Cargando…

In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)

Industrial hemp is a diploid (2n = 20), dioecious plant, and an essential source of various phytochemical productions. More than 540 phytochemicals have been described, some of which proved helpful in the remedial treatment of human diseases. Therefore, further study of hemp phytochemicals in medici...

Descripción completa

Detalles Bibliográficos
Autores principales: Deguchi, Michihito, Dhir, Seema, Potlakayala, Shobha, Dhir, Sarwan, Curtis, Wayne R., Rudrabhatla, Sairam
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9356322/
https://www.ncbi.nlm.nih.gov/pubmed/35941940
http://dx.doi.org/10.3389/fpls.2022.921970
_version_ 1784763491878436864
author Deguchi, Michihito
Dhir, Seema
Potlakayala, Shobha
Dhir, Sarwan
Curtis, Wayne R.
Rudrabhatla, Sairam
author_facet Deguchi, Michihito
Dhir, Seema
Potlakayala, Shobha
Dhir, Sarwan
Curtis, Wayne R.
Rudrabhatla, Sairam
author_sort Deguchi, Michihito
collection PubMed
description Industrial hemp is a diploid (2n = 20), dioecious plant, and an essential source of various phytochemical productions. More than 540 phytochemicals have been described, some of which proved helpful in the remedial treatment of human diseases. Therefore, further study of hemp phytochemicals in medicine is highly anticipated. Previously, we developed the vacuum agroinfiltration method, which allows the transient gene expression in hemp tissues including female flowers, where cannabinoids are produced and accumulated. In this study, we attempted to alter the composition of total CBD and THC. The RT-PCR and sanger sequence identified eleven copies of the CBDAS gene, two copies of the THCAS gene, and one CBCAS gene. Binary vectors were constructed to overexpress the CBDAS gene and silence the THCAS gene via RNA interference. The Transcript level of the CBDAS gene was increased by more than 10 times than the plants used as a control, which led to a 54% higher total CBD content. The silencing of the THCAS gene led to downregulation of the THCAS gene, with an 80% reduction in transcript levels, and total THC content was reduced to 43% compared with mock plant. These results suggest that hemp vacuum infiltration is highly effective for metabolic engineering of cannabinoids in hemp.
format Online
Article
Text
id pubmed-9356322
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-93563222022-08-07 In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.) Deguchi, Michihito Dhir, Seema Potlakayala, Shobha Dhir, Sarwan Curtis, Wayne R. Rudrabhatla, Sairam Front Plant Sci Plant Science Industrial hemp is a diploid (2n = 20), dioecious plant, and an essential source of various phytochemical productions. More than 540 phytochemicals have been described, some of which proved helpful in the remedial treatment of human diseases. Therefore, further study of hemp phytochemicals in medicine is highly anticipated. Previously, we developed the vacuum agroinfiltration method, which allows the transient gene expression in hemp tissues including female flowers, where cannabinoids are produced and accumulated. In this study, we attempted to alter the composition of total CBD and THC. The RT-PCR and sanger sequence identified eleven copies of the CBDAS gene, two copies of the THCAS gene, and one CBCAS gene. Binary vectors were constructed to overexpress the CBDAS gene and silence the THCAS gene via RNA interference. The Transcript level of the CBDAS gene was increased by more than 10 times than the plants used as a control, which led to a 54% higher total CBD content. The silencing of the THCAS gene led to downregulation of the THCAS gene, with an 80% reduction in transcript levels, and total THC content was reduced to 43% compared with mock plant. These results suggest that hemp vacuum infiltration is highly effective for metabolic engineering of cannabinoids in hemp. Frontiers Media S.A. 2022-07-21 /pmc/articles/PMC9356322/ /pubmed/35941940 http://dx.doi.org/10.3389/fpls.2022.921970 Text en Copyright © 2022 Deguchi, Dhir, Potlakayala, Dhir, Curtis and Rudrabhatla. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Deguchi, Michihito
Dhir, Seema
Potlakayala, Shobha
Dhir, Sarwan
Curtis, Wayne R.
Rudrabhatla, Sairam
In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)
title In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)
title_full In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)
title_fullStr In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)
title_full_unstemmed In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)
title_short In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)
title_sort in planta female flower agroinfiltration alters the cannabinoid composition in industrial hemp (cannabis sativa l.)
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9356322/
https://www.ncbi.nlm.nih.gov/pubmed/35941940
http://dx.doi.org/10.3389/fpls.2022.921970
work_keys_str_mv AT deguchimichihito inplantafemalefloweragroinfiltrationaltersthecannabinoidcompositioninindustrialhempcannabissatival
AT dhirseema inplantafemalefloweragroinfiltrationaltersthecannabinoidcompositioninindustrialhempcannabissatival
AT potlakayalashobha inplantafemalefloweragroinfiltrationaltersthecannabinoidcompositioninindustrialhempcannabissatival
AT dhirsarwan inplantafemalefloweragroinfiltrationaltersthecannabinoidcompositioninindustrialhempcannabissatival
AT curtiswayner inplantafemalefloweragroinfiltrationaltersthecannabinoidcompositioninindustrialhempcannabissatival
AT rudrabhatlasairam inplantafemalefloweragroinfiltrationaltersthecannabinoidcompositioninindustrialhempcannabissatival