In vitro transformation of pearl millet (Pennisetum glaucum (L). R. BR.): Selection of chlorsulfuron-resistant plants and long term expression of the gus gene under the control of the emu promoter

The plasmids pULGU1 and pEmuGN were introduced by biolistics in embryogenic cell suspensions of pearl millet, Pennisetum glaucum. The plasmid pULGU1 contained the mutant acetolactate synthase (ALS) gene of Arabidopsis thaliana, responsible for resistance to the chlorsulfuron herbicide. The plasmid pEmuGN carried the reporter β-glucuronidase (GUS) gene of Escherichia coli. Two months after bombardment of the cells, transformed calli were selected on Murashige and Skoog medium containing 30 nM chlorsulfruron, a completely concentration inhibitory for non-transformed cells. A concentration of 200 nM chlorsufuron did not affect the growth of these selected resistant calli. Genomic DNA of resistant cells presented an electrophoretic pattern revealing the integration of the mutant ALS gene into the genome of Pennisetum glaucum. Expression of the GUS reporter gene was revealed by histochemical assay of the blue coloration of the calli in the presence of the substrate Xgluc (5-bromo-4-chloro-3-indolyl-β-D-Glucuronic acid). The presence of GUS gene was furthermore confirmed, by southern blot hybridization of non-radioactive probes, labelled with digoxigenin, on genomic DNA extracted from selected chlorsulfuron-resistant calli. GUS gene activity observed in all selected calli was high during the first 6 months after bombardment and then decreased. Pearl millet plants were regenerated from cell lines derived from chlorsulfuron resistant calli. Southern blot hybridization of non-radioactive probes with genomic DNA extracted from these regenerated plants showed the presence of the GUS and ALS transgenes, confirming the stable transformation of these plants.Key words: Pennisetum glaucum, stable transformation, chlorsulfuron (ALS), β-glucuronidase (GUS), CaMV35S and Emu promoters

Influence de l’âge des explants primaires sur la régénération des vitroplants de deux espèces d’ignames en côte d’ivoire: Dioscorea alata et Dioscorea cayenensis-rotundata (Dioscoreacea)

Objectif: En Côte d’Ivoire la production annuelle de Dioscorea spp est insuffisante à cause de la conservation des tubercules qui réduisent fortement les rendements des tubercule-semences. Il s’avère nécessaire de conserver tous les cultivars d’igname sous forme de vitroplant. Cette étude vise à évaluer l’effet du milieu de culture, de la désinfection et du type d’explants primaires sur la régénération et la conservation in vitro.Méthodologie et résultats: Les explants primaires prélevés sur les tiges âgées de 14, 21, 35 et 60 jours ont été désinfectés et mis en culture sur deux milieux de culture MS et MS modifié. Les explants primaires issus des tiges de 14 et 21 jours ont permis d’obtenir 99 à 100 % de vitroplants sains, 100 % de régénération au bout de 2 à 4 jours dans le milieu MS modifié avec en moyenne 3,82 à 5,14 noeuds et 1 à 2 tiges au bout de 60 jours. Tandis que ceux de 35 et 60 jours ont donné 35 à 74 %.de vitroplants sains, 1 à 4 noeuds et 1 à 2 tiges. Le milieu MS modifié et Les explants primaires issus des tiges de 14 et 21 jours permettant de régénérer les vitroplants au bout de 3 à 4 jours en moyenne ont été retenus.Conclusions et application des résultats: La conservation de toutes les variétés d’igname de la Côte d’Ivoire sous forme de vitroplants par la technique de régénération de tiges aériennes in vitro nécessite la satisfaction de certaines conditions. Ainsi pour la régénération de tous les cultivars les explants primaires prélevés sur les tiges âgées de 14 et 21 jours et le milieu de culture MS modifié ont été meilleurs. Tous les cultivars dans le milieu MS modifié ont été régénérés en moins d’une semaine et les vitroplants sains ont eu une bonne croissance offrant beaucoup de noeuds qui ont servi d’explants secondaires. La présente étude indique les  possibilités de production à grande échelle de semences des variétés d’igname pour répondre aux impératifs de sauvegarder et d’enrichir les ressources génétiques de l’igname en Côte d’Ivoire.Mots clés: Igname, explants primaires, cultivars, régénération, vitroplants, milieu de culture, âge, Côte d’IvoireEnglish Title: Influence of age to primary explants on the regeneration of vitroplant of two yam species in Côte d’Ivoire: Dioscorea alata and Dioscorea cayenensis-rotundata (Dioscoreacea)English AbstractObjectives: The annual production of Dioscorea spp in Côte d'Ivoire is insufficient because of the conservation of tubers, which greatly reduce the tuber yields. It is necessary to keep all of yam cultivars as vitroplant. This study aims to evaluate the effect of the culture medium, disinfection and type of primary explants on regeneration and in vitro conservation.Methodology and Results: Primary explants taken on older stems 14, 21, 35 and 60 days were disinfected and cultured on MS culture media and modified MS media. Primary explants from the stems of 14 and 21 days have resulted in 99 to 100% of healthy vitroplants, 100% of regeneration after 2 to 4 days in MS medium amended with an average of 3 to 5 nodes and 1 to 2 stems after 60 days. While those of 35 and 60 days gave 35-74% healthy vitroplants, from 1 to 4 nodes and 1 to 2 stems. The modified MS medium and primary explants from stems 14 and 21 days to regenerate the vitroplants after 3 to 4 days on average were selected.Conclusions and application of findings: The conservation of all varieties of yam from Côte d'Ivoire as vitroplants by aerial stems regeneration technique requires the satisfaction of certain conditions.So for the regeneration of all cultivars primary explants taken on older stems of 14 and 21 days and the modified MS culture medium were better. All cultivars in the modified MS medium were regenerated in less than a week and healthy vitroplants had good growth with many knots that served as secondary explants. This study shows the large-scale possibilities of seeds production for yam varieties to meet the requirements to safeguard and enrich the genetic resources of yams in Côte d'Ivoire.Keywords: yam, primary explants, cultivars, regeneration, vitroplants, culture medium, age, Côte d'Ivoir

Evaluation of agro-morphologic diversity of some accessions of eggplant (Solanum melongena), some wild and cultivated relatives and their interspecific hybrid progenies

[EN] Commercial varieties of eggplant, Solanummelongena, are vulnerable to future challenges related to the effects of climate change. It is therefore important to provide plant breeders and farmers with plant material that can meet these new challenges by exploiting some of the natural abilities of wild relatives.The aim of this study was to appreciate the transmissibility of interesting traits to the descendants by evaluating the agro-morphologic diversity of 8 accessions of S. melongena, 8 accessions of 6 wild or cultivated relatives and 36 interspecific progenies. Mean values of agro-morphologic traits revealed heterosis effects for vegetative growth characteristics in interspecific hybrids between S. melongena and wild relatives. A Principle Component Analysis identified three morpho-groups that discriminate S. insanum, S. anguivi and S. dasyphyllum, the respective wild ancestors of the three main cultivated eggplant species, S. melongena, S. aethiopicum and S. macrocarpon. In addition, interspecific hybrids belong to the same morpho-groups as the wild parental accessions, which could therefore carry dominant alleles of genes controlling the expression of these agro-morphologic traits. In general, fertile interspecific hybrids have values of production characteristics intermediate between those of parental accessions, suggesting the involvement of genes with additive effects in the expression of these traits.This work was undertaken as part of the initiative Adapting Agriculture to Climate Change: Collecting, Protecting and Preparing Crop Wild Relatives, which is supported by the Government of Norway. The project is managed by the Global Crop Diversity Trust with the Millennium Seed Bank of the Royal Botanic Gardens, Kew and implemented in partnership with national and international gene banks and plant breeding institutes around the world. For further information, see the project website: http://www.cwrdiversity.org/.Kouassi, KBA.; Kouassi, AB.; Prohens Tomás, J.; N'gbesso, MFDP.; Akanvou, L.; N'guetta, AS.; N Zi, JC.... (2019). Evaluation of agro-morphologic diversity of some accessions of eggplant (Solanum melongena), some wild and cultivated relatives and their interspecific hybrid progenies. Global Journal of Advanced Research. 6(1):10-19. http://hdl.handle.net/10251/156953S10196

Effet de la Période de Pollinisation Manuelle sur la Fructification de l’igname (Dioscorea rotundata Poir; Dioscoreaceae) en Côte d’Ivoire

D. rotundata is the most popular species among consumers in Côte d'Ivoire. However, its production is stagnating due to the many constraints associated with its cultivation, particularly virus diseases. The use of improved varieties from sexual reproduction could effectively contribute to sustainable production. This work aimed to evaluate the crossing period in order to increase fruiting rates in this species. A total of 28 intra-specific crosses were carried out using 4 female and 7 male clones from the CNRA-Bouaké yam collection. Two time slots H1 (from 7:30 to 11:00) and H2 (from 11:00 to 13:30), selected for crossings, were compared to determine the optimal pollination period. Out of 30061 pollinated flowers, 2249 fruits were obtained i.e 7.48% with a variation range from 1.16 to 51.06% as the average fruiting rate depending on the clone combinations. Fruiting rates were significantly higher in H1 (8.53%) compared to 6.22% in H2. The female clone, Cnraigr09/00001, has recorded the highest fruiting rate especially in H1 (18.10%). Similarly, the male clones CIVCDr072, TDr00/00380 and TDr95/18555 have got the highest rates in H1 with 9.3; 9.8 and 16.3% respectively compared to those in H2. The couple Cnraigr09/00001 x TDr99/18555 recorded fruiting rates of 35.41% and 8.78% statistically different at schedule H1 and H2, respectively. A strong variation in the fruiting rate in favor of the H1 schedule was obtained between the different clone combinations.L’igname D. rotundata est l’espèce la plus appréciée des consommateurs en Côte d’Ivoire. Cependant, sa production stagne du fait des nombreuses contraintes liées à sa culture notamment les viroses. L’utilisation de variétés améliorées issues de reproduction sexuelle, pourrait contribuer efficacement à une production durable. Ce travail visait à évaluer la période des croisements afin d’accroitre les taux de fructifications chez cette espèce. Au total, 28 croisements intra-spécifiques ont été réalisés en utilisant 4 clones femelles et 7 clones mâles issus de la collection d’ignames du CNRA-Bouaké. Deux plages horaires H1 (de 7h30 à 11h) et H2 (de 11h à 13h30), retenues pour les croisements ont été comparées en vue de déterminer la période optimale de pollinisation. Sur 30061 fleurs pollinisées, 2249 fruits ont été obtenus soit 7,48% avec une variation allant de 1,16 à 51,06% de taux moyen de fructification selon les combinaisons de clones. Les taux de fructifications ont été significativement plus élevés en H1 (8,53%) contre 6,22% en H2. Le clone femelle, Cnraigr09/00001 a eu le taux de fructification le plus élevé et spécialement en H1 (18,10%). De même les clones mâles CIVCDr072, TDr00/00380 et TDr95/18555 ont eu les meilleurs taux en H1 avec respectivement 9,3 ; 9,8 et 16,3% par rapport à H2. Le couple Cnraigr09/00001 x TDr99/18555 a donné des taux de fructifications 35,41% et 8,78% statistiquement différents aux horaires H1 et H2, respectivement. Une forte variation du taux de fructification en faveur de l’horaire H1 a été obtenue entre les différentes combinaisons de clones

International Peer-Reviewed Journal ISSN 2321 3655 Oct

Abstract: To study the diversity in the in vitro and phenological responses, the model discriminant was used. Six new hybrid cocoa genotypes namely L120-A2, L126-A3, L231-A4, L232-A9, L233-A4 and L330-A9 were analyzed. Three culture media namely PCG1, PCG3 and PCG4, based on DKW basal medium, only differing in hormonal concentration, were sown with staminodes and petals extracted from floral buds. SCA6 and C151-61 were used as controls. The 6 cocoa hybrids were assessed 3 months later by scoring and calculating of the callogenesis and embryogenesis variables. Parameters from variable means were processed via the Principal Component (PCA), Hierarchical Cluster (HCA) and Factorial Discriminant Analyses (FDA). For the PCA, the number of embryogenic explants, embryos number yielded per embryogenic explants, embryogenesis percentage, flowering level, fructification level and leaves flush were revealed to be pertinent. It was the same in the first year of the study. However, for the FDA, only the number of callogenic explants and leaves flush were showed to be pertinent. This indicates that the pertinentness appears to vary from analysis to analysis. Cocoa hybrids known as L120-A2, L126-A3, L231-A4 and L330-A9 yielded the highest number of callogenic explants and expressed the highest leaf flush. The first 3 are half sibs of common father IMC67, justifying their belonging to the same group. The discriminant model associated with the analysis is spelt Z1 = -29.123 + 0.201*Flush + 1.710*Ncal. It significantly and completely discriminated the clusters in the proportion of 96.20%. The predicting of the membership cluster of new observations, from their number of callogenic explants and leaves flush, was discussed

Diversity of Water Yam (<i>Dioscorea alata</i> L.) Accessions from Côte d’Ivoire Based on SNP Markers and Agronomic Traits

Dioscorea alata (L.), also referred to as water, winged, or greater yam, is one of the most economically important staple food crops in tropical and subtropical areas. In Côte d’Ivoire, it represents, along with other yam species, the largest food crop and significantly contributes to food security. However, studies focusing on better understanding the structure and extent of genetic diversity among D. alata accessions, using molecular and phenotypic traits, are limited. This study was, therefore, conducted to assess the pattern of genetic variability in a set of 188 D. alata accessions from the National Agronomic Research Centre (CNRA) genebank using 11,722 SNP markers (generated by the Diversity Arrays Technology) and nine agronomic traits. Phylogenetic analyses using hierarchical clustering, admixture, kinship, and Discriminant analysis of principal component (DAPC) all assigned the accessions into four main clusters. Genetic diversity assessment using molecular-based SNP markers showed a high proportion of polymorphic SNPs (87.81%). The analysis of molecular variance (AMOVA) showed low molecular variability within genetic groups. In addition, the agronomic traits evaluated for two years in field conditions showed a high heritability and high variability among D. alata accessions. This study provides insights into the genetic diversity among accessions in the CNRA genebank and opens an avenue for sustainable resource management and the identification of promising parental clones for water yam breeding programs in Côte d’Ivoire

Genetic parameters of drought tolerance for agromorphological traits in eggplant, wild relatives, and interspecific hybrids

[EN] Crop wild relatives (CWRs) have barely been used in eggplant (Solanum melongena) breeding. However, introgression breeding may help in coping with the challenges posed by climate change. During the rainy and dry seasons, we evaluated nine accessions of eggplant, nine accessions of seven related species (of which six are CWRs and one is a cultivated relative), and 12 interspecific hybrids. Drought tolerance scores, calculated as ratios of growing rates and ratios of mean values during the rainy and dry seasons, were obtained for vegetative growth and yield-related traits. Considering the vegetative growth and yield-related traits evaluated, the F-1 hybrid progenies of eggplant with S. insanum MEL1 x INS2 and MEL4 x INS3, S. anguivi MEL6 x ANG1, and S. dasyphyllum MEL6 x DAS1, plus the S. sysimbriifolium accession SIS1, displayed drought tolerance. Small to large gaps were observed between phenotypic and genotypic CVs of drought tolerance scores, reflecting variable influence of environmental factors on drought tolerance of the traits evaluated. In addition, the narrow-sense heritability was generally moderate, indicating that genes with dominance and/or epistasis effects may be involved in the expression of drought tolerance. High genotypic correlations for drought tolerance scores between pairs of traits such as branching index and leaf length, leaf petiole diameter and plant height, number of stamens per flower, and fruit pedicel length suggest that drought tolerance of these traits is controlled by genes that are in linkage disequilibrium and/or have pleiotropic effects. The results are relevant for the development of drought-tolerant cultivars of eggplant.Universitat Politecnica de Valencia, Grant/Award Number: PAID06-18; Ministerio de Ciencia e Innovacion, Grant/Award Number: RTI-2018-094592-B-I00; Generalitat Valenciana, Grant/Award Number: APOSTD/2018/014; Global Crop Diversity Trust, Grant/Award Number: GS20001 This work was undertaken as part of the initiative ¿Adapting Agriculture to Climate Change: Collecting, Protecting and Preparing Crop Wild Relatives,¿ which is supported by the Government of Norway. The project is managed by the Global Crop Diversity Trust with the Millennium Seed Bank of the Royal Botanic Gardens, Kew, and implemented in partnership with national and international gene banks and plant breeding institutes around the world. For further information, see the project website: http://www.cwrdiversity.org/. Funding was also received from Ministerio de Ciencia, Innovación y Universidades, Agencia Estatal de Investigación and Fondo Europeo de Desarrollo Regional (Grant RTI-2018-094592-B-I00 from MCIU/AEI/FEDER, UE) and from Vicerrectorado de Investigación, Innovación y Transferencia de la Universitat Politecnica de Valencia (Ayuda a Primeros Proyectos de Investigación; PAID-06-18), Mariola Plazas is grateful to Generalitat Valenciana and Fondo Social Europeo for a postdoctoral contract (APOSTD/2018/014).Kouassi, AB.; Kouassi, KBA.; Sylla, Z.; Plazas Ávila, MDLO.; Fonseka, RM.; Kouassi, A.; Fonseka, H.... (2021). Genetic parameters of drought tolerance for agromorphological traits in eggplant, wild relatives, and interspecific hybrids. Crop Science. 61(1):55-68. https://doi.org/10.1002/csc2.20250556861