F.H.Machate Poster ID 682
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Poster ID: 682 DNA Barcoding of Invasive Succulent Plants in the Horticultural Trade Industry in South Africa Future H. Machate, Bezeng S. Bezeng, Michelle van der Bank African Centre for DNA Barcoding (ACDB), University of Johannesburg, P.O. Box 524, Auckland Park, 2006, Johannesburg, South Africa ABSTRACT Globally, the ornamental horticultural trade has been identified as a key pathway through which non-native species are introduced into new environments. Amongst the non-native- species, succulent plants have been increasingly prioritised despite the negative ecological and economic impacts they pose to local native diversity. Although strict laws exist in South Africa governing the trade in biological materials, current measures to regulate this trade are insufficient. This is due in part to challenges intrinsic to species identification, and the lack of a reliable standard with which traded species can be compared. We seek to use molecular sequencing techniques employing the core DNA barcode regions (rbcLa and matK) to (i) create a reference library of all traded succulent species in South Africa; in order to aid rapid and accurate species identification (ii) evaluate the potential risk of ornamental horticultural trade in the spread of alien species in South Africa. Results: Of the 73 succulent plant species commonly listed in trade in South Africa, 38% of the species had DNA barcode data on BOLD. To complement this list, we additionally generated DNA barcode data for 45 species representing 62% of the total succulent species in our list. Additionally, we found that 63% and 1% of the total succulent species have been formally categorized as either invasive or prohibited. AN extra 36% of the species are in need of formal categorization. Significance: The results show the efficacy of DNA barcoding techniques as an appropriate tool to correctly assign traded unknown succulent to species. We recommend the use of this technique to ensure traders comply with legislature governing the trade in biological materials, which is largely being violated in the horticultural trade industry. If implemented, this will help to preventing further environmental and economic challenges posed by non-native species. INTRODUCTION RESULTS The introduction of non native species into an ecosystem can 87 Echinocereus pentalophus FM008 have detrimental ecological and economic impacts (Figure 1) 59 Echinocereus pentalophus GBVP6099 73 Myrtillocactus geometrizans KF783461 Peniocereus lazaro cardenasii KF783464 (Lowe et al., 2000). Succulent plants in particular have been 99 Hylocereus costaricensis JQ590992 Hylocereus costaricensis JQ590993 introduced globally (with South Africa being no exception) Peniocereus chiapensis KF783472 82 Echinopsis aurea FM002 65 Echinopsis aurea FR853367 through pathways such as the horticultural trade (Nkambule Weingartia kargliana FM006 Weingartia kargliana GBVS3440 et al., 2017). Despite current legislation governing the trade 67 Austrocylindropuntia subulata AY875235 Tephrocactus articulatus AY875248 Cylindropuntia spinosior JN796947 in biological material (e.g. National Environmental Opuntia ficus indica JX572807 Opuntia ficus indica KR736851 Management: Biodiversity Act (NEM:BA)) in South Africa; 91 Opuntia ficus indica KR737121 Opuntia humifusa EF590552 81 Opuntia humifusa GQ248660 measures to regulate and enforce these laws have been 62 Opuntia humifusa KJ773703 Opuntia stricta JQ412402 limited due in part to species identification challenges. 63 Opuntia stricta KU877485 Opuntia stricta KU877486 Opuntia stricta KU877487 Opuntia fragilis EF590551 63 Opuntia fragilis GQ248659 Opuntia fragilis KX678636 Opuntia guatemalensis JQ590994 Opuntia guatemalensis JQ590995 55 Opuntia guatemalensis JQ590996 51 Pereskia aculeata AF206805 Pereskia aculeata JX905965 Pereskia aculeata AY875229 100 Basella alba MF135399 Basella alba MF135428 Calandrinia ciliata KX677816 87 Portulaca grandiflora JN114835 98 Portulaca grandiflora M62568 88 Portulaca oleracea AY875249 100 Portulaca oleracea HQ621340 89 Talinum paniculatum AY875214 92 Talinum paniculatum GQ436529 Talinum paniculatum HM850388 95 Phytolacca dioica JX571876 Phytolacca dioica JX572856 Figure 1: A) Cylindropuntia fulgida B) Opuntia microdasys 92 Phytolacca icosandra JQ593127 67 Phytolacca icosandra JQ593128 70 Phytolacca icosandra JQ593129 that are amongst the problematic invasive succulents in RSA. 84 Phytolacca octandra MF135317 Phytolacca octandra MF135319 Phytolacca octandra MF135407 100 Tetragonia tetragonioides AF132094 Tetragonia tetragonioides HM850395 100 Plectranthus barbatus JQ230987 METHODOLOGY Plectranthus barbatus U28882 92 Euphorbia cactus FM012 52 95 Euphorbia cactus GBVP7287 Euphorbia tithymaloides AY794825 96 Euphorbia tithymaloides JQ591504 Euphorbia tithymaloides KX527246 95 Jatropha curcas KP898351 Jatropha curcas KP898353 Jatropha curcas KP898362 50 Jatropha gossypiifolia GU441785 Jatropha gossypiifolia KP898356 87 Jatropha gossypiifolia KF432063 84 Jatropha integerrima KP898354 86 Jatropha multifida KP898357 Jatropha platyphylla KP898358 Jatropha podagrica GQ436323 Jatropha podagrica KP898355 Pilea microphylla KF138219 100 Pilea microphylla KF138220 Pilea microphylla KF724293 97 Kalanchoe delagoensis FM001 Kalanchoe delagoensis KX783840 74 62 Kalanchoe tomentosa FM014 Kalanchoe tomentosa KX783909 99 84 Kalanchoe beharensis JQ412375 Kalanchoe beharensis KX783905 Aeonium cuneatum KM360615 99 Gasteria vlokii FM009 Gasteria vlokii SAFH2560 85 Agave americana JQ412308 100 Agave angustifolia JQ590090 100 58 Agave angustifolia JQ590091 50 100 Sansevieria trifasciata HM640471 Sansevieria trifasciata JQ273927 100 Callisia repens AF312247 Figure 2: Nurseries from, which succulent plants were 95 Tradescantia fluminensis GU135256 Tradescantia fluminensis HM850406 Acorus tatarinowii JN089956 purchased A) “Pick me” nursery, B) Lifestyle nursery, and C) Acorus tatarinowii JN089957 Sterlig nursery. Acorus tatarinowii JN089959 Figure 3: Phylogeny based on rbcLa locus of traded species in nurseries around Johannesburg. Inventory of succulent plants in South Africa Samples identified as A) Echinocereus pentalophus B) Echinopsis aurea C) Kalanchoe A list of succulent plants was compiled using NEM:BA 2016 delagoensis and D) Kalanchoe tomentosa. and Walters et al. (2011). Table 1: Species identification and their NEM:BA categories. Generating a DNA barcode reference library Corresponding core barcoding sequences were mined from GenBank and compiled into a matrix. Genetic and Sample ID Species ID NEM:BA Category Native Range morphological lookalikes were included. FM001 Kalanchoe delagoensis 1b Madagascar FM002 Echinopsis aurea 1b Argentina Nursery Survey FM009 Gasteria vlokii Native South Africa We surveyed nurseries in the West Rand (Figure 2) and all succulent species were collected for molecular analyses. FM008 Echinocereus pentalophus Uncategorised North America FM012 Euphorbia cactus Uncategorised West Africa DNA Barcoding FM014 Kalanchoe tomentosa Uncategorised Madagascar Standard procedures as set out by the Consortium for the Barcode of Life Plant Working Group (CBOL, 2009) were followed. DISCUSSION & CONCLUSION Preliminary results point towards a mix of both native and non-native species. Of particular Data Analyses interest was the prohibited Echinopsis aurea, which is currently being traded. Once completed, Two approaches were followed to assign query samples to the reference library for succulent species will go a long way to avert the potential negative species. First, NCBI’s MegaBlast algorithm was used. impacts introduced succulent species might have on the rich native flora of South Africa. Thereafter, a Maximum Parsimony (MP) tree was generated Additionally, this library will aid biosecurity officials in regulating this industry and enable to verify the BLAST results. future species identification. REFERENCES ACKNOWLEDGEMENTS CBOL plant working group. 2009. A DNA barcode for land plants. Proceedings of the National Academy of Science USA 106: 12794-12797. Government Gazette. 2016. NEM:BA act 10 of 2004: Alien and Invasive Species. Pg1-74. Available online via:www.gov.ac.za. Lowe, S., Browne, M., Boudjelas, S. and De Poorter, M., 2000. 100 of the world's worst invasive alien species: a selection from the global invasive species database (Vol. 12). Auckland: Invasive Species Specialist Group. Nkambule, N.P., Blignaut, J.N., Vundla, T., Morokong, T. and Mudavanhu, S., 2017. The benefits and costs of clearing invasive alien plants in northern Zululand, South Africa. Ecosystem Services. Walters, M., Figueiredo, E., Crouch, N.R., Winter, P.J.D., Smith, G.F., Zimmermann, H.G., and Mashope, B.K. 2011. Naturalised and invasive succulents of southern Africa (Vol. 11). ABC Taxa. Pretoria. Pg1-305..