Assessment of genetic stability on in vitro and ex vitro plants of Ficus carica var. black jack using ISSR and DAMD markers

Ankita Rajendra Parab,Chew Bee Lynn,Sreeramanan Subramaniam

doi:10.1007/s11033-021-06714-1

Ankita Rajendra Parab, Chew Bee Lynn + Show 1 more

Open Access

https://doi.org/10.1007/s11033-021-06714-1

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Abstract

Clonal propagation is one of the attributes of plant tissue culture. Therefore, analysis of genetic stability among the in vitro cultured plants is a crucial step. It helps to signify the clonal propagation of the micropropagated plants. Regenerated Ficus carica var. Black Jack plantlets were established using woody plant medium supplemented with 20μM 6-Benzylaminopurine and 8μM Indole-3-acetic acid under different light treatments such as normal fluorescent white light (60μmolm-2s-1), and four different LED spectra, white (400-700nm), blue (440nm), red (660nm) and blue + red (440nm + 660nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of Ficus carica var. Black Jack. Ten primers of each, ISSR and DAMD molecular markers, were used to assess the genetic stability of the eight samples of Ficus carica var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR-DNA primer which was negated under the genetic similarity index analysis for the eight samples. The findings of this study revealed that ISSR and DAMD markers are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var. Black Jack. Monomorphism of 100% obtained using DAMD markers and more than 95% of monomorphism obtained using ISSR markers indicate that the regenerated plants are significantly genetically stable. These molecular markers can be used to test the genetic stability of in vitro regenerated plants. It is recommended that genetic stability analysis should be performed for long-term maintenance of such micropropagated plants.

Highlights

Micropropagation is a technique widely used for the commercialization of many woody plant species
The findings of this study revealed that inter simple sequence repeats (ISSR) and directed amplification of minisatellite Deoxyribonucleic acid (DNA) (DAMD) markers (DNA primers) are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var
Polymorphism of 2.13% was observed for the UBC840 ISSR – DNA primer which was negated under the genetic similarity index analysis for the eight samples

Summary

Introduction

Micropropagation is a technique widely used for the commercialization of many woody plant species. Producing a whole plat using an explant is termed micropropagation. The application of micropropagation includes growing any plant species on a large scale commercially, clonal propagation of endangered species of plants, somaclonal variations, producing transgenic plant cell lines and sterile hybrids with enhanced properties, and acquiring essential secondary metabolites. The micropropagation system has a huge advantage in the commercial market of plant tissue culture, owing to its excellent multiplication techniques and viability. Using highly dividing meristematic tissues of the apical buds, leaves, or roots are used as explants to achieve plant regeneration through various techniques such as organogenesis or somatic embryogenesis. Using highly dividing meristematic tissues of the apical buds, leaves, or roots are used as explants to achieve plant regeneration through various techniques such as organogenesis or somatic embryogenesis. (Rathore et al 2005; Iliev et al 2010; Ružićet al. 2012)

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