High multiplication frequency and genetic stability analysis of Ceropegia panchganiensis, a threatened ornamental plant of Western Ghats: Conservation implications
2013
Chavan, Jaykumar J. | Gaikwad, Nikhil B. | Yadav, Shrirang R.
The aim of the present study was to develop a protocol for in vitro propagation and genetic stability analysis of Ceropegia panchganiensis Blatter and McCann, a threatened ornamental plant of Western Ghats, India. Axillary shoots were generated from nodal explants on MS medium supplemented with different concentrations of plant growth regulators. The best frequencies of shoot multiplication (100±0.0%), maximum numbers of shoots (13.2±0.5) and longest shoot lengths (10.1±0.8cm) were obtained on MS media containing BAP (13.31μM) and NAA (2.69μM). Optimum callus induction response (95±2.3%) was observed on MS medium supplemented with 2,4-D (9.05μM). Half-strength MS medium supplemented with BAP (4.44μM) in combination with sucrose (175mM) produced an average of 8.2±0.4 flower buds with 80±2.9% flowering response. Cultures treated with BAP (17.74μM) along with sucrose (175mM) showed the highest percentage (85%) of microtuber formation. Microshoots rooted best (96±1.9%) in ½MS medium supplemented with IBA (7.36μM), produced an average of 9.3±0.9 roots/shoot with 3.6±0.5cm root length. The rooted plantlets survived best (85%) in a mixture of sterile soil, sand and coco peat (1:2:1). The molecular status/genetic stability of micropropagated plants was assessed by random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers. A total of 1149 bands were generated with 10 RAPD and 10 ISSR primers in a mother plant and 10 micropropagated plants, out of which 1134 (98.69%) bands were monomorphic and rest 15 (1.31%) were polymorphic. The present study has established the rapid micropropagation protocol for the first time and will be of great use in conservation of C. panchganiensis with low risk of genetic instability.
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