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Micropropagation through excised root culture of Clitoria ternatea and comparison between in vitro –regenerated plants and seedlings
Author(s) -
Shahzad A.,
Faisal M.,
Anis M.
Publication year - 2007
Publication title -
annals of applied biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.677
H-Index - 80
eISSN - 1744-7348
pISSN - 0003-4746
DOI - 10.1111/j.1744-7348.2007.00132.x
Subject(s) - shoot , kinetin , clitoria ternatea , biology , explant culture , murashige and skoog medium , micropropagation , botany , in vitro , biochemistry , medicine , alternative medicine , pathology
An efficient protocol has been developed for high‐frequency shoot regeneration and plant establishment of Clitoria ternatea – a potential medicinal legume. Adventitious shoots were regenerated from young excised root segments of aseptic seedlings on Murashige and Skoog (MS) medium supplemented with various concentrations of 6‐benzyladenine (BA), kinetin, α‐naphthalene acetic acid (NAA) or 2,4‐dichlorophenoxy acetic acid (2,4‐D) either singly or in various combinations. The highest frequency (100%) of shoot regeneration and maximum number (16.4 ± 0.24) of shoots per explant was obtained on MS medium supplemented with 20 μ m BA and 2.0 μ m NAA. Organogenic calli were produced on a medium containing 2,4‐D (10 or 20 μ m ) and BA (5.0 μ m ). The calli were differentiated into multiple shoots on MS medium supplemented with 2.5–10 μ m BA and 2.0 μ m NAA. The microshoots were rooted on half‐strength MS medium supplemented with 5.0 μ m indole‐3‐butyric acid and transplanted successfully in field conditions. After 12 months of transfer to ex vitro conditions, the performance of micropropagated plants were evaluated on the basis of some physiological and biochemical parameters and compared with the in vivo –grown plants of the same age. The sodium dodecyl sulphate polyacrylamide gel electrophoresis protein profile was same between regenerated and naturally growing shoots. Total soluble protein in aerial part as well as in seeds of in vitro –regenerated and in vivo –grown plants was almost the same. The mitotic study showed normal chromosomal movement and numbers (2 x = 16).