Assessment of genetic stability in somatic embryo derived plantlets of Pterocarpus marsupium Roxb. using inter-simple sequence repeat analysis
Authors
Radhika Tippani
Department of Biotechnology, Kakatiya University, Warangal, Telangana 506 009 India
Rama Nanna
Department of Biotechnology, Kakatiya University, Warangal, Telangana 506 009 India
Praveen Mamidala
Department of Biotechnology, Telangana University, Nizamabad, Telangana India
Christopher Thammidala
Department of Biotechnology, Kakatiya University, Warangal, Telangana 506 009 India
Keywords:
Indian Kino tree, Immature zygotic embryos, Somatic embryogenesis, Plantlet conversion, Genetic stability
Abstract
An efficient plantlet regeneration protocol using immature zygotic embryos (IZEs) via somatic embryogenesis has been developed in Pterocarpus marsupium Roxb. The regenerated plantlets were evaluated for their genetic stability. IZEs were incubated on Murashige and Skoog (MS) media augmented with 1.07–16.11 μM naphthalene acetic acid (NAA) or 0.90–13.97 μM 2,4-dichlorophenoxyacetic acid. The optimum callus induction (96.6%) was observed on MS medium augmented with 5.37 μM NAA. Induction of somatic embryos (SEs) was observed after sub-culture of calli on medium with decreased concentrations of NAA (0.54–5.37 μM), either alone or 2.69 μM NAA in combination with 2.22–8.90 μM benzyladenine (BA) or 2.32–9.30 μM Kinetin. Maximum number (33.4 ± 0.85) of SEs occurred on MS medium augmented with 2.69 μM NAA + 4.40 μM BA + 3% sucrose. Highest percentage (67.3 ± 0.37) of SEs matured and developed into cotyledonary stage by subsequent subculture on the same medium. SE formation and maturation decreased when sucrose concentrations were higher than 3%. Seventy percent of mature somatic embryos developed into plantlets on half strength MS medium augmented with 5.80 µM gibberellic acid. The various stages of development during somatic embryogenesis include globular, heart, torpedo and mature stages as revealed by the stereomicroscopic and histological studies of explants. Plantlets derived from SEs were successfully acclimatized in the greenhouse with a survival rate of 78%. Among the survived plantlets, 9 plantlets were randomly selected for inter-simple sequence repeat (ISSR) analysis. Of the 13 primers used, 8 produced reproducible and monomorphic bands. ISSR analysis revealed a homogenous amplification profile for all regenerated plantlets analyzed validating the genetic stability of somatic embryo derived plantlets.
Electronic supplementary material
The online version of this article (10.1007/s12298-018-0602-8) contains supplementary material, which is available to authorized users.
Keywords: Indian Kino tree, Immature zygotic embryos, Somatic embryogenesis, Plantlet conversion, Genetic stability
Author Biography
Christopher Thammidala, Department of Biotechnology, Kakatiya University, Warangal, Telangana 506 009 India
Department of Botany, Kakatiya University, Warangal, Telangana 506 009 India
Click on "Archives" to access the full archive of scientific preprints. You may use the categories and the search functionality to find select preprints you're interested in.
