scholarly journals Delineating the genetic regulation of cannabinoid biosynthesis during female flower development in Cannabis sativa

2021 ◽  
Author(s):  
Peter Apicella ◽  
Lauren Sands ◽  
Yi Ma ◽  
Gerald A Berkowitz

Cannabinoids are predominantly produced in the glandular trichomes on cannabis female flowers. There is little known on how cannabinoid biosynthesis is regulated during female flower development. We aim to understand the rate-limiting step(s) in the cannabinoid biosynthetic pathway. We investigated the transcript levels of cannabinoid biosynthetic genes as well as cannabinoid contents during 7 weeks of female flower development. We demonstrated that the enzymatic steps for producing CBG, which involve genes GPPS, PT and OAC, could be rate limit cannabinoid biosynthesis. Our findings further suggest that cannabinoid synthases, CBDAS and THCAS in a hemp and medical marijuana variety respectively, are not critical for cannabinoid biosynthesis. The cannabinoid biosynthetic genes are generally upregulated during flower maturation, which indicate glandular trichome development. MeJA can potentially increase cannabinoid production. We propose that biweekly application of 100 μM MeJA staring from flower initiation would be efficacious for promoting cannabinoid biosynthesis.

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1194
Author(s):  
Limeng Zhou ◽  
Yingzhang Huang ◽  
Qi Wang ◽  
Dianjing Guo

Glandular trichome (GT) is the dominant site for artemisinin production in Artemisia annua. Several critical genes involved in artemisinin biosynthesis are specifically expressed in GT. However, the molecular mechanism of differential gene expression between GT and other tissue types remains elusive. Chromatin accessibility, defined as the degree to which nuclear molecules are able to interact with chromatin DNA, reflects gene expression capacity to a certain extent. Here, we investigated and compared the landscape of chromatin accessibility in Artemisia annua leaf and GT using the Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) technique. We identified 5413 GT high accessible and 4045 GT low accessible regions, and these GT high accessible regions may contribute to GT-specific biological functions. Several GT-specific artemisinin biosynthetic genes, such as DBR2 and CYP71AV1, showed higher accessible regions in GT compared to that in leaf, implying that they might be regulated by chromatin accessibility. In addition, transcription factor binding motifs for MYB, bZIP, C2H2, and AP2 were overrepresented in the highly accessible chromatin regions associated with artemisinin biosynthetic genes in glandular trichomes. Finally, we proposed a working model illustrating the chromatin accessibility dynamics in regulating artemisinin biosynthetic gene expression. This work provided new insights into epigenetic regulation of gene expression in GT.


2020 ◽  
Author(s):  
Lee J. Conneely ◽  
Ramil Mauleon ◽  
Jos Mieog ◽  
Bronwyn J. Barkla ◽  
Tobias Kretzschmar

AbstractCannabis sativa has been cultivated since antiquity as a source of fibre, food and medicine. The recent resurgence of Cannabis as a cash crop is mainly driven by the medicinal and therapeutic properties of its resin, which contains compounds that interact with the human endocannabinoid system. Compared to other medicinal crops of similar value, however, little is known about the biology of C. sativa. Glandular trichomes are small hair-like projections made up of stalk and head tissue and are responsible for the production of the resin in C. sativa. Trichome productivity, as determined by Cannabis sativa resin yield and composition, is only beginning to be understood at the molecular level. In this study the proteomes of glandular trichome stalks and heads, were investigated and compared to the proteome of the whole flower tissue, to help elucidate Cannabis sativa glandular trichome biochemistry. The data suggested that the floral tissue acts as a major source of carbon and energy to the glandular trichome head sink tissue, supplying sugars which drive secondary metabolite biosynthesis in the glandular trichome head; the location of the secretory cells. The trichome stalk seems to play only a limited role in secondary metabolism and acts as both source and sink.


PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0242633
Author(s):  
Lee James Conneely ◽  
Ramil Mauleon ◽  
Jos Mieog ◽  
Bronwyn J. Barkla ◽  
Tobias Kretzschmar

Cannabis sativa has been cultivated since antiquity as a source of fibre, food and medicine. The recent resurgence of C. sativa as a cash crop is mainly driven by the medicinal and therapeutic properties of its resin, which contains compounds that interact with the human endocannabinoid system. Compared to other medicinal crops of similar value, however, little is known about the biology of C. sativa. Glandular trichomes are small hair-like projections made up of stalk and head tissue and are responsible for the production of the resin in C. sativa. Trichome productivity, as determined by C. sativa resin yield and composition, is only beginning to be understood at the molecular level. In this study the proteomes of glandular trichome stalks and heads, were investigated and compared to the proteome of the whole flower tissue, to help further elucidate C. sativa glandular trichome biochemistry. The data suggested that the floral tissue acts as a major source of carbon and energy to the glandular trichome head sink tissue, supplying sugars which drive secondary metabolite biosynthesis. The trichome stalk seems to play only a limited role in secondary metabolism and acts as both source and sink.


Euphytica ◽  
2021 ◽  
Vol 217 (3) ◽  
Author(s):  
Joris Santegoets ◽  
Marcella Bovio ◽  
Wendy van’t Westende ◽  
Roeland E. Voorrips ◽  
Ben Vosman

AbstractThe greenhouse whitefly Trialeurodes vaporariorum is a major threat in tomato cultivation. In greenhouse grown tomatoes non-trichome based whitefly resistance may be better suited than glandular trichome based resistance as glandular trichomes may interfere with biocontrol, which is widely used. Analysis of a collection of recombinant inbred lines derived from a cross between Solanum lycopersicum and Solanum galapagense showed resistance to the whitefly T. vaporariorum on plants without glandular trichomes type IV. The resistance affected whitefly adult survival (AS), but not oviposition rate. This indicates that S. galapagense, in addition to trichome based resistance, also carries non-trichome based resistance components. The effectiveness of the non-trichome based resistance appeared to depend on the season in which the plants were grown. The resistance also had a small but significant effect on the whitefly Bemisia tabaci, but not on the thrips Frankliniella occidentalis. A segregating F2 population was created to map the non-trichome based resistance. Two Quantitative trait loci (QTLs) for reduced AS of T. vaporariorum were mapped on chromosomes 12 and 7 (explaining 13.9% and 6.0% of the variance respectively). The QTL on chromosome 12 was validated in F3 lines.


Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 930
Author(s):  
Xu Yu ◽  
Xiwu Qi ◽  
Shumin Li ◽  
Hailing Fang ◽  
Yang Bai ◽  
...  

Light is a key environmental aspect that regulates secondary metabolic synthesis. The essential oil produced in mint (Mentha canadensis L.) leaves is used widely in the aromatics industry and in medicine. Under low-light treatment, significant reductions in peltate glandular trichome densities were observed. GC-MS analysis showed dramatically reduced essential oil and menthol contents. Light affected the peltate glandular trichomes’ development and essential oil yield production. However, the underlying mechanisms of this regulation were elusive. To identify the critical genes during light-regulated changes in oil content, following a 24 h darkness treatment and a 24 h recovery light treatment, leaves were collected for transcriptome analysis. A total of 95,579 unigenes were obtained, with an average length of 754 bp. About 56.58% of the unigenes were annotated using four public protein databases: 10,977 differentially expressed genes (DEGs) were found to be involved in the light signaling pathway and monoterpene synthesis pathway. Most of the TPs showed a similar expression pattern: downregulation after darkness treatment and upregulation after the return of light. In addition, the genes involved in the light signal transduction pathway were analyzed. A series of responsive transcription factors (TFs) were identified and could be used in metabolic engineering as an effective strategy for increasing essential oil yields.


Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1420
Author(s):  
Takahiro Ueda ◽  
Miki Murata ◽  
Ken Yokawa

Environmental light conditions influence the biosynthesis of monoterpenes in the mint plant. Cyclic terpenes, such as menthol, menthone, pulegone, and menthofuran, are major odor components synthesized in mint leaves. However, it is unclear how light for cultivation affects the contents of these compounds. Artificial lighting using light-emitting diodes (LEDs) for plant cultivation has the advantage of preferential wavelength control. Here, we monitored monoterpene contents in hydroponically cultivated Japanese mint leaves under blue, red, or far-red wavelengths of LED light supplements. Volatile cyclic monoterpenes, pulegone, menthone, menthol, and menthofuran were quantified using the head-space solid phase microextraction method. As a result, all light wavelengths promoted the biosynthesis of the compounds. Remarkably, two weeks of blue-light supplement increased all compounds: pulegone (362% increase compared to the control), menthofuran (285%), menthone (223%), and menthol (389%). Red light slightly promoted pulegone (256%), menthofuran (178%), and menthol (197%). Interestingly, the accumulation of menthone (229%) or menthofuran (339%) was observed with far-red light treatment. The quantification of glandular trichomes density revealed that no increase under light supplement was confirmed. Blue light treatment even suppressed the glandular trichome formation. No promotion of photosynthesis was observed by pulse-amplitude-modulation (PAM) fluorometry. The present result indicates that light supplements directly promoted the biosynthetic pathways of cyclic monoterpenes.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Peina Zhou ◽  
Mengjiao Yin ◽  
Shilin Dai ◽  
Ke Bao ◽  
Chenglin Song ◽  
...  

Abstract Background Perilla frutescens (L.) Britt is a medicinal and edible plant widely cultivated in Asia. Terpenoids, flavonoids and phenolic acids are the primary source of medicinal ingredients. Glandular trichomes with multicellular structures are known as biochemical cell factories which synthesized specialized metabolites. However, there is currently limited information regarding the site and mechanism of biosynthesis of these constituents in P. frutescens. Herein, we studied morphological features of glandular trichomes, metabolic profiling and transcriptomes through different tissues. Results Observation of light microscopy and scanning electron microscopy indicated the presence of three distinct glandular trichome types based on their morphological features: peltate, capitate, and digitiform glandular trichomes. The oil of peltate glandular trichomes, collected by custom-made micropipettes and analyzed by LC–MS and GC–MS, contained perillaketone, isoegomaketone, and egomaketone as the major constituents which are consistent with the components of leaves. Metabolomics and transcriptomics were applied to explore the bioactive constituent biosynthesis in the leaves, stem, and root of P. frutescens. Transcriptome sequencing profiles revealed differential regulation of genes related to terpenoids, flavonoids, and phenylpropanoid biosynthesis, respectively with most genes expressed highly in leaves. The genes affecting the development of trichomes were preliminarily predicted and discussed. Conclusions The current study established the morphological and chemical characteristics of glandular trichome types of P. frutescens implying the bioactive constituents were mainly synthesized in peltate glandular trichomes. The genes related to bioactive constituents biosynthesis were explored via transcriptomes, which provided the basis for unraveling the biosynthesis of bioactive constituents in this popular medicinal plant.


Author(s):  
KU Ahamed ◽  
B Akhter ◽  
MR Islam ◽  
MR Humaun ◽  
MJ Alam

Genetic divergence of 110 lentil germplasm with checks was assessed based on morphological traits using multivariate analysis. Mahalanobis generalized distance (D2) analysis was used to group the lentil genotypes. Significant variations among lentil genotypes were observed in respect of days to 1st flowering, days to 50% flowering, days to maturity, plant height, and number of pods per peduncle, number of pods per plant, number of seeds per plant, 100 seed weight and yield per plant. Considering the mean values, the germplasm were grouped into ten clusters. The highest number of genotypes (17) was in cluster X and lowest (5) both in cluster II and IV. Cluster IV had the highest cluster mean for number of pods per plant (297.08), number of seeds per plant (594.16), 100 seed weight (1.44 g) and yield per plant (8.53 g). Among them, the highest inter-cluster distance was obtained between the cluster IV and I (24.61) followed by IV and III (22.33), while the lowest was between IX and II (1.63). The maximum value of inter-cluster distance indicated that genotypes belonging to cluster IV were far diverged from those of cluster I. The first female flower initiation was earlier in BD-3812 (49 days) in cluster I and cluster IV had highest grain yield per plant (8.53). BD-3807 produced significant maximum number of pods per plant (298.40) in cluster IV. DOI: http://dx.doi.org/10.3329/ijarit.v4i1.21095 Int. J. Agril. Res. Innov. & Tech. 4 (1): 70-76, June, 2014


1997 ◽  
Vol 122 (3) ◽  
pp. 373-379 ◽  
Author(s):  
Richard Grazzini ◽  
Donald Walters ◽  
Jody Harmon ◽  
David J. Hesk ◽  
Diana Cox-Foster ◽  
...  

Diploid zonal geraniums (Pelargonium ×hortorum) are able to resist attack by small arthropod pests such as the two-spotted spider mite (Tetranychus urticae Koch) when exudate produced by tall glandular trichomes contains a high percentage of ω5-unsaturated anacardic acids. Trichomes of susceptible plants exude primarily saturated anacardic acids. Inbred mite-resistant and -susceptible geraniums were reciprocally crossed and the F1, F2, and backcross generations were examined for anacardic acid composition and trichome density. Selected F2 plants were bioassayed for resistance to two-spotted spider mites. High concentrations of ω5-unsaturated anacardic acids in resistant plants are conditioned by a single dominant allele. We propose that inheritance of tall glandular trichome density can be controlled by a small number of loci (possibly as few as one) exhibiting codominance. F2, with low densities of tall glandular trichomes and producing ω5-unsaturated anacardic acids, displayed effective resistance to two-spotted spider mites as measured by mite mortality and fecundity. A genetic model for the biosynthesis of anacardic acids is proposed.


2003 ◽  
Vol 128 (4) ◽  
pp. 479-485 ◽  
Author(s):  
Erik S. Runkle ◽  
Royal D. Heins

Plastics that selectively reduce the transmission of far-red light (FR, 700 to 800 nm) reduce extension growth of many floricultural crops. However, FR-deficient (FRd) environments delay flowering in some long-day plants (LDPs), including `Crystal Bowl Yellow' pansy (Viola ×wittrockiana Gams). Our objective was to determine if FR light could be added to an otherwise FRd environment to facilitate flowering with minimal extension growth. In one experiment, plants were grown under a 16-hour FRd photoperiod, and FR-rich light was added during portions of the day or night. For comparison, plants were also grown with a 9-hour photoperiod [short-day (SD) control] or under a neutral (N) filter with a 16-hour photoperiod (long day control). Flowering was promoted most (i.e., percent of plants that flowered increased and time to flower decreased) when FR-rich light was added during the entire 16-hour photoperiod, during the last 4 hours of the photoperiod, or during the first or second 4 hours after the end of the photoperiod. In a separate experiment, pansy was grown under an FRd or N filter with a 9-hour photoperiod plus 0, 0.5, 1, 2, or 4 hours of night interruption (NI) lighting that delivered a red (R, 600 to 700 nm) to FR ratio of 0.56 (low), 1.28 (moderate), or 7.29 (high). Under the N filter, the minimum NI duration that increased percent flowering was 2 hours with a moderate or low R:FR and 4 hours with a high R:FR. Under the FRd filter, 2 or 4 hours of NI lighting with a moderate or low R:FR, respectively, was required to increase percent flowering, but a 4-hour NI with a high R:FR failed to promote flowering. Pansy appears to be day-neutral with respect to flower initiation and a quantitative LDP with respect to flower development. The promotion of reproductive development was related linearly to the promotion of extension growth. Therefore, it appears that in LDPs such as pansy, light duration and quality concomitantly promote extension growth and flowering, and cannot readily be separated with lighting strategies.


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