Abstract
Background and objectiveA simple, rapid, efficient, and reproducible protocol for callus induction and regeneration of plantlets from callus of Jatropha curcas plant was established.Materials and methodsRandomly amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses were used to determine the genetic variation between the regenerated, micropropagated, and mother plants.ResultsThe highest callus induction percentage from leaf explant was recorded with MS medium containing 2.5 mg/l BA (6-benzylaminopurine) + 1.0 mg/l NAA (1-naphthaleneacetic acid). Leaf-derived callus was grown on medium containing 2.0 mg/l BA + 0.2 mg/l IBA (indole-3-butyric acid) for adventitious shoot regeneration. In addition, using five random RAPD primers with the tested samples produced 117 amplified products out of which 25 were polymorphic bands resulting in 21.37% polymorphism, whereas the five ISSR primers used yielded 116 scorable bands out of which 22 were polymorphic bands producing a polymorphism pecentage of 18.96.ConclusionAn optimized protocol for large-scale production of J. curcas plants using plant biotechnology tools was achieved. RAPD and ISSR techniques would introduce an alternative system for large-scale production and establishment of genetically stable plants.
Highlights
Background and objectiveA simple, rapid, efficient, and reproducible protocol for callus induction and regeneration of plantlets from callus of Jatropha curcas plant was established
Regeneration of plantlets from callus cultures Effect of different concentrations of BA and 2,4-D or Indole3-butyric acid (IBA) on regeneration From Table 2, it could be observed that the highest regeneration percentage was 83.33 recorded by MS medium supplemented with 2.0 mg/l BA + 0.2 mg/l IBA (Fig. 2a), followed by 66.67% that was reported by MS medium supplemented with 1.0 mg/l BA + 0.1 mg/l IBA (Fig. 2b), whereas MS medium supplemented with 0.5 mg/l BA + 1.0 mg/l 2,4-D gave the lowest regeneration percentage (21.67) (Fig 2c)
MS medium free of growth regulators did not show any response for regeneration of adventitious shoots (Fig 2d)
Summary
A simple, rapid, efficient, and reproducible protocol for callus induction and regeneration of plantlets from callus of Jatropha curcas plant was established. Jatropha curcas L. belongs to family Euphorbiaceae and grows as a large shrub or small tree. The plant is found in tropical and subtropical areas, where it is used for reclaiming land and feedstuff; besides, the plant has different medicinal properties and many other uses. Due to the high oil content of J. curcas seeds (up to 60% oil) and low production cost, the plant attracted global attention for the development as a source for bio-fuel (Li et al 2007). Recent interest in promotion of J. curcas as a biodiesel crop demands genetic improvement of the crop for increased seed yield and oil content. A large number of polymorphic markers are required to
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