A b s t r a c t A r t i c l e I n f oThe impact of the number of subcultures on the stability of yam genotype maintained in in vitro genebank was investigated. Six accessions of yam (Ala, Bagri Kogan, Kokoro, Sossou, and Tankpanou), belonging to the complex Dioscorea cayenensis-rotundata were initiated on free-hormone MS medium and were micropropagated each four months during five subcultures. DNA was extracted both from mother plant and plantlets provided from the subcultures for each accession and was exhibited Random Amplified Polymorphic DNA (RAPD) analysis using four selected primers to detect somaclonal variation. No phenotypic variation was observed during the fifth subcultures. From the RAPD analysis of both mother plants and in vitro plantlets, no significant variation of DNA profiles was observed with the highest of the coefficient of similarity (85% to 100%) for all accessions, thus ensuring the genetic stability of the plants and regeneration of true to type plants for at least five subcultures.
Bananas and plantains are among the most important food crops in Central and West Africa. Their plantation is lead to many problems. In the recent decades, biotechnology tools using in vitro culture technics are used for the mass and free disease plantlets production in order to increase the bananas production and the yield. The main way of in vitro tissue culture at this end is the direct organogenesis i.e., the ability of plant tissues to form various organs de novo by shoots or roots induction to differentiate from a cell or cell clusters. This review aims to summarize the main results obtained in the organogenesis of bananas and plantains (Musa spp.) under in vitro conditions and to identify the challenges during the process. The research articles used in this review show that micropropagation is a reliable alternative to conventional production system of bananas and plantains planting material. However, the use of the in vitro micropropagation for bananas and plantains entails choosing the optimal explant type and size according to objectives. Benzylaminopurine remains the preferred cytokinin for in vitro banana and plantain shoot proliferation, while the use of thidiazuron appears to be more and more common. Whichever cytokinin used, the optimal cytokinin concentration for shoot proliferation is genotype dependent. This review also focuses on the causes and control measures of the two major banana and plantain micropropagation constraints: lethal tissues browning/darkening and microbial contaminations. It showed that applying the suitable and available control measure, according to the evolution of culture, is necessary. All this available information on the in vitro conditions makes banana and plantain cultivars in vitro organogenesis possible.
Objective: Cryopreservation is one of the biotechnological methods currently used for long term conservation of plant genetic resources. It requires many steps such as a pretreatment, which involves cells dehydration in order to make them tolerant to desiccation and freezing in nitrogen liquid using high sucrose concentration. The present study aims to establish a protocol for long term conservation of yam germplasm in Benin using encapsulation/dehydration technique. Methodology and Results: The effects of different sucrose concentrations and immersion durations on encapsulated apices of two yams genotypes (Da93G1 from Dioscorea alata and Dcr164 from complex Dioscorea cayenensis/ D. rotundata) were tested. The apices were excised from six months old vitroplants on stereoscope and transferred in M1 medium (MS+2g.L-1 activated charcoal) prior to encapsulation in alginate (3%) beads with calcium chloride (1.32M). Then, the apices were exposed to osmotic dehydration with two concentrations of sucrose (0.75M and 1.25M) at two durations (24h and 40h) before their culture in M2 medium (MS + 2mg.L-1 BAP, 100µg.L-1 d'ANA and 2g.L-1 activated charcoal). Conclusion and application: The results showed that 0.75M sucrose increased high survival rate (80%) and high rate of regenerated plant (70%) at 24h of immersion duration. Furthermore, significant difference was observed between the two genotypes response. This work has allowed adopting in further experiments 0.75M sucrose and 24hours of immersion duration in pretreatment of yam apices for the development of cryopreservation techniques for yam conservation in Benin.
Aims: This study aims to identify the best surface sterilization and evaluate the effect of haustorium suppression on in vitro germination of coconut palm (Cocos nucifera L.) zygotic embryos. Study Design: Survival rate and contamination rate of zygotic embryos after different surface sterilization treatments, regeneration rate and organogenesis through the number of leaves and the length of shoots after haustorium suppression were determined. For data processing, the Analysis of Variance was used to compare the means which were separated according to Tukey test (P = 0.05). Place and Duration of Study: Coconut fruits (hybrid PB121) were collected 12 to 14 months after controlled pollination from CRAPP (Centre de Recherches Agricoles Plantes Pérennes), station of Sèmè-kpodji in Benin. Experiments were done in Central Laboratory of Plant Biotechnology and Plant Improvement, University of Abomey-Calavi and conducted from june to december in 2019. Methodology: For the zygotic embryos surface sterilization, four treatments combining three concentrations (3%, 6% and 15%) of commercial bleach (Javel la Croix© containing 12° active chlorine) and immersion durations (5 min, 10 min and 20 min) were tested and the survival rate were determined for each treatment after two months culture. The zygotic embryos were then divided in two sets (haustorium excised embryos set and the whole embryos set) and cultured in modified Y3 medium supplemented with 7 g L-1 agar, 2.5 g L-1 activated charcoal, 5% sucrose, 6.10-3 mM 2.4 D (2.4-dichlorophonoxyacetic acid), gibberellic acid and 0.3 mM BAP(6-benzylaminopurine). After five months culture, the regeneration rate, the number of leaves and the length of shouts were recorded. Results: The high survival rate (80%) was obtained with 6% of bleach and 20 min for the immersion duration without pre-disinfection. The suppression of haustorium have significantly increased the number of leaves (4.3 ± 0.02) and the length of shoots (16.2 ±0.7cm) compared to the whole zygotic embryos. Conclusion: This protocol can help to ensure better surface sterilization of zygotic embryos before their in vitro culture and the development of vigorous plantlets in order to improve the slow growth of plantlets, when transferred to the greenhouse or field.
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