Genetic and phytochemical analysis of the in vitro regenerated Pilosocereus robinii by ISSR, SDS–PAGE and HPLC.Gene 533 (2014) 313–321
• 2013
Publication Information
Authors
Salah Khattab a, Fadia El Sherif a, Hoda A. El-Garhy b, Safwat Ahmedc, Amany Ibrahim
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publication.type
International
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Abstract
Pilosocereus robinii is a rare specieswhich is experiencing sudden population collapse. Identifying and developing
effective conservation and management strategies to halt the forestall extinction of this species is crucial. The
present study was conducted to assess the best conditions for in vitro propagation of this plant in regard to its
morphogenic, genetic as well as the chemical potentials. A successful in vitro propagation system of P. robinii
has been developed. MS hormone-free medium induced the best root morphogenic potential. The plants were
acclimatized in the greenhouse at 100% survival rate. Besides, the somaclonal variations between the in vitro
raised plants were analyzed using PCR-ISSR markers and SDS–PAGE protein, where the regenerated explants
onMSmediumsupplementedwith TDZwere the highest in inducing newspecificmarker bands.Sh6ISSRprimer
showed the highest polymorphismvalue, 81.8%with 33 totalamplified fragments,while Sh3 ISSR primer showed
the lowest value with polymorphic percentage of 14.3%. Furthermore, SDS–PAGE protein analysis showed no
variation in protein pattern of the studied treatments. On the other side, HPLC analysis of the in vitro plantlets extracts
has shown that 2iP based treatments were the highest in organic acids accumulation, while the phenolic
constituents' accumulation was found to reach its peak in the BA based treatments
effective conservation and management strategies to halt the forestall extinction of this species is crucial. The
present study was conducted to assess the best conditions for in vitro propagation of this plant in regard to its
morphogenic, genetic as well as the chemical potentials. A successful in vitro propagation system of P. robinii
has been developed. MS hormone-free medium induced the best root morphogenic potential. The plants were
acclimatized in the greenhouse at 100% survival rate. Besides, the somaclonal variations between the in vitro
raised plants were analyzed using PCR-ISSR markers and SDS–PAGE protein, where the regenerated explants
onMSmediumsupplementedwith TDZwere the highest in inducing newspecificmarker bands.Sh6ISSRprimer
showed the highest polymorphismvalue, 81.8%with 33 totalamplified fragments,while Sh3 ISSR primer showed
the lowest value with polymorphic percentage of 14.3%. Furthermore, SDS–PAGE protein analysis showed no
variation in protein pattern of the studied treatments. On the other side, HPLC analysis of the in vitro plantlets extracts
has shown that 2iP based treatments were the highest in organic acids accumulation, while the phenolic
constituents' accumulation was found to reach its peak in the BA based treatments
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