Cargando…
Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp
There has been significant interest in researching the pharmaceutical applications of Industrial hemp since its legalization three years ago. The crop is mostly dioecious and known for its production of phytocannabinoids, flavonoids, and terpenes. Although many scientific reports have showed gene ex...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8687539/ https://www.ncbi.nlm.nih.gov/pubmed/34928958 http://dx.doi.org/10.1371/journal.pone.0260660 |
_version_ | 1784618192517201920 |
---|---|
author | Deguchi, Michihito Potlakayala, Shobha Spuhler, Zachary George, Hannah Sheri, Vijay Agili, Ruba Patel, Aayushi Rudrabhatla, Sairam |
author_facet | Deguchi, Michihito Potlakayala, Shobha Spuhler, Zachary George, Hannah Sheri, Vijay Agili, Ruba Patel, Aayushi Rudrabhatla, Sairam |
author_sort | Deguchi, Michihito |
collection | PubMed |
description | There has been significant interest in researching the pharmaceutical applications of Industrial hemp since its legalization three years ago. The crop is mostly dioecious and known for its production of phytocannabinoids, flavonoids, and terpenes. Although many scientific reports have showed gene expression analysis of hemp through OMICs approaches, unreliable reference genes for normalization of qRT-PCR data make it difficult to validate the OMICs data. Four software packages: geNorm, NormFinder, BestKeeper, and RefFinder were used to evaluate the differential gene expression patterns of 13 candidate reference genes under osmotic, heavy metal, hormonal, and UV stresses. EF-1α ranked as the most stable reference gene across all stresses, TUB was the most stable under osmotic stress, and TATA was the most stable under both heavy metal stress and hormonal stimuli. The expression patterns of two cannabinoid pathway genes, AAE1 and CBDAS, were used to validate the reliability of the selected reference genes. This work provides useful information for gene expression characterization in hemp and future research in the synthesis, transport, and accumulation of secondary metabolites. |
format | Online Article Text |
id | pubmed-8687539 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-86875392021-12-21 Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp Deguchi, Michihito Potlakayala, Shobha Spuhler, Zachary George, Hannah Sheri, Vijay Agili, Ruba Patel, Aayushi Rudrabhatla, Sairam PLoS One Research Article There has been significant interest in researching the pharmaceutical applications of Industrial hemp since its legalization three years ago. The crop is mostly dioecious and known for its production of phytocannabinoids, flavonoids, and terpenes. Although many scientific reports have showed gene expression analysis of hemp through OMICs approaches, unreliable reference genes for normalization of qRT-PCR data make it difficult to validate the OMICs data. Four software packages: geNorm, NormFinder, BestKeeper, and RefFinder were used to evaluate the differential gene expression patterns of 13 candidate reference genes under osmotic, heavy metal, hormonal, and UV stresses. EF-1α ranked as the most stable reference gene across all stresses, TUB was the most stable under osmotic stress, and TATA was the most stable under both heavy metal stress and hormonal stimuli. The expression patterns of two cannabinoid pathway genes, AAE1 and CBDAS, were used to validate the reliability of the selected reference genes. This work provides useful information for gene expression characterization in hemp and future research in the synthesis, transport, and accumulation of secondary metabolites. Public Library of Science 2021-12-20 /pmc/articles/PMC8687539/ /pubmed/34928958 http://dx.doi.org/10.1371/journal.pone.0260660 Text en © 2021 Deguchi et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Deguchi, Michihito Potlakayala, Shobha Spuhler, Zachary George, Hannah Sheri, Vijay Agili, Ruba Patel, Aayushi Rudrabhatla, Sairam Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp |
title | Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp |
title_full | Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp |
title_fullStr | Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp |
title_full_unstemmed | Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp |
title_short | Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp |
title_sort | selection and validation of reference genes for normalization of qrt-pcr data to study the cannabinoid pathway genes in industrial hemp |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8687539/ https://www.ncbi.nlm.nih.gov/pubmed/34928958 http://dx.doi.org/10.1371/journal.pone.0260660 |
work_keys_str_mv | AT deguchimichihito selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT potlakayalashobha selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT spuhlerzachary selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT georgehannah selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT sherivijay selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT agiliruba selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT patelaayushi selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp AT rudrabhatlasairam selectionandvalidationofreferencegenesfornormalizationofqrtpcrdatatostudythecannabinoidpathwaygenesinindustrialhemp |