Development of in-vitro protocol for direct regeneration from thalamus ex-plant of Tagetes patula L. var. Pusa Deep

Abstract

The effect of growth hormones such as BAP, IAA and KINETIN in different treatment combinations on direct differentiation of shoots from thalamus ex-plants of French marigold (Tagetes patula L. var. “Pusa Deep”) have been studied. A total of nine treatments was used to screen thalamus ex-plants to determine the capability for plant regeneration and subsequently to find out the optimum medium conditions for high potentiality of direct shoot formation. Thalamus ex-plants behaved differently in all over nine treatments and all the treatments are highly significant over the (T0) control-MS devoid of hormones. The best medium found for direct, shoot organogenesis from the thalamus of Pusa Deep was treatment (T7) MS + BAP 1.0 mg/l + KIN 0.5 mg/l + IAA 1.0 mg/. Pre-treatment with Carbendazim (0.2%) + Mancozeb (0.2%) + 8-HQC (200 mg/l) for 1.5hr resulted in minimum fungal (28.33%), bacterial (25.33%) contamination. Surface sterilization with 0.1% HgCl2 for 5 min reduced microbial contamination up to (44.33%) simultaneously increased the survival percentage upto (55.67%). This technique can be utilized in gene transfer technology, in vitro mutagenesis, production of lutein rich pharmaceutical compounds and secondary metabolites using tissue culture techniques.