In vitro PLANT REGENERATION FROM PETIOLE EXPLANTS AND ASSESSMENT OF GENETIC FIDELITY USING ISSR MARKERS IN Momordica charantia L.
YASHODHARA VELIVELA *
Department of Biotechnology, Kakatiya University, Warangal, Telangana, 506009, India
MURALIKRISHNA NARRA
Department of Biotechnology, Kakatiya University, Warangal, Telangana, 506009, India
RAGHU ELLENDULA
Department of Biotechnology, Kakatiya University, Warangal, Telangana, 506009, India
BHARATH KUMAR KALVA
Department of Biotechnology, Kakatiya University, Warangal, Telangana, 506009, India
RATHNAPRABHA DHARAVATH
Department of Biotechnology, Kakatiya University, Warangal, Telangana, 506009, India
SADANANDAM ABBAGANI
Department of Biotechnology, Kakatiya University, Warangal, Telangana, 506009, India
*Author to whom correspondence should be addressed.
Abstract
An in vitro protocol for organogenesis was achieved from petiole explants of Momordica charantia L., a very important vegetable and medicinal plant of Cucurbitaceae. Morphogenetic response of these explants has been studied with different concentrations of various plant growth regulators alone and in combinations (BAP, TDZ, IAA, NAA and IBA). Shoot buds from petiole explants were initiated after three weeks of culture on Murashige and Skoog (MS) medium with 30 g/L sucrose, 8.9 μM BAP, 1.14 μM IAA along with 0.1 μM glutamine through intervening callus phase. Multiple shoots (10.92±1.32) were induced after four weeks of culture on same medium and their elongation was achieved on MS medium supplemented with 8.9 μM BAP along with 1.14 μM IAA and 0.34 μM GA3. The elongated shoots were rooted on MS medium supplemented with 0.49 μM IBA. Rooted plants were acclimatized in the greenhouse and subsequently transplanted in soil and successfully transferred to natural conditions with maximum survival rate of 80%. The clonal fidelity of in vitro raised plants was determined using Inter Simple Sequence Repeat (ISSR) analysis. The present investigation showed a good opportunity to raise in vitro plantlets from petiole explants and successful assessment of their genetic stability.
Keywords: 6-Benzyl amino purine, indole-3-aceticacid, glutamine, inter simple sequence repeat