Micropropagation of <i>Clerodendrum aculeatum </i>through adventitious shoot induction and production of consistent amount of virus resistance inducing protein

Abstract

Rapid micropropagation through adventitious shoot induction from <i>in vitro </i>raised leaf ex plants of <i>Clerodendrum aculeatum </i>(Verbenaceae), was successfully achieved for the first time. Basal portion of the leaves showed highest regeneration<i> </i>potential when grown on MS medium supplemented with BA (5.0 mg/l) and NAA and IBA (0.5 mg/l of each). Shoots after<i> </i>elongation in growth regulator-free medium, were rooted in MS medium containing 0.5 mg/l of NAA and IBA. Aqueous<i> </i>leaf extract of <i>in vitro </i>raised plants, induced high degree of resistance against viruses in susceptible healthy hosts when applied<i> </i>prior to virus inoculation. Upon purification from leaves of cultured plants, the resistance inducing protein, showed

molecular mass of 34 kDa. Amount of resistance inducing protein obtained from leaves of cultured plants, was consistent throughout the year, as compared to the protein isolated from leaves of field grown plants, which showed marked seasonal fluctuation. The purified 34 kDa protein from <i>in vitro </i>raised plants, was serologically related to field grown plants and possessed similar characteristics. The micropropagated plants were successfully established in earthen pots under greenhouse conditions.