ABSTRACT
AbstractSpatiotemporal environmental change can produce phenotypic differences within and between populations. For scyphozoans, the effect of environmental variation on phenotype has been unclear because of multiple challenges, including difficulties delimiting populations. Marine lakes, bodies of seawater entirely surrounded by land, provide an opportunity to study discrete populations and capture responses to perturbations. We use this opportunity to compare Mastigias papua (Lesson, 1830) medusae before and after a demographic and environmental perturbation. We reconstructed mitochondrial DNA haplotype networks, measured morphological variation, and assessed swimming behavior of pre- and post-perturbation samples to evaluate two hypotheses about the source of variation: recolonization from an alternate location or endemic phenotypic variation. We found significant differences between samples in morphology (F > 9.5, P < 0.001) and in two of three behaviors (F > 8.45, P < 0.005) but no substantial genetic differentiation (ΦST = 0.03, P = 0.09). We reject the hypothesis of recolonization because pre- and post-perturbation lake medusae were genetically similar to each other and also significantly different from any potential source locations (ΦST > 0.48, P > 0.001). We could not distinguish the source of endemic variation; this will require genomic or experimental analyses. Increasing climatic variability emphasizes the need for understanding population-level responses to environmental change and how responses may be modified by sources of intraspecific variation.
Subject(s)
Cnidaria , Scyphozoa , Animals , Demography , Genetic Variation , Lakes , PhenotypeABSTRACT
The inclusion of next-generation sequencing technologies in population genetic and phylogenetic studies has elevated the need to balance time and cost of DNA extraction without compromising DNA quality. We tested eight extraction methods - ranging from low- to high-throughput techniques - and eight phyla: Annelida, Arthropoda, Cnidaria, Chordata, Echinodermata, Mollusca, Ochrophyta and Porifera. We assessed DNA yield, purity, efficacy and cost of each method. Extraction efficacy was quantified using the proportion of successful polymerase chain reaction (PCR) amplification of two molecular markers for metazoans (mitochondrial COI and nuclear histone 3) and one for Ochrophyta (mitochondrial nad6) at four time points - 0.5, 1, 2 and 3 years following extraction. DNA yield and purity were quantified using NanoDrop absorbance ratios. Cost was estimated in terms of time and material expense. Results show differences in DNA yield, purity and PCR success between extraction methods and that performance also varied by taxon. The traditional time-intensive, low-throughput CTAB phenol-chloroform extraction performed well across taxa, but other methods also performed well and provide the opportunity to reduce time spent at the bench and increase throughput.