Response of sugarcane (Saccharum sp.) varieties to embryogenic callus induction and in vitro salt stress

Response of three varieties of sugarcane (Saccharum sp.) to callus induction, embryogenic callus production and in vitro salt tolerance was studied. For callus induction and embryogenic callus production, leaf bases segments were subjected to in vitro culture on Murashige and Skoog (MS) medium supplemented with 3 mg 1-1 2,4 Dichlorophenoxyacetic acid for 4 weeks. To evaluate salt tolerance of the varieties, growing calli were exposed after two subsequent subcultures (4 weeks each) to different concentrations of NaCl (0, 17, 34, 68 and 102 mM) added to the culture medium for 4 weeks. Comparision of genotypes was based on callus induction percentage, embryogenic callus production percentage and relative fresh weight growth (RFWG). For salt tolerance, necrosis percentage and relative fresh weight growth of callus were used. The three varieties responded well to callus induction with a percentage of induction about 82, 84 and 100% for CP70-321, NCo310 and CP65-357, respectively. The high percentages of embryogenic callus obtained for the three varieties indicated that these varieties have a high capacity for embryogenic callus production. Relative fresh weight growth of callus was about 1.076, 1.282 and 0.925 for CP70-321, NCo310 and CP65-357, respectively. NaCl effect resulted in calli necrosis and a reduction of their growth. However, growing calli derived from varieties CP70-321 and NCo310 showed less necrosis percentages and less relative fresh weight growth reduction under salt stress. They appeared to be more salt tolerant in vitro than CP65-357.African Journal of Biotechnology Vol. 4 (4), pp. 350-354, 200

Effect of two cytokinins in combination with acetic acid α-naphthalene on yams (Dioscorea spp.) genotypes’ response to in vitro morphogenesis

The effect of two growth regulator combinations was studied on the morphogenesis in vitro of 3 genotypes of yams (Kounondakou, Gnon-boya and RB 89579). Benzyl aminopurin (BAP) and zeatin (ZEAT) were tested, respectively at a concentration of 0.5 mg/l with the galzy glutamine basic medium containing naphtalene acetic acid (NAA) (0.5 mg/l). Stem fragments were used as explants. The number of stripped buds and explants having stem and roots are sampled after 2 weeks in culture. The dry matter content, the number of roots and leaves and the height of each young sprout were determined after 5 weeks in culture. The results obtained indicated that no break in leaf growth was observed on the control medium (without cytokinin) but media with BAP and zeatin presented a good plants aerial part development. A significant interaction (p < 0.05) was observed between the genotypes and the typeof cytokinin. However, the highest bud sprouting and shoot development were obtained with BAP. Thus, BAP can be considered as cytokinin having a good morphogenic aptitude when compared to zeatin for yam micropropagation.Keywords: Cytokinin, auxin, morphogenesis, yam, Beni

Micropropagation in vitro de la variété locale « Aloga » du bananier plantain (Musa x paradisiaca L.) au Bénin

Le bananier plantain (Musa x paradisiaca L.) est une banane consommée sous forme cuite qui contribue à la sécurité alimentaire et aux revenus des populations de l'Afrique sub-saharienne. Le développement de la culture de cette plante est limité par de nombreuses maladies dont la dissémination est facilitée par la multiplication végétative par rejets qui est le principal mode de reproduction de cette plante. Dans ce travail, la multiplication in vitro de la variété locale Aloga de bananier plantain (Musa x paradisiaca L.) a été étudiée. Le milieu de culture de base utilisé a été additionné de différentes concentrations de l’acide naphtylacétique (ANA), de l’acide indole-3-acétique (AIA) et de benzylaminopurine (BAP). Le milieu d’induction et de multiplication contient l’AIA (10-6 N) et la BAP (0.02 N), tandis que le milieu d’enracinement est composé de BAP (0.001 N) et de l’ANA (10-6 N). Les explants sont constitués des bourgeons apicaux prélevés à partir de rejets fraîchement récoltés. Nous avons réussi à multiplier in vitro la variété Aloga avec un taux de multiplication de 3 à 8 rejets par explant et par mois. Par ailleurs, les vitroplants obtenus se sont bien enracinés et ont bien répondu à l’acclimatation. Leur transfert en plein champ a permis d’avoir des plantes avec une taille moyenne d’environ 20 cm, un diamètre à la base de la tige avoisinant 10 cm et un nombre de feuilles vertes de l’ordre de 6 après 3 à 4 mois de culture en plein champ. Cette étude ouvre la voie au développement de la micropropagation du plantain au Bénin dans le but de fournir aux producteurs du matériel de plantation indemne de parasites et en grande quantité.Mots clés: Banane plantain, variété locale, Aloga, culture in vitro, micropropagation, acclimatatio

Effect of genotype on callus induction and plant regeneration from leaf explants of sugarcane (Saccharum sp.)

Nine sugarcane genotypes (CP59-73, CP63-588, CP80-314, SP71-1081, F160, L62-96, CP70-321, CP57- 614 and Clone III) were evaluated for their callus induction capacity, embryogenic callus production and plant regeneration ability. Leaf cylinders were used as explants using Murashige and Skoog (MS) based medium supplemented with 3 mg l-1 2,-4 dichlorophenoxyacetic acid. Plant regeneration was accomplished on hormone free modified MS medium supplemented with casein hydrolyzate. The genotypes tested showed high callus induction percentage (69 to 95%) and high embryogenic callus percentage (60 to 100%). These genotypes also showed excellent regeneration capacities, with regeneration percentages ranged between 88 and 100%. Significant differences were observed between genotypes for callus induction capacity, embryogenic response and plant regeneration ability indicating that these criteria are genotype dependent. Plant regeneration ability is highly correlated with embryogenic callus production. The in vitro regenerated plants were successfully rooted and well acclimatised in growth cabinet conditions

Auxin pretreatment promotes regeneration of sugarcane (Saccharum spp. hybrids) midrib segment explants

We have developed a new, simple, quick and genotype-independent method for direct regeneration of sugarcane using novel midrib segment explants. Our protocol involves two steps: the pretreatment of starting material on MS (Murashige and Skoog (1962) Physiol Plant 15:473–497) medium containing 3.0 mg/l 2,4- dichlorophenoxyacetic acid (2,4-D) for 8 days under continuous dark and subsequent transfer of the explants to MS medium augmented with 0.1 mg/l benzyladenine (BA) and 0.1 mg/l naphthaleneacetic acid (NAA) under light-dark conditions. On the regeneration medium, numerous globular structures appeared from the explants and subsequently differentiated into shoots. Regenerated shoots attained 2–5 cm height within 30 days of culture initiation and readily rooted on MS basal medium. Hardened plants were successfully established in the greenhouse. The regulation of sugarcane morphogenesis by auxin pretreatment is discussed

In vitro culture techniques as a tool of sugarcane bud germination study under salt stress

Germination was the first stage confronted to soil salinity and it is important to determine salt effects on this stage. In this study, we reported an in vitro procedure for studying sugarcane bud germinationand shoot growth under salt stress with different NaCl concentrations (0, 17, 34, 68 and 102 mM) using cultivar NCo310. Germination percentage of control was about 92% after 8 days. Germination of buds,plant fresh and dry mass decreased with increasing salinity. Data indicated that in vitro culture techniques could be used to evaluate salt stress effects in sugarcane at the germination stage

Response of Chili Pepper (Capsicum spp.) Cultivars Cultivated in Benin to Salt Stress at Germination Stage

Aims: In this study, salt resistance level of six chili (Capsicum spp.) cultivars including five local cultivars (Adologbo, Gbatakin, Pili-pili, Gbatakin d’Agbédranfo, TPS0251) and one imported variety (Démon) grown in Benin was evaluated at the germination stage. Study Design: The experiment was laid out as a completely randomized design with four replications. Place and Duration of Study: The experiment was carried out in the Laboratory of Plant Physiologyand Abiotic Stresses Study of University of Abomey-Calavi, Republic of Benin from September through October, 2016. Methodology: Seeds were submitted to treatment with five NaCl concentrations (0; 30; 60; 90 and 120 mM NaCl) in petri dishes.Seed germination was checked every day during sixteen days incubation period. Four replicates of 40 seeds each were used. Results: From day 2 to day 16, NaCl delayed seed germination rate proportionately to NaCl concentration except for cultivars TPS0251 and Démon. At the end of the 16 days, NaCl stress effects on seed germination of cultivars were significantly variable. No significant reduction was observed for cultivars Démon and TPS0251 whereas a significant decrease was observed for the four other cultivars with a significant difference among them. The average reduction due to NaCl stress was lower for cultivars Démon (0%) and TPS0251 (4.31%) and higher for cultivar Pili-pili (63.61%). Salt Tolerance Index was significantly variable according to the cultivar with the highest values for cultivar Démon (1.227) and TPS0251 (1.127) and the weakest values for cultivars Pili-pili (0.374) and Gbatakin d’Agbédranfo (0.46). Conclusion: NaCl stress delayed seed germination and reduced the rate of final germination. Salt Tolerance Index was variable among the six cultivars: cultivars Démon and TPS0251 appeared to be the most salt resistant whereas Pili-pili and Gbatakin d’Agbédranfo appeared as the most salt sensitive at germination stage. For the first time, we demonstrated a variability of relative salinity resistance among local chili pepper cultivars at germination stage

Initiation of embryogenic cell suspensions of taro (Colocasia esculenta var. esculenta) and plant regeneration

Embryogenic callus was initiated by culturing in vitro taro corm slices on agar-solidified half-strength MS medium containing 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for 20 days followed by transfer to 1.0 mg/L thidiazuron (TDZ). Callus was subsequently proliferated on solid medium containing 1.0 mg/L TDZ, 0.5 mg/L 2,4- D and 800 mg/L glutamine before transfer to liquid medium containing the same components but with reduced glutamine (100 mg/L). After 3 months in liquid culture on an orbital shaker, cytoplasmically dense cell aggregates began to form. Somatic embryogenesis was induced by plating suspension cells onto solid media containing reduced levels of hormones (0.1 mg/L TDZ, 0.05 mg/L 2,4-D), high concentrations of sucrose (40–50 g/L) and biotin (1.0 mg/L). Embryo maturation and germination was then induced on media containing 0.05 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-acetic acid (IAA). Histological studies of the developing embryos revealed the presence of typical shoot and root poles suggesting that these structures were true somatic embryos. The rate of somatic embryos formation was 500–3,000 per mL settledcell volume while approximately 60% of the embryos regenerated into plants

Pretreatments, conditioned medium and co-culture increase the incidence of somatic embryogenesis of different Cichorium species

Somatic embryogenesis (SE) in Cichorium involves dedifferentiation and redifferentiation of single cells and can be induced by specific in vitro culture conditions. We have tested the effect of various treatments on the incidence of SE (ISE) of an interspecific embryogenic hybrid (C. endivia x C. intybus) and of different commercial chicories (C. endivia and C. intybus) that are typically recalcitrant to SE in standard culture conditions. We found that the ISE of the hybrid is significantly increased by pretreatment of tissues by submersion in solutions of glycerol, abscisic acid, spermine, putrescine or of combinations of these compounds. Interestingly, the most efficient of these pretreatments also had an unexpectedly high effect on the ISE of the C. intybus cultivars. The ISE of the hybrid and of the commercial chicories were increased when explants were co-cultured with highly embryogenic chicory explants or when they were cultured in conditioned medium. These observations established that unidentified SE-promoting factors are released in the culture medium. HPLC analyses of secreted Arabino-Galactan Proteins (AGPs), which are known to stimulate SE, did not allow identifying a fraction containing differentially abundant AGP candidates. However, pointing to their role in promoting SE, we found that the hybrid had a drastically higher ISE when amino sugars and L-Proline, the putative precursors of secreted AGPs, were both added to the medium