The role of waterholes as 'refugia' in sustaining genetic diversity and variation of two freshwater species in dryland river systems (Western Queensland, Australia)

1. Episodic floods and extended low or no flow periods characterise dryland river systems in Western Queensland, Australia. During protracted intervals between floods, rivers consist of a series of isolated waterholes, which serve as ‘refugia’ for aquatic species and much of the channel is dry. We categorised these waterholes into ‘main waterholes’, which are located in the main part of the river channel and ‘satellite waterholes’, which are located in distributary river channels. 2. We used mitochondrial sequences and allozymes to investigate levels of genetic diversity and patterns of connectivity among waterholes for two obligate freshwater species: Macrobrachium australiense (Decapoda: Palaemonidae) and Notopala sublineata (Gastropoda: Viviparidae). 3. We sampled 31 waterholes for M. australiense and 12 for N. sublineata. Based on a 505‐bp fragment of cytochrome oxidase subunit I, we identified 54 haplotypes in a sample of 232 individuals for M. australiense and based on a 457‐bp fragment of the same gene, 36 haplotypes in a sample of 145 individuals for N. sublineata. 4. Both nuclear and mitochondrial genetic data sets indicated that estimates of genetic diversity were not different in populations inhabiting main and satellite waterholes for either species. Also, there was generally very limited genetic differentiation among populations at any site. 5. We suggest that levels of connectivity among populations inhabiting waterholes at most sites are higher than expected. High levels of connectivity may help to maintain overall high levels of genetic diversity as well as low levels of genetic differentiation among waterholes within sites.     

Genetic evidence for historical continuity between populations of the Australian freshwater fish Craterocephalus stercusmuscarum (Atherinidae) east and west of the Great Dividing Range

The freshwater Craterocephalus stercusmuscarum (Atherinidae) has a broad northern and south-eastern Australian distribution, and has been divided into two sub-species. Craterocephalus stercusmuscarum fulvus occurs in eastern and western flowing drainages of the southeast, while C. s. stercusmuscarum occurs in north eastern and northern flowing drainages. Four populations of each sub-species were sampled from different river systems to examine if genetic diversity was consistent with this nomenclature or a previously proposed vicariance hypothesis. Allozyme data did not support the notion that the subspecies were genetically distinct, but the mtDNA data showed that haplotypes from one sub-species, regardless of geography, were reciprocally monophyletic to haplotypes of the other subspecies. Thus, mtDNA genetic diversity was partitioned by prevailing taxonomy and the data suggest that C. s. fulvus populations in eastern and western flowing drainages may have had a relatively recent connection subsequently interrupted by geological events.     

Genetic variation and population subdivision of the endangered Iberian cyprinid Chondrostoma lusitanicum

Products of 24 presumptive enzyme loci were used to analyse the consequences on genetic structure and variation of the Chondrostoma lusitanicum population decline within the Tejo basin. This rare cyprinid is endemic to the Iberian Peninsula and has a very restricted distribution. Five samples from the Tejo catchment were compared with a sample from a small basin, the Samarra, that has not suffered obvious anthropogenic pressures and where the fish is still abundant. Heterozygosity and polymorphism were higher overall in the Samarra. Several lines of evidence indicate a high degree of population subdivision within the Tejo basin. In fact, about half of the total gene diversity detected in the Tejo population was due to differences among samples. This differentiation appeared to be caused by genetic drift and possibly differential local selection, coupled with reduced gene flow among localities. The accelerated process of habitat degradation occurring in the lowland streams of the Tejo basin will lead to the inevitable reduction of intraspecies genetic diversity.     

Allozyme comparison of two populations of Rineloricaria (Siluriformes, Loricariidae) from the Ivaí River, upper Paraná River basin, Brazil

Two allopatric morphotypes of the genus Rinelocaria were compared through the allozyme electrophoresis technique: one morphotype, R. pentamaculata, from the Keller River in the middle stretch of the Ivaí River basin and the other, R. aff. pentamaculata, from the São João River in the upper portion of the Ivaí River basin. The morphotype from the São João River was collected upstream from the São João waterfall, which is about 80 m deep. Twelve enzymatic systems (AAT, ADH, EST, GCDH, G3PDH, GPI, IDH, LDH, MDH, ME, PGM and SOD) were analyzed, which allowed to score 22 loci. Only lociAat-2, Est-3 and Mdh-C showed polymorphism. The two samples differed in allele frequencies at the three polymorphic loci. The average expected heterozygosity for all loci was 0.0806 ± 0.0447 in the Keller River sample. For the São João River morphotype, this value was 0.0489 ± 0.0350. Nei'' s genetic identity and distance between the two populations were respectively 0.9789 and 0.0213. Wright''s F_IS , F_IT and F_ST over all loci were estimated as 0.3121, 0.4021 and 0.1309, respectively. We consider that the two morphotypes represent species in statunascendi.     

Are populations of mayflies living in adjacent fish and fishless streams genetically differentiated?

1. Conspecific populations living in habitats with different risks of predation often show phenotypic variation in defensive traits. Traits of two species of mayflies (Baetidae: Baetis bicaudatus and Baetis sp. nov.) differ between populations living in fish and fishless streams in a high altitude drainage basin in western Colorado, U.S.A. We tested for genetic differentiation between mayfly populations in these two habitat types, assuming that lack of genetic differentiation would be consistent with the hypothesis that those traits are phenotypically plastic. 2. Previous work has shown that larvae of both species behave differently and undergo different developmental pathways in adjacent fish and fishless streams. These phenotypic differences in behaviour and development have been induced experimentally, suggesting that populations from fishless streams have the genetic capability to respond to fish. 3. During summer 2001 we collected Baetis larvae from several fish and fishless streams, and from fish and fishless sections of the same streams. We used allozymes and a fragment of the cytochrome oxidase subunit 1 mitochondrial gene to examine genetic variation of Baetis individuals within and among streams. 4. Results showed that genetic variation exists among populations of the same species of Baetis from different streams, but none of that variation was associated with the presence or absence of fish. These data confirm that populations of Baetis living in fish and fishless streams are not genetically distinct, and are consistent with the hypothesis that traits associated with environments of different risk are phenotypically plastic.     

Constraints on recovery: using molecular methods to study connectivity of aquatic biota in rivers and streams

1. The ‘Field of Dreams Hypothesis’ states ‘if we build it, they will come’, referring to the assumption that if habitats are restored, species will recolonise them. However, the ability of a species to recolonise a restored site will depend not only on the appropriate habitat being present, but also on the ability to get there. This is likely to depend on both the species’ dispersal behaviour and the position of a site in the landscape. 2. Animals with good potential for dispersal are more likely to be able to disperse to newly restored sites. Similarly, sites in lowland streams with limited altitudinal differences between sites may be easier to reach than upstream sites. This is because upstream sites are connected to one another via lowland streams that have different characteristics and therefore may be difficult for animals to traverse. 3. In this paper, genetic data from a range of freshwater species that have been analysed in my laboratory are used to assess the importance of life cycle and position in the landscape (i.e. upland versus lowland streams) on connectivity patterns (and thus recolonisation potential) among populations. 4. In general, contemporary dispersal across catchment boundaries is negligible, except for aquatic insects with an adult flight stage. Dispersal among streams within catchments appears to be more limited than was predicted from knowledge on life histories, except for fish in lowland rivers and streams. 5. As predicted, dispersal of fish, crustaceans and molluscs among streams within catchments is significantly greater in lowland rivers than in upland streams. 6. Overall, these analyses suggest that, with the exception of most insects, and fishes in lowland rivers, natural recolonisation of restored sites is only likely from sites within the same stream. If a species has disappeared from the whole stream, then restoration of habitat alone may not be sufficient for its re‐establishment.     

Does the Mediterranean killifish Aphanius fasciatus (Teleostei: Cyprinodontidae) fit the one-dimensional stepping-stone model of gene flow?

Five samples (30 individuals each) of the Mediterranean cyprinodontid fish, Aphanius fasciatus, were analysed by allozyme electrophoresis for a study concerning genetic structure and gene flow among populations. Forty-three loci, twelve of which were polymorphic at P_0.99 level, were analysed. A. fasciatus showed low levels of genetic polymorphism, with expected heterozygosity values ranging from 0.027 (SE = 0.013) to 0.064 (SE = 0.023). The modified Rogers' genetic distances ranged from 0.053 to 0.202. The observed pattern of genetic differentiation among populations, obtained by multidimensional scaling of the modified Rogers' genetic distances, was consistent with geographic distribution. Coancestry coefficient ( = 0.302, SE = 0.045) indicated a very high degree of genetic subdivision among populations and the estimate of gene flow (the effective number of migrants per generation, Nm = 0.445), computed by Wright's method, showed very restricted gene flow among populations. The regression analysis carried out using log-transformed values of geographic distance against M^ (pairwise values of Nm) gave a regression coefficient, b = –1.03, indicating that a restricted migration rate among populations can occur according to the one-dimensional stepping-stone model. The amount and modality of gene flow in A. fasciatus inferred from the results of the present study are consistent with the absence of dispersal stages and with the fragmented nature of brackish-water habitats along the coasts.