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Abstract Plants regenerated from seedling explants (hypocotyls and cotyledons) of the Solanaceae family membersPhysalis grisea(groundcherry),Solanum lycopersicum(tomato), andSolanum prinophyllum(forest nightshade) were used to determine the in vitro culture parameters that contribute to the incidence in polyploidization of tissue culture-derived plants (regenerants) from these species. We examined the possible effects of zeatin concentration in the plant regeneration medium, explant source, and species. Plants were grown to maturity under greenhouse conditions, pollen was collected and germinated. Flow cytometry analysis verified the utility of the pollen germination method for determining differences in ploidy, which was based on the number of pollen tubes produced with one tube representing diploid and two indicating polyploid. As for zeatin concentration, we assessed the effect of our standard method of initiation on medium containing 2 mg/l followed by 1 mg/l 2 weeks after culture initiation in comparison with 0.25, 0.5, and 1 mg/l throughout the culture lifetime. There were no major correlations for zeatin concentration on ploidy status across the species except for plants regenerated fromS. lycopersicumhypocotyl explants where the percentage of polyploid regenerants increased with increasing concentrations. As for species and explant effects,P. griseaplants regenerated from hypocotyl explants had the highest percentage of polyploid plants at 81% compared to 43% and 35% forS. lycopersicumandS. prinophyllum, respectively. From cotyledons, 8% ofS. lycopersicumand 20% ofS. prinophyllumwere polyploid. A comparison withP. griseacould not be made because cotyledon explants do not regenerate on zeatin-containing medium. The results indicated the incidence of polyploidization cannot be generalized for zeatin concentration, however, an influence of explant type and species was observed. Effects of increased ploidy on plant morphology were primarily larger flower and seed size; however, no significant differences were observed in plant or fruit size. 

Abstract This report provides an overview of the content and data collected from the “Successes, Challenges, and Opportunities Plant Transformation Research in Africa” panel discussion. Organized by PlantGENE, this event brought together scientists and stakeholders across the globe to examine the complex challenges and emerging opportunities in plant transformation research in laboratories across Africa. The discussion, rooted in insights from a panel of six leading scientists, highlights critical issues including restrictive regulatory environments, prohibitive costs, and the inconsistent availability of essential research materials. Additionally, the pervasive “brain drain” phenomenon, where skilled researchers leave the continent for better opportunities, exacerbates the difficulties faced by African scientists. Despite these challenges, the report also identifies significant advancements, particularly in the growing recognition of African leadership within universities and national agricultural research systems (NARS). These institutions, supported by highly skilled faculty and motivated graduate students, are producing high-quality research that contributes to global scientific knowledge. The panelists emphasized the necessity of creating an environment that encourages African scientists to remain on the continent and address local challenges through innovative research. Strengthening intra-African networks and fostering collaborations with the global scientific community are proposed as essential strategies to achieve this. This report underscores the critical need for substantial investments from both global and African organizations, working with African governments, to support these efforts. Furthermore, it calls for science-based decision-making and fair regulatory frameworks to align with unique opportunities and risks associated with technological advancements in Africa. This paper details the observations of six panelists and analyzes the results of attendee surveys in order to document these challenges and opportunities while advocating for sustained investment and strategic partnerships to build a thriving bioeconomy across Africa. 

Societal Impact StatementGroundcherry (Physalis grisea) is a plant species grown for its flavorful fruit. The fruit drops from the plant, hence the common name groundcherry. This makes harvest cumbersome and puts the fruit at risk for carrying soil‐borne pathogens, therefore making them unsellable. Furthermore, insects often damage the plants, reducing yield. Advances in gene editing offer promise for addressing these issues and aiding home gardeners and farmers. Improvement will expand access to this nutritious fruit, rich in potassium, vitamin C, and antioxidants. Additionally, studies of its biology could serve as a model for improving other fruiting plants, particularly underutilized species. SummaryP. griseais an underutilized, semidomesticated fruit crop with rising agronomic value. Several resources have been developed for its use in fundamental biological research, including a plant transformation system and a high‐quality reference genome. Already,P. griseahas been used as a model to investigate biological phenomena including inflated calyx syndrome and gene compensation.P. griseahas also been used to demonstrate the potential of fast‐tracking domestication trait improvement through approaches such as CRISPR/Cas9 gene editing. This work has led to thePhysalisImprovement Project, which relies on reverse genetics to understand the mechanisms that underlie fruit abscission and plant–herbivore interactions to guide approaches for improvement of undesirable characteristics. CRISPR/Cas9 gene editing has been used to targetP. griseagenes that are suspected to act in fruit abscission, particularly orthologs of those that are reported in tomato abscission zone development. A similar approach is being taken to targetP. griseagenes involved in the withanolide biosynthetic pathway to determine the impact of withanolides on plant–herbivore interactions. Results from these research projects will lead to a greater understanding of important biological processes and will also generate knowledge needed to develop cultivars with reduced fruit drop and increased resistance to insect herbivory. 

Abstract Pan-genomics and genome-editing technologies are revolutionizing breeding of global crops^1,2. A transformative opportunity lies in exchanging genotype-to-phenotype knowledge between major crops (that is, those cultivated globally) and indigenous crops (that is, those locally cultivated within a circumscribed area)^3–5to enhance our food system. However, species-specific genetic variants and their interactions with desirable natural or engineered mutations pose barriers to achieving predictable phenotypic effects, even between related crops^6,7. Here, by establishing a pan-genome of the crop-rich genusSolanum⁸and integrating functional genomics and pan-genetics, we show that gene duplication and subsequent paralogue diversification are major obstacles to genotype-to-phenotype predictability. Despite broad conservation of gene macrosynteny among chromosome-scale references for 22 species, including 13 indigenous crops, thousands of gene duplications, particularly within key domestication gene families, exhibited dynamic trajectories in sequence, expression and function. By augmenting our pan-genome with African eggplant cultivars⁹and applying quantitative genetics and genome editing, we dissected an intricate history of paralogue evolution affecting fruit size. The loss of a redundant paralogue of the classical fruit size regulatorCLAVATA3(CLV3)^10,11was compensated by a lineage-specific tandem duplication. Subsequent pseudogenization of the derived copy, followed by a large cultivar-specific deletion, created a single fusedCLV3allele that modulates fruit organ number alongside an enzymatic gene controlling the same trait. Our findings demonstrate that paralogue diversifications over short timescales are underexplored contingencies in trait evolvability. Exposing and navigating these contingencies is crucial for translating genotype-to-phenotype relationships across species. 

Abstract Hornworts are a deeply diverged lineage of bryophytes and a sister lineage to mosses and liverworts. Hornworts have an array of unique features that can be leveraged to illuminate not only the early evolution of land plants, but also alternative paths for nitrogen and carbon assimilation via cyanobacterial symbiosis and a pyrenoid-based CO2-concentrating mechanism (CCM), respectively. Despite this, hornworts are one of the few plant lineages with limited available genetic tools. Here we report an efficient biolistics method for generating transient expression and stable transgenic lines in the model hornwort, Anthoceros agrestis. An average of 569 (±268) cells showed transient expression per bombardment, with green fluorescent protein expression observed within 48–72 h. A total of 81 stably transformed lines were recovered across three separate experiments, averaging six lines per bombardment. We followed the same method to transiently transform nine additional hornwort species, and obtained stable transformants from one. This method was further used to verify the localization of Rubisco and Rubisco activase in pyrenoids, which are central proteins for CCM function. Together, our biolistics approach offers key advantages over existing methods as it enables rapid transient expression and can be applied to widely diverse hornwort species. 

SUMMARY Plant transformation is an important part of plant research and crop improvement. Transformation methods remain complex, labor intensive, and inefficient. PlantGENE is a community of scientists from academia, industry, non‐profit research institutes, and government organizations working to improve plant transformation. PlantGENE hosts virtual training, interactive webinars, and a website with career opportunities, directories, and more. The plant science community has shown great interest and support for PlantGENE. 

An enduring question in evolutionary biology concerns the degree to which episodes of convergent trait evolution depend on the same genetic programs, particularly over long timescales. In this work, we genetically dissected repeated origins and losses of prickles—sharp epidermal projections—that convergently evolved in numerous plant lineages. Mutations in a cytokinin hormone biosynthetic gene caused at least 16 independent losses of prickles in eggplants and wild relatives in the genusSolanum. Homologs underlie prickle formation across angiosperms that collectively diverged more than 150 million years ago, including rice and roses. By developing newSolanumgenetic systems, we leveraged this discovery to eliminate prickles in a wild species and an indigenously foraged berry. Our findings implicate a shared hormone activation genetic program underlying evolutionarily widespread and recurrent instances of plant morphological innovation. 

Abstract An enduring question in evolutionary biology concerns the degree to which episodes of convergent trait evolution depend on the same genetic programs, particularly over long timescales. Here we genetically dissected repeated origins and losses of prickles, sharp epidermal projections, that convergently evolved in numerous plant lineages. Mutations in a cytokinin hormone biosynthetic gene caused at least 16 independent losses of prickles in eggplants and wild relatives in the genusSolanum. Strikingly, homologs promote prickle formation across angiosperms that collectively diverged over 150 million years ago. By developing newSolanumgenetic systems, we leveraged this discovery to eliminate prickles in a wild species and an indigenously foraged berry. Our findings implicate a shared hormone-activation genetic program underlying evolutionarily widespread and recurrent instances of plant morphological innovation. 

Summary Despite their key phylogenetic position and their unique biology, hornworts have been widely overlooked. Until recently there was no hornwort model species amenable to systematic experimental investigation.Anthoceros agrestishas been proposed as the model species to study hornwort biology.We have developed anAgrobacterium‐mediated method for the stable transformation ofA. agrestis, a hornwort model species for which a genetic manipulation technique was not yet available.High transformation efficiency was achieved by using thallus tissue grown under low light conditions. We generated a total of 274 transgenicA. agrestislines expressing the β‐glucuronidase (GUS), cyan, green, and yellow fluorescent proteins under control of the CaMV 35S promoter and several endogenous promoters. Nuclear and plasma membrane localization with multiple color fluorescent proteins was also confirmed.The transformation technique described here should pave the way for detailed molecular and genetic studies of hornwort biology, providing much needed insight into the molecular mechanisms underlying symbiosis, carbon‐concentrating mechanism, RNA editing and land plant evolution in general.