Selection and Optimization of Reference Genes for MicroRNA Expression Normalization by qRT-PCR in Chinese Cedar (Cryptomeria fortunei) under Multiple Stresses

被引:11
|
作者
Zhang, Yingting [1 ,2 ,3 ]
Xue, Jinyu [1 ,2 ,3 ]
Zhu, Lijuan [1 ,2 ,3 ]
Hu, Hailiang [1 ,2 ,3 ]
Yang, Junjie [1 ,2 ,3 ]
Cui, Jiebing [1 ,2 ,3 ]
Xu, Jin [1 ,2 ,3 ]
机构
[1] Nanjing Forestry Univ, Key Lab Forest Genet & Biotechnol, Minist Educ, Nanjing 210037, Peoples R China
[2] Nanjing Forestry Univ, Coinnovat Ctr Sustainable Forestry Southern China, Nanjing 210037, Peoples R China
[3] Nanjing Forestry Univ, Coll Forestry, Nanjing 210037, Peoples R China
关键词
miRNAs; reference gene; qRT-PCR; abiotic stress; hormone treatment; tissue; SUITABLE REFERENCE GENES; INNER REFERENCE GENES; REAL-TIME PCR; ABIOTIC STRESS;
D O I
10.3390/ijms22147246
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
MicroRNA (miRNA) expression analysis is very important for investigating its functions. To date, no research on reference genes (RGs) for miRNAs in gymnosperms, including Cryptomeria fortunei, has been reported. Here, ten miRNAs (i.e., pab-miR159a, cln-miR162, cas-miR166d, pab-miR395b, ppt-miR894, cln-miR6725, novel1, novel6, novel14 and novel16) and three common RGs (U6, 5S and 18S) were selected as candidate RGs. qRT-PCR was used to analyse their expressions in C. fortunei under various experimental conditions, including multiple stresses (cold, heat, drought, salt, abscisic acid and gibberellin) and in various tissues (roots, stems, tender needles, cones and seeds). Four algorithms (delta Ct, geNorm, NormFinder and BestKeeper) were employed to assess the stability of candidate RG expression; the geometric mean and RefFinder program were used to comprehensively evaluate RG stability. According to the results, novel16, cln-miR6725, novel1 and U6 were the most stable RGs for studying C. fortunei miRNA expression. In addition, the expression of three target miRNAs (aly-miR164c-5p, aly-miR168a-5p and smo-miR396) was examined to verify that the selected RGs are suitable for miRNA expression normalisation. This study may aid further investigations of miRNA expression/function in the response of C. fortunei to abiotic stress and provides an important basis for the standardisation of miRNA expression in other gymnosperm species.
引用
收藏
页数:21
相关论文
共 50 条
  • [41] Identification of Accurate Reference Genes for qRT-PCR Analysis of Gene Expression in Eremochloa ophiuroides under Multiple Stresses of Phosphorus Deficiency and/or Aluminum Toxicity
    Chen, Ying
    He, Qingqing
    Li, Xiaohui
    Zhang, Yuan
    Li, Jianjian
    Zhang, Ling
    Yao, Xiang
    Zhang, Xueli
    Liu, Chuanqiang
    Wang, Haoran
    PLANTS-BASEL, 2023, 12 (21):
  • [42] Selection and validation of reference genes for qRT-PCR in cultivated octoploid strawberry
    Mao, Jianxin
    Li, Jiqi
    Wang, Yan
    Zhang, Zhihong
    FRUIT RESEARCH, 2024, 4
  • [43] Reference gene selection for qRT-PCR normalization of gene expression analysis in Melaleuca bracteata F. Muell. under abiotic stresses and hormonal stimuli
    Yang, Chao
    Lin, Yongsheng
    Qiu, Ziwen
    Xiang, Xuwen
    Shao, Dandan
    Li, Yongyu
    Wu, Shaohua
    SCIENTIA HORTICULTURAE, 2023, 319
  • [44] Selection and Validation of Reference Genes in Sudan Grass (Sorghum sudanense (Piper) Stapf) under Various Abiotic Stresses by qRT-PCR
    Wang, Fangyan
    Li, Peng
    Liu, Qiuxu
    Nie, Gang
    Zhu, Yongqun
    Zhang, Xinquan
    GENES, 2024, 15 (02)
  • [45] Selection of Reference Genes for Normalization of qRT-PCR Analysis in the Soybean Aphid Aphis glycines Matsumura (Hemiptera: Aphididae)
    Wang, Zhengbing
    Zhang, Hongfei
    Zhang, Ziyi
    Zhao, Jiayu
    Ma, Feilong
    Zheng, Manman
    Yang, Mingsheng
    Sang, Xiaoyue
    Ma, Keshi
    Li, Lili
    JOURNAL OF ECONOMIC ENTOMOLOGY, 2022, 115 (06) : 2083 - 2091
  • [46] Selection and validation of reference genes for real-time qRT-PCR normalization in different tissues of Eucalyptus tereticornis
    Sundari, B. Karpaga Raja
    Dasgupta, M. Ghosh
    SILVAE GENETICA, 2012, 61 (06) : 280 - 286
  • [47] Selection and Validation of Appropriate Reference Genes for qRT-PCR Analysis of Iris germanica L. Under Various Abiotic Stresses
    Yuan, Yuan
    Liu, Chungui
    Bao, Jianzhong
    Li, Fengtong
    FOOD SCIENCE & NUTRITION, 2025, 13 (01):
  • [48] Selection of suitable reference genes for qRT-PCR analysis of Begonia semperflorens under stress conditions
    Yanman Li
    Ying Qu
    Yang Wang
    Xue Bai
    Geng Tian
    Zhirou Liu
    Yonghua Li
    Kaiming Zhang
    Molecular Biology Reports, 2019, 46 : 6027 - 6037
  • [49] Selection of suitable reference genes for qRT-PCR analysis of Begonia semperflorens under stress conditions
    Li, Yanman
    Qu, Ying
    Wang, Yang
    Bai, Xue
    Tian, Geng
    Liu, Zhirou
    Li, Yonghua
    Zhang, Kaiming
    MOLECULAR BIOLOGY REPORTS, 2019, 46 (06) : 6027 - 6037
  • [50] Selection and Validation of Reference Genes for qRT-PCR Analysis of Gene Expression in Tropaeolum majus (Nasturtium)
    Tang, Qing
    Zhou, Guang-Can
    Liu, Si-Jie
    Li, Wen
    Wang, Yi-Lei
    Xu, Gao-Ying
    Li, Teng-Fei
    Meng, Guo-Qing
    Xue, Jia-Yu
    HORTICULTURAE, 2023, 9 (11)