In vitro propagation of spine gourd (Momordica dioica Roxb.) and assessment of genetic fidelity of micropropagated plants using RAPD analysis

Abstract

An efficient protocol for rapid in vitro clonal propagation of spine gourd (Momordica dioica Roxb.) genotype RSR/DR15 (female) and DR/NKB-28 (male) was developed through enhanced axillary shoot proliferation from nodal segments. Maximum shoot proliferation of 6.2 shoots per explant with 100 % shoot regeneration frequency was obtained from the female genotype on Murashige and Skoog's (1962) medium supplemented with 0.9μM N6-benzyladenine (BA) and 200 mg l -1 casein hydrolysate (CH). While from the male genotype the optimum shoot regeneration frequency (86.6 %) and 6.4 shoots per explant was obtained on MS medium supplemented with 2.2μM BA. CH induced vigorous shoots, promoted callus formation, and proved inhibitory for shoot differentiation and shoot length, especially in explants from male genotype. Rooting was optimum on half-strength MS medium (male 92.8 %, female 74.6 %) containing 4.9μM indole-3-butyric acid (IBA). Plantlets were transferred to plastic cups containing a mixture of cocopit and perlite (1:1 ratio) and then to soil after 2-3 weeks. 84 % female and 81 % male regenerated plantlets survived and grew vigorously in the field. Genetic stability of the regenerated plants was assessed using random amplified polymorphic DNA (RAPD). The amplification products were monomorphic in the in vitro propagated plants and similar to those of mother plant. No polymorphism was detected revealing the genetic integrity of in vitro propagated plants. This micropropagation procedure could be useful for raising genetically uniform planting material of known sex for commercial cultivation or build-up of plant material of a specific sex-type.

Plant regeneration from direct and indirect organogenesis and assessment of genetic fidelity in Saccharum officinarum using DNA-based markers

“…2. Similar results were reported in micro-propagated shoots of Curcuma longa (Salvi et al 2001), sugarcane (Khan et al 2009), Gerbera jasmesonii (Bhatia et al 2011), Momordica dioica (Rai et al 2012), Terminallia bellerica (Dangi et al 2014), and Dendrocalamus strictus (Goyal et al 2015). Zucchi et al (Zucchi et al 2002) reported that polymorphism occurred in sugarcane shoots regenerated from meristem cultures because of the number of subcultures and RAPD analysis was used for detection of polymorphism.…”

Clonal Propagation and Karyomorphological Study of Solanum torvum Swartz: An Ethnobotanical Species of Tripura, India

“…This observation is in corroboration with previous records on several micropropagation studies (Rai et al 2012, Zakia et al 2013, Mehdi et al 2014. Presence of cytokinin at higher concentration in the medium lead to reduced number of shoot formation.…”

A genetically stable rooting protocol for propagating a threatened medicinal plant--Celastrus paniculatus

“…3 D) clearly suggest that the micropropagated plants were genetically identical to the mother plant and no variation was induced during in vitro propagation. Molecular markers, mainly RAPD, have been widely used for testing the genetic fidelity among in vitro regenerated plantlets in a number of plant species ( Ahmad and Anis 2011 ; Ramesh et al 2011 ; Sharma et al 2011 ; Rai, G. K. et al 2012 ; Savita et al 2012 ; Singh et al 2012 ), owing to their simplicity, cost-effectiveness and lower DNA requirement for analysis ( Asthana et al 2011 ).…”