Hermaphroditism in Marijuana (Cannabis sativa L.) Inflorescences - Impact on Floral Morphology, Seed Formation, Progeny Sex Ratios, and Genetic Variation.
TL;DR: The morphological features of anthers, pollen production and germination in hermaphroditic flowers and in staminate inflorescences on male plants were compared and Copia-like retrotransposons within the C. sativa genome which may be associated with the expression of male or female phenotype were revealed.
Abstract: Cannabis sativa L (hemp, marijuana) produces male and female inflorescences on different plants (dioecious) and therefore the plants are obligatory out-crossers In commercial production, marijuana plants are all genetically female; male plants are destroyed as seed formation reduces flower quality Spontaneously occurring hermaphroditic inflorescences, in which pistillate flowers are accompanied by formation of anthers, leads to undesired seed formation; the mechanism for this is poorly understood We studied hermaphroditism in several marijuana strains with three objectives: (i) to compare the morphological features of this unique phenotype with normal male flowers; (ii) to assess pollen and seed viability from hermaphroditic flowers; and (iii) to assess the effect of hermaphroditism on progeny male:female (sex) ratios and on genetic variation using molecular methods The morphological features of anthers, pollen production and germination in hermaphroditic flowers and in staminate inflorescences on male plants were compared using light and scanning electron microscopy Seeds produced on hermaphroditic plants and seeds derived from cross-fertilization were germinated and seedlings were compared for gender ratios using a PCR-based assay as well as for the extent of genetic variation using six ISSR primers Nei's index of gene diversity and Shannon's Information index were compared for these two populations The morphology of anthers and pollen formation in hermaphroditic inflorescences was similar to that in staminate flowers Seedlings from hermaphroditic seeds, and anther tissues, showed a female genetic composition while seedlings derived from cross-fertilized seeds showed a 1:1 male:female sex expression ratio Uniquely, hermaphroditic inflorescences produced seeds which gave rise only to genetically female plants In PCR assays, a 540 bp size fragment was present in male and female plants, while a 390 bp band was uniquely associated with male plants Sequence analysis of these fragments revealed the presence of Copia-like retrotransposons within the C sativa genome which may be associated with the expression of male or female phenotype In ISSR analysis, the percentage of polymorphic loci ranged from 44 to 72% in hermaphroditic and cross-fertilized populations Nei's index of gene diversity and Shannon's Information index were not statistically different for both populations The extent of genetic variation after one generation of selfing in the progeny from hermaphroditic seed is similar to that in progeny from cross-fertilized seeds
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TL;DR: The current manuscript reviews the advances in Cannabis biotechnology, including molecular markers, microRNA, omics-based methods, and functional genes related to the terpene and cannabinoid biosynthesis as well as fiber quality.
52 citations
TL;DR: A review of the important diseases currently affecting the cannabis and hemp industries in North America is presented in this article, where the authors discuss various mitigation strategies, such as establishing clean planting stock, modifying environmental conditions to reduce pathogen development, implementing sanitation measures, and applying fungal and bacterial biological control agents.
Abstract: Cultivation of cannabis plants (Cannabis sativa L., marijuana) has taken place worldwide for centuries. In Canada, legalization of cannabis in October 2018 for the medicinal and recreational markets has spurned interest in large-scale growing. This increased production has seen a rise in the incidence and severity of plant pathogens, causing a range of previously unreported diseases. The objective of this review is to highlight the important diseases currently affecting the cannabis and hemp industries in North America and to discuss various mitigation strategies. Progress in molecular diagnostics for pathogen identification and determining inoculum sources and methods of pathogen spread have provided useful insights. Sustainable disease management approaches include establishing clean planting stock, modifying environmental conditions to reduce pathogen development, implementing sanitation measures, and applying fungal and bacterial biological control agents. Fungicides are not currently registered for use and hence there are no published data on their efficacy. The greatest challenge remains in reducing microbial loads (colony-forming units) on harvested inflorescences (buds). Contaminating microbes may be introduced during the cultivation and postharvest phases, or constitute resident endophytes. Failure to achieve a minimum threshold of microbes deemed to be safe for utilization of cannabis products can arise from conventional and organic cultivation methods, or following applications of beneficial biocontrol agents. The current regulatory process for approval of cannabis products presents a challenge to producers utilizing biological control agents for disease management. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
35 citations
TL;DR: This work aims at supporting the development of the cannabis industry in a sustainable way, by exploiting the many beneficial attributes of Pseudomonas spp.
Abstract: Among the oldest domesticated crops, cannabis plants (Cannabis sativa L., marijuana and hemp) have been used to produce food, fiber, and drugs for thousands of years. With the ongoing legalization of cannabis in several jurisdictions worldwide, a new high-value market is emerging for the supply of marijuana and hemp products. This creates unprecedented challenges to achieve better yields and environmental sustainability, while lowering production costs. In this review, we discuss the opportunities and challenges pertaining to the use of beneficial Pseudomonas spp. bacteria as crop inoculants to improve productivity. The prevalence and diversity of naturally occurring Pseudomonas strains within the cannabis microbiome is overviewed, followed by their potential mechanisms involved in plant growth promotion and tolerance to abiotic and biotic stresses. Emphasis is placed on specific aspects relevant for hemp and marijuana crops in various production systems. Finally, factors likely to influence inoculant efficacy are provided, along with strategies to identify promising strains, overcome commercialization bottlenecks, and design adapted formulations. This work aims at supporting the development of the cannabis industry in a sustainable way, by exploiting the many beneficial attributes of Pseudomonas spp.
10 citations
TL;DR: The development of a phytocannabinoid-specific accurate-mass MSn fragmentation spectral library and gene pool representative metabolome has the potential to improve small molecule compound annotation and accelerate understanding of metabolic variation underlying phenotypic diversity in Cannabis.
Abstract: Cannabis is a mostly dioecious multi-use flowering plant genus. Sexual dimorphism is an important characteristic in Cannabis-based commercial production systems, which has consequences for fibre, seed, and the yield of secondary metabolites, such as phytocannabinoid and terpenes for therapeutic uses. Beyond the obvious morphological differences between male and female plants, metabolic variation among dioecious flowers is largely undefined. Here, we report a pilot metabolomic study comparing staminate (male) and pistillate (female) unisexual flowers. Enrichment of the α-linolenic acid pathway and consensus evaluation of the jasmonic acid (JA) related compound 12-oxo-phytodienoicacid (OPDA) among differentially abundant metabolites suggests that oxylipin signalling is associated with secondary metabolism and sex expression in female flowers. Several putative phytocannabinoid-like compounds were observed to be upregulated in female flowers, but full identification was not possible due to the limitation of available databases. Targeted analysis of 14 phytocannabinoids using certified reference standards (cannabidiolic acid (CBDA), cannabidiol (CBD), Δ9-tetrahydrocannabinolic acid A (Δ9-THCAA), Δ9-tetrahydrocannabinol (Δ9-THC), cannabichromenic acid (CBCA), cannabichromene (CBC), cannabigerolic acid (CBGA), cannabigerol (CBG), cannabinolic acid (CBNA), cannabinol (CBN), cannabidivarinic acid (CBDVA), cannabidivarin (CBDV), tetrahydrocannabivarinic acid (THCVA), and tetrahydrocannabivarin (THCV)) showed a higher total phytocannabinoid content in female flowers compared with the male flowers, as expected. In summary, the development of a phytocannabinoid-specific accurate-mass MSn fragmentation spectral library and gene pool representative metabolome has the potential to improve small molecule compound annotation and accelerate understanding of metabolic variation underlying phenotypic diversity in Cannabis.
10 citations
TL;DR: In this paper, the authors evaluated variables that can influence the success of shoot growth and plantlet production in tissue cultures of drug-type Cannabis sativa L. (marijuana) and showed significant differences in response to shoot growth from meristems and nodal explants on Murashige and Skoog (MS) medium containing thidiazuron (1 μM) and naphthaleneacetic acid plus 1% activated charcoal.
Abstract: Tissue culture approaches are widely used in crop plants for the purposes of micropropagation, regeneration of plants through organogenesis, obtaining pathogen-free plantlets from meristem culture, and developing genetically modified plants. In this research, we evaluated variables that can influence the success of shoot growth and plantlet production in tissue cultures of drug-type Cannabis sativa L. (marijuana). Various sterilization methods were tested to ensure shoot development from nodal explants by limiting the frequency of contaminating endophytes, which otherwise caused the death of explants. Seven commercially grown tetrahydrocannabinol (THC)-containing cannabis genotypes (strains) showed significant differences in response to shoot growth from meristems and nodal explants on Murashige and Skoog (MS) medium containing thidiazuron (1 μM) and naphthaleneacetic acid (0.5 μM) plus 1% activated charcoal. The effect of Driver and Kuniyuki Walnut (DKW) or MS basal salts in media on shoot length and leaf numbers from nodal explants was compared and showed genotype dependency with regard to the growth response. To obtain rooted plantlets, shoots from meristems and nodal explants of genotype Moby Dick were evaluated for rooting, following the addition of sodium metasilicate, silver nitrate, indole-3-butyric acid (IBA), kinetin, or 2,4-D. Sodium metasilicate improved the visual appearance of the foliage and improved the rate of rooting. Silver nitrate also promoted rooting. Following acclimatization, plantlet survival in hydroponic culture, peat plugs, and rockwool substrate was 57, 76, and 83%, respectively. The development of plantlets from meristems is described for the first time in C. sativa and has potential for obtaining pathogen-free plants. The callogenesis response of leaf explants of 11 genotypes on MS medium without activated charcoal was 35% to 100%, depending on the genotype; organogenesis was not observed. The success in recovery of plantlets from meristems and nodal explants is influenced by cannabis genotype, degree of endophytic contamination of the explants, and frequency of rooting. The procedures described here have potential applications for research and commercial utility to obtain plantlets in stage 1 tissue cultures of C. sativa.
9 citations
References
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TL;DR: A method is presented by which the gene diversity (heterozygosity) of a subdivided population can be analyzed into its components, i.e., the gene diversities within and between subpopulations.
Abstract: A method is presented by which the gene diversity (heterozygosity) of a subdivided population can be analyzed into its components, i.e., the gene diversities within and between subpopulations. This method is applicable to any population without regard to the number of alleles per locus, the pattern of evolutionary forces such as mutation, selection, and migration, and the reproductive method of the organism used. Measures of the absolute and relative magnitudes of gene differentiation among subpopulations are also proposed.
8,465 citations
TL;DR: The evidence that the evolution of breeding systems of animals and plants has been significantly influenced by the occurrence of inbreeding depression is reviewed, and the contemporary genetic theory of inmarriage depression and heterosis and the experimental data concerning the strength of in breeding depression are considered.
Abstract: The harmful effects of close inbreeding have been noticed for many centuries (34, 35, 165). With the rise of Mendelian genetics, it was realized that the main genetic consequence of inbreeding is homozygosis (165, Ch. 2). Two main theories were early proposed to account for inbreeding depression and its converse, heterosis (the increase in vigor observed in an F1 between two inbred lines). These are the overdominance and partial dominance hypotheses, discussed in more detail below. Research into this question has continued up to the present, and this is one of the topics that we discuss. Darwin (35, 36) was the first to point out that the evident adaptations of many plants for ensuring outcrossing could be understood in terms of the selective advantage of avoiding inbreeding depression. We review the evidence that the evolution of breeding systems of animals and plants has been significantly influenced by the occurrence of inbreeding depression. In order to do this, we consider the contemporary genetic theory of inbreeding depression and heterosis, and the experimental data concerning the strength of inbreeding depression. We emphasize data and theory relevant to natural, rather than domesticated, populations as we are chiefly concerned to evaluate the evolutionary significance of inbreeding depression. We do not attempt to give a complete bibliography of this very extensive field but try to concentrate on what seem to be the most significant findings in relation to this aim.
3,135 citations
"Hermaphroditism in Marijuana (Canna..." refers background in this paper
...Inbreeding can reduce the fitness of the inbred relative to outbred offspring, due to an increase of homozygous loci in the former (Charlesworth and Charlesworth, 1987)....
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TL;DR: This work proposes the first unified hierarchical classification system, designed on the basis of the transposition mechanism, sequence similarities and structural relationships, that can be easily applied by non-experts.
Abstract: Our knowledge of the structure and composition of genomes is rapidly progressing in pace with their sequencing. The emerging data show that a significant portion of eukaryotic genomes is composed of transposable elements (TEs). Given the abundance and diversity of TEs and the speed at which large quantities of sequence data are emerging, identification and annotation of TEs presents a significant challenge. Here we propose the first unified hierarchical classification system, designed on the basis of the transposition mechanism, sequence similarities and structural relationships, that can be easily applied by non-experts. The system and nomenclature is kept up to date at the WikiPoson web site.
2,425 citations
"Hermaphroditism in Marijuana (Canna..." refers background in this paper
...Both domains are features of Class I LTR retrotransposons (Wicker et al., 2007; Llorens et al., 2011)....
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...In addition, the placement of these domains is characteristic of the Copia Superfamily of LTR retrotransposons (Wicker et al., 2007)....
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TL;DR: An analysis of twelve plant families indicated that species within families with predominately outcrossing, woody species had more genetic diversity and less interpopulation differentiation than species withinfamilies with predominate herbaceous species.
Abstract: Seven two-trait combinations (e.g. breeding system and seed dispersal mechanism) of five life history characteristics were used to analyse interspecific variation in the level and distribution of allozyme genetic diversity in seed plants. Highly significant differences were seen among categories for all seven comparisons. Life form and breeding system had highly significant influences on genetic diversity and its distribution. Regardless of other traits, outcrossing species tended to be more genetically diverse and had less genetic differentiation among their populations. Similarly, woody plants have less among population differentiation and somewhat more genetic diversity than non-woody species with similar life history traits. An analysis of twelve plant families indicated that species within families with predominately outcrossing, woody species had more genetic diversity and less interpopulation differentiation than species within families with predominately herbaceous species.
2,090 citations
"Hermaphroditism in Marijuana (Canna..." refers background in this paper
...It has been proposed that dioecy is a basic evolutionary mechanism to ensure cross-fertilization and, as a consequence, results in maintenance of high genetic diversity and heterozygosity (Dellaporta and Calderon-Urrea, 1993; Hamrick and Godt, 1996; Ainsworth, 2000)....
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TL;DR: The conclusion is that, although FST and rare‐alleles methods are expected to be equally effective in analyzing ideal data, practical problems in estimating the frequencies of rare alleles in electrophoretic studies suggest that FST is likely to be more useful under realistic conditions.
Abstract: Three methods for estimating the average level of gene flow in natural population are discussed and compared. The three methods are FST , rare alleles, and maximum likelihood. All three methods yield estimates of the combination of parameters (the number of migrants [Nm] in a demic model or the neighborhood size [4πDσ2 ] in a continuum model) that determines the relative importance of gene flow and genetic drift. We review the theory underlying these methods and derive new analytic results for the expectation of FST in stepping-stone and continuum models when small sets of samples are taken. We also compare the effectiveness of the different methods using a variety of simulated data. We found that the FST and rare-alleles methods yield comparable estimates under a wide variety of conditions when the population being sampled is demographically stable. They are roughly equally sensitive to selection and to variation in population structure, and they approach their equilibrium values at approximately the same rate. We found that two different maximum-likelihood methods tend to yield biased estimates when relatively small numbers of locations are sampled but more accurate estimates when larger numbers are sampled. Our conclusion is that, although FST and rare-alleles methods are expected to be equally effective in analyzing ideal data, practical problems in estimating the frequencies of rare alleles in electrophoretic studies suggest that FST is likely to be more useful under realistic conditions.
1,349 citations
"Hermaphroditism in Marijuana (Canna..." refers methods in this paper
...Nm was also calculated according to the formula, Nm = (1 - Gst)/4Gst (Slatkin and Barton, 1989)....
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