An Integrated Approach for Biofortification of Carotenoids in Cowpea for Human Nutrition and Health.

Stress-resilient and highly nutritious legume crops can alleviate the burden of malnutrition and food security globally. Here, we focused on cowpea, a legume grain widely grown and consumed in regions at a high risk of micronutrient deficiencies, and we discussed the past and present research on carotenoid biosynthesis, highlighting different knowledge gaps and prospects for increasing this micronutrient in various edible parts of the crop. The literature survey revealed that, although carotenoids are important micronutrients for human health and nutrition, like in many other pulses, the potential of carotenoid biofortification in cowpea is still underexploited. We found that there is, to some extent, progress in the quantification of this micronutrient in cowpea; however, the diversity in content in the edible parts of the crop, namely, grains, pods, sprouts, and leaves, among the existing cowpea genetic resources was uncovered. Based on the description of the different factors that can influence carotenoid biosynthesis and accumulation in cowpea, we anticipated that an integrated use of omics in breeding coupled with mutagenesis and genetic engineering in a plant factory system would help to achieve a timely and efficient increase in carotenoid content in cowpea for use in the food systems in sub-Saharan Africa and South Asia.

Habitat range shift and prediction of the potential future distribution of Ricinodendron heudelotii (Baill.) Heckel in Benin (West Africa).

Ricinodendron heudelotii (Baill.) Heckel is an important nutraceutical reservoir. Its Sustainable exploitation requires information on its potential distribution in the current context of rapid population growth and climate change threats. This study aimed to map the suitable areas for its domestication and conservation under current and future climate conditions in Benin. Occurrence data were recorded and combined with the environmental layers of two climatic scenarios (optimistic RCP 4.5 and pessimistic RCP 8.5) following the biodiversity modelling approach (biomod2). Currently, about four percent (5082 Km2) of the country's area mainly located in the sub-humid and the humid zones were potentially suitable for R. heudelotii distribution. Under future climatic conditions the potentially suitable areas were mainly in the sub-humid zone, but almost all the highly suitable areas located in the humid zone will become medium suitable areas by the years 2055 and 2085 horizons. This study shows that, whatever the future climatic scenarios, R. heudelotii will substantially maintain the size of its range across the country. These findings allow undertaking anticipated actions to better adapt to the potential effects of climate change and to better guide policies for the conservation and development of forest resources.

Genetic and morphological diversity in populations of Annona senegalensis Pers. occurring in Western (Benin) and Southern (Mozambique) Africa.

Background: Understanding morpho-genetic diversity and differentiation of species with relatively large distributions is crucial for the conservation and sustainable management of their genetic resources. The present study focused on Annona senegalensis Pers., an important multipurpose wild plant, distributed exclusively in natural ecosystems but facing several threats. The study assessed the genetic and morphological diversity, structure, and differentiation of the species in populations from Western (Benin) and Southern (Mozambique) Africa. The material was evaluated to ascertain the environmental (climatic) determinants of the variation within this species. Methods: Four sub-populations comprised of 154 individuals were phenotyped based on nineteen plant, fruit, and leaf morphological traits and further genotyped using ten polymorphic nuclear microsatellite (nSSR) markers. Results: The results indicated strong differences in plant, fruit, and leaf morphological traits between Western and Southern populations. Furthermore, the studied populations were characterized by high genetic diversity, with an average genetic diversity index of 1.02. Western populations showed higher heterozygosity values (0.61-0.71) than Southern populations (0.41-0.49). Western and Southern populations were clearly differentiated into two different genetic groups, with further genetic subdivisions reflecting four sub-populations. Genetic variation between regions (populations) was higher (69.1%) than among (21.3%) and within (9.6%) sub-populations. Four distinct morphological clusters were obtained, which were strongly associated with the four genetic groups representing each sub-population. Climate, mainly precipitation and temperature indexes, explained the relatively higher variation found in morphological traits from Western (40.47%) in relation to Southern (27.98%) populations. Our study suggests that both environmental and genetic dynamics play an important role in the development of morphological variation in A. senegalensis.

Modeling current and future distribution patterns of Uvaria chamae in Benin (West Africa): Challenges and opportunities for its sustainable management.

Uvaria chamae is a wild shrub species widely used as a source for traditional medicine, food and fuel in West Africa. The species is threatened by uncontrolled harvesting of its roots for pharmaceutical applications and by the extension of agricultural land. This study assessed the role of environmental variables for the current distribution and the potential impact of climate change on the future spatial distribution of U. chamae in Benin. We used data related to climate, soil, topography and land cover to model the distribution of the species. Occurrence data were combined with six least correlated bioclimatic variables derived from the WorldClim database, data on soil layers (texture and pH) and topography (slope) obtained from the FAO world database and land cover from the DIVA-GIS site. Random Forest (RF), Generalized Additive Models (GAM), Generalized Linear Models (GLM) and the Maximum Entropy (MaxEnt) algorithm were used to predict the current and future (2050-2070) distribution of the species. Two climate change scenarios (SSP245 and SSP585) were considered for the future predictions. The results showed that climate (i.e., water availability) and soil type are the key predictors of the distribution of the species. Based on future climate projections, RF, GLM and GAM models predict that the Guinean-Congolian and Sudano-Guinean zones of Benin will remain suitable for U. chamae, while it will decline in these zones according to the MaxEnt model. These results call for a timely management effort for the species in Benin through its introduction into agroforestry systems to ensure the continuity of its ecosystem services.

Genetic Diversity and Association Analysis for Carotenoid Content among Sprouts of Cowpea (Vigna unguiculata L. Walp).

The development and promotion of biofortified foods plants are a sustainable strategy for supplying essential micronutrients for human health and nutrition. We set out to identify quantitative trait loci (QTL) associated with carotenoid content in cowpea sprouts. The contents of carotenoids, including lutein, zeaxanthin, and ß-carotene in sprouts of 125 accessions were quantified via high-performance liquid chromatography. Significant variation existed in the profiles of the different carotenoids. Lutein was the most abundant (58 ± 12.8 mg/100 g), followed by zeaxanthin (14.7 ± 3.1 mg/100 g) and ß-carotene (13.2 ± 2.9 mg/100 g). A strong positive correlation was observed among the carotenoid compounds (r ≥ 0.87), indicating they can be improved concurrently. The accessions were distributed into three groups, following their carotenoid profiles, with accession C044 having the highest sprout carotenoid content in a single cluster. A total of 3120 genome-wide SNPs were tested for association analysis, which revealed that carotenoid biosynthesis in cowpea sprouts is a polygenic trait controlled by genes with additive and dominance effects. Seven loci were significantly associated with the variation in carotenoid content. The evidence of variation in carotenoid content and genomic regions controlling the trait creates an avenue for breeding cowpea varieties with enhanced sprouts carotenoid content.

Food and medicinal uses of Annona senegalensis Pers.: a country-wide assessment of traditional theoretical knowledge and actual uses in Benin, West Africa.

BACKGROUND: The growing interest for more natural products in food and health industries has led to increasing research on traditional knowledge related to plants. While theoretical knowledge (TK) on the uses of a species informs on the wide spectrum of potential uses of that species, actual uses (AU) highlight their potential being actually used. Distinguishing between the two is important when reporting ethnobotanical studies. However, studies often equated AU and TK, sometimes misleading conclusions, and decision-making. This study assessed TK, AU, and difference between TK and AU of Annona senegalensis and how each is related to factors such as age, sex, sociolinguistic group, and main activity in Benin republic. METHODS: Data were collected through semi-structured individual interviews (n = 755) and analyzed using among others, relative frequency of citation (RFC), and use-value (UV). RESULTS: A total of 168 theoretical uses were recorded but only 92 were "actually" practiced, of which four were food and 88 medicinal uses. TK and AU were positively correlated. As expected, TK was also significantly higher than AU, indicating that some potential uses of the species are still not valued. Sociolinguistic group and main activity, not age and sex, were the main factors influencing TK, AU, and difference between TK and AU. The highest TK was found with Bariba sociolinguistic group and the highest AU with Otamari. Fruits (100%) and flowers (10%) were the most used organs for food, while leaves (40%) and roots (7%) were mostly used for medicinal purposes. The most common food uses were consumption of the ripe fruits (100%), and food seasoning with flowers (10%). The most cited diseases were malaria (28%) and intestinal worms (8%). CONCLUSIONS: The study illustrated the importance of differentiating between TK and AU. It documented  the wide range of the uses of A. senegalensis, while highlighting its most common uses, and the need to better valorize and sustainably manage the species.

Predicting the potential impacts of climate change on the endangered Caesalpinia bonduc (L.) Roxb in Benin (West Africa).

Caesalpinia bonduc (L.) Roxb is a medicinal plant with high therapeutic values but declared extinct in the wild in Benin. This study explored the potential distribution and climatic suitability of the species under the present-day and future conditions in Benin, based on two Representative Concentration Pathways (RCP4.5 and RCP8.5) at the 2055-time horizon. The occurrence data were recorded in the distribution area of the species in Benin. These data were supplemented with those from the Global Biodiversity Information Facility (GBIF, www.gbif.org) website and the literature. A total of 23 environmental variables (15 bioclimatic data and 8 biophysical data) were used. The Bioclimatic variables for temperature and humidity were downloaded from Africlim site at 1 km resolution. The biophysical variables concern population, elevation, slope, landcover, wetland, distance to river, soil and distance to dwellings data that are downloaded respectively from DIVA-GIS, ISRIC and SEDAC website at different resolution. A correlation test has been applied to eliminate the highly correlated variables (r ≥ 0.9) using Pearson correlation coefficient. Species distribution modelling data were processed using five algorithms namely Random Forest (RF), Boosted Regression Trees (BRT), Maximum entropy (MAXENT), Generalized Linear Models (GLM) and Generalized Additive Models (GAM). The results showed that all models performed well with the area under the curve (AUC) values greater than 0.9. The RF, GLM, and GAM models predicted an increase in the suitable areas for the cultivation of the species. BRT and MaxEnt showed a substantial decrease in the suitable areas based on the two scenarios but this reduction is more observed with the MaxEnt model. These results show that climate change and human pressures will have significant effects on the distribution of C. bonduc throughout Benin. Sustainable management measures are necessary for C. bonduc and should be integrated in development policies to preserve the population of the species from total extinction in Benin.

Effects of seed provenance, pre-treatment and mass on germinability and seedling growth of Balanites aegyptiaca (L.) Delile and Ricinodendron heudelotii (Bail.) Pierre in Benin (West Africa).

Balanites aegyptiaca (L.) Delile and Ricinodendron heudelotii (Bail.) Pierre are socioeconomically important species in sub-Saharan Africa. This study was conducted to assess the seed germinability and seedling growth of those species based on several treatments and to define proper conservation and domestication strategies in Benin. The seeds were randomly collected in their natural habitats. The experiment was conducted using a split-split plot design and the data was analyzed using the generalized linear mixed and survival models. The heaviest seeds (B. aegyptiaca seed mass ≥3 g and R. heudelotii ≥ 1.50 g) provided the highest germination rates (73.60 ± 5.19% and 62.50 ± 5.71%, respectively) when seeds were scarified with a hammer. For B. aegyptiaca seedlings, the seeds from the phytodistrict of North Borgou scarified with a hammer and the heaviest seeds showed the highest total height (36.43 ± 1.03 cm), basal diameter (2.84 ± 0.03 mm), the greatest number of leaves (32), and ramifications. The heaviest seeds of R. heudelotii had also the highest value for total height at the day-28 after sowing (26.73 ± 13.56 cm) until the day-105 (151.97 ± 6.37 cm). The heaviest seeds of R. heudelotii from the phytodistrict of Pobe showed the highest basal diameter (12.53 ± 1.47 mm) and the greatest number of leaves (14), with almost no ramification during the trial period. These findings constitute a step forward in upscaling the reproduction of these species for better contribution to economies while serving in restoration plans.

Diversity, population structure, and linkage disequilibrium among cowpea accessions.

Cowpea [Vigna unguiculata (L.) Walp] is a globally important food security crop. However, it is susceptible to pest and disease; hence, constant breeding efforts based on its diversity are required for its improvement. The present study aims to investigate the genetic diversity, population structure, and linkage disequilibrium (LD) among 274 cowpea accessions from different origins. A total of 3,127 single nucleotide polymorphism (SNP) markers generated using diversity array technology (DArT) was used. Population structure, neighbor-joining clustering, and principal component analyses indicated three subpopulations within the germplasm. Results of STRUCTURE analysis and discriminant analysis of principal components (DAPC) were complementary in assessing the structuration of the diversity among the germplasm, with the grouping of the accessions improved in DAPC. Genetic distances of 0.005-0.44 were observed among accessions. Accessions from western and central Africa, eastern and central Africa, and Asia were predominant and distributed across all subpopulations. The subpopulations had fixation indexes of 0.48-0.56. Analysis of molecular variance revealed that within subpopulation variation accounted for 81% of observed genetic variation in the germplasm. The subpopulations mainly consisted of inbred lines (inbreeding coefficient = 1) with common alleles, although they were from different geographical regions. This reflects considerable seed movement and germplasm exchange between regions. The LD was characterized by low decay for great physical distances between markers. The LD decay distance varied among chromosomes with the average distance of 80-100 kb across the genome. Thus, crop improvement is possible, and the LD will facilitate genome-wide association studies on quality attributes and critical agronomic traits in cowpea.

Species richness, cultural importance, and prioritization of wild spices for conservation in the Sudano-Guinean zone of Benin (West Africa).

BACKGROUND: Spices have always been used for their flavor-enhancement characteristics and for their medicinal properties. In Benin, scientific research on spices is scarce, despite their importance in the local population's daily needs. This study investigated the diversity of wild spices and documented the associated traditional knowledge that can be used for their valuation, domestication, and sustainable management in the Sudano-Guinean Zone of Benin. METHODS: Data were collected during field expeditions using semi-structured interviews in ten localities across the three phytodistricts of the zone. Species richness and Shannon's diversity index were estimated using species accumulation curves. Use report (UR), cultural importance, use value (UV) index, and informant consensus factor (Fic) were used to assess traditional knowledge on wild species, their local importance, and informants' agreement among sociolinguistic groups. Priority wild spices were finally identified using an approach combining eight criteria (native status, economic value, ethnobotanical value, global distribution, national distribution, in-situ and ex-situ conservation status, legislation, and threats assessment) in four prioritization methods (point scoring procedure, point scoring procedure with weighting, compound ranking system, and binomial ranking system). RESULTS: A total of 14 species, belonging to 12 genera and 9 families, were inventoried. The most prominent families were Zingiberaceae (21.43%), Annonaceae (21.43%), and Rutaceae (14.29%). More than 200 specific uses were reported, with the Tchabè people holding the greatest level of knowledge (70 uses; UR = 5.70 ± 0.33). The culturally most important spices differed among sociolinguistic groups. Most of the informants agree on the use of the species among (Fic = 0.72-0.98) and across the considered use categories (Fic = 0.88-0.99). The highest UV were registered for Aframomum alboviolaceum (UV = 0.93), Lippia multiflora (UV = 0.76), and Aframomum angustifolium (UV = 0.18). Overall, people perceived wild spices as declining due to agriculture, grazing, and drought. Five species, A. alboviolaceum, L. multiflora, Monodora tenuifolia, Xylopia aethiopica, and Z. zanthoxyloides, were the most prioritized for conservation. CONCLUSIONS: This study provides information relevant for the implementation of conservation and domestication actions of wild spices in Benin. Priority species could be integrated into traditional agroforestry systems (e.g., home gardens). However, for this to be effective, further research should be undertaken on morphological and genetic diversity and propagation methods of priority wild spices.

Traditional knowledge and cultural importance of Borassus aethiopum Mart. in Benin: interacting effects of socio-demographic attributes and multi-scale abundance.

BACKGROUND: Eliciting factors affecting distribution of traditional knowledge (TK) and cultural importance of plant resources is central in ethnobiology. Socio-demographic attributes and ecological apparency hypothesis (EAH) have been widely documented as drivers of TK distribution, but their synergistic effect is poorly documented. Here, we focused on Borassus aethiopum, a socio-economic important agroforestry palm in Africa, analyzing relationships between the number of use-reports and cultural importance on one hand, and informant socio-demographic attributes (age category and gender) on the other hand, considering the EAH at multi-scale contexts. Our hypothesis is that effects of socio-demographic attributes on use-reports and cultural importance are shaped by both local (village level) and regional (chorological region level) apparency of study species. We expected so because distribution of knowledge on a resource in a community correlates to the versatility in the resource utilization but also connections among communities within a region. METHODS: Nine hundred ninety-two face-to-face individual semi-structured interviews were conducted in six villages of low versus high local abundance of B. aethiopum spanning three chorological regions (humid, sub-humid and semi-arid) also underlying a gradient of increasing distribution and abundance of B. aethiopum. Number of use-reports and score of importance of uses of B. aethiopum were recorded in six use-categories including medicine, food, handcraft, construction, firewood, and ceremonies and rituals. Data were analyzed using Poisson and ordered logistic models. RESULTS: Informants listed 121 uses for B. aethiopum: medicine (66 uses), handcraft (16 uses), food (16 uses), construction (12 uses), firewood (6 uses), and ceremonies and rituals (5 uses); but food use was the most culturally important use (2.45 ± 0.03), followed by construction (0.61 ± 0.03), medicinal (0.57 ± 0.03) and handcraft (0.56 ± 0.03), firewood (0.29 ± 0.02), and ceremonies and rituals (0.03 ± 0.01). Food use was the most important for women who were specialized in hypocotyls and fruits collection for commercialization. Men valued more the species for handcrafting, construction, and medicine. The number of use-reports was significantly dependent on age category and gender, and differences between age categories (young, adult, and old) in particular were dependent upon local and regional apparency. In particular, discrepancies among age categories were higher in areas of low abundance and distribution, which may be linked to different speed in the process of knowledge acquisition. In areas of low abundance, the species past abundance was also found instrumental in understanding current knowledge distribution. CONCLUSION: Findings suggest that studies aiming at understanding relationship between current TK and cultural importance of a resource on one hand and socio-demographic attributes on the other hand should consider the resource current local and regional apparency but further its local past abundance. The study also confirms that B. aethiopum is a socio-economic important species in Benin.

Functional diversity of home gardens and their agrobiodiversity conservation benefits in Benin, West Africa.

BACKGROUND: Understanding the functional diversity of home gardens and their socio-ecological determinants is essential for mainstreaming these agroforestry practices into agrobiodiversity conservation strategies. This paper analyzed functional diversity of home gardens, identified the socio-ecological drivers of functions assigned to them, and assessed the agrobiodiversity benefits of home gardens functions. METHODS: Using data on occurring species in home garden (HG) and functions assigned to each species by the gardeners, the study combined clustering and discriminant canonical analyses to explore the functional diversity of 360 home gardens in Benin, West Africa. Next, multinomial logistic models and chi-square tests were used to analyze the effect of socio-demographic characteristics of gardeners (age, gender, and education level), agro-ecological zones (humid, sub-humid, and semi-arid), and management regime (single and multiple managers) on the possession of a functional type of home gardens. Generalized linear models were used to assess the effect of the functions of home gardens and the determinant factor on their potential in conserving agrobiodiversity. RESULTS: Seven functional groups of home gardens, four with specific functions (food, medicinal, or both food and medicinal) and three with multiple functions (more than two main functions), were found. Women owned most of home gardens with primarily food plant production purpose while men owned most of home gardens with primarily medicinal plant production purposes. Finding also showed that multifunctional home gardens had higher plant species diversity. Specifically, crops and crop wild relatives occurred mainly in home gardens with food function while wild plant species were mostly found in home gardens with mainly medicinal function. CONCLUSIONS: Home gardening is driven by functions beyond food production. These functions are mostly related to direct and extractive values of home gardens. Functions of home gardens were gendered, with women mostly involved in home food gardens, and contribute to maintenance of crops and crop wild relatives while men were mostly home medicinal gardeners and contribute to the maintenance of wild plant species in home gardens. Although multiple functional home gardens were related to higher plant diversity, there was no guarantee for long-term maintenance of plant species in home gardens.

Factors affecting home gardens ownership, diversity and structure: a case study from Benin.

BACKGROUND: Home gardens (HGs) provide perspectives for conservation of plant genetic resources while contributing to improving livelihoods. However, knowledge of local factors shaping their ownership, plant diversity (PD) and structure is still limited especially in West-Africa, where food insecurity is acute. This is critical to ensure effective mainstreaming of HGs into future biodiversity conservation and food production policies. METHODS: Socio-economic and PD data were obtained from individual interviews (n = 470) and gardens inventories (n = 235) spanning humid, sub-humid and semi-arid zones of Benin. Generalised Linear Models, Hierarchical Cluster Analysis, Principal Component Analysis and Simple Correspondence Analysis were performed to examine socio-economic characteristics (age, gender, education level and main economic activity) affecting HGs ownership, and their effect coupled with intrinsic HGs characteristics (size, age) on PD and structure within HGs, across contrasting bio-geographical regions. RESULTS: HG ownership was significantly dependent upon a complex relationship between age, gender and education level of the farmers. The probability to own HG increased with age with an early involvement in home gardening for women. Similarly, with increasing age, it was more likely to find a male owner than a female owner among the uneducated informants and those of primary school. Inversely, it was more likely to find female owner than a male owner among secondary school level or more. PD increased with increasing owner age and size of the HG. Larger and more diversified HGs were found in sub-humid and semi-arid zones while smaller and less diversified HGs were encountered in the humid zone. HGs were multi-layered. Based on the prevailing plant groups, three categories of HG were distinguished: Herb based gardens, Herb and Shrub/Trees based gardens, and Palm and Liana based gardens. Their prevalence was dependent upon bio-geographical zones and HG owner socio-economic characteristics, with herbs based HGs being mainly associated to women. CONCLUSION: Results suggest effects of complex interactions between socio-economic factors on HG ownership, and influence of these effects combined with intrinsic characteristics of HGs on PD. The early involvement of women in home gardening and their particular interest in herbs and shrubs are important assets for future conservation strategies based on HG and food production. Interventions are required to interfere with declining PD in HG across generations to accommodate multiple ecosystem service benefits.