Abstract
Molecular phylogenetics in general, particularly DNA barcoding, has revolutionized the systematic botany for exact identification of the species with respect to its possible affinities. Pakistan being blessed with a wide range of rare and endemic plants is devoid of critical data on its native flora mostly threatened to the verge of extinction. This paper explores the applicability of DNA barcodes, viz. rbcLa, matK and nrITs for the reconstruction of the phylogeny of Haplophyllum gilesii (Hemsel.) C. C. Townsend, a critically endangered, narrow endemic medicinal plant in Pakistan. The DNA sequences after retrieving from NCBI, for maximum likelihood, near neighbor joining and maximum parsimony, close match and best close match analyses, were performed using MUSCLE, BioEdit, MEGA-7 and TAXONDNA softwares and revealed that the universal plastid barcodes, rbcLa and nrlTs carrying 593 bp and 458 bp, respectively, are effective candidates for precise molecular description of this species, while matK remained as a challenge in this regard. BlastN algorithm successfully matched giving 99% and 96% identity with the reference datasets. The results further confirmed 75% best and best close match for rbcLa 12% while 0.0% for nrITs. Our findings are the first of its nature for Pakistan, which provides a sound foundation for molecular biosystematics of the threatened flora of the region.
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Acknowledgements
The Higher Education Commission of Pakistan (HEC) graciously sponsored the project under HEC 5000 Ph.D. program, which is admirable at all the levels. Prof. Dr. Anjum Parveen, Director, Karachi University Herbarium (KUH) and Dr. Jan Alam Assistant Professor, Department of Botany, Hazara University donated the plant material, which is acknowledged with thanks.
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BK (Principal author) designed and performed the experiments, prepared figures and/or tables, authored, reviewed and approved the final draft; HA (Supervisor) conceptualized the project and designed the work plan and approved the final version of the manuscript for publication; KM (Co-Supervisor) contributed reagents/materials/analysis tools, bioinformatic analysis, interpretation of data, drafting of article and final approval of the version to be published; IU helped in the final approval of the version to be published; MSN and AU reviewed and approved the final draft; JA provided the plant research material and assisted in correct plant identification; and SR performed the experiments.
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Khan, B., Ahmad, H., Alam, J. et al. DNA barcoding of Haplophyllum gilesii (Hemsel.) C. C. Townsend from the Gilgit-Baltistan Province of Pakistan. Braz. J. Bot 42, 651–659 (2019). https://doi.org/10.1007/s40415-019-00568-5
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DOI: https://doi.org/10.1007/s40415-019-00568-5