16The invasive silver carp Hypophthalmichthys molitrix escaped from southern U.S. aquaculture 17 during the 1970s to spread throughout the Mississippi River basin and steadily moved northward, 18 now reaching the threshold of the Laurentian Great Lakes. The silver carp is native to eastern 19 Asia and is a large, prolific filter-feeder that decreases food availability for fisheries. The present 20 study evaluates its population genetic variability and differentiation across the introduced range 21 using 10 nuclear DNA microsatellite loci, sequences of two mitochondrial genes (cytochrome b 22 and cytochrome c oxidase subunit 1), and a nuclear gene (ribosomal protein S7 gene intron 1). 23Populations are analyzed from two invasion fronts threatening the Great Lakes (the Illinois River 24 outside Lake Michigan and the Wabash River, leading into the Maumee River and western Lake 25 Erie), established areas in the southern and central Mississippi River, and a later Missouri River 26 colonization. Results discern considerable genetic diversity and some significant population 27 differentiation, with greater mtDNA haplotype diversity and unique microsatellite alleles 28 characterizing the southern populations. Invasion fronts significantly differ, diverging from the 29 southern Mississippi River population. About 3% of individuals contain a unique and very 30 divergent mtDNA haplotype (primarily the southerly populations and the Wabash River), which 31 may stem from historic introgression in Asia with female largescale silver carp H. harmandi. 32Nuclear microsatellites and S7 sequences of the introgressed individuals do not significantly 33 differ from silver carp. MtDNA variation is used in a high-throughput sequence assay that 34 identifies and distinguishes invasive carp species and their population haplotypes (including H. 35 molitrix and H. harmandi) at all life stages, in application to environmental (e)DNA water and 36 plankton samples. We discerned silver and bighead carp eDNA from four bait and pond stores in 37 the Great Lakes watershed, indicating that release from retailers comprises another likely vector. 38Our findings provide key baseline population genetic data for understanding and tracing the 39 invasion's progression, facilitating detection, and evaluating future trajectory and adaptive 40 success. 41 42 43 Populations at the expansion fronts of an invasion front frequently possess less genetic 68 variability than those at the core of the invasion, which is termed the "leading edge" hypothesis 69 [20, 21]. The phenotypes of individuals at these front populations are postulated to be adapted 70 for dispersal and high reproductive output [22], where they experience low population density, 71 and may benefit from greater resource availability and less intraspecific competition, 72 enhancing reproductive success [23, 24]. These processes may lead to genetic differences in 73 expansion fronts versus longer-established populations, which may result from drift and/or 74 4 selection. However, documentation...