧光定量PCR 内参基因 筛选 

分 类 号：S646[农业科学—蔬菜学]