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Highly efficient in vitro proliferation and genetic stability analysis of micropropagated Ceropegiaevansii by RAPD and ISSR markers: A critically endangered plant of Western Ghats
Authors:J. J. Chavan  N. B. Gaikwad  P. R. Kshirsagar  S. D. Umdale  K. V. Bhat  G. B. Dixit
Affiliation:1. Department of Botany, Shivaji University, Kolhapur, Maharashtra416 004, India;2. Department of Botany, Yashavantrao Chavan Institute of Science, Satara, Maharashtra415 001, Indiajaychavansu@gmail.com;4. Division of Genomic Resources, National Research Centre on DNA Fingerprinting, NBPGR, New Delhi110 012, India;5. Division of Genomic Resources, National Research Centre on DNA Fingerprinting, NBPGR, New Delhi110 012, India
Abstract:Ceropegiaevansii McCann (family: Asclepiadaceae), a critically endangered plant of Western Ghats has acquired significant importance due to its medicinal implications, edible tubers, and ornamental flowers. This study deals with the optimization of axillary bud proliferation using nodal explants followed by genetic stability analysis of regenerants. Maximum number of shoots (11.6 ± 1.1) was observed on the Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (4.0 mg/l) and indole-3-acetic acid (0.3 mg/l) with 85% shoot multiplication frequency. In vitro-grown shoots were rooted best in 1/2 MS medium supplemented with indole-3-butyric acid (1.0 mg/l) with an average of 10.3 ± 0.9 roots per shoot and 92% rooting frequency. Plantlets were acclimatized best (90%) in a mixture of sterile soil, sand, and coco peat (1:2:1). Micropropagated plants were subjected to random amplified polymorphic DNA and inter simple sequence repeat markers analyses. Collectively, 759 bands were generated which were monomorphic and similar to the mother plant. Findings of this study are the first report on micropropagation and assessment of genetic stability of micropropagated plantlets in C. evansii which suggests that axillary shoot proliferation can safely be used as an effective tool for propagation and conservation of C. evansii.
Keywords:Acclimatization  Ceropegia evansii  genetic stability  ISSR  micropropagation  plant growth regulators  RAPD
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