In Vitro Cloning of Cissus Quadrangularis Linn an Important Medicinal Plant

Abstract

The present investigation was carried out on an important vulnerable medicinal plant Cissus quadrangularis Linn. belonging to family Vitaceae. This plant is commonly known as “Hadjod” in Hindi because of its ability to promote healing of the fractured bones. The different vegetative parts i.e. nodal explant, shoot apices, stem and leaves were excised from an elite field grown mature plant and thereafter planted on variously supplemented Murashige and Skoog’s medium for multiple shoot proliferation, callus induction and organogenesis. Cissus exhibits a good degree of propensity of multiple shoot proliferation from nodal and shoot apex segments. Multiple shoot proliferation from nodal explants was observed on MS medium supplemented with Zn (2.28μM-9.12μM) either alone or in combination with BAP (4.4μM-8.8μM). Best results were however obtained on MS medium supplemented with Zn (4.56μM), where 10-12 shoots were obtained after 8 weeks of culturing. Shoot apices also exhibited multiple shoot proliferation when cultured on MS medium supplemented with Zn (2.28μM-9.12μM) either alone or in supplement with BAP (4.4μM-8.8μM). BAP (8.8μM- 26.4μM) in combination with Kn (4.65μM) or AS (1.35μM-21.9μM) also exhibited the formation of multiple shoots from shoot apices. Regenerated shoots were rooted on a separate root inducing medium which consisted of full strength basal MS medium. Well developed, long white roots were formed in about 100% of cultures after 3 weeks of culturing. The plantlets with elongated root and shoot systems were subjected to hardening and attempts are underway to establish these plantlets in the soil. Callus formation occurred from stem and leaf segments when planted on different concentrations and combinations of auxins and cytokinins. Stem segments callused on MS medium supplemented with different concentrations of NAA (7.35μM-14.7μM) and Kn (2.325 μM -9.3μM). Murashige and Skoog’s medium supplemented with NAA (7.35μM) and Kn (2.325μM), hereby, designated as NK medium turned out to be optimal medium for initiation and good growth of callus. Further, addition of 15% CM to the NK medium considerably enhanced the callus formation and extending its browning period. The callus formed from the stem explant was highly friable and heterogenous in nature being composed of cells of different sizes and shapes and showed the presence of significant amount of starch granules. Formation of leaf callus was observed on MS supplemented with NAA (7.35 μM) and Kn (2.325μM). Root differentiation from leaf calli occurred on NK medium after 6-7 weeks of culturing. The roots were thick, small and white in colour. A low frequency of shoot differentiation from the leaf callus was observed in 2% of the cultures on NAA (14.7μM) and Formation of leaf callus was observed on MS supplemented with NAA (7.35 μM) and Kn (2.325μM). Root differentiation from leaf calli occurred on NK medium after 6-7 weeks of culturing. The roots were thick, small and white in colour. A low frequency of shoot differentiation from the leaf callus was observed in 2% of the cultures on NAA (14.7μM) and Formation of leaf callus was observed on MS supplemented with NAA (7.35 μM) and Kn (2.325μM). Root differentiation from leaf calli occurred on NK medium after 6-7 weeks of culturing. The roots were thick, small and white in colour. A low frequency of shoot differentiation from the leaf callus was observed in 2% of the cultures on NAA (14.7μM) and kn(2.325μM) after 8 weeks of culturing.