Morphological and Molecular Characterization of Gnetum africanum (Welw.) Germplasm Using DNA Barcoding Method

Abstract

Gnetum africanum is a twisted woody climbing wild plant i.e.liana found growing naturally in dense equatorial rainforest but currently domesticated in tropical areas of the world. There has been taxonomic confusion in classifying this liana due to its observable diverse morphological characters. Indeed, morphological characterization alone has not offered the needed solution. However, there is dearth of information on the molecular characterization of Gnetum africanum. Morphological and molecular descriptors, could help offers the needed phenotypic and genotypic characterization of this plant much better. This study is aimed at characterizing Gnetum africanum land races to identify species that could be used for the development of high yielding hybrids. Three different land races or accessions of Gnetum africanum was collected from different locations in Nigeria. High quality genomic DNA was isolated from the different samples; it was checked for purity and quantified as appropriate. The DNA samples were subjected to PCR amplification and DNA barcoding studies (chloroplast large subunit of ribulose -1, 5-bisphophate carboxylase rbcL gene). The amplicons were gel eluted, sequenced and checked for homology by using Basic local alignment search tool -BLAST. Identification was obtained from the top similar outcomes of the samples. For Ekim Fig. 1b sample, the forward blast showed 95% similarity with Basella alba voucherID: NC041293.1 and the reverse had 88% similarity with Talinum fruticosum cultivar ID: MK598685.1. For Etinan Fig. 1a sample, the forward blast had 86% similarity with Rhabdothamnussolandri voucher ID: JQ933464.1 and for reverse, there was no similarity founded.

Evaluation methods for development and selection of novel probiotics

“…This is followed by characterization of the isolates, usually using morphological, biochemical and molecular characterization methods leading to rapid identification of the probiotic organisms. The most common molecular characterization for identification of unknown organisms is the DNA Barcoding Method 14 and Simple Sequence Repeat-SSR marker 15 for sequencing of conserved regions within the genome, such as 16S ribosomal DNA in bacteria and ITS regions for yeasts and fungi. 16 The identification is done by comparing the sequences with data stored in biotechnology data-bank or GenBank and in addition with phylogenetic analysis.…”