农业生物技术学报2024，Vol.32Issue(12)：2915-2925,11.DOI:10.3969/j.issn.1674-7968.2024.12.020
实时荧光定量PCR筛选荷叶离褶伞的内参基因及验证
Screening and Validation of Reference Genes in Lyophyllum decastes by qRT-PCR
摘要
Abstract
Choosing an appropriate internal reference gene is an essential requirement for qRT-PCR.However,Lyophyllum decastes has no internal reference genes reported.In this study,mycelia at different after-ripening times,fruiting bodies of different developmental stages and different cap morphologies were selected as the experimental materials.qRT-PCR was used to quantitatively amplify 6 commonly internal reference genes which were cytochrome c oxidase subunit 1(Cox1),ATPase,glucose-6-phosphate isomerase(PGI),protein phosphatase 2A(PP2A),DNA-directed RNA polymerase subunit 2(Rpb2),and ubiquitin-conjugating enzyme(UBC)and 4 rarely internal reference genes which were chaperonin containing TCP1,subunit 2(CCT2),cytochrome b560 subunit(Cyb),17 beta-hydroxysteroid dehydrogenase type 3(HSD17B3)and copper/zinc superoxide dismutase(SODC).The stability of 10 internal reference genes were evaluated by ΔCt,geNorm,NormFinder,BestKeeper and RefFinder and validated by β-1,3-glucanases,which was the key enzyme in starch and sucrose metabolism.The results showed that HSD17B3 were the most stable gene in different after-ripening times and different developmental stages;Rpb2 was the most stable gene in different cap morphologies.However,PP2A was the most unstable gene in different after-ripening times and different cap morphologies samples.Moreover,PGI was the most unstable in different developmental stages.The study evaluate the stability of internal reference genes in L.decastes and expect to offer reference for researches about different stages and different cap morphologies in L.decastes.关键词
荷叶离褶伞/内参基因/实时荧光定量PCR/稳定性Key words
Lyophyllum decastes/Internal reference gene/qRT-PCR/Stability分类
农业科技引用本文复制引用
梁力丹,李华君,兰玉菲,臧西哲,林嘉龙,韦勇琪,张佩锦,任鹏飞,孟丽..实时荧光定量PCR筛选荷叶离褶伞的内参基因及验证[J].农业生物技术学报,2024,32(12):2915-2925,11.基金项目
山东省重点研发计划项目(2022CXGC010610 （）
2022LZGC023) （）
山东省食用菌产业技术体系(SDAIT-07-06) （SDAIT-07-06）
