Selection of Reference Genes for microRNA Real-time PCR Under Salt Stress in Armillaria gallica
10.13422/j.cnki.syfjx.20202062
- VernacularTitle:盐胁迫蜜环菌Real-time PCR内参microRNA的筛选
- Author:
Qi-rui YU
1
;
Zhong-yi HUA
2
;
Jun-hui ZHOU
2
;
Qi-tiao HU
2
;
Yuan YUAN
2
;
Quan YANG
1
;
Lu-qi HUANG
1
Author Information
1. School of Traditional Chinese Medicine，Guangdong Pharmaceutical University，Guangzhou 510006，China
2. State Key Laboratory Breeding Base of Dao-di Herbs，National Resource Center for Chinese Materia Medica，China Academy of Chinese Medical Sciences，Beijing 100700，China
- Publication Type:Research Article
- Keywords:
Armillaria gallica;
salt stress;
miRNA reference;
Real-time PCR
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2020;26(19):35-42
- CountryChina
- Language:Chinese
-
Abstract:
Objiective： In the process of microRNA expression analysis by quantitative Real-time polymerase chain reaction（Real-time PCR），the selection of miRNA plays an important role in data standardization. Method:In this paper，13 Armillaria gallica.Candidate miRNAs were selected for bioinformatics analysis of their precursors，and the PMRD was used to predict similar sequences of their precursors，and the RNAfold was used to predict the secondary structure of the candidate miRNAs and their similar sequences. Real-time PCR was used to detect miRNAs expression in two genotypes of Armillaria gallica（genotype A，genotype B） before and after salt stress，and geNorm，NormFinder and BestKeeper were used to analyze the stability of miRNAs expression. Result:Secondary structure prediction and characterization of 9 candidate miRNA precursors showed that the miRNA predicted belonged to the miR family with typical stem-loop structure and the mature miRNAs were at the 5' or 3' end of the miRNA precursors.geNorm analysis showed that genotype A Armillaria gallica could select Novel-4* and Novel-9 as reference gene，genotype B could select Novel-9 and Novel-16 as its reference gene.NormFinder analysis showed that Novel-9 was stable in both genotype A and B Armillaria gallica.BestKeeper analysis showed that Novel-12* was stable in genotype A Armillaria gallica and Novel-2* was stable in genotype B Armillaria gallica. Conclusion:miRNA Novel-9 is the best stable reference gene，which lays a foundation for further research on the regulation mechanism of miRNA in Armillaria gallica.
