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Cannabis sativa: Advances in Biology and Cultivation—2nd Edition

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A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Physiology and Metabolism".

Deadline for manuscript submissions: closed (20 March 2025) | Viewed by 16492

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Dr. Youbin Zheng

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School of Environmental Sciences, University of Guelph, Guelph, ON, Canada
Interests: cannabis cultivation; controlled environment agriculture; lighting; rootzone management
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Dear Colleagues,

The rapidly growing trend in legalization of Cannabis sativa (cannabis) cultivation and utilization for medical and recreational purposes in regions across the planet has increased the intensity of scientific research in cannabis biology and cultivation technologies. These research activities are essential to the cannabis cultivation industry for efficiently producing cannabis with high yield and consistent quality. This Special Issue of Plants provides a platform for researchers to publish their research results on cannabis biology and cultivation and, at the same time, provides easy access for all stakeholders for information exchange. This Special Issue encourages authors to submit well-designed original research papers in all aspects of cannabis biology and cultivation, such as cannabis genetics, breeding, morphogenesis, physiology and biochemistry, tissue culture and propagation, atmosphere and rootzone management, lighting, pathogen and insect pest control, plant training, postharvest, etc. Research regarding cannabis grown in all types of cultivation environments (e.g., outdoors, greenhouse, indoors) is welcome; however, only papers involving drug-type cannabis (including hemp cultivated for medicinal or recreational purposes) will be included in this Issue.

Dr. Youbin Zheng
Guest Editor

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Keywords

biocontrol
biostimulant
breeding
Cannabis sativa
cannabis
cannabinoids
environmental control
fertigation
flavonoids
genetics
growing media
insect pest
lighting
light spectrum
light intensity
morphogenesis
morphology
tissue culture
pathogens
plant nutrition
photoperiod
plant training
postharvest
rootzone management
terpenes
terpenoids
VPD

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Related Special Issue

Cannabis sativa: Advances in Biology and Cultivation in Plants (12 articles)

Published Papers (9 papers)

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Research

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22 pages, 2034 KiB

Open AccessArticle

Subcanopy and Inter-Canopy Supplemental Light Enhances and Standardizes Yields in Medicinal Cannabis (Cannabis sativa L.)

by José Garrido, Carolina Corral, María Teresa García-Valverde, Jesús Hidalgo-García, Carlos Ferreiro-Vera and Juan José Martínez-Quesada

Plants 2025, 14(10), 1469; https://doi.org/10.3390/plants14101469 - 14 May 2025

Viewed by 291

Abstract

Light supplementation within the canopy is an effective method to improve light distribution throughout the whole plant, ensuring the inner canopies receive adequate light exposure to maximize overall growth. This approach is gaining interest among cannabis growers looking for more efficient lighting strategies to enhance their valuable production for medicinal purposes. We compared the traditional top lighting (TL) approach with two light supplementation methods: subcanopy lighting (SCL), which adds extra light to the inner canopies from below, and inter-canopy lighting (ICL), providing dedicated light at the basal and middle levels. Both SCL and ICL resulted in a more uniform light distribution throughout the plants and increased the yields of inflorescences, cannabinoids, and terpenes. The ICL treatment achieved the highest yield increases, showing a 29.95% increase in dry inflorescence yield, a 24.4% higher accumulation of THC, and a 12.5% increase in total terpene concentration. Notably, both SCL and ICL reduced the coefficients of variation, yielding more standardized products by decreasing the variability of the dry inflorescences yield, which also had more consistent chemical profiles, with reductions in variability for both THC and total terpene yields of over 50%. Although using more energy for lighting, SCL was more power-efficient for inflorescence and cannabinoid yields, while ICL was more efficient in achieving yield enhancements. In conclusion, adding supplemental light to the inner canopies enhances the profitability of medical cannabis cultivation, resulting in higher yields, improved energy efficiency, and more standardized products for research and medical purposes. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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18 pages, 1854 KiB

Open AccessArticle

Water Stress Effects on Biomass Allocation and Secondary Metabolism in CBD-Dominant Cannabis sativa L.

by Maddalena Cappello Fusaro, Irene Lucchetta and Stefano Bona

Plants 2025, 14(8), 1267; https://doi.org/10.3390/plants14081267 - 21 Apr 2025

Viewed by 523

Abstract

Water availability is a key factor affecting both morphological development and secondary metabolite production in Cannabis sativa L. This study evaluated the effects of water stress applied during the vegetative and flowering stages on plant performance, cannabinoid concentration, and terpene composition in two Chemotype III (cannabidiol-dominant) varieties. Plants were subjected to moderate and severe water stress, and responses were assessed through biomass measurements, GC-MS analyses, and multivariate statistics. Water stress significantly influenced biomass allocation, with increased dry biomass but reduced harvest index, particularly under flowering-stage stress. Cannabidiol (CBD) content declined with increasing stress, while tetrahydrocannabinol (THC) levels increased under vegetative stress, indicating a stress-induced shift in cannabinoid biosynthesis. Cannabinol (CBN) levels also increased, suggesting enhanced THC degradation. Terpene composition was predominantly genotype-driven. PCA-MANOVA showed significant effects of variety, stress level, and their interaction, yet only minor volatiles were modulated by stress, while the most abundant terpenes remained stable across treatments, preserving the varietal aroma profile. These results underline the importance of genetic background and irrigation timing in determining cannabis yield and quality. Optimized water management is essential to ensure phytochemical consistency and sustainable production, especially in high-value medicinal and aromatic applications. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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49 pages, 14633 KiB

Open AccessArticle

Transmission, Spread, Longevity and Management of Hop Latent Viroid, a Widespread and Destructive Pathogen Affecting Cannabis (Cannabis sativa L.) Plants in North America

by Zamir K. Punja, Cameron Scott, Heather H. Tso, Jack Munz and Liam Buirs

Plants 2025, 14(5), 830; https://doi.org/10.3390/plants14050830 - 6 Mar 2025

Cited by 1 | Viewed by 2858

Abstract

Hop latent viroid (HLVd), a 256-nucleotide RNA strand with complementary base-pairing and internal stem loop structures, forms circular or rod-shaped molecules within diseased plants. RT-PCR/RT-qPCR was used to assess HLVd transmission, spread and longevity. The viroid was detected in asymptomatic stock plants and in rooted vegetative cuttings, as well as in recirculated nutrient solution sampled from propagation tables and nozzles. Plant-to-plant spread through root infection in hydroponic cultivation was demonstrated. The viroid survived for 7 days and 4 weeks, respectively, in crushed leaf extracts (sap) or dried leaves/roots at room temperature. Following stem inoculation with infectious sap, HLVd was detected in root tissues within 2–3 weeks and in the foliage within 4–6 weeks. Plants grown under a 12:12 h photoperiod to induce inflorescence development showed more rapid spread of HLVd compared to 24 h lighting. The viroid was subsequently detected in inflorescence tissues, in trichome glands, in dried cannabis flowers and in crude resinous oil extracts. Anthers and pollen from infected male plants and seeds from infected female plants contained HLVd, giving rise to up to 100% infected seedlings. Artificially inoculated tomato and tobacco plants supported viroid replication in roots and leaves. Infected cannabis leaf and root tissues treated with UV-C for 3–5 min or temperatures of 70–90 °C for 30 min contained amplifiable HLVd-RNA. Infectious plant extract treated with 5–10% bleach (0.825% NaOCl) or 1000 ppm hypochlorous acid yielded no RT-PCR bands, suggesting the RNA was degraded. Meristem tip culture from HLVd-infected plants yielded a high frequency of pathogen-free plants, depending on the genotype. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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25 pages, 6438 KiB

Open AccessArticle

An In Vitro Phytohormone Survey Reveals Concerted Regulation of the Cannabis Glandular Trichome Disc Cell Proteome

by Nicolas Dimopoulos, Qi Guo, Lei Liu, Matthew Nolan, Rekhamani Das, Lennard Garcia-de Heer, Jos C. Mieog, Bronwyn J. Barkla and Tobias Kretzschmar

Plants 2025, 14(5), 694; https://doi.org/10.3390/plants14050694 - 24 Feb 2025

Cited by 1 | Viewed by 684

Abstract

Cannabis (Cannabis sativa L.) flower glandular trichomes (GTs) are the main site of cannabinoid synthesis. Phytohormones, such as jasmonic acid (JA) and salicylic acid (SA), have been shown to increase cannabinoid content in cannabis flowers, but how this is regulated remains unknown. This study aimed to understand which biological processes in GT disc cells phytohormones control by using an in vitro assay. Live GT disc cells were isolated from a high-tetrahydrocannabinol cannabis cultivar and incubated on basal media plates supplemented with either kinetin (KIN), JA, SA, abscisic acid, ethephon, gibberellic acid, brassinolide, or sodium diethyldithiocarbamate. Quantitative proteomic analysis revealed that KIN, JA, and SA caused the greatest number of changes in the GT disc cell proteome. Surprisingly, none of the treatments concertedly increased cannabinoid content or the abundance of related biosynthetic proteins in the GT, suggesting that cannabinoid increases in previous in planta phytohormone studies are likely due to other processes, such as increased GT density. As well, KIN-, JA-, and SA-treated GTs had numerous differentially abundant proteins in common. Several were key proteins for leucoplast differentiation, cuticular wax and fatty acid metabolism, and primary metabolism regulation, denoting that cytokinin, JA, and SA signalling are likely important for coordinating cannabis GT differentiation and development. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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24 pages, 4685 KiB

Open AccessArticle

Flowering Synchronization Using Artificial Light Control for Crossbreeding Hemp (Cannabis sativa L.) with Varied Flowering Times

by Gergő Somody and Zoltán Molnár

Plants 2025, 14(4), 594; https://doi.org/10.3390/plants14040594 - 15 Feb 2025

Viewed by 622

Abstract

Hemp (Cannabis sativa L.), one of the earliest domesticated crops, has diverse applications in textiles, construction, nutrition, and medicine. Breeding advancements, including speed breeding, accelerate genetic improvements in crops by optimizing environmental conditions for reduced generation times. This study employed greenhouse and field experiments to develop a proprietary yellow-stemmed hemp germplasm with a unique stem trait. Initial crossbreeding between the late Eletta Campana (medium green stems) and the early Chamaeleon (yellow stems) demonstrated the recessive monogenic inheritance of the yellow-stem trait and fast and safe stabilization even in the case of parent varieties with different flowering times. Controlled flowering in the case of photoperiod-sensitive genotypes, manual pollination, and successive backcrossing stabilized the yellow-stem trait over six cycles, with 100% trait consistency achieved by the fifth cycle within just 12 months in total. Open-field trials validated greenhouse results, showing strong correlations between visual stem color assessments and visible atmospherically resistant index (VARI) obtained through remote sensing imagery. Cannabinoid analyses indicated significant reductions in tetrahydrocannabinol (THC) content while maintaining optimal cannabidiol (CBD) levels. Accumulated growing degree days (GDDs) optimized flowering and maturity, ensuring consistency in phenological traits. This research highlights the utility of speed breeding and chemical analysis to accelerate trait stabilization and improve industrial hemp’s agronomic potential for fiber and CBD production while adhering to regulatory THC limits. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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16 pages, 8749 KiB

Open AccessArticle

Characterization of Male Flower Induction by Silver Thiosulfate Foliar Spray in Female Cannabis at the Middle Reproductive Stage for Breeding

by Juyoung Kim, Dong-Gun Kim, Woon Ji Kim, Ye-Jin Lee, Seung Hyeon Lee, Jaihyunk Ryu, Jae Hoon Kim and Sang Hoon Kim

Plants 2024, 13(17), 2429; https://doi.org/10.3390/plants13172429 - 30 Aug 2024

Cited by 2 | Viewed by 2227

Abstract

Cannabis (Cannabis sativa) is a versatile crop belonging to the Cannabaceae family, and is dioecious, typically with separate male and female plants. The flowers of female plants, especially the trichomes, accumulate relatively higher contents of cannabinoids compared with those of male plants. For this reason, to obtain seeds that are genetically female, it is desirable to induce the development of male flowers on a female plant that produces genetically female haploid gametes. Silver thiosulfate (STS) is a highly effective chemical for male flower induction. We investigated male flower induction in three commercial cultivars of female cannabis (Spectrum303, SuperwomanS1, and CBGambit) regarding the treatment frequency, stage of application, and concentration of STS applied as a foliar spray. All three cultivars showed adequate induction of male flowers in response to 1.5 mM STS applied at the early reproductive stage. In particular, SuperwomanS1 was most highly responsive to induction of male flowers, even when treated with 0.3 mM STS at the early reproductive stage. Treatment with three applications of STS was more effective compared with a single application, but a single application of 1.5 mM STS at the early reproductive stage was sufficient for male flower induction. A single STS application during the middle stage of reproductive growth was inadequate for induction of male flowers. However, 6 weeks after three applications of STS, CBGambit exhibited approximately 54% male flower induction at 0.3 mM STS, Spectrum303 showed approximately 56% induction at 3 mM STS, and SuperwomanS1 yielded approximately 26% induction at 1.5 mM (expressed as percentage of total number of individuals with the induced male flowers). Pollen stainability tests using KI-I₂ solution and Alexander’s staining showed high pollen viability with over 65% at different single STS concentrations, indicating that pollen grains induced by STS have sufficient viability for the self-pollination. This study demonstrated that different cultivars of cannabis respond diversely to different STS concentrations and highlighted the potential benefits of three STS applications during the middle reproductive stage for cannabis breeding. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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14 pages, 869 KiB

Open AccessArticle

Somatic Mutation Accumulations in Micropropagated Cannabis Are Proportional to the Number of Subcultures

by Kristian Adamek, Andrew Maxwell Phineas Jones and Davoud Torkamaneh

Plants 2024, 13(14), 1910; https://doi.org/10.3390/plants13141910 - 11 Jul 2024

Cited by 1 | Viewed by 2276

Abstract

Advancements in micropropagation techniques have made it easier to produce large numbers of cannabis clones, but these methods may also introduce genetic instability over successive generations. This instability often manifests as somaclonal variation, characterized by the progressive accumulation of genetic mutations or epigenetic alterations with each subculture. In this study, we examined how mutations accumulate in cannabis clones subjected to 6–11 subcultures. Using genotyping-by-sequencing, we identified 9405 polymorphic variants across 70 clones. The analysis revealed a correlation between the number of subcultures and the frequency of these mutations, revealing that genetic changes accumulate over successive subcultures despite clones sharing the same chronological age. Furthermore, we evaluated the functional impacts of accumulated mutations, with particular attention to implications on gene function and overall plant health. While rare, 14 high-impact variants were identified in genes that are important for plant development. Notably, six variants were also found in genes related to cannabinoid and terpene synthesis pathways, potentially affecting the plant’s biochemical composition. These findings highlight the need for genetic assessments in micropropagation protocols, impacting plant breeding and conservation. Understanding genetic variations in clonally propagated plants optimizes practices for stability. Crucial for cannabis and horticultural plants, it emphasizes techniques to prevent genetic decay and ensure viability. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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Review

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39 pages, 17649 KiB

Open AccessReview

Endophytes in Cannabis sativa: Identifying and Characterizing Microbes with Beneficial and Detrimental Effects on Plant Health

by Liam Buirs and Zamir K. Punja

Plants 2025, 14(8), 1247; https://doi.org/10.3390/plants14081247 - 19 Apr 2025

Viewed by 646

Abstract

The roles of endophytes in Cannabis sativa (cannabis, hemp) remain poorly explored. While in vitro studies suggest that there can be several benefits, such as plant growth promotion and protection against pathogens, more in planta studies are needed. This review summarizes the bacterial and fungal endophytes previously reported in tissues of C. sativa and discusses the factors influencing their presence, as well as their potential beneficial and detrimental effects. Using genome sequencing and culture-based approaches, we describe the microbial diversity in hydroponically cultivated cannabis plants at several developmental stages. These include mother plants, cuttings, vegetative and flowering plants, and tissue-cultured plantlets. Microbes that were present include fungal, yeast, and bacterial endophytes found in roots, stems, leaves, inflorescences, and seeds. These may have originated from the growing substrate or be transmitted through vegetative propagation. Notable endophytes included Rhizophagus irregularis (a mycorrhizal fungus), Penicillium chrysogenum (an antibiotic producer), and various endophytic yeast species not previously described in C. sativa. Endophytes representing potential plant pathogens, such as Fusarium oxysporum, are also present within cannabis tissues, which can negatively impact plant health. Using scanning electron microscopy, we observed that fungal propagules are present within pith parenchyma cells and xylem vessel elements in stem tissues, illustrating for the first time the in situ localization and distribution of endophytes in cannabis vascular tissues. The mechanism of spread through xylem vessels likely contributes to the spread of endophytes within cannabis and hemp plants. Further research is required to validate the roles of endophytes in cannabis and hemp plants grown under commercial production conditions. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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30 pages, 2467 KiB

Open AccessReview

Illuminating Cannabis sativa L.: The Power of Light in Enhancing C. sativa Growth and Secondary Metabolite Production

by S.M. Ahsan, Md. Injamum-Ul-Hoque, Shifa Shaffique, Akhtar Ayoobi, Md Atikur Rahman, Md. Mezanur Rahman and Hyong Woo Choi

Plants 2024, 13(19), 2774; https://doi.org/10.3390/plants13192774 - 3 Oct 2024

Cited by 6 | Viewed by 4907

Abstract

Light is crucial for higher plants, driving photosynthesis and serving as a powerful sensory signal that profoundly modulates growth, development, physiological functions, hormone activation, and biochemical pathways. Various light parameters—quality, intensity, composition, and photoperiod—exert a tremendous influence on plant growth and development, particularly in industrial hemp (Cannabis sativa L.). C. sativa, a crop of historical significance and unparalleled versatility, holds immense value in the food, fiber, and medicinal industries. The cultivation of medicinal cannabis is burgeoning in controlled environments due to evolving healthcare regulations. Optimal light conditions significantly enhance both yield and harvest quality, notably increasing the density of apical inflorescences and the ratio of inflorescence to total aboveground biomass. C. sativa metabolites, especially phenolic and terpene compounds and Phytocannabinoids like CBD (cannabidiol), THC (tetrahydrocannabinol), and CBG (cannabigerol), possess immense medicinal value. Secondary metabolites in C. sativa predominantly accumulate in the trichomes of female flowers and surrounding sugar leaves, underscoring the critical need to boost inflorescence weight and metabolite concentrations while ensuring product consistency. Different light parameters distinctly impact C. sativa’s metabolic profile, providing a robust foundation for understanding the optimal conditions for synthesizing specific secondary metabolites. While the effects of light measurement on various crops are well-established, scientific evidence specifically relating to light quality effects on C. sativa morphology and secondary metabolite accumulation remains scarce. In this review, we critically summarized how different light properties can alter cannabis growth (vegetative and reproductive), physiology and metabolism. Furthermore, the mechanisms by which specific wavelengths influence growth, development, and secondary metabolite biosynthesis in C. sativa are not fully elucidated, which could be a prospective task for future researchers. Our review paves the way for a profound understanding of light’s influence on C. sativa growth and advancements in greenhouse settings to maximize metabolite production for commercial use. Full article

(This article belongs to the Special Issue Cannabis sativa: Advances in Biology and Cultivation—2nd Edition)

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