IJCOT-book-cover International Journal of Biotech Trends and Technology  (IJBTT)          
 
© 2023 by IJBTT Journal
Volume - 13 Issue - 2
Year of Publication : 2023
Authors : Jyotsana Khushwaha, Alpana Joshi
DOI :   10.14445/22490183/IJBTT-V13I2P604

How to Cite?

Jyotsana Khushwaha, Alpana Joshi, "In Silico Characterization and Phylogenetic Analysis of Elaeocarpus Ganitrus based on ITS2 Barcode Sequence" International Journal of Biotech Trends and Technology  vol. 13, no. 2, pp. 26-37, 2023. Crossref, https://doi.org/10.14445/22490183/IJBTT-V13I2P604 

Abstract

Plant molecular systematics relies on using DNA barcodes for studying the evolutionary relationship between species Sequences of the nuclear internal transcribed spacer (ITS) regions have been used widely in molecular phylogenetic studies because of their high variability compared to plastid sequences. Elaeocarpus is a diverse genus within the family Elaeocarpaceae and is widely distributed worldwide among tropical and subtropical climatic zones. Elaeocarpus ganitrus has important medicinal and religious values in India. However, Elaeocarpus ganitrus's evolutionary relationship with other Elaeocarpus species is not much explored, especially at the molecular and phylogenetic levels. The present research successfully amplified the nuclear gene ITS2, sequenced and submitted it to NCBI Genbank after using Basic Local Alignment Search Tool (BLAST). Automatic Barcode Gap Discovery (ABGD) and Assemble Species by Automatic Partitioning (ASAP) resulted in different numbers of molecular operational taxonomic units (MOTUs). The lowest score of ASAP (4.5) segregated the sequences into 31 MOTUs with the Threshold dist. value of 0.003524. This study establishes an evolutionary relationship between Elaeocarpus ganitrus and other species belonging to the same genus through the neighbor-joining method. The 38 Elaeocarpus samples were clustered into seven major groups based on ITS2 sequence: Group I is represented by Elaeocarpus ganitrus along with Elaeocarpus sylvestris, Elaeocarpus glabripetalus, Elaeocarpus duclouxii, Elaeocarpus decipiens, and Elaeocarpus zollingeri. Group II is characterized by Elaeocarpus austroyunnanensis and Elaeocarpus glaber. Group III comprises Elaeocarpus sphaericus, Elaeocarpus angustifolius, Elaeocarpus grandis, Elaeocarpus ptilanthus, and Elaeocarpus sphaerocarpus. Three accessions of Elaeocarpus hookerianus are placed in group IV. Elaeocarpus largiflorens and Elaeocarpus thelmae represent group V. Groupr VI contains three species: Elaeocarpus sylvestris, Elaeocarpus dubius, and Elaeocarpus johnsonii. Group VII comprises five species which include Elaeocarpus glabripetalus, Elaeocarpus rugosus, Elaeocarpus tuberculatus, Elaeocarpus hainanensis, and Elaeocarpus angustifolius. The study concludes with the possibility of correctly using the ITS2 gene to identify, discriminate, and document Elaeocarpus ganitrus and other species of the same genus.

Keywords

DNA barcoding, Elaeocarpus ganitrus, Internal Transcribed Spacer (ITS), Molecular Identification, Molecular Operational Taxonomic Units (MOTUs).

References

[1] M. J. E. Coode, "Elaeocarpus for Flora Malesiana: New Taxa and Understanding in the Ganitrus Group," Kew Bulletin, vol. 65, no. 3, pp. 355–399, 2010.
[CrossRef] [Google Scholar] [Publisher Link]
[2] Sook-Ngoh Phoon, "Systematics and Biogeography of Elaeocarpus (Elaeocarpaceae)," PhD Thesis, James Cook University, 2015.
[CrossRef] [Google Scholar] [Publisher Link]
[3] D. J. Maynard, "A Molecular Phylogeny for Australian Elaeocarpus (Elaeocarpaceae) and the Affinities of a Putative New Taxon," Honours Thesis, University of New South Wales, 2004.
[Google Scholar]
[4] Darren M. Crayn, Maurizio Rossetto, and David J. Maynard, "Molecular Phylogeny and Dating Reveals an Oligo-Miocene Radiation of Dry-Adapted Shrubs (former Tremandraceae) from Rainforest Tree Progenitors (Elaeocarpaceae) in Australia," American Journal of Botany, vol. 93, no. 9, pp. 1328–1342, 2006.
[CrossRef] [Google Scholar] [Publisher Link]
[5] Yumiko Baba, "Evolution, Systematics and Taxonomy of Elaeocarpus (Elaeocarpaceae) in Australasia," PhD Thesis, James Cook University, Queensland, 2013.
[CrossRef] [Google Scholar] [Publisher Link]
[6] Ning Yu et al., "Barcode ITS2: a Useful Tool for Identifying Trachelospermum Jasminoides and a Good Monitor for Medicine Market," Scientific Reports, vol. 7, no. 5037, 2017.
[CrossRef] [Google Scholar] [Publisher Link]
[7] Paul D. N. Hebert et al., "Biological Identifications through DNA Barcodes," Proceedings of the Biological Science, The Royal Society B, vol. 270, no. 1512, pp. 313–321, 2003.
[CrossRef] [Google Scholar] [Publisher Link]
[8] Huili Li et al., “The Specific DNA Barcodes Based on Chloroplast Genes for Species Identification of Orchidaceae Plants,” Scientific Reports, vol. 11, no. 1424, 2021.
[CrossRef] [Google Scholar] [Publisher Link]
[9] Zulkifli Ahmad Seman et al., "Optimization of High-Yielding Protocol for DNA Extraction From-Leaves of Asam Gelugor (Garcinia Atroviridis), Malaysian High Economic Value of Medicinal Plant," SSRG International Journal of Agriculture & Environmental Science, vol. 9, no. 3, pp. 62-68, 2022.
[CrossRef] [Publisher Link]
[10] W. John Kress et al., "DNA Barcodes for Ecology, Evolution, and Conservation," Trends Ecology and Evolution, vol. 30, no. 1, pp. 25– 35, 2015.
[CrossRef] [Google Scholar] [Publisher Link]
[11] Pierre Taberlet et al., "Power and Limitations of the Chloroplast trn L (UAA) Intron for Plant DNA Barcoding," Nucleic Acids Research, vol. 35, no. 3, pp. e14, 2007.
[CrossRef] [Google Scholar] [Publisher Link]
[12] Aron J. Fazekas et al., DNA Barcoding Methods for Land Plants, Methods in Molecular Biology, vol. 858, pp. 223–252, 2012.
[CrossRef] [Google Scholar] [Publisher Link]
[13] Conrad L. Schoch et al., "Nuclear Ribosomal Internal Transcribed Spacer (ITS) Region as a Universal DNA Barcode Marker for Fungi," PNAS, vol. 109, no. 16, pp. 6241–6246, 2012.
[CrossRef] [Google Scholar] [Publisher Link]
[14] Gobinda Chandra Acharya et al., "Molecular Phylogeny, DNA Barcoding, and ITS2 Secondary Structure Predictions in the Medicinally Important Eryngium Genotypes of East Coast Region of India," Genes, vol. 13, no. 9, pp. 1678, 2022.
[CrossRef] [Google Scholar] [Publisher Link]
[15] Aekkhaluck Intharuksa et al., "The Combination of ITS2 and psbA-trnH Region is Powerful DNA Barcode Markers for Authentication of Medicinal Terminalia Plants from Thailand," Journal of Natural Medicines, vol. 74, no. 1, pp. 282–293, 2020.
[CrossRef] [Google Scholar] [Publisher Link]
[16] Mayengbam Premi Devi et al., “DNA Barcoding and ITS2 Secondary Structure Predictions in Taro (Colocasia esculenta L. Schott) from the North Eastern Hill Region of India,” Genes, vol. 13, no. 12, p. 2294, 2022.
[CrossRef] [Google Scholar] [Publisher Link]
[17] J. J. Doyle, "A Rapid DNA Isolation Procedure for Small Quantities of Fresh Leaf Tissue," Phytochemical Bulletin, vol. 19, pp. 11–15, 1987.
[Google Scholar]
[18] Nadia Aboul-Ftooh Aboul-Maat, and Hanaa Abdel-Sadek Oraby, "Extraction of High-Quality Genomic DNA from Different Plant Orders Applying A Modified CTAB-Based Method," Bulletin of the National Research Centre, vol. 43, no. 25, 2019.
[CrossRef] [Google Scholar] [Publisher Link]
[19] Alpana Joshi et al., “Phylogenetic Relationships Among Low-Ploidy Species of Poa using Chloroplast Sequences,” Genome, vol. 60, no. 5, pp. 384–392, 2017.
[CrossRef] [Google Scholar] [Publisher Link]
[20] Koichiro Tamura, Glen Stecher, and Sudhir Kumar, "MEGA11: Molecular Evolutionary Genetics Analysis Version 11," Molecular Biology and Evolution, vol. 38, no. 7, pp. 3022–3027, 2021.
[CrossRef] [Google Scholar] [Publisher Link]
[21] Motoo Kimura, "A Simple Method for Estimating Evolutionary Rates of Base Substitutions through Comparative Studies of Nucleotide Sequences," Journal of Molecular Evolution, vol. 16, pp. 111–120, 1980.
[CrossRef] [Google Scholar] [Publisher Link]
[22] Julio Rozas et al., "DnaSP 6: DNA Sequence Polymorphism Analysis of Large Data Sets," Molecular Biology and Evolution, vol. 34, no. 12, pp. 3299–3302, 2017.
[CrossRef] [Google Scholar] [Publisher Link]
[23] Nicolas Puillandre et al., “ABGD, Automatic Barcode Gap Discovery for Primary Species Delimitation,” Molecular Ecology, vol. 21, no. 8, pp. 1864–1877, 2012.
[CrossRef] [Google Scholar] [Publisher Link]
[24] Nicolas Puillandre, Sophie Brouillet, and Guillaume Achaz, "ASAP: Assemble Species by Automatic Partitioning," Molecular Ecology Resources, vol. 21, no. 2, pp. 609– 620, 2020.
[CrossRef] [Google Scholar] [Publisher Link]
[25] Tanakorn Suesatpanit et al., "Should DNA Sequence be Incorporated with Other Taxonomical Data for Routine Identifying of Plant Species?," BMC Complementary and Alternative Medicine, vol. 17, no. 437, 2017.
[CrossRef] [Google Scholar] [Publisher Link]
[26] K. Tamura, "Estimation of the Number of Nucleotide Substitutions When There are Strong Transition-Transversion and G+C-content Biases," Molecular Biology and Evolution, vol. 9, no. 4, pp. 678–687, 1992.
[CrossRef] [Google Scholar] [Publisher Link]
[27] Ling-zhen Cao, and Kong-ming Wu, "Genetic Diversity and Demographic History of Globe Skimmers (Odonata: Libellulidae) in China Based on Microsatellite and Mitochondrial DNA Markers," Scientific Reports, vol. 9, no. 8619, 2019.
[CrossRef] [Google Scholar] [Publisher Link]
[28] Dewi Indriyani Roslim, Siti Khumairoh, and Herman Herman, "Confirmation of Tuntun Angin (Elaeocarpus floribundus) Taxonomic Status Using matK and ITS Sequences," Biosaintifika: Journal of Biology & Biology Education, vol. 8, no. 3, pp. 393–400, 2016.
[CrossRef] [Google Scholar] [Publisher Link]
[29] CBOL Plant Working Group, "A DNA Barcode for Land Plants," PNAS, vol. 106, no. 31, pp. 12794–12797, 2009.
[CrossRef] [Google Scholar] [Publisher Link]