The Gynandropsis gynandra genome provides insights into whole-genome duplications and the evolution of C4 photosynthesis in Cleomaceae.

Gynandropsis gynandra (Cleomaceae) is a cosmopolitan leafy vegetable and medicinal plant, which has also been used as a model to study C4 photosynthesis due to its evolutionary proximity to C3 Arabidopsis (Arabidopsis thaliana). Here, we present the genome sequence of G. gynandra, anchored onto 17 main pseudomolecules with a total length of 740 Mb, an N50 of 42 Mb and 30,933 well-supported gene models. The G. gynandra genome and previously released genomes of C3 relatives in the Cleomaceae and Brassicaceae make an excellent model for studying the role of genome evolution in the transition from C3 to C4 photosynthesis. Our analyses revealed that G. gynandra and its C3 relative Tarenaya hassleriana shared a whole-genome duplication event (Gg-α), then an addition of a third genome (Th-α, +1×) took place in T. hassleriana but not in G. gynandra. Analysis of syntenic copy number of C4 photosynthesis-related gene families indicates that G. gynandra generally retained more duplicated copies of these genes than C3T. hassleriana, and also that the G. gynandra C4 genes might have been under positive selection pressure. Both whole-genome and single-gene duplication were found to contribute to the expansion of the aforementioned gene families in G. gynandra. Collectively, this study enhances our understanding of the polyploidy history, gene duplication and retention, as well as their impact on the evolution of C4 photosynthesis in Cleomaceae.

Phenotypic variation in biomass and related traits among four generations advanced lines of Cleome (Gynandropsis gynandra L. (Briq.)).

Gynandropsis gynandra (spider plant) is an African traditional leafy vegetable rich in minerals, vitamins and health-promoting compounds with potential for health promotion, micronutrients supplementation and income generation for stakeholders, including pharmaceutical companies. However, information on biomass productivity is limited and consequently constrains breeders' ability to select high-yielding genotypes and end-users to make decisions on suitable cultivation and production systems. This study aimed to assess the phenotypic variability in biomass and related traits in a collection of G. gynandra advanced lines to select elite genotypes for improved cultivar development. Seventy-one advanced lines selected from accessions originating from Asia, West Africa, East Africa and Southern Africa were evaluated over two years with two replicates in a greenhouse using a 9 x 8 alpha lattice design. Significant statistical differences were observed among lines and genotype origins for all fourteen biomass and related traits. The results revealed three clusters, with each cluster dominated by lines derived from accessions from Asia (Cluster 1), West Africa (Cluster 2), and East/Southern Africa (Cluster 3). The West African and East/Southern African groups were comparable in biomass productivity and superior to the Asian group. Specifically, the West African group had a low number of long primary branches, high dry matter content and flowered early. The East/Southern African group was characterized by broad leaves, late flowering, a high number of short primary branches and medium dry matter content and was a candidate for cultivar release. The maintenance of lines' membership to their group of origin strengthens the hypothesis of geographical signature in cleome diversity and genetic driver of the observed variation. High genetic variance, broad-sense heritability and genetic gains showed the potential to improve biomass yield and related traits. Significant and positive correlations among biomass per plant, plant height, stem diameter and leaf size showed the potential of simultaneous and direct selection for farmers' desired traits. The present results provide insights into the diversity of spider plant genotypes for biomass productivity and represent key resources for further improvement in the species.

Leaf elemental composition analysis in spider plant [Gynandropsis gynandra L. (Briq.)] differentiates three nutritional groups.

Understanding the genetic variability within a plant species is paramount in implementing a successful breeding program. Spider plant (Gynandropsis gynandra) is an orphan leafy vegetable and an extraordinary source of vitamins, secondary metabolites and minerals, representing an important resource for combatting malnutrition. However, an evaluation of the leaf elemental composition, using a worldwide germplasm collection to inform breeding programs and the species valorization in human nutrition is still lacking. The present study aimed to profile the leaf elemental composition of G. gynandra and depict any potential geographical signature using a collection of 70 advanced lines derived from accessions originating from Asia and Eastern, Southern and West Africa. The collection was grown in a greenhouse using a 9 × 8 alpha lattice design with two replications in 2020 and 2021. Inductively coupled plasma-optical emission spectrometry was used to profile nine minerals contents. A significant difference (p < 0.05) was observed among the lines for all nine minerals. Microelements such as iron, zinc, copper and manganese contents ranged from 12.59-430.72, 16.98-166.58, 19.04-955.71, 5.39-25.10 mg kg-1 dry weight, respectively, while the concentrations of macroelements such as potassium, calcium, phosphorus and magnesium varied in the ranges of 9992.27-49854.23, 8252.80-33681.21, 3633.55-14216.16, 2068.03-12475.60 mg kg-1 dry weight, respectively. Significant and positive correlations were observed between iron and zinc and calcium and magnesium. Zinc, calcium, phosphorus, copper, magnesium, and manganese represented landmark elements in the genotypes. Eastern and Southern African genotypes were clustered together in group 1 with higher phosphorus, copper and zinc contents than Asian and West African lines, which clustered in group 2 and were characterized by higher calcium, magnesium and manganese contents. An additional outstanding group 3 of six genotypes was identified with high iron, zinc, magnesium, manganese and calcium contents and potential candidates for cultivar release. The genotype × year interaction variance was greater than the genotypic variance, which might translate to phenotypic plasticity in the species. Broad-sense heritability ranged from low to high and was element-specific. The present results reveal the leaf minerals diversity in spider plant and represent a baseline for implementing a minerals-based breeding program for human nutrition.

Natural variation in specialised metabolites production in the leafy vegetable spider plant (Gynandropsis gynandra L. (Briq.)) in Africa and Asia.

The improvement and promotion of leafy vegetables, used for both food and medicine, benefits greatly from detailed knowledge of their health-promoting specialised metabolites. In the present study, we investigated the global metabolite variation in the leaves of 48 accessions of the leafy vegetable Gynandropsis gynandra using two complementary analytical platforms: liquid-chromatography mass spectrometry (LC-MS) for an untargeted comparison of non-volatile semi-polar metabolites and gas-chromatography mass spectrometry (GC-MS) for an untargeted comparison of volatile metabolites. Our results revealed large variation in 936 semi-polar compounds including flavonoids, terpene glycosides, glucosinolates and various phenolic compounds. Unsupervised multivariate analysis indicated the variation in levels of the semi-polar metabolites was mainly driven by geography, with accessions from both West Africa and Asia forming a group clearly separated from East African accessions. Detected volatile metabolites included various sesquiterpenes, aldehydes, ketones, and sulphur-containing isothiocyanates. Variation in these compounds was however not geographically specific, but most likely linked to the taste and aroma of the leaves. The relative abundance in glucosinolates and isothiocyanates in the leaves allowed the clustering of accessions into two main groups that could be used for further plant-herbivore interaction studies. This study revealed both the broad spectrum of phytochemicals present in Gynandropsis gynandra leaves and the substantial variation in metabolite profiles among accessions from different regions of the world. Our results provide a basis for the development of breeding programs aiming at improving the levels of specialised metabolites in this tropical leafy vegetable for increased resistance against pests and diseases and improved human health.

Origin-dependence of variation in seed morphology, mineral composition and germination percentage in Gynandropsis gynandra (L.) Briq. accessions from Africa and Asia.

BACKGROUND: Spider plant [Gynandropsis gynandra (L.) Briq.], an economically promising African leafy vegetable, characterized for leaf yield components and nutritive quality, exhibits poor seed germination that hinders a wider expansion of the crop in urban and periurban horticultural systems. So far, there is little information pertaining to seed morphological traits and mineral elements content that may be associated with higher seed germination. This research investigated the hypothesis that spider plants from different geographical areas exhibited differences in seed mineral composition, morphological traits, and germination capacity. To this end, twenty-nine accessions of Gynandropsis gynandra from West and East-Southern Africa, and Asia were screened for variation in seed size (area, perimeter, length, width), 10-seed weight, mean germination time, germination percentage and mineral content variations. The scanning electron microscopy (SEM), light microscopy and energy dispersive spectroscopy (EDS) solution were used to study seed morphology and mineral composition. RESULTS: We show for the first time the external and internal structure of the seeds of Gynandropsis gynandra and measured eight mineral elements, including carbon (C), oxygen (O), magnesium (Mg), aluminium (Al), phosphorus (P), sulphur (S), potassium (K) and calcium (Ca). The accessions differed significantly (p < 0.001) with respect to seed size (area, perimeter, length, width), 10-seed weight, mean germination time and germination percentage. The hierarchical cluster analysis based on fourteen variables grouped the accessions into three distinct clusters, partially dependent on their geographical origin. Asian accessions exhibited smaller seeds and recorded higher values in terms of germination percentage. West African accessions had bigger seeds but with lower germination percentage. Variation in minerals such as potassium, carbon, and calcium content showed different patterns according to geographical origins. CONCLUSION: Smaller seeds in G. gynandra exhibited better germination capacity. The Asian germplasm is a potential source of cultivars with a higher germination percentage for improving seed quality in the species.

Association between vitamin content, plant morphology and geographical origin in a worldwide collection of the orphan crop Gynandropsis gynandra (Cleomaceae).

MAIN CONCLUSION: The variability in nutrient content and morphology in Gynandropsis gynandra is associated with the geographic origin of the accessions and provides a basis for breeding for higher levels of vitamin C, carotenoids or tocopherols in higher-yielding cultivars. We examined the variation in carotenoids, tocopherols and ascorbic acid as well as morphological traits in a worldwide germplasm of 76 accessions of the orphan leafy vegetable Gynandropsis gynandra (Cleomaceae) using greenhouse experiments and high-performance liquid chromatography analysis. The levels of carotenoids and tocopherols accumulating in the leaves varied significantly across accessions and were linked with the geographical origin and morphological variation. The main carotenoids included lutein, ß-carotene, α-carotene and violaxanthin. A twofold to threefold variation was observed for these compounds. The main tocopherols detected were α-tocopherol and γ-tocopherol with a 20-fold variation. A ninefold variation in vitamin C concentration and independent of geographical origin was observed. Overall, the accessions were grouped into three clusters based on variation in nutrient content and morphology. West African accessions were short plants with small leaves and with high tocopherol contents and relatively low carotenoid contents, Asian accessions were short plants with broad leaves and with relatively low carotenoid and high tocopherol contents, while East-Southern African plants were tall with high contents of both carotenoids and chlorophylls and low tocopherol contents. Carotenoids were positively correlated with plant height as well as foliar and floral traits but negatively correlated with tocopherols. The absence of a significant correlation between vitamin C and other traits indicated that breeding for high carotenoids or tocopherols content may be coupled with improved leaf yield and vitamin C content. Our study provides baseline information on the natural variation available for traits of interest for breeding for enhanced crop yield and nutrient content in Gynandropsis gynandra.

Natural Variation within a Species for Traits Underpinning C4 Photosynthesis.

Engineering C4 photosynthesis into C3 crops could substantially increase their yield by alleviating photorespiratory losses. This objective is challenging because the C4 pathway involves complex modifications to the biochemistry, cell biology, and anatomy of leaves. Forward genetics has provided limited insight into the mechanistic basis of these properties, and there have been no reports of significant quantitative intraspecific variation of C4 attributes that would allow trait mapping. Here, we show that accessions of the C4 species Gynandropsis gynandra collected from locations across Africa and Asia exhibit natural variation in key characteristics of C4 photosynthesis. Variable traits include bundle sheath size and vein density, gas-exchange parameters, and carbon isotope discrimination associated with the C4 state. The abundance of transcripts encoding core enzymes of the C4 cycle also showed significant variation. Traits relating to water use showed more quantitative variation than those associated with carbon assimilation. We propose that variation in these traits likely adapted the hydraulic system for increased water use efficiency rather than improving carbon fixation, indicating that selection pressure may drive C4 diversity in G. gynandra by modifying water use rather than photosynthesis. The accessions analyzed can be easily crossed and produce fertile offspring. Our findings, therefore, indicate that natural variation within this C4 species is sufficiently large to allow genetic mapping of key C4 traits and regulators.

A roadmap for breeding orphan leafy vegetable species: a case study of Gynandropsis gynandra (Cleomaceae).

Despite an increasing awareness of the potential of "orphan" or unimproved crops to contribute to food security and enhanced livelihoods for farmers, coordinated research agendas to facilitate production and use of orphan crops by local communities are generally lacking. We provide an overview of the current knowledge on leafy vegetables with a focus on Gynandropsis gynandra, a highly nutritious species used in Africa and Asia, and highlight general and species-specific guidelines for participatory, genomics-assisted breeding of orphan crops. Key steps in genome-enabled orphan leafy vegetables improvement are identified and discussed in the context of Gynandropsis gynandra breeding, including: (1) germplasm collection and management; (2) product target definition and refinement; (3) characterization of the genetic control of key traits; (4) design of the 'process' for cultivar development; (5) integration of genomic data to optimize that 'process'; (6) multi-environmental participatory testing and end-user evaluation; and (7) crop value chain development. The review discusses each step in detail, with emphasis on improving leaf yield, phytonutrient content, organoleptic quality, resistance to biotic and abiotic stresses and post-harvest management.