Dataset Information

Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata

ABSTRACT: The desert pioneer plant Stipagrostis pennata plays an important role in sand fixation, wind prevention, and desert ecosystem recovery. An absence of reference genes greatly limits investigations into the regulatory mechanism by which S. pennata adapts to adverse desert environments at the molecular and genetic levels. In this study, eight candidate reference genes were identified from rhizosheath development transcriptome data from S. pennata, and their expression stability in the rhizosheaths at different development stages, in a variety of plant tissues, and under drought stress was evaluated using four procedures, including geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that GAPDH and elF were the most stable reference genes under drought stress and in rhizosheath development, and ARP6 and ALDH were relatively stable in all plant tissues. In addition, elF was the most suitable reference gene for all treatments. Analysis of the consistency between the reverse transcription-quantitative PCR (RT-qPCR) and RNA sequencing data showed that the identified elF and GAPDH reference genes were stable during rhizosheath development. These results provide reliable reference genes for assuring the accuracy of RT-qPCR and offer a foundation for further investigations into the genetic responses of S. pennata to abiotic stress.

SUBMITTER: Li R

PROVIDER: S-EPMC8571334 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:Selection of reference genes has become an integral step in any real time quantitative PCR (RT-qPCR) based expression studies. The importance of this study stems from the fact that riverine buffaloes are major dairy species of Indian sub-continent and the information generated here will be of great interest to the investigators engaged in functional genomic studies of this important livestock species. In this study, an effort was made to evaluate a panel of 10 candidate reference genes (glyceraldehyde 3-phosphate dehydrogenase (GAPDH), beta- actin (ACTB), ubiquitously expressed transcript (UXT), ribosomal protein S15 (RPS15), ribosomal protein L-4 (RPL4), ribosomal protein S9 (RPS9), ribosomal protein S23 (RPS23), hydroxymethylbilane synthase (HMBS), ?2 Microglobulin (?2M) and eukaryotic translation elongation factor 1 alpha 1 (EEF1A1) across 12 tissues (mammary gland, kidney, spleen, liver, heart, intestine, ovary, lung, muscle, brain, subcutaneous fat and testis) of riverine buffaloes. In addition to overall analysis, tissue wise evaluation of expression stability of individual RG was also performed. Three different algorithms provided in geNorm, NormFinder and BestKeeper softwares were used to evaluate the stability of 10 potential reference genes from different functional classes. The M-value given by geNorm ranged from 0.9797 (RPS9 and UXT) to 1.7362 (RPS15). From the most stable to the least stable, genes were ranked as: UXT/RPS9> RPL4> RPS23> EEF1A1> ACTB> HMBS> GAPDH> B2M> RPS15. While NormFinder analysis ranked the genes as: UXT> RPS23> RPL4> RPS9> EEF1A1> HMBS> ACTB> ?2M> GAPDH> RPS15. Based on the crossing point SD value and range of fold change expression, BestKeeper analysis ranked the genes as: RPS9> RPS23/UXT> RPL4> GAPDH> EEF1A1> ACTB> HMBS> ?2M> RPS15. Overall the study has identified RPS23, RPS9, RPL4 and UXT genes to be the most stable and appropriate RGs that could be utilized for normalization of transcriptional data in various tissues of buffaloes. This manuscript thus provide useful information on panel of reference genes that could be helpful for researchers conducting functional genomic studies in riverine buffaloes.

Dataset Information

Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets