Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration
ABSTRACT Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5 mg L−1 TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca2+ concentrations to 0.22 g L−1 in the germination medium increased the number of responding explants.Induced shoot buds, upon transfer to MS medium containing 2 mg L−1 Kn (Kinetin) and 1 mg L−1 BAP (6-benzylamino purine) elongated. These elongated shoots were further proliferated on MS medium supplemented with 1.5 mg L−1 IAA (indole-3-acetic acid) and 0.5 mg L−1 BAP and 3.01–3.91 cm elongation was achieved after 6 weeks. No genotype specific variance in shoot elongation was observed among the toxic genotypes except the CSMCRI-JC2, which showed reduced response. And for proliferation among the toxic genotypes, CSMCRI-JC4 showed highest number of shoots formed. Among the rest, no significant differences were observed. The elongated shoot could be rooted by pulse treatment on half-strength MS medium supplemented with 2% sucrose, 3 mg L−1 IBA (indole-3-butyric acid), 1 mg L−1 IAA, 1 mg L−1 NAA (α-naphthalene acetic acid) and subsequent transfer on 0.25 mg L−1 activated charcoal medium. The rooted plants could be established in soil with more than 90% success. No significant differences were observed in rooting of shoots in the different toxic genotypes. However, rooting response was reduced in non-toxic genotype as compared to toxic genotypes.Highlights► Efficient and reproducible in vitro propagation protocol in Jatropha curcas via hypocotyls explants was developed. ► Regeneration was found to be genotype dependent. ► In vitro source of explants led to higher regeneration efficiency as compared to ex vitro source. ► Number of shoot buds induced were affected by the explants position on seedling axis. ► Age of seedling also influenced the regeneration potential which decreased with increase in age. ► Transient decrease of Ca2+ concentrations in germination medium increased percentage response.
SourceAvailable from: Fardous Mohammad Safiul Azam[Show abstract] [Hide abstract]
ABSTRACT: Wedelia calendulacea (Family Asteraceae) is a rare medicinal herb, used in Asian countries including Bangladesh that serves in traditional medicine and contains varied pharmacological properties including hepatoprotective, cytotoxicity, antibacterial, neuroprotective, and antiosteoporotic activity; importantly demethylwedelolactone and wedelolactone, reported to have anti-cancer potency. Our objective was to study the effect of phytohormones on mass propagation of this species in vitro. Young shoot tips and nodes were cultured onto MS (Murashige and Skoog) medium fortified with different concentrations of phytohormones alone or in combinations, sucrose and agar, following surface sterilization with 0.1% HgCl2. Good result was found in MS + 6-benzyleaminopurine [0.5 mg/l] where shoots regenerated up to 1.11±0.04 cm at 6th week. During synergism of cytokinins, highest multiplication (33.50±6.50 shoots/explant) was resulted as clumps at 6th week in MS + 6-benzyleaminopurine [1.5 mg/l] + Kinetin [1.0 mg/l]. The best root induction (18.90±2.47 roots/shoot) was seen when micro-shoots were implanted in MS + indole-3-butyric acid [1.0 mg/l]. After hardening and acclimatization into soil, 90% plantlets were survived after three weeks with no morphological variation with donors. This study could play a significant role for plantlets regeneration in vitro to meet commercial demands and as well as for conservation of this species.American-Eurasian Journal of Sustainable Agriculture 06/2014; 8(5):18-25.
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ABSTRACT: In this study to evaluate the effectiveness of two auxins 2, 4-D and NAA and two cytokinin TDZ and BAP on callus induction from leaf explants of Ziziphus spina christti (L.) Willd in vitro conditions, carried out in factorial adopted on completely randomized design with 12 treatment and 4 replications. Explants on the medium MS contain a level 0.5, 1 and 2 mg/l of four hormones were cultured and the percentage of callus creation and degree of formation callus was measured. The results showed that among four cytokines and auxin used, the level of 0.5 and 1.0 mg/L 2, 4-D and TDZ in both media had the highest amount of callus induction. All concentration of BAP was the lowest than that of other treatments in explants induced callus parameter. Among treatments, in 0.5, 1 mg/l TDZ medium recorded the highest degree of callus formation parameter. As compared to all concentration of BAP medium and 0.5 mg/l NAA recorded the lowest degree of callus formation parameter. Also in this study it was observed that leaf explants cultured on NAA medium in root hair (rhizoid) production.3 th international congress climate change & dendrochronology, IRAN; 05/2012
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ABSTRACT: Jatropha curcas L. is attaining worldwide interest as an important biofuel crop. Experiments were conducted to improve the prevailing micropropagation technique as well as to develop a new ex vitro rooting method for J. curcas plant regeneration. Regeneration and ex vitro rooting efficiency was enhanced by augmenting the culture medium with abscisic acid (ABA). Different concentrations of 6-benzylaminopurine (BAP) and indole-3-butyric acid (IBA) were tested for callus generation from both in vitro and in vivo explants (leaf and petiole) on Murashige and Skoog (MS) medium. The best regenerative callus was achieved on MS medium supplement-ed with BAP (4.44 μM) and IBA (2.45 μM) from in vitro-cultured petioles. Highest regeneration (91%) was achieved by culturing petiole callus on MS medium supplemented with BAP (8.88 μM), IBA (0.49 μM), and ABA (1.9 μM), whereas 61% regeneration was obtained from in vitro leaf callus. Shoot proliferation and elongation was achieved on BAP (2.22 μM) and IAA (8.56 μM) with 10–13 shoots per explants. Highest rooting (65%) was achieved from M1 shoots (BAP, IAA, and ABA) on MS medium supplemented with IBA (2.45 μM), naphthaleneacetic acid NAA (0.54 μM), and 0.02% activated charcoal. Ex vitro rooting of 1-mo-old M1 shoots obtained from the charcoal-containing medium resulted optimum rooting (>72%) when transferred to polybags containing ster-ile sand. The plantlets were successfully acclimatized in soil with more than 98% survival rate in the greenhouse.In Vitro Cellular & Developmental Biology - Plant 06/2014; 50(5). DOI:10.1007/s11627-014-9628-y/fulltext.html · 1.16 Impact Factor