Abstract
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.
Highlights
Industrial hemp (Cannabis sativa L.) has a rich history in the civilization of humans because it can provide both phytochemicals and lignocellulosic biomass
Transcript abundances of 13 candidate reference genes were assessed by quantitative real-time PCR (qRT-PCR) for each gene, tested in triplicates across all 11 treatments and a control, which was 36 biological samples (Fig 2)
Industrial hemp is from the plant species Cannabis sativa and has gained importance as a medicinal crop because of its potential to produce secondary metabolites such as cannabinoids, terpenes, and phenolic compounds [33]
Summary
Industrial hemp (Cannabis sativa L.) has a rich history in the civilization of humans because it can provide both phytochemicals and lignocellulosic biomass. This crop originated in Eurasia and is useful all over the world, largely as a fiber crop [1]. Following the emergence of more economically helpful fiber crops, the demand for hemp reduced, and the purpose shifted to usage as a food additive. Hemp seed contains essential fatty acids and proteins and gamma-linolenic acid, which has many health benefits [2]. Hemp seeds and oils are used to produce nutritional supplements and cosmetics. Hemp produces a diverse array of phytocannabinoids, terpenes, and phenolic compounds
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