For two decades, the combination 2,4-dichlorophenoxyacetic acid (2,4-D)/thidiazuron (TDZ) has been the most used to induce somatic embryogenesis in cocoa. The aim of this study was to compare the effect of combination systems, auxin/TDZ and auxin/kinetin (Kin) systems, as well as the addition of polyvinylpirrolidone (PVP) to these combinations on the induction of embryogenic calli in cocoa (Theobroma cacao L.). To this end, eight induction media combining auxin 2,4-dichlorophenoxyacetic acid (2,4-D) or 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) at 4,5 mM with cytokinin thidiazuron (TDZ) (22.8 nM) or kinetin (Kin) (1.125 mM) supplemented or not with PVP (300 mg / L) were evaluated on induction of embryogenic callus with petals or staminodes in three genotypes of cocoa. The basal salts medium was DKW (Driver and Kuniyuki Walnut medium). This study showed that the effect of 2,4-D combinations with Kinetin or TDZ was statistically identical on the induction of embryogenic callus. On the other hand, with 2,4,5-T, the association with Kin was more embryogenic than with TDZ. Furthermore, addition of PVP improved the induction of embryogenic callus in both combination systems. In the presence of PVP, media containing 2,4,5-T exhibited higher levels of embryogenic callus induction than their counterparts containing 2,4-D. This study allows the expansion of the possibilities of induction of embryogenic callus in cocoa.
The improvement of pineapple (Ananas comosus var. Smooth Cayenne) by means of in vitro culture is less studied in Côte d'Ivoire despite the importance of this plant for this country’s economy. Our work consisted in highlighting nature and concentration effects of carbohydrates on the proliferation of calli in pineapple as a prelude to efficient embryogenesis. Callus proliferation was carried out from the base of pineapple vitroplants leaves. Thirty (30) explants were cultured on the tested culture medium. MS medium (micro- and macro elements of Murashige and Skoog) supplemented with vitamin Gamborg B5 was used as base medium to which were added 0.05 mg/L BAP, 3 mg/L picloram, 2 mg/L glycine, 1,000 mg/L glutamine, 100 mg/L casein hydrolyzate and 30 g/L carbohydrate. Sucrose was tested at different concentrations (20, 25, 30, 35 and 40 g/L). The results revealed that callus proliferation is strongly influenced (p ˂ 0.0001) by nature and concentration of carbohydrate. Sucrose with the highest dry matter content (61.34 mg) has a higher callogenic potential than the other studied carbohydrates. The concentration of 30 g/L sucrose significantly improved the calli proliferation in pineapple. Galactose and maltose were less favorable to proliferation.
A highly reproducible and efficient in vitro shoot regeneration system was developed for two banana varieties (FHIA-21 and PITA-3) using shoot tip as explant. Shoot tip was inoculated onto Murashige and Skoog (MS, 1962) medium supplemented with cytokinins [benzylaminopurine (BAP), kinetin (Kin) and 2-isopentenyl (2-iP)] and additives [Adenine sulphate (Ads), spermidine (Spd) and casein hydrolysate (CH)] for shoot multiplication. In all varieties, the maximum number of shoots and shoot length was obtained with 3 and 4 mg/L BAP, respectively. This rate was further enhanced by adding Ads (25 mg/L), CH (25 or 50 mg/L) and Spd (100 or 200 mg/L). In vitro raised shoots were successfully rooted on 1 mg/L 2-iP in combination with 0.5 mg/L naphthaleneacetic acid (NAA). Rooting was significantly enhanced by adding casein hydrolysate (25 or 50 mg/L). The well rooted plantlets were successfully acclimatized on different substrates (compost, forest soil, sand, forest soil + sawdust from dead tree and sand + sawdust from dead tree). The compost substrate was found to be preferable. Finally, after one month in the greenhouse, the hardened plants were transfered to the field environment for utmost survivability.
The cultivation of pineapple contributes 1.6% of the gross Ivorian national product (GDP). However, this crop is facing a severe production crisis due to the aging of the orchards. Revising this sector requires the rejuvenation of orchards with healthy and improved planting material. This work was conducted to study the conditions for the efficient in vitro production of restorative pineapple planting material by somatic embryogenesis. The effects of seven culture media consisting of a different combination of nitrogen sources (casein hydrolyzate, glutamine, and glycine), cytokinins (kinetin or BAP), and auxins (2,4-D or picloram) were tested on somatic embryos induction and maturation in pineapple. Results of the study revealed that EIM1 (EIM added with 3 mg.L-1 picloram, 0.05 mg.L-1BAP, 2 mg.L-1 glycine, 1000 mg.L-1glutamine, 100 mg.L-1casein hydrolyzate) and EIM5 (EIM added with 2 mg.L-1glycine, 100 mg.L-1casein hydrolyzate, 0.2 mg.L-1kinetin) media induced the highest numbers of embryogenic cells, i.e., 154 and 149 cells respectively. Further, the EIM5 medium was more embryogenic, with the most significant number of mature embryos (66 mature embryos), and allowed the observation of all embryonic maturation stages. Embryogenic cell induction in pineapple is thought to be controlled by a low NH4+/NO3- ratio in interactions with phytohormones. In the presence of 2,4-D, embryogenic cell maturation was improved by kinetin addition to the culture medium containing glycine and casein hydrolyzate.
Ginger (Zingiber officinale Rosc.) is a spice, considered as a food medicine because of its numerous beneficial actions on health. However, its production faces several constraints, including the lack of efficient planting material. This is a limiting factor for industrial ginger production. The present study therefore aimed at developing an effective in vitro regeneration protocol for ginger. First, three disinfecting agents (sodium hypochlorite, calcium hypochlorite and mercury chloride) were tested. Then, different combinations of naphthalene acetic acid (0.5 mg/L NAA) and/or benzyl amino purine (1, 3 and 5 mg/L BAP) were evaluated on in vitro shoots regeneration. The results revealed that 3.6% sodium hypochlorite, used for 20 min, induced the best disinfection rates (100%) and healthy buds (84.66%). Furthermore, this study showed that the tested hormonal combinations significantly influenced shoots proliferation in ginger. However, the Proliferation Medium 4 (PM4) [Murashige and Skoog medium including vitamin B5 (MSB) + 0.5 mg/L of Naphthalene Acetic Acid (ANA) + 5 mg/L of Benzyl Amino Purine (BAP)] was the most effective. It induced the highest average number of shoots (22.83 shoots) with an induction rate of 80.50%. As a result of this study, 3.6% sodium hypochlorite used for 20 min and MP4 medium (MSB + 0.5 mg/L ANA + 5 mg/L BAP) were selected for in vitro ginger regeneration.
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