Articles
OPTIMIZATION OF SPILANTHES ACMELLA L. CULTIVATION BY IN VITRO NODAL SEGMENT CULTURE
Article number
865_13
Pages
109 – 114
Language
English
Abstract
An efficient protocol for clonal propagation of Spilanthes acmella L., a medicinally valuable plant, has been developed.
Nodal explants were collected at monthly intervals to initiate in vitro cultures.
More than 60% aseptic cultures were established during the months from January to April.
Long, multinodal shoots were developed from the nodes on full strength Murashige and Skoog (1962; MS) basal medium or basal medium supplemented with at least one cytokinin.
Irrespective of the treatments, excessive adventitious root proliferation was observed from all over the surface of the in vitro developed shoots and on the explants as well.
Of all the treatments, 6-benzylaminopurine (BAP) supported maximum percentage of cultures showing shoot proliferation.
Interestingly, higher concentration of BAP resulted in slow growth of shoots while at lower concentration excessive adventitious roots were observed in the cultures, in addition to axillary shoot proliferation.
However, intermediate concentration of BAP at 5 µM favored maximum rate of shoot multiplication with no adventitious roots.
On MS + 5.0 µM BAP, 10 fold shoot multiplication was achieved every 5 weeks by cutting the solitary in vitro shoot into single node segments and culturing them onto the fresh medium of the same composition.
The shoots were successfully rooted on half strength MS medium (major salts reduced to half strength) with 50 g/L sucrose; roots developed directly at the base of the shoot in 100% cultures.
Plantlets were hardened and successfully established in the soil.
Regeneration protocol developed in this study would provide a basis for germplasm conservation and mass cultivation of the plant for later biotechnological use.
Nodal explants were collected at monthly intervals to initiate in vitro cultures.
More than 60% aseptic cultures were established during the months from January to April.
Long, multinodal shoots were developed from the nodes on full strength Murashige and Skoog (1962; MS) basal medium or basal medium supplemented with at least one cytokinin.
Irrespective of the treatments, excessive adventitious root proliferation was observed from all over the surface of the in vitro developed shoots and on the explants as well.
Of all the treatments, 6-benzylaminopurine (BAP) supported maximum percentage of cultures showing shoot proliferation.
Interestingly, higher concentration of BAP resulted in slow growth of shoots while at lower concentration excessive adventitious roots were observed in the cultures, in addition to axillary shoot proliferation.
However, intermediate concentration of BAP at 5 µM favored maximum rate of shoot multiplication with no adventitious roots.
On MS + 5.0 µM BAP, 10 fold shoot multiplication was achieved every 5 weeks by cutting the solitary in vitro shoot into single node segments and culturing them onto the fresh medium of the same composition.
The shoots were successfully rooted on half strength MS medium (major salts reduced to half strength) with 50 g/L sucrose; roots developed directly at the base of the shoot in 100% cultures.
Plantlets were hardened and successfully established in the soil.
Regeneration protocol developed in this study would provide a basis for germplasm conservation and mass cultivation of the plant for later biotechnological use.
Publication
Authors
M. Singh , R. Chaturvedi
Keywords
axillary shoot proliferation, clonal propagation, cytokinins, shoot multiplication
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