Optimization of quality DNA isolation protocol from various mucilage rich cultivated and wild Abelmoschus sp. and its validation through PCR amplification

Abstract

Okra is such a genomically less explored crop that, many
labs are still trying for the optimization of quality DNA
isolation protocol for the efficient use in the contemporary
genomic studies. In this study, we have optimized a quick
and reproducible DNA isolation protocol for the isolation
of quality genomic DNA from different tissues of two
cultivated (Abelmoschus esculentus and A. caillei) and seven
wild okra species (A. manihot, A. moschatus, A. tetraphyllus,
A. tuberculatus, A. ficulneus, A. rugosus and A. angulosus),
including related species Hibiscus cannabinus. The quality
of isolated DNA was also confirmed using PCR amplification
of various DNA markers like Inter Simple Sequence Repeat
(ISSR), Random Amplified Polymorphic DNA (RAPD), Start
Codon Targeted (SCoT) and Expressed Sequence Tag-Simple
Sequence Repeat (EST-SSR). This is probably the first report
wherein the DNA isolation protocol has been optimized for
nine Abelmoschus species along with a related species
Hibiscus. Thus, good quality and quantity of DNA can be
isolated in okra, with the care that it should be performed
using appropriate plant tissue, either very young leaves or
etiolated seedlings, and well optimized DNA isolation
protocol as prescribed in this study.