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bioRxiv2020

A recombineering pipeline to clone large and complex genes in Chlamydomonas

The ability to clone genes has driven fundamental advances in cell and molecular biology, enabling researchers to introduce precise mutations, generate fluorescent protein fusions for localization and to confirm genetic causation by mutant complementation. Most gene cloning is PCR or DNA synthesis dependent, which can become costly and technically challenging as genes increase in size and particularly if they contain complex regions. This has been a long-standing challenge for the Chlamydomonas reinhardtii research community, with a high percentage of genes containing complex sequence structures, an average genomic GC content of 64% and gene expression requiring regular introns for stable transcription. Here we overcome these challenges via the development of a recombineering pipeline that enables the rapid parallel cloning of genes from a Chlamydomonas BAC collection. We show the method can successfully retrieve large and complex genes that PCR-based methods have previously failed to clone, including genes as large as 23 kilobases, thus making previously technically challenging genes to study now amenable to cloning. We initially applied the pipeline to 12 targets with a 92% cloning success rate. We then developed a high-throughput approach and targeted 191 genes relating to the Chlamydomonas CO2 concentrating mechanism (CCM) with an overall cloning success rate of 77% that is independent of gene size. Localization of a subset of CCM targets has confirmed previous mass spectrometry data and identified new pyrenoid components. To expand the functionality of our system, we developed a series of localization vectors that enable complementation of Chlamydomonas Library Project mutants and enable protein tagging with a range of fluorophores. Vectors and detailed protocols are available to facilitate the easy adoption of this method by the Chlamydomonas research community. We envision that this technology will open up new possibilities in algal and plant research and be complementary to the Chlamydomonas mutant library.

plant biology↗

bioRxiv2022

Curation of the Deep Green list of unannotated green lineage proteins to enable structural and functional characterization

An explosion of sequenced genomes and predicted proteomes enabled by low cost deep sequencing has revolutionized biology. Unfortunately, protein functional annotation is more complex, and has not kept pace with the sequencing revolution. We identified unannotated proteins in three model organisms representing distinct parts of the green lineage (Viridiplantae); Arabidopsis thaliana (dicot), Setaria viridis (monocot), and Chlamydomonas reinhardtii (Chlorophyte alga). Using similarity searching we found the subset of unannotated proteins that were conserved between these species and defined them as Deep Green proteins. Informatic, genomic, and structural predictions were leveraged to begin inferring functional information about Deep Green genes and proteins. The Deep Green set was enriched for proteins with predicted chloroplast targeting signals that are predictive of photosynthetic or plastid functions. Strikingly, structural predictions using AlphaFold and comparisons to known structures show that a significant proportion of Deep Green proteins may possess novel protein tertiary structures. The Deep Green genes and proteins provide a starting resource of high value targets for further investigation of potentially new protein structures and functions that are conserved in the green lineage.

plant biology↗

bioRxiv2021

High-quality Arabidopsis thaliana genome assembly with Nanopore and HiFi long reads

Arabidopsis thaliana is an important and long-established model species for plant molecular biology, genetics, epigenetics, and genomics. However, the latest version of reference genome still contains significant number of missing segments. Here, we report a high-quality and almost complete Col-0 genome assembly with two gaps (Col-XJTU) using combination of Oxford Nanopore Technology ultra-long reads, PacBio high-fidelity long reads, and Hi-C data. The total genome assembly size is 133,725,193 bp, introducing 14.6 Mb of novel sequences compared to the TAIR10.1 reference genome. All five chromosomes of Col-XJTU assembly are highly accurate with consensus quality (QV) scores > 60 (ranging from 62 to 68), which are higher than those of TAIR10.1 reference (QV scores ranging from 45 to 52). We have completely resolved chromosome (Chr) 3 and Chr5 in a telomere-to-telomere manner. Chr4 has been completely resolved except the nucleolar organizing regions, which comprise long repetitive DNA fragments. The Chr1 centromere (CEN1), reportedly around 9 Mb in length, is particularly challenging to assemble due to the presence of tens of thousands of CEN180 satellite repeats. Using the cutting-edge sequencing data and novel computational approaches, we assembled about 4 Mb of sequence for CEN1 and a 3.5-Mb-long CEN2. We investigated the structure and epigenetics of centromeres. We detected four clusters of CEN180 monomers, and found that the centromere-specific histone H3-like protein (CENH3) exhibits a strong preference for CEN180 cluster 3. Moreover, we observed hypomethylation patterns in CENH3-enriched regions. We believe that this high-quality genome assembly, Col-XJTU, would serve as a valuable reference to better understand the global pattern of centromeric polymorphisms, as well as genetic and epigenetic features in plants.

genomics↗

bioRxiv2021

Water stress-associated isolation barriers between two sympatric oak species

Drought and waterlogging impede tree growth and may even lead to tree death. With climate change, these environmental factors are a growing source of concern, particularly for temperate forests. Oaks, an emblematic group of tree species, have evolved a range of adaptations to cope with these constraints. The two most widely distributed European species -- pedunculate oak (PO) and sessile oaks (SO) -- have overlapping ranges, but are highly constrained locally by soil water content variation. These differences in local ecological requirements provide a powerful biological model for studying the role of ecological barriers in speciation. We used an experimental set-up mimicking the ecological preferences of these species, in which seedlings were subjected to waterlogging and drought. We studied gene expression in roots by RNA-seq and identified genes differentially expressed between treatments with different outcomes depending on species. These "species x environment"-responsive genes revealed adaptive molecular strategies involving adventitious and lateral root formation, aerenchyma formation in PO, and osmoregulation and ABA regulation in SO. With this experimental design, we also identified genes with expression profiles presenting a "species" effect regardless of imposed constraints with important roles in intrinsic reproductive barriers. Finally, we compared our findings with those for a genome scan of species divergence and found that the candidate genes were enriched in highly differentiated SNPs. This suggests that many of the genes involved in the contrasting transcriptomic responses are subject to natural selection and that gene regulation helps to maintain these two different oak species in sympatry.

plant biology↗

bioRxiv2022

Interaction between photoperiod and variation in circadian rhythms in tomato

Many biological processes follow circadian rhythmicity and are controlled by the circadian clock. Predictable environmental changes such as seasonal variation in photoperiod can modulate circadian rhythms, allowing organisms to adjust to the time of the year. Modification of circadian clocks is especially relevant in crops to enhance their cultivability in specific regions by changing their sensibility to photoperiod. In tomato, the appearance of mutations in EMPFINDLICHER IM DUNKELROTEN LICHT 1 (EID1, Solyc09g075080) and NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED GENE 2 (LNK2, Solyc01g068560) during domestication delayed its circadian rhythms, and allowed its expansion outside its equatorial origin. Here we study how variation in circadian rhythms in tomato affects its perception of photoperiod. To do this, we create near isogenic lines carrying combinations of wild alleles of EID1 and LNK2 and perform transcriptomic profiling under two different photoperiods. We observe that EID1, but not LNK2, has a large effect on the tomato transcriptome and its response to photoperiod. This large effect of EID1 is likely a consequence of the global phase shift elicited by this gene in tomatos circadian rhythms.

plant biology↗

bioRxiv2023

Exploring the family feud: a fungal endophyte induces local cell wall-mediated resistance in wheat roots against the closely related "take-all" pathogen

Take-all disease, caused by the ascomycete fungus Gaeumannomyces tritici, is one of the most important root diseases of wheat worldwide. The fungus invades the roots and destroys the vascular tissue, hindering the uptake of water and nutrients. Closely related non-pathogenic species in the Magnaporthaceae family, such as Gaeumannomyces hyphopodioides, occur naturally in arable and grassland soils and have previously been reported to reduce take-all disease in field studies. However, the mechanism of take-all protection has remained unknown. Here, we characterise the root infection biologies of G. tritici and G. hyphopodioides in wheat. We investigate the ultrastructure of previously described "subepidermal vesicles" (SEVs), produced in wheat roots by non-pathogenic G. hyphopodioides, but not by pathogenic G. tritici. We show that G. hyphopodioides SEVs share key characteristics of fungal resting structures; containing a greater number of putative lipid bodies and a significantly thickened cell wall compared to infection hyphae. We demonstrate that take-all control is achieved via local but not systemic host changes in response to prior G. hyphopodioides root colonisation. A time-course wheat RNA sequencing analysis revealed extensive transcriptional reprogramming in G. hyphopodioides colonised tissues, characterised by a striking downregulation of key cell-wall related genes, including cellulose synthase (CESA), and xyloglucan endotransglucosylase/hydrolase (XTH) genes. In the absence of take-all resistant wheat cultivars or non-virulent G. tritici strains, studying closely related non-pathogenic G. hyphopodioides provides a much-needed avenue to elucidate take-all resistance mechanisms in wheat.

plant biology↗

bioRxiv2026

Potential of Bacillus subtilis as a biological control agent against three rot-causing pathogens in seed yams

Ghana is the largest exporter of yams in West Africa. However, yam production, particularly seed yam production, is constrained by storage rot during the off-season. Farmers seldom use synthetic pesticides to control seed yam rot. However, these are costly and pose adverse health risks to farmers. Biological antagonists offer a sustainable, relatively cost-effective, and safe alternative to synthetic pesticides. Therefore, this study aimed to test the efficacy of Bacillus subtilis as an alternative to synthetic pesticides. Bacillus subtilis supplied through the biofungicide Serenade ASO (Bayer) was assayed against three storage-rot pathogens: Lasiodiplodia theobromae, Aspergillus niger, and Rhizopus sp. These pathogens were previously isolated from the tissues of rotten seed yams. The efficacy of the bacterium was tested at concentrations of 17 %, 33 %, and 50 % in both in vitro and in vivo bioassays. B. subtilis (50 %) completely inhibited the growth (100 %) of L. theobromae in the in vitro studies. In contrast, there was little to no growth inhibition of the other two test fungi. In the in vivo assay, B. subtilis (50%) significantly (P < 0.01) inhibited L. theobromae, resulting in minimal rot lesions. However, B. subtilis (50 %) was ineffective against the other two test pathogens, resulting in large rot lesions on the seed yams. This suggests that B. subtilis could be an ideal alternative to synthetic pesticides for controlling L. theobromae on seed yams.

plant biology↗

bioRxiv2025

The Rapid Mechanically Activated (RMA) channel transduces increases in plasma membrane tension into transient calcium influx

O_LIPlants respond to mechanical stimuli by a rapid increase in cytosolic calcium. The intensity and kinetics of the calcium changes define calcium signatures important for biological responses . In this study, we determine the properties of a calcium permeable force-gated channel localized at the plasma membrane called Rapid Mechanically Activated (RMA). C_LIO_LIUsing patch-clamp and pressure-clamp, we characterized the kinetics of activation and inactivation of RMA channel upon stimulation by pulses of pressure applied onto the plasma membrane. Combining repetitive pressure pulse protocols at different frequencies with modeling, we investigated the channels capacity to transduce high frequency mechanical stimuli. C_LIO_LIRMA channel rapidly activates in response to membrane tension, then it inactivates during prolonged stimulation. Upon repeated stimulations, RMA current amplitude decreases irreversibly indicating that undergoes adaptation. The channel kinetics may be modeled with four chemical states and the model predicts that it behaves as a pass band filter in the 10 Hz - 1 kHz range. C_LIO_LIIn conclusion, due to its activation/inactivation characteristics, RMA channel is a candidate to mediate cytosolic calcium signaling in response to mechano-stimulation. Its adaptation and filtering properties suggest its involvement in the transduction of high frequency mechanical stimulation such as those produced by insects vibrations. C_LI

plant biology↗

bioRxiv2026

Integrated genomic analyses reveal hierarchical organisation of quantitative resistance to biologically distinct soil-borne pathogens in wheat

Quantitative disease resistance is essential for durable crop protection, yet how numerous small-effect loci are organised across genomic and functional scales remains poorly understood. We investigated resistance to four distinct soil-borne pathogens in bread wheat (Triticum aestivum) by integrating genome-wide association mapping, linkage disequilibrium (LD)-defined hotspot analysis, comparative genomics, epistatic network analysis, haplotype analysis, functional annotation, transcriptomics and experimental evaluation. Resistance was highly polygenic and pathogen- and environment-dependent, with associated loci distributed across all three wheat subgenomes. Approximately two-thirds of hotspot components were shared among pathosystems, although evidence for excess sharing varied among comparisons and null models. Epistatic hubs were enriched within shared components, while interaction networks were dominated by interchromosomal and cross-subgenome interactions. Haplotype effects depended on local genomic context. Despite limited sharing of individual genes, convergence increased at the putative gene-family and broader defence-related functional levels. These findings support a hierarchical model in which pathogen- and environment-dependent resistance architectures are organised through partially shared genomic regions, local haplotypes and genetic interaction networks, with increasing convergence at broader functional levels. Limited transferability of individual marker associations in the independent panel further highlights the potential of multilocus breeding strategies that integrate favourable haplotypes, recurrent genomic regions, and diverse resistance mechanisms.

plant biology↗

bioRxiv2021

ESD1 Affects Seed Setting Rate in Rice by Controlling Embryo Sac Development

Seed setting rate is one of the critical factors that determine rice yield. Grain formation is a complex biological process, whose molecular mechanism is yet to be improved. Here we investigated the function of an OVATE family protein, Embryo Sac Development 1 (ESD1), in the regulation of seed setting rate in rice (Oryza sativa) by examining its loss-of-function mutants generated via CRISPR/Cas9 technology. ESD1 was predominantly expressed at stage 6 of panicle development, especially in the ovules. esd1 mutants displayed reduced seed setting rates with normal stamen development and pollen tube growth but abnormal pistil group. Investigation of embryo sacs revealed that during the mitosis of functional megaspores, some egg cells degraded during differentiation in esd1 mutants, thereby hindering subsequent fertilization process and reducing seed setting rate. In addition, the transcriptional level of OsAPC6, a reported embryo sac developing gene, was found to be significantly reduced in esd1 mutants. These results support that ESD1 is an important modulator of embryo sac development and seed setting rate in rice. Together, this finding demonstrates that ESD1 positively regulates the seed setting rate by controlling embryo sac development in rice, and has implications for the improvement of rice yield.

plant biology↗

bioRxiv2018

Chromatin immunoprecipitation-sequencing and RNA-sequencing for complex and low-abundance tree buds

Chromatin immunoprecipitation-sequencing (ChIP-seq) is a robust technique to study interactions between proteins, such as histones or transcription factors, and DNA. This technique in combination with RNA-sequencing (RNA-seq) is a powerful tool to better understand biological processes in eukaryotes. We developed a combined ChIP-seq and RNA-seq protocol for tree buds (Prunus avium L., Prunus persica L Batch, Malus x domestica Borkh.) that has also been successfully tested on Arabidopsis thaliana and Saccharomyces cerevisiae. Tree buds contain phenolic compounds that negatively interfere with ChIP and RNA extraction. In addition to solving this problem, our protocol is optimised to work on small amounts of material. Furthermore, one of the advantages of this protocol is that samples for ChIP-seq are cross-linked after flash freezing, making it possible to work on trees growing in the field and to perform ChIP-seq and RNA-seq on the same starting material. Focusing on dormant buds in sweet cherry, we explored the link between expression level and H3K4me3 enrichment for all genes, including a strong correlation between H3K4me3 enrichment at the DORMANCY-ASSOCIATED MADS-box 5 (PavDAM5) loci and its expression pattern. This protocol will allow analysis of chromatin and transcriptomic dynamics in tree buds, notably during its development and response to the environment.

plant biology↗

bioRxiv2022

A happy accident: a novel turfgrass reference genome

Poa pratensis, commonly known as Kentucky bluegrass, is a popular cool-season grass species used as turf in lawns and recreation areas globally. Despite its substantial economic value, a reference genome had not previously been assembled due to the genomes relatively large size and biological complexity that includes apomixis, polyploidy, and interspecific hybridization. We report here a fortuitous de novo assembly and annotation of a P. pratensis genome. Instead of sequencing the genome of a C4 grass, we accidentally sampled and sequenced tissue from a weedy P. pratensis whose stolon was intertwined with that of the C4 grass. The draft assembly consists of 6.09 Gbp with an N50 scaffold length of 65.1 Mbp, and a total of 118 scaffolds, generated using PacBio long reads and Bionano optical map technology. We annotated 256K gene models and found 58% of the genome to be composed of transposable elements. To demonstrate the applicability of the reference genome, we evaluated population structure and estimated genetic diversity in P. pratensis collected from three North American prairies, two in Manitoba, Canada and one in Colorado, USA. Our results support previous studies that found high genetic diversity and population structure within the species. The reference genome and annotation will be an important resource for turfgrass breeding and study of bluegrasses.

plant biology↗

bioRxiv2024

Integrated Dual-Channel Retrograde Signaling Directs Stress Responses by Degrading the HAT1/TPL/IMPalpha-9 Suppressor Complex and Activating CAMTA3

The intricate communication between plastids and the nucleus, shaping stress-responsive gene expression, has long intrigued researchers. This study combines genetics, biochemical analysis, cellular biology, and protein modeling to uncover how the plastidial metabolite MEcPP activates the stress-response regulatory hub known as the Rapid Stress Response Element (RSRE). Specifically, we identify the HAT1/TPL/IMP- 9 suppressor complex, where HAT1 directly binds to RSRE and its activator, CAMTA3, masking RSRE and sequestering the activator. Stress-induced MEcPP disrupts this complex, exposing RSRE and releasing CAMTA3, while enhancing Ca2+ influx and raising nuclear Ca2+levels crucial for CAMTA3 activation and the initiation of RSRE- containing gene transcription. This coordinated breakdown of the suppressor complex and activation of the activator highlights the dual-channel role of MEcPP in plastid-to- nucleus signaling. It further signifies how this metabolite transcends its expected biochemical role, emerging as a crucial initiator of harmonious signaling cascades essential for maintaining cellular homeostasis under stress. SummaryThis study uncovers how the stress-induced signaling metabolite MEcPP disrupts the HAT1/TPL/IMP-9 suppressor complex, liberating the activator CAMTA3 and enabling Ca2+ influx essential for CAMTA3 activation, thus orchestrating stress responses via repressor degradation and activator induction.

plant biology↗

bioRxiv2025

Bacillus velezensis EU07 suppresses Fusarium graminearum via transcriptomic reprogramming

Fusarium graminearum, the causal agent of Fusarium head blight, is a devastating pathogen of cereals worldwide. Biological control using Bacillus species has emerged as a sustainable strategy to suppress this pathogen, but the molecular basis of antagonism remains poorly understood. Here, we investigated the interaction between Bacillus velezensis EU07 and F. graminearum strain K1-4 through morphological assays and RNA-seq profiling. Microscopy revealed severe hyphal distortions including swelling and branching abnormalities, following exposure to EU07 cell pellets. Transcriptomic analysis after 6 h of treatment identified 1,264 differentially expressed genes (DEGs), with 732 downregulated and 532 upregulated. Genes encoding secondary metabolite biosynthesis enzymes, including trichothecene (TRI) cluster genes, cytochrome P450s, and transporters, were strongly repressed. Key metabolic pathways, such as amino acid catabolism and mitochondrial transporters (e.g., 2-oxoglutarate/malate carrier protein), also showed reduced expression. Conversely, genes associated with oxidative stress responses, detoxification, and membrane transport were induced, reflecting a compensatory survival strategy. These results demonstrate that EU07 disrupts F. graminearum both morphologically and at the transcriptional level, suppressing virulence-associated pathways while triggering stress adaptation. This dual impact highlights B. velezensis EU07 as a promising biocontrol agent and provides candidate fungal genes for targeted RNAi-based crop protection strategies.

plant biology↗

bioRxiv2026

Evaluating cacao pod resistance to black pod disease caused by three Phytophthora species

Black pod disease is a major threat to global cacao (Theobroma cacao L.) production. It is caused by multiple Phytophthora species with geographic ranges from regionally restricted to pantropical, and developing resistant cultivars requires screening against the diversity of these pathogens. This study evaluated the response of 25 cacao clones to 13 isolates of P. palmivora (eight), P. megakarya (two), and P. theobromicola (three) by nonwounding inoculation of detached pods. Lesion size and lesion expansion status were evaluated 4 days after inoculation. The linear mixed model included 178 combinations among 17 clones and 12 isolates, each represented by at least two biological replicate pods. Lesion size differed among clones (F(16, 449.2) = 25.16) and among isolates (F(11, 416.0) = 32.90), and the clone by isolate interaction was significant (F(150, 465.4) = 1.72; all P < 0.001). PENTAGONA-17 had the smallest estimated marginal lesion size (9.3 mm) and fitted values of 3.8 to 16.9 mm across 11 isolates representing all three species. ARF-31, ICS-47, POUND-7, and PMCT-73 had the next four smallest estimates, whereas NA-246 had the largest (36.7 mm). Isolate estimates varied more than fourfold, from 9.6 to 42.0 mm. Despite a marginal estimate of 12.9 mm, POUND-7 had a fitted estimate of 49.2 mm with P. megakarya GH34. The probability of lesion expansion also differed among clones and isolates (both P < 0.001). Together, these findings identify candidates for further resistance evaluation and show that multi-isolate screening provides a stronger basis for selecting materials for black pod resistance breeding.

plant biology↗

bioRxiv2022

Functional analysis of V2 protein of Beet curly top Iran virus

The geminivirus beet curly top Iran virus (BCTIV) is one of the main causal agents of the beet curly top disease in Iran and the newly established Becurtovirus genus type species. Although the biological features of known becurtoviruses are similar to those of curtoviruses, they only share a limited sequence identity, and no information is available on the function of their viral genes. In this work, we demonstrate that BCTIV V2, as the curtoviral V2, is also a strong local silencing suppressor in Nicotiana benthamiana and can delay the systemic silencing spreading, although it cannot block the cell-to-cell movement of the silencing signal to adjacent cells. BCTIV V2 shows the same subcellular localization as curtoviral V2, being detected in the nucleus and perinuclear region, and its ectopic expression from a PVX-derived vector also causes the induction of necrotic lesions in N. benthamiana like the ones produced during the HR, both at local and systemic levels. The results from the infection of N. benthamiana with a V2 BCTIV mutant showed that V2 is required for systemic infection but not for viral replication in a local infection. Considering all these results, we can conclude that BCTIV V2 is a functional homologue of curtoviral V2 and plays a crucial role in viral pathogenicity and systemic movement.

plant biology↗

bioRxiv2021

Auxin-responsive (phospho)proteome analysis reveals regulation of cell cycle and ethylene signaling during rice crown root development

The rice root system, which primarily consists of adventitious/crown roots (AR/CR) developed from the coleoptile base, is an excellent model system for studying shoot-to-root trans-differentiation process. We reveal global changes in protein and metabolite abundance, and protein phosphorylation in response to an auxin stimulus during CR development. Global proteome and metabolome analyses of developing crown root primordia (CRP) and emerged CRs uncovered that the biological processes associated with chromatin conformational change, gene expression, and cell cycle were translationally regulated by auxin signaling. Spatial gene expression pattern analysis of differentially abundant proteins disclosed their stage-specific dynamic expression pattern during CRP development. Further, our tempo-spatial gene expression and functional analyses revealed that auxin creates a regulatory feedback module during CRP development and activates ethylene biosynthesis exclusively during CRP initiation. Ethylene signaling promotes CR formation by repressing the cytokinin response regulator, OsRR2. Additionally, the (phospho)proteome analysis identified differential phosphorylation of the Cyclin-dependent kinase G-2 (OsCDKG;2), and cell wall proteins, in response to auxin signaling, suggesting that auxin-dependent phosphorylation may be required for cell cycle activation, and cell wall synthesis during root organogenesis. Thus, our study provides evidence for the translational and post-translational regulation during CRP trans-differentiation downstream of the auxin signaling pathway. HighlightGlobal (phospho)proteome and metabolic profiling of rice CRP and CRs uncover differential proteins and metabolites associated with gene expression, cell cycle, ethylene signaling and cell wall synthesis during CR development.

plant biology↗

bioRxiv2023

Optimizing ex-vitro one-step RUBY-equipped hairy root transformation in drug- and hemp-type Cannabis

Using synthetic biology techniques to engineer secondary metabolic pathways through hairy root transformation is one of the most advanced approaches used in research. In this study, we optimized an ex-vitro one-step hairy root transformation of the RUBY system in both drug- and hemp-type cannabis, shedding light on its potential applications in secondary metabolite production. Three different strains of A. rhizogenes including (A4, ARqual, and K599) were used. Significant variation in HR induction and transformation efficiency (TE) was observed based on A. rhizogenes strains and seed types. Drug-type seedlings exhibited the highest hairy root induction, increasing by 58.8% compared to hemp-type seedlings. Also, the A4 strain consistently demonstrated the highest transformation efficiency (75%) irrespective of genotype, while the ARqual strain yielded the lowest one (8.33%). In conclusion, our study is the first to present an ex-vitro one-step transformation of both hemp- and drug-type cannabis. In comparison to the in vitro method, our ex-vitro method is simpler, faster, and has a lower risk of contamination, making it an excellent choice for the efficient production of secondary metabolites in cannabis using the CRISPR/Cas system.

plant biology↗