DNA Barcoding, Identification and Validation of the Genus Phyllanthus in Nigeria using Plastid rbcL and matK Genetic Markers

Chiedozie Joel Uka; Daniel Awomukwu; Bio Louis Nyananyo

Authors

Chiedozie Joel Uka
Daniel Awomukwu
Bio Louis Nyananyo

Phyllanthus species, DNA extraction, PCR amplification, nucleotide sequences, BLAST, Bold Systems, rbcL, matk, Barcoding

Abstract

Mull-Arg. and P. muellerianus (O. Ktze) Excel belonging to the family of Phyllanthaceae were extracted, purified, PCR amplified and sequenced using plastid Ribulose -1,5 bisphosphate carboxylase large chain (rbcL a) and Maturase K (matK) genetic marker to identify unknown Phyllanthus species. The plastid region revealed that the Phyllanthus species were able to be amplified optimally for sequencing. The results of the nucleotide sequences were further compared on Basic Local Alignment Sequence Tool (BLAST) on Gen Bank and Bold Systems for validation. Results revealed that the closely related species, P. niruroides Mull_Arg. and P. odontadenius Mull-Arg. had no DNA record to separate them on both GenBank and Bold Systems using both rbcL and matK gene regions while P. amarus Schum and Thonn and P. urinaria Linn. were clearly compatible with other works. P. muellerianus (O. Ktze) Excel was only compatible with other works using matK gene region but none for rbcL.

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DNA Barcoding, Identification and Validation of the Genus Phyllanthus in Nigeria using Plastid rbcL and matK Genetic Markers. (2015). Global Journal of Science Frontier Research, 15(36), 1-12. https://journalofscience.org/index.php/GJSFR/article/view/

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References

D. Awomukwu, B. Nyananyo, N. Onukwube, C. Uka, C. Okeke, A. Ikpeama (2014) Comparative Phytochemical Constituents and Pharmacognistic Importance of the Vegetative Organs of Some Phyllanthus species in South Eastern Nigeria. 4(2), 29-39.

D. Awomukwu, B. Nyananyo, P. Spies, B. Sizani (2015) Identification, Validation and Classification of the genus Phyllanthus in Nigeria using ITS Genetic Marker and the Taxonomic Implication. 3(1), 1-7.

L. Bancilhon (1971) Contribution a´ l'e´tude taxonomique du genre Phyllanthus (Euphorbiace´es). 18, 9-81.

M. Blaxter (2004) The promise of a DNA taxonomy. Philosophical Transactions of the Royal Society of London. 359, 669-679.

CBOL Plant Working Group (2009) A DNA barcode for land plants. Proceedings of the National Academy of Sciences. 106, 2794-1279.

J. Doyle, J. Doyle (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. 19, 11115.

J. Doyle, J. Doyle (1990) Isolation of plant DNA from fresh tissue. 12, 13-15.

H. Edeoga, G. Omosun, G. Osuagwu, D. Awomukwu (2007) Anatomical Features of Vegetative Organs of Phyllanthus species. 25(3), 548-553.

Geneious 7.0.4. http://www.geneious.com

Genbank ( http://www.ncbi.nlm.nih.gov/Blast.cgi

R. Henry (2001) Plant genotyping: the DNA fingerprinting of plants. https://doi.org/10.1079/9780851995151.0000

J. Hutchinson, M. Dalziel (1963) Flora of West Tropical Africa. 2.

H. Kathriarachchi, P. Hoffmann, R. Samuel, K. Wurdack, M. Chase (2005) Molecular phylogenetics of Phyllanthaceae inferred from five genes (plastid atpB, matK, 30ndhF, rbcL, and nuclear PHYC). 36, 112-134.

W. Kress, David Erickson (2007) A Two-Locus Global DNA Barcode for Land Plants: The Coding rbcL Gene Complements the Non-Coding trnH-psbA Spacer Region. 2(6), e508. https://doi.org/10.1371/journal.pone.0000508

W. Kress, Kenneth Wurdack, Elizabeth Zimmer, Lee Weigt, Daniel Janzen (2005) Use of DNA barcodes to identify flowering plants. 102(23), 8369-8374. https://doi.org/10.1073/pnas.0503123102

E. Ko¨ hler (1965) Die Pollenmorphologie der Biovulaten Euphorbiaceae und ihre Bedeutung für die Taxonomie. 6(1), 26-120. https://doi.org/10.1080/00173136509429138

E. Ko¨ hler (1967) Über Beziehungen zwischen Pollenmorphologie und Polyploidiestufen im Verwandtschaftsbereich der Gattung Phyllanthus. (Euphorbiaceae)Mit 1 Abbildung und 3 Tafeln. 74(3), 159-165. https://doi.org/10.1002/fedr.19670740304

V. Kumari, A. Bansal, R. Aminedi, D. Taneja, N. Das (2012) Simplified extraction of good quality genomic DNA from a variety of plant materials. 11, 6420-6427.

D. Paul, A. Herbert, L. Shelly, R. Jeremy (2003) Biological Identification through DNA barcodes. 270, 313-321.

W. Punt (1967) Pollen morphology of the genus Phyllanthus (Euphorbiaceae). 3(1-4), 141-150. https://doi.org/10.1016/0034-6667(67)90047-4

W. Punt (1987) A survey of pollen morphology in Euphorbiaceae with special reference to Phyllanthus. 94(1-2), 127-142. https://doi.org/10.1111/j.1095-8339.1987.tb01042.x

M. Rahman (2007) DNA fingerprinting in Utricularia L. Section Utricularia as revealed by PCR based assay. 3(1), 56-68.

(2003) Pollen morphology of the Flueggeinae (Euphorbiaceae, Phyllanthoideae). 42(4), 193-219. https://doi.org/10.1080/00173130310016464

Rosabelle Samuel, Hashendra Kathriarachchi, Petra Hoffmann, Michael Barfuss, Kenneth Wurdack, Charles Davis, Mark Chase (2005) Molecular phylogenetics of Phyllanthaceae: evidence from plastid MATK and nuclear PHYC sequences. 92(1), 132-141. https://doi.org/10.3732/ajb.92.1.132

M. Stoeckle, D. Janzen, W. Hallwachs, J. Hanken, J. Baker (2003) Taxonomy, DNA, and the Barcode of Life.

M. Stoeckle, P. Waggoner, J. Ausubel (2004) Consortium for the Barcode of Life, Inaugural Meeting.

N. Sucher, M. Charles (2008) Genome-Based Approaches to the Authentication of Medicinal Plants. 74(6), 603-623. https://doi.org/10.1055/s-2008-1074517

C. Uka, C. Okeke, D. Awomukwu, B. Aziagba, R. Muoka (2014) Taxonomic Significance of Foliar Epidermis of some Phyllanthus species in South Eastern Nigeria. 9(4), 1-6.

G. Webster (1956) A monographic study of the West Indian species of Phyllanthus. 37, 217-268. https://doi.org/10.5962/bhl.part.25737

Grady Webster, Kevin Carpenter (2002) Pollen morphology and phylogenetic relationships in neotropical Phyllanthus (Euphorbiaceae). 138(3), 325-338. https://doi.org/10.1046/j.1095-8339.2002.00009.x

Grady Webster, J. Ellis (1962) CYTOTAXONOMIC STUDIES IN THE EUPHORBIACEAE, SUBTRIBE PHYLLANTHINAE. 49(1), 14-18. https://doi.org/10.1002/j.1537-2197.1962.tb11740.x

K. Weisling, H. Nybom, K. Wolff, G. Kahl (2005) DNA fingerprinting in plants: Principles, methods and applications.

Kenneth Wurdack, Petra Hoffmann, Rosabelle Samuel, Anette De Bruijn, Michelle Van Der Bank, Mark Chase (2004) Molecular phylogenetic analysis of Phyllanthaceae (Phyllanthoideae pro parte, Euphorbiaceae sensu lato) using plastid RBCL DNA sequences. 91(11), 1882-1900. https://doi.org/10.3732/ajb.91.11.1882