Cannabis sativa L. photoautotrophic micropropagation: a powerful tool for industrial scale in vitro propagation

Zarei, Adel; Behdarvandi, Behrang; Dinani, Elham Tavakouli; Maccarone, Jennifer

doi:10.1007/s11627-021-10167-3

Cited by 13 publications

(15 citation statements)

References 32 publications

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“…This method, however, did not use rockwool blocks as a substrate. Moreover, the results of this study converged with those of Zarei et al [ 70 ], who applied a similar in vitro propagation method using cuttings in rockwool blocks, using only the cuttings of similar size (ca 3 cm). They reported that the cutting length influenced the rooting percentage and the mean root length, i.e., the longer cuttings exhibited better performance.…”

Section: Discussionsupporting

confidence: 87%

An Alternative In Vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) Presenting Efficient Rooting, for Commercial Production

Ioannidis¹,

Tomprou²,

Mitsis³

2022

Plants

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An alternative in vitro propagation protocol for medical Cannabis sativa L. cultivars for pharmaceutical industrial use was established. The aim of the protocol was to reduce the culture time, offering healthy and aseptic propagating material, while making the whole process more economic for industrial use. The propagation procedure was performed using plastic autoclavable vented and non-vented vessels, containing porous rooting fine-milled sphagnum peat moss-based sponges, impregnated in ½ Murashige and Skoog liquid growth medium, supplemented with indole-3-butyric acid (IBA) at various concentrations (0, 2.46, 4.92, and 9.84 µM) or by dipping nodal cuttings into 15 mM IBA aqueous solution. The highest average root numbers per cutting, 9.47 and 7.79 for high cannabidiol (H_CBD) and high cannabigerol (H_CBG) varieties, respectively, were achieved by dipping the cuttings into IBA aqueous solution for 4 min and then placing them in non-vented vessels. The maximum average root length in H_CBD (1.54 cm) and H_CBG (0.88 cm) was ascertained using 2.46 μM filter sterilized IBA in non-vented vessels. Filter-sterilized IBA at concentrations of 2.46 μM in vented and 4.92 μM in non-vented vessels displayed the maximum average rooting percentages in H_CBD (100%) and H_CBG (95.83%), respectively. In both varieties, maximum growth was obtained in non-vented vessels, when the medium was supplemented with 4.92 μM filter-sterilized IBA. Significant interactions between variety and vessel type and variety and IBA treatments were observed in relation to rooting traits. Approximately 95% of plantlets were successfully established and acclimatized in field. This culture system can be used not only for propagating plant material at an industrial scale but also to enhance the preservation and conservation of Cannabis genetic material.

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Section: Discussionsupporting

confidence: 87%

An Alternative In Vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) Presenting Efficient Rooting, for Commercial Production

Ioannidis¹,

Tomprou²,

Mitsis³

2022

Plants

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“…Another important advantage of plant culture is the possibility of increasing the scale of the entire culture process, and a number of new culture systems for plant in vitro cultures have been developed [ 10 , 70 , 71 , 72 , 73 , 74 ]. Combining these culture strategies with the use of special procedures, such as elicitation with biotic or abiotic elicitors, allows for the large-scale production of the target biologically active compound [ 75 ].…”

Section: Plant Tissue and Organ Cultures In Bioreactorsmentioning

confidence: 99%

Genetic Manipulation and Bioreactor Culture of Plants as a Tool for Industry and Its Applications

et al. 2022

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In recent years, there has been a considerable increase in interest in the use of transgenic plants as sources of valuable secondary metabolites or recombinant proteins. This has been facilitated by the advent of genetic engineering technology with the possibility for direct modification of the expression of genes related to the biosynthesis of biologically active compounds. A wide range of research projects have yielded a number of efficient plant systems that produce specific secondary metabolites or recombinant proteins. Furthermore, the use of bioreactors allows production to be increased to industrial scales, which can quickly and cheaply deliver large amounts of material in a short time. The resulting plant production systems can function as small factories, and many of them that are targeted at a specific operation have been patented. This review paper summarizes the key research in the last ten years regarding the use of transgenic plants as small, green biofactories for the bioreactor-based production of secondary metabolites and recombinant proteins; it simultaneously examines the production of metabolites and recombinant proteins on an industrial scale and presents the current state of available patents in the field.

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“…Nowadays, the influence of physical conditions that may control organogenetic responses in tissue cultured plants of Cannabis, especially those that are associated with light conditions, is becoming more apparent. Ventilation of the culture vessels for micropropagation in Cannabis and other plant genera is thought to be important in producing true-to-type plantlets that are can be readily acclimatised [67,78]. The benefits of investigating physical factors are many.…”

Section: Indirect Organogenesis Of Cannabis Sativamentioning

confidence: 99%

“…Many studies that have investigated micropropagation and other biotechnologies of Cannabis are not easy to adopt for large-scale industrial application as they require lengthy periods for plantlets production and are plagued by high costs whilst being inefficient as plantlet regenerants are often showing symptoms of plantlet hyperhydration, poor rooting and acclimatisation frequencies [78]. For these reasons, it thus becomes important to further develop protocols that are more amenable to a commercial setup.…”

Section: Commercial Micropropagation Of Cannabis Sativamentioning

confidence: 99%

“…For these reasons, it thus becomes important to further develop protocols that are more amenable to a commercial setup. The recent study by Zarei et al [78]. provides a route of micropropagation that could easily be adopted with the objective of generating plants en masse, in a commercial tissue culture setting, as rooting capabilities using rockwool improved significantly.…”

Section: Commercial Micropropagation Of Cannabis Sativamentioning

confidence: 99%

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Cannabis sativa: From Therapeutic Uses to Micropropagation and Beyond

Adams

Malatsi

Makunga

2021

Plants

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The development of a protocol for the large-scale production of Cannabis and its variants with little to no somaclonal variation or disease for pharmaceutical and for other industrial use has been an emerging area of research. A limited number of protocols have been developed around the world, obtained through a detailed literature search using web-based database searches, e.g., Scopus, Web of Science (WoS) and Google Scholar. This article reviews the advances made in relation to Cannabis tissue culture and micropropagation, such as explant choice and decontamination of explants, direct and indirect organogenesis, rooting, acclimatisation and a few aspects of genetic engineering. Since Cannabis micropropagation systems are fairly new fields, combinations of plant growth regulator experiments are needed to gain insight into the development of direct and indirect organogenesis protocols that are able to undergo the acclimation stage and maintain healthy plants desirable to the Cannabis industry. A post-culture analysis of Cannabis phytochemistry after the acclimatisation stage is lacking in a majority of the reviewed studies, and for in vitro propagation protocols to be accepted by the pharmaceutical industries, phytochemical and possibly pharmacological research need to be undertaken in order to ascertain the integrity of the generated plant material. It is rather difficult to obtain industrially acceptable micropropagation regimes as recalcitrance to the regeneration of in vitro cultured plants remains a major concern and this impedes progress in the application of genetic modification technologies and gene editing tools to be used routinely for the improvement of Cannabis genotypes that are used in various industries globally. In the future, with more reliable plant tissue culture-based propagation that generates true-to-type plants that have known genetic and metabolomic integrity, the use of genetic engineering systems including “omics” technologies such as next-generation sequencing and fast-evolving gene editing tools could be implemented to speed up the identification of novel genes and mechanisms involved in the biosynthesis of Cannabis phytochemicals for large-scale production.

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Cannabis sativa L. photoautotrophic micropropagation: a powerful tool for industrial scale in vitro propagation

Cited by 13 publications

References 32 publications

An Alternative In Vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) Presenting Efficient Rooting, for Commercial Production

An Alternative In Vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) Presenting Efficient Rooting, for Commercial Production

Genetic Manipulation and Bioreactor Culture of Plants as a Tool for Industry and Its Applications

Cannabis sativa: From Therapeutic Uses to Micropropagation and Beyond

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