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
  • [1] Optimization of reference genes for qRT-PCR analysis of microRNA expression under abiotic stress conditions in sweetpotato
    Liu, Xiayu
    Liu, Shifang
    Zhang, Jie
    Wu, Yuhao
    Wu, Wanyi
    Zhang, Yi
    Liu, Baoling
    Tang, Ruimin
    He, Liheng
    Li, Runzhi
    Jia, Xiaoyun
    PLANT PHYSIOLOGY AND BIOCHEMISTRY, 2020, 154 : 379 - 386
  • [2] Selection and validation of reference genes for normalization of qRT-PCR gene expression in wheat (Triticum durum L.) under drought and salt stresses
    Kiarash, Jamshidi Goharrizi
    Wilde, Henry Dayton
    Amirmahani, Farzane
    Moemeni, Mohammad Mehdi
    Zaboli, Maryam
    Nazari, Maryam
    Moosavi, Sayyed Saeed
    Jamalvandi, Mina
    JOURNAL OF GENETICS, 2018, 97 (05) : 1433 - 1444
  • [3] Selection and validation of reference genes for normalization of qRT-PCR gene expression in wheat (Triticum durum L.) under drought and salt stresses
    Jamshidi Goharrizi Kiarash
    Henry Dayton Wilde
    Farzane Amirmahani
    Mohammad Mehdi Moemeni
    Maryam Zaboli
    Maryam Nazari
    Sayyed Saeed Moosavi
    Mina Jamalvandi
    Journal of Genetics, 2018, 97 : 1433 - 1444
  • [4] Determination of the reference genes for qRT-PCR normalization and expression levels of MAT genes under various conditions in Ulocladium
    Ma, Li-Guo
    Geng, Yun
    PEERJ, 2020, 8
  • [5] Identification of Suitable Reference Genes for qRT-PCR Normalization in Kiwifruit
    Zhou, Yuanjie
    Xia, Hui
    Liu, Xinling
    Lin, Zhiyi
    Guo, Yuqi
    Deng, Honghong
    Wang, Jin
    Lin, Lijin
    Deng, Qunxian
    Lv, Xiulan
    Xu, Kunfu
    Liang, Dong
    HORTICULTURAE, 2022, 8 (02)
  • [6] Selection of Reference Genes for qRT-PCR Analysis of Gene Expression in Stipa grandis during Environmental Stresses
    Wan, Dongli
    Wan, Yongqing
    Yang, Qi
    Zou, Bo
    Ren, Weibo
    Ding, Yong
    Wang, Zhen
    Wang, Ruigang
    Wang, Kai
    Hou, Xiangyang
    PLOS ONE, 2017, 12 (01):
  • [7] Identification of Suitable Reference Genes for Gene Expression Normalization in qRT-PCR Analysis in Watermelon
    Kong, Qiusheng
    Yuan, Jingxian
    Gao, Lingyun
    Zhao, Shuang
    Jiang, Wei
    Huang, Yuan
    Bie, Zhilong
    PLOS ONE, 2014, 9 (02):
  • [8] Selection and Verification of Appropriate Reference Genes for Expression Normalization in Cryptomeria fortunei under Abiotic Stress and Hormone Treatments
    Zhang, Yingting
    Zhu, Lijuan
    Xue, Jinyu
    Yang, Junjie
    Hu, Hailiang
    Cui, Jiebing
    Xu, Jin
    GENES, 2021, 12 (06)
  • [9] Selection of reference genes for normalization of qRT-PCR analysis of differentially expressed genes in soybean exposed to cadmium
    Wang, Yi
    Yu, Kangfu
    Poysa, Vaino
    Shi, Chun
    Zhou, Yonghong
    MOLECULAR BIOLOGY REPORTS, 2012, 39 (02) : 1585 - 1594
  • [10] Selection of reference genes for normalization of qRT-PCR analysis of differentially expressed genes in soybean exposed to cadmium
    Yi Wang
    Kangfu Yu
    Vaino Poysa
    Chun Shi
    Yonghong Zhou
    Molecular Biology Reports, 2012, 39 : 1585 - 1594