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Molecular Genetic Variation in Cogongrass Near the Point of Initial Introduction into the Southeastern United States Ludovic J.A.Capo-chichi W.H. Faircloth,A.G. Williamson M. G. Patterson, J.H. Miller S. L. Noffsinger, and Edzard van Santen Auburn University Botanical Characteristics of I. cylindrica May-17 2 Inflorescences of I. cylindrica May-17 3 Morphological differences May-17 4 Infestation with I. cylindrica Year Infested (ha) Location Reference 1954 200 Mobile Cty Tabor (1952) 1974 4,000 Mobile Cty Dickens (1974) 2003 200,000 SW Alabama Faircloth et al. (2003) 20-fold increase from 1954 to 1974 50-fold increase from 1974 to 2003 Most likely an underestimate of the total area currently infested Survey concentrated on highway right-of-ways May-17 5 Sampling Cogongrass in Mobile Cty May-17 6 Amplified Fragment Length Polymorphism (AFLP) A PCR-based molecular technique that can provide An estimate of genetic diversity in introduced species, Evidence of multiple introductions, Evidence of zones of hybridization, and Identify compatible relationships between biocontrol agent and weed host May-17 7 AFLP protocol Extract total genomic DNA (CTAB protocol) Quantify and standardize DNA concentrations Standard AFLP protocol Pre-amplification Selective amplification Banding patterns visualized with IR dyes Manual scoring May-17 8 Statistical Analysis Cluster analysis Relationship based on the banding profile of individuals Assigns these individuals to artificial clusters Canonical discriminant analysis Analysis based on preexisting groups (sampling sites) Maximizes among group differences based on the common profile of members of a group May-17 9 Statistical Analysis (cont.) Gene diversity estimates A measure of genetic variance of a population equal to the probability of identity of randomly chosen genes Gene flow (Nm) estimates Refers to all mechanisms resulting in the movement of genes from one population to another May-17 10 Polymorphism and Gene Diversity Gene Polymorhic markers diversity HT Population n % P1 59 43 0.10 P2 55 40 0.14 P3 68 50 0.16 P4 58 42 0.13 P5 48 35 0.09 P6 59 43 0.10 P7 44 32 0.11 P8 45 33 0.09 Pi 27 20 0.07 May-17 11 AFLP Fingerprint - Grouping by Location May-17 12 Analysis of Molecular Variance (AMOVA) Source of Variance Fixation variation df components percentage Index ФST Among populations 8 5.8 44 0.44 Within populations 126 7.5 56 Total 13.3 100 May-17 134 13 Canonical Discriminant Analysis Can3 P1 P5 13.59 P2, P4 P6, P7, P8 2.41 Pi -8.78 10.44 P3 1.53 -19.97 25.85 -7.39 Can2 6.69 Can1 May-17 -12.47 -31.63 -16.30 14 Gene Flow Estimates R = 0.09 6.0 P2/P3 5.0 Nm 4.0 3.0 P2/P3 2.0 P4/Pi P8/Pi Nm > 1 P4/P5 P7/Pi 1.0 Nm < 1 P2/Pi 0.0 0.1 P5/Pi P3/Pi 0.3 1 P6/Pi P1/Pi 3 10 30 100 log Distance (km) May-17 15 Why are Gene Flow Estimates N.S. Close geographic proximity Statistics are not very sensitive May-17 16 Summary AFLPs work well in weedy species Populations of I. cylindrica near the point of initial introduction contain quite a bit of genetic variation 74 % of all AFLP markers showed polymorphism May-17 17 Summary (cont.) Founder effect. The founder source contains the lowest gene diversity. Something that was not well-known 56 % of the molecular variance was contained within populations Evidence that populations are differentiated May-17 18 Summary (cont.) In both cases (molecular and historical) there is no evidence for private alleles Creation of genetic diversity Genetic recombination Variation created at vegetative level Anthropogenic dispersal is one of the powerful agents for local dispersal of I. cylindrica May-17 19 “ Invasive Plants – Arming to Defend and Win” Southeast Exotic Pest Plant Council’s 7th Annual Conference Alabama Invasive Plant Council’s 3rd Annual Conference May-17 20