Establishment and optimization of a hemp (Cannabis sativa L.) agroinfiltration system for gene expression and silencing studies
| Autor: | Takumasa Kondo, Zhanyuan J. Zhang, Zachary Spuhler, Michihito Deguchi, Sairam Rudrabhatla, Shobha Potlakayala, Daniel Bogush, Hannah Weeden |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Phytoene desaturase Agroinfiltration Plant molecular biology lcsh:Medicine Agrobacterium Flowers Poloxamer Biology 01 natural sciences Article Small hairpin RNA 03 medical and health sciences RNA interference Gene Expression Regulation Plant Botany Gene expression Gene silencing RNA Small Interfering lcsh:Science Cannabis Plant Proteins Regulation of gene expression Multidisciplinary beta-Glucosidase lcsh:R Ascorbic acid Plant Leaves 030104 developmental biology lcsh:Q Plant biotechnology RNA Interference Oxidoreductases 010606 plant biology & botany Plasmids |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-11 (2020) |
| ISSN: | 2045-2322 |
| Popis: | Industrial hemp (Cannabis sativa L.) is a high-yielding annual crop primarily grown for fiber, seeds, and oil. Due to the phytochemical composition of hemp, there has been an increased interest in the market for nutraceuticals and dietary supplements for human health. Recent omics analysis has led to the elucidation of hemp candidate genes involved in the syntheses of specialized metabolites. However, a detailed study of these genes has not been undertaken due to the lack of a stable transformation system. We report for the first time an agroinfiltration system in hemp utilizing vacuum infiltration, which is an alternative method to stable transformation. A combination of 0.015% Silwett L-77, 5 mM ascorbic acid, and thirty second sonication followed by a 10-minute vacuum treatment resulted in the highest β-glucuronidase expression in the leaf, male and female flowers, stem, and root tissues. The phytoene desaturase gene was silenced with a transient hairpin RNA expression, resulting in an albino phenotype in the leaves and the male and female flowers. This agroinfiltration system would be useful for overexpression and silencing studies of target genes to regulate the yield of specialized metabolites in hemp. |
| Databáze: | OpenAIRE |
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