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Impacts of cytokinins and auxins on organogenesis and somatic embryogenesis of Vitex negundo var. Negundo L. and evaluation of genetic fidelity

M.Johnson, D.Sonali, N.Yasmin, A.Babu

The present study was aimed to produce an efficient direct and indirect micro propagation systemfor Vitex negundo var. negundo Linn. a medicinally important plant using nodal, leaves and intermodal segments as explants. Adventitious proliferation was obtained from Vitex negundo var. negundo nodal segments inoculated on Murashige and SkoogÂ’s basal medium with 3% sucrose and augmented with 6-Benzyl Amino purine. Highest frequency of shoot proliferation (75.3  0.03) was observed in Murashige and SkoogÂ’s medium augmented with 6.66M of 6-Benzyl Amino purine and the maximumnumber per explants (8.2  0.79)was also obtained in very same concentration. Maximum percentage of callus formation (stem 75.2  0.90; leaves 72.3  0.38) was obtained onMurashige and SkoogÂ’s basal medium supplemented with 3% and 2,4-Dichlorophenoxy acetic acid 2.26M.Maximumpercentage (80.2  0.2) of shoot proliferation fromthe inter-nodal derived calli was achieved onMurashige and SkoogÂ’s mediumaugmented with 3%sucrose and 6-BenzylAmino purine 4.44M in combinationwith á-NaphthaleneAceticAcid 2.69M. Threemonth old leaves calli cultures were produced the pro-embryogenic calli on the surface of the callus tissue. The maximum percentage (69.8  0.96) of proembryogenic calli obtained in Murashige and SkoogÂ’s medium supplemented with 2,4-Dichlorophenoxy acetic acid (4.52M) combination with Kinetin (2.32M).Murashige and SkoogÂ’s medium supplemented with Kinetin (4.65µM) combination with 2,4-Dichlorophenoxy acetic acid (2.26M) showed themaximumproliferation of embryo like structure.Maximumpercentage of shootlet proliferation (60.8  0.58) obtained onMurashige and SkoogÂ’s medium augmented with 6-Benzyl Amino purine 6.66M alone. Half strengthMurashige and SkoogÂ’s medium with 3% sucrose augmented with Indole 3-ButyricAcid 4.92Mshowed themaximumfrequency (85.3  0.03) of root formation fromthe in vitro derived shootlets. Themicro propagated plantlets genetic uniformity was confirmed through the isozyme analysis. The in vitro raised plants were hardened then transferred to field for re-establishment.