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Plant tissue culture techniques offer a great opportunity to overcome the limitations associated with the large scale cultivation of spine gourd. Present study was carried out to formulate the best possible media for large scale production of spine gourd and result of the study revealed that highest percentage (85%) of embryogenic callus was obtained from MS medium supplemented with 2.0 mg/L each of 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and 6-Benzylamino Purine (BAP) in leaf explants of spine gourd. Maximum number of shoots (12.15 ± 1.51 shoots) were observed on MS medium augmented with BAP (4.0 mg/L) in combination with L-glutamine (2.0 mg/L) from leaf derived embryogenic callus of spine gourd. Identification of sex by using morphological characters in the newly regenerated plantlets of spine gourd at fourth leaf stage is another problem for large scale propagation of female plants. PCR based molecular marker OPA-15, a Random Amplified Polymorphic DNA (RAPD) primer can be used as a differential marker to identify female plants form male plants at pre-flowering stage in newly regenerated plantlets (in vitro) and as well as in field plants (in vivo) of spine gourd. A unique amplification band (700 bp) in size appeared only in female samples, but not in male samples of spine gourd.

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