MOLECULAR IDENTIFICATION OF Juniperus GENOTYPES FROM GILGIT-BALTISTAN USING CHLOROPLAST DNA BARCODES

Authors: Rohma Nazir, Sajid ul Ghafoor, Aziz Uddin,  Hamid Ali, Sajjad, Samman Iqbal, Muhammad Haris, Irfan Ullah
Journal: Journal of Animal and Plant Sciences (JAPS)
ISSN: 1018-7081 (Print), 2309-8694 (Online)
Volume: 36  Issue: 4  
Year: 2026
DOI: https://doi.org/10.36899/JAPS.2026.4.0096
URL: https://doi.org/https://doi.org/10.36899/JAPS.2026.4.0096
Publisher: Pakistan Agricultural Scientists Forum

Abstract:
<p style="text-align: justify;"><span lang="EN-GB" style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-GB; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">DNA barcoding is a molecular identification technique that utilizes a standardized region of the genome to distinguish organisms at the genus and species levels. The current study aims to confirm and identify <em>Juniperus</em> species using chloroplast DNA Barcodes. A total of 16 samples were collected from various biodiversity-rich locations in the Hunza Valley, Gilgit-Baltistan (GB), Pakistan. Among them, 4 samples were selected based on variable morphological traits for molecular confirmation. A total gDNA was isolated by CTAB method and confirmed through gel electrophoresis. Chloroplast markers (<em style="mso-bidi-font-style: normal;">mat</em>K<em style="mso-bidi-font-style: normal;"> </em><span style="mso-bidi-font-style: italic;">and</span><em style="mso-bidi-font-style: normal;"> rbc</em>L) were used to amplify and sequence the targeted regions to confirm homology and resolving taxonomic uncertainties. Both the selected markers showed good results in amplification, and sequencing. While in molecular data analysis, various types of sites and domains were recorded in the form of conserved, variable, parsimony-informative and singleton sites. In phylogenetic tree analysis, the query species sequences shared a clade with <em>Juniperus </em><em style="mso-bidi-font-style: normal;">chinensis,</em> <em>Juniperus</em><em style="mso-bidi-font-style: normal;"> </em></span><em style="mso-bidi-font-style: normal;"><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">communis,</span></em><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-GB; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> <em><span lang="EN-GB">Juniperus</span></em></span><em style="mso-bidi-font-style: normal;"><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> excelsa, </span></em><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">and <em>Juniperus</em><em style="mso-bidi-font-style: normal;"> squamata </em>with bootstrap values greater than 90%. In conclusion, this study provides one of the first molecular characterizations studies of <em>Juniperus </em>species specifically from the high-altitude zones of Hunza, GB. The findings contribute to accurate species identification and conservation efforts for these ecologically important </span><em><span lang="EN-GB" style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-GB; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">Juniperus </span></em><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 107%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-font-kerning: 0pt; mso-ligatures: none; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">species.</span></p>

Keywords: Genetic diversity; Himalayan flora; Conifer identification; Chloroplast genome