2 research outputs found
<i style="mso-bidi-font-style:normal">In vitro</i> regeneration and ploidy level analysis of <i style="mso-bidi-font-style:normal">Eulophia ochreata </i>Lindl
1112-1121Various parameters including explant-type,
medium compositions, use of phytohormones and additives were optimized for
direct and indirect regeneration of E.
ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs)
proved to be the best explants for shoot initiation, proliferation and callus induction.
Murashige and Skoog’s (MS) medium containing 2.5
mg L-1 6-benzylaminopurine (BAP),
1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine,
citric acid, 30 mg
L-1
each and 50 mg L-1 ascorbic
acid) was optimal for shoot multiplication (12.1 shoots
and 7.1 PLBs per explant with synchronized growth), which also produced callus.
Shoot number was further increased with three successive subcultures on same
media and ~40 shoots per explant were achieved after 3 cycles of 30 days each.
Additives and
casein hydrolysate (CH) showed advantageous effects on indirect shoot
regeneration via protocorm-derived callus. Optimum indirect regeneration was
achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP
and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size).
The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1
activated charcoal and additives. The rooted plantlets were hardened and
transferred to greenhouse with 63% survival rate. Flow-cytometry
based DNA content analysis
revealed that the ploidy levels were maintained in <i style="mso-bidi-font-style:
normal">in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.<i style="mso-bidi-font-style:
normal"></i