Symbiosis WP2: Impacts of Road Landscaping on Gene Flow in Plants
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1 Symbiosis WP2: Impacts of Road Landscaping on Gene Flow in Plants Investigation of genetic diversity of hawthorn (Crataegus monogyna) populations in Ireland in context of road landscaping schemes. Angela Mina-Vargas 1,*, Kenneth Dinan 1,*, Reetu Tuteja 1,*, Grace Hoysted 1,*, Tatsuo Kanno 1, Peter McKeown 1, Padraig Whelan 2, Charles Spillane 1 1 Genetics & Biotechnology Lab, Plant & AgriBiosciences Centre, National University Ireland Galway (NUIG) 2 Zoology, Ecology & Plant Science (ZEPS), University College Cork (UCC) * Researchers contributed equally
2 Project Aims Develop molecular tools for investigation of genetic diversity of hawthorn materials used for roadside landscaping, whether of introduced or Irish provenance. Investigate use of hawthorn as an indicator of the effect of roadside landscape planting on intra-species genetic diversity Determine whether introduced hawthorn planting materials can impact on genetic diversity of pre-existing populations of hawthorn in Ireland
3 What plant species to choose for genetic diversity analysis? Why hawthorn (Crateagus monogyna)? Species with a broad distribution across Ireland Planted populations along newly constructed Hawthorn, Crataegus monogyna. Family: Rosaceae roads are of foreign origin (mainly from nurseries in the Netherlands) Possible polymorphism between the Irish stands of C. monogyna and the introduced C. monogyna genotypes Indicator species to assess hybridization and introgression between Irish and introduced genotypes
4 Methodology Sampling strategy Collection of berries and/or leaves from individual hawthorn trees (within each population) DNA extraction Nuclear molecular marker development (novel nuclear microsatellite SSR markers for hawthorn) Chloroplast genome molecular marker development Use of molecular markers to investigate genetic diversity relationships within and between all populations of hawthorn Data analysis - determination of molecular diversity within and between the populations
5 Hawthorn flowers, berries and tree
6 Sampling of populations & samples across Ireland, UK and continental Europe Collection Sites - N22/N25 Road (Tralee to Wexford) Uragh Wood Nature Reserve
7 Road Sampling Strategy Pre-existing Irish provenance Hawthorn germplasm Non- Roadside 5 KM Roadside Introduced Hawthorn germplasm (from nurseries in Holland)
8 Key to the hawthorn populations analyzed Cork RS = Roadside Samples, Carrigrohane, Cork Cork IS = Inside Road Samples, Glunishine, Cork East RS = Roadside Samples, Cork, Waterford, Wexford East IS = Inside Road Samples, Cork, Waterford, Wexford West RS = Roadside Samples, Tralee, Kerry West IS = Inside Road Samples, Tralee, Kerry U-F = Fairy trees & Uragh Wood Nature Reserve, Kerry OI = Other parts of Ireland (e.g. Offaly, Galway, Clare, Monaghan, Meath) INT = e.g. UK, Germany, Greece, Austria, Denmark Sweden = Swedish samples
9 Development of novel molecular markers for hawthorn genetic diversity assessment Prior to EPA project no molecular markers (SSRs, microsatellites) available for hawthorn (Crataegus monogyna) EPA Project identifies six nuclear microsatellites marker loci (SSRs) that can be used for genetic diversity analysis of hawthorn
10 Excess homozygosity in inbred Irish hawthorn populations Population Na Ne Na Freq. >=5% No. P.A. Ho He F N Cork RS East RS West RS Cork IS West IS U-F OI INT Sweden Na- total number of alleles Ne- effective number of alleles Na freq>5%- number of alleles freq >0.05 N- Sample size Ho- observed heterozygosis He- expected heterozygosis F- Coefficient of inbreeding
11 Most of the genetic variation detected is within hawthorn populations rather than between populations Amova performed to detect the sources of variation within and between hawthorn population groups Source of variation df SS MS Est. Var. (%) Variance (%) P Value Among C. monogyna Within C. monogyna Total Stat PhiPT Value P(rand>=data) At the resolution of the molecular markers used there are no significant differences in genetic diversity between any of the populations of hawthorn analysed in this study (i.e. roadside vs inside road, foreign vs Irish)
12 Low level of differentiation between the hawthorn population groups analyzed
13 Chloroplast loci across all hawthorn populations analysed display lack of genetic diversity -Founder event at European scale? -Evolution: recent radiation of hawthorn? -Irish populations are not distinct HW046-Wexford RS C.maximowiczii HW107-Sweden HW022-Glunishine IS HW035-Tralee IS HW133-Meath HW039-Wexford IS HW058-Tralee IS HW102-Greece HW033-Uragh HW084-Offaly HW078-Tralee RS HW075-Tralee IS HW001-Cork RS HW040-Tallow RS HW003-Cork RS HW031-Uragh HW134-Meath HW061-Tralee RS HW044-Tallow RS HW087-The Burren HW032-Uragh HW023-Cork IS HW077-Tralee IS HW118-Monaghan HW111-Germany HW028-Uragh HW128-Monaghan HW113-Sweden C.clorosarca HW016-Cork RS HW055-Tralee IS HW131-Monaghan HW053-Tralee RS HW037-Youghal HW086-Offaly HW095-Sweden HW014-Cork IS HW074-Tralee IS HW088-The Burren HW100-Austria HW054-Uragh HW124-Monaghan HW112-Sweden C.laevigata- HW132-Meath HW038-Tallow RS HW008-Cork IS HW076-Tralee IS HW079-Tralee RS HW085-Offaly HW094-UK HW006-Cork RS HW071-Tralee IS HW083-Offaly HW099-Germany HW064-Fairy HW122-Monaghan HW108-Sweden C.laevigata HW109-Sweden HW127-Monaghan HW010-Cork RS HW012-Cork RS HW101-Crete HW015-Cork RS Neighbour Joining, derives from sequences of rps16 and LSC region of the chloroplast of 128 individuals of hawthorn C. monogyna, two individuals of C. laevigata and C. chlorosarca
14 Conclusions Successful development of novel molecular markers (microsatellites, SSRs) for hawthorn (C. monogyna) Genetic diversity analysis of Irish hawthorn populations indicates significant excess of homozygotes & indicated that populations were inbred and displayed low genetic variability No significant genetic diversity differences between any of the population groups analysed i.e. roadside vs inside and road and foreign (planted or sourced) versus Irish provenance Genetic diversity does not provide a basis for limiting roadside management to planting hawthorn of Irish provenance
15 Acknowledgements Research Prof. Charles Spillane Dr. Padraig Whelan Angela Mina-Vargas Lisa Dolan Kenneth Dinan Reetu Tuteja Grace Hoysted Dr. Tatsuo Kanno Dr. Peter McKeown Pat O Rafferty Dr. Colin Kelleher Funding/Support EPA SIMBIOSYS NUIG Botany & Plant Science
16 Thank You
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