Analysis of gene flow in North American face fly (Musca autumnalis) populations

Abstract. Vertical polyacrylamide gel electrophoresis was used to resolve enzymes at ten putative loci in face flies Musca autumnalis De Geer, a colonizing, Palaearctic species established in North America for at least 210 generations. Flies were sampled in 1991 from six locations in Iowa, two in Maryland, two in Minnesota, two in Tennessee, three in New York, and three in Missouri. Nondirectional temporal variation in gene frequencies over a 4-year interval was detected at farms in central Iowa. Heterogeneity in allele frequencies was detected among farms in Iowa, Maryland, New York and Minnesota but not in Tennessee and Missouri. There were no consistent departures from random mating. Partitioning variances in gene frequencies showed that 58% of the variation occurred in populations among states and 43% between populations within states. Mean reproducing immigrants per population per generation was estimated to be eighteen flies. No regional genetic differentiation was detected, and there were no barriers to gene flow within or among diverse populations in the six states. Earlier data bearing on gene flow among and between Nearctic and Palearctic face fly samples were analysed and significant differentiation was not detected.     

Allozyme variation in stable flies (Diptera: Muscidae)

Polyacrylamide gel electrophoresis was used to resolve allozymes in the cosmopolitan blood-feeding stable fly,Stomoxys calcitrans (L.). Nineteen of 38 loci were polymorphic (53%). Mean heterozygosities among all loci and among only polymorphic loci were 0.096 and 0.182, respectively. These gene diversity measures are about half those among other muscid Diptera. Variation in gene frequencies was examined in 10 natural stable fly populations from Iowa and Minnesota. Gene frequencies were homogeneous at five of eight loci among six populations in 1990 and eight of eight loci among four populations in 1992. Wright'sF statistics showed no significant departure from random mating among stable flies. It was concluded that gene flow compensates for any local differentiation in stable fly populations.     

Electrophoretic analysis of genetic variability in the house fly (Musca domestica L.)

Methods are described for the resolution of house fly, Musca domestica L., enzymes by vertical polyacrylamide gel electrophoresis. An electrophoretic survey in Ames, Iowa, of 51 loci distributed among 26 enzyme systems revealed that 40% of the loci are polymorphic. Observed and expected heterozygosities measured at 33 loci were 0.0981 and 0.1148, respectively. A significant deficiency of heterozygotes was noted at certain loci.Journal Paper No. J-11423 of the Iowa Agriculture and Home Economics Experiment Station, Ames. Project No. 2411.     

Genetic diversity at electrophoretic loci in the house fly,Musca domestica L.

Vertical polyacrylamide gel electrophoresis was used to separate enzyme proteins at 73 putative loci in natural house fly populations sampled in central Iowa. Thirty-nine of the loci were polymorphic (53%). The mean effective number of alleles per polymorphic locus was 1.93 and 1.47 alleles among 68 scored loci. Observed and expected heterozygosities at 34 house fly loci were 0.1628 and 0.1834, respectively. No statistically significant differentiation was detected among nine central Iowa fly populations in 1989 or among nine Iowa and three Minnesota populations in 1990. Journal Paper No. J-14125 of the Iowa Agriculture and Home Economics Experiment Station, Ames. Project No. 2949.     

A FORTRAN program for the calculation and analysis of two-locus linkage disequilibrium coefficients

Summary A FORTRAN program was written that calculates composite linkage disequilibrium coefficients from genotypic data. Chi-square tests determine whether coefficients calculated for allele and locus pairs are significantly greater than zero. A subroutine is provided that partitions the variance in linkage disequilibrium into within- and between-subpopulation components. Output obtained from analysis of allozyme data collected from natural subpopulations of the house fly (Musca domestica L.) are included to illustrate features of the program.Journal Paper No. J-11345 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 2411     

Allozyme genotype-environment relationships in natural populations of Drosophila buzzatii

Allozyme frequency data from natural populations of Drosophila buzzatii were analyzed for genotype-environment relationships. Allele frequency and heterozygosity at six loci polymorphic throughout eastern Australia and a number of environmental factors (both means and variabilities) were examined by a variety of multivariate techniques. Significant genotype-environment associations were found for five of the six loci, and after correcting for geographic location significant associations remained for Est-2 and Adh-1 gene frequencies and heterozygosities and for Pgm gene frequencies. The results are discussed in relation to selection and gene flow and provide the basis for laboratory studies to disentangle confounded effects of (1) environmental means and environmental variabilities and (2) allele frequency and heterozygosity, and thus to further test for and determine the nature of any natural selection at particular allozyme loci.This work was supported by a grant from the Australian Research Grants Committee to J. S. F. B.     

Microsatellite Variation in Two Populations of Free-Ranging Yellow Baboons (Papio hamadryas cynocephalus)

We investigated genetic variation at six microsatellite (simple sequence repeat) loci in yellow baboons (Papio hamadryas cynocephalus) at two localities: the Tana River Primate Reserve in eastern Kenya and Mikumi National Park, central Tanzania. The six loci (D1S158, D2S144, D4S243, D5S1466, D16S508, and D17S804) were all originally cloned from and characterized in the human genome. These microsatellites are polymorphic in both baboon populations, with the average heterozygosity across loci equal to 0.731 in the Tana River sample and 0.787 in the Mikumi sample. The genetic differentiation between the two populations is substantial. Kolmogornov–Smirnov tests indicate that five of the six loci are significantly different in allele frequencies in the two populations. The mean F _ST across loci is 0.069, and Shriver's measure of genetic distance, which was developed for microsatellite loci (Shriver et al., 1995), is 0.255. This genetic distance is larger than corresponding distances among human populations residing in different continents. We conclude that (a) the arrays of alleles present at these six microsatellite loci in two geographically separated populations of yellow baboons are quite similar, but (b) the two populations exhibit significant differences in allele frequencies. This study illustrates the potential value of human microsatellite loci for analyses of population genetic structure in baboons and suggests that this approach will be useful in studies of other Old World monkeys.     

Repeatability and heritability of divergent recombination frequencies in the Iowa Stiff Stalk Synthetic (Zea mays L.)

Variability in recombination frequency was reported in the Iowa Stiff Stalk Synthetic. The objectives of the present research were to verify the differences in recombination frequency among individuals in the Iowa Stiff Stalk Synthetic maize population and to determine if the recombination frequency differences persisted among the S₁ progeny. Testcrosses to measure male recombination frequency on three chromosomes (4, su1-c2; 5, a2-bt1-pr1; 9, sh1-bz1-wx1) were repeated for eight S₀ individuals. Recombination frequencies were repeatably divergent among those individuals which were selected based on high or low recombination frequencies on specific chromosomes. Individuals which had been selected for long and short total map distances across the three chromosome regions produced repeatably divergent recombination frequencies only at the su1-c2 region. The recombination frequencies of the S₁ lines, derived from the S₀ individuals which had the most divergent recombination frequencies on a single chromosome, were significantly different. The broadsense heritability estimates derived from the regression of six S₁ lines on six S₀ individuals ranged from 0.69 to 0.20 for the five chromosome regions. We conclude that genetic differences for recombination frequency exist in this population and that modification by selection should be possible.     

Molecular and biochemical characterization of puroindoline a and b alleles in Chinese landraces and historical cultivars

Kernel hardness that is conditioned by puroindoline genes has a profound effect on milling, baking and end-use quality of bread wheat. In this study, 219 landraces and 166 historical cultivars from China and 12 introduced wheats were investigated for their kernel hardness and puroindoline alleles, using molecular and biochemical markers. The results indicated that frequencies of soft, mixed and hard genotypes were 42.7, 24.3, and 33.0%, respectively, in Chinese landraces and 45.2, 13.9, and 40.9% in historical cultivars. The frequencies of PINA null, Pinb-D1b and Pinb-D1p genotypes were 43.8, 12.3, and 39.7%, respectively, in hard wheat of landraces, while 48.5, 36.8, and 14.7%, respectively, in historical hard wheats. A new Pinb-D1 allele, designated Pinb-D1t, was identified in two landraces, Guangtouxianmai and Hongmai from the Guizhou province, with the characterization of a glycine to arginine substitution at position 47 in the coding region of Pinb gene. Surprisingly, a new Pina-D1 allele, designated Pina-D1m, was detected in the landrace Hongheshang, from the Jiangsu province, with the characterization of a proline to serine substitution at position 35 in the coding region of Pina gene; it was the first novel mutation found in bread wheat, resulting in a hard endosperm with PINA expression. Among the PINA null genotypes, an allele designed as Pina-D1l, was detected in five landraces with a cytosine deletion at position 265 in Pina locus; while another novel Pina-D1 allele, designed as Pina-D1n, was identified in six landraces, with the characterization of an amino acid change from tryptophan-43 to a ‘stop’ codon in the coding region of Pina gene. The study of puroindoline polymorphism in Chinese wheat germplasm could provide useful information for the further understanding of the molecular basis of kernel hardness in bread wheat.     

Genetics of tassel branch number in maize and its implications for a selection program for small tassel size

Summary Tassel branch numbers of six crosses of maize (Zea mays L.) were analyzed to determine inheritance of this trait. Generation mean analyses were used to estimate genetic effects, and additive and nonadditive components of variance were calculated and evaluated for bias due to linkage. Both narrow-sense and broad-sense heritabilities were estimated. Additive genetic variance estimates were significant in five of the six crosses, whereas estimates of variance due to nonadditive components were significant in only three crosses. Additionally, estimates of additive variance components usually were larger than corresponding nonadditive components. There was no evidence for linkage bias in these estimates. Estimates of additive genetic effects were significant in four of six crosses, but significant dominance, additive × additive and additive × dominance effects also were detected. Additive, dominance, and epistatic gene action, therefore, all influenced the inheritance of tassel branch number, but additive gene action was most important. Both narrow-sense and broadsense heritability estimates were larger than those reported for other physiological traits of maize and corroborated conclusions concerning the importance of additive gene action inferred from analyses of genetic effects and variances. We concluded that selection for smalltasseled inbreds could be accomplished most easily through a mass-selection and/or pedigree-selection system. Production of a small-tasseled hybrid would require crossing of two small-tasseled inbreds. We proposed two genetic models to explain unexpected results obtained for two crosses. One model involved five interacting loci and the other employed two loci displaying only additive and additive × additive gene action.Journal Paper No. J-9231 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011. Project No. 2152     

Gregarious foraging in barn swallows after the breeding season

Aerial vertebrate foragers, e.g. insectivorous bats, martins and swallows, often show gregarious behavior such as colonial breeding, communal roosting and aggregating behavior during foraging. Studies of gregariousness in aerial foragers have mostly focused on colonial breeding or communal roosting, and only a few intensive studies have dealt with gregariousness during foraging. Here, we report on large and stable aggregations of barn swallows, Hirundo rustica, observed during daytime after the breeding season in Japan. Relatively large aggregations of 20 or more birds were located around poultry, pig and cattle farms and mounds of manure. The aggregations were stable in size both within-days and between-days. Their activities consisted only of repeated cycles of foraging and resting around the farms where their prey, the large black soldier fly Hermetia illuceus (>10 mm) and other medium (5–10 mm) flies, was dense. Distributions of swallows around the farms overlapped with the distributions of prey, and the size of the aggregations significantly correlated with the abundance of prey.     

Identification of Encephalitozoon cuniculi genotype III and two novel genotypes of Enterocytozoon bieneusi in swine

Samples of intestinal content from thirty fattened pigs of six farms slaughtered at an abattoir in North-Western Germany, and faecal samples of four pigs kept as laboratory animals at the Federal Institute for Risk Assessment (BfR, Berlin, Germany) were investigated for the occurrence of microsporidia by light microscopy, PCR and sequencing. A modified Webers trichrome staining and the immunohistochemistry (the Avidin-Biotin-Peroxidase-Complex technique with a polyclonal anti-Encephalitozoon cuniculi-serum and monoclonal antibodies against Encephalitozoon intestinalis and Enterocytozoon bieneusi) was used as a screening method for the light microscopical detection of these pathogenic eukaryotes. By this light microscopically methods microsporidia suspected organisms were found in all samples (100%). By the use of PCR, microsporidia were identified in fourteen samples (41.2%). The prevalence of microsporidia infections among the farms diversifies from 0 to 80% as considered by PCR. E. bieneusi was the most prevalent species and was identified in twelve fattened pigs (40%) from five of the six tested farms (83.3%) and in two of the four laboratory animals (50%). Three of the E. bieneusi species belonged to the genotype O, one to the genotype E, and one to the genotype F. Two isolates were identified as novel genotypes and two samples showed a mixed infection of different genotypes. In three faecal samples of the pigs from two farms E. cuniculi genotype III was identified. One sample contained both microsporidia species. To our knowledge, this is the first time that the genotype III of E. cuniculi was identified in swine.     

Measures of gene flow in the Columbian ground squirrel

From analyses of published data and a review of the literature, I studied indirect and direct measures of gene flow among populations of Columbian ground squirrels, Spermophilus columbianus. New analyses were used to examine an allozyme data set (seven polymorphic loci) that had been collected by Zammuto and Millar (1985a) from six populations of ground squirrels that were spread over 183 km. G-tests indicated significant variation in allele frequencies among populations, but F-statistics revealed relatively little population differentiation (average F _ST=0.026). F _ST values were used to estimate rates of gene flow indirectly and indicated fairly high rates of gene flow (average N _e m=13.5). Recorded dispersal distances of individual ground squirrels were fairly short (most<4 km, maximum recorded distance was 8.5 km), and the minimum distance between populations used to create the allozyme data set was about 25 km. Thus, direct dispersal among the populations in the allozyme data set was highly unlikely. Small genetically effective populations may have experienced high rates of migration over short distances (about 43% of adults in local populations were immigrants), however, resulting in homogeneous allele frequencies over the geographic range. This explanation provides an alternative to invoking gene flow in the recent past to explain discrepancies between dispersal distances in the field and homogenization of allele frequencies over large ranges, Mammalian species that have virtually complete dispersal of subadult males from the natal area might be expected to exhibit relatively high rates of gene flow, regardless of actual dispersal distances. Genetically effective populations may be much smaller than more extensive ecological populations and experience higher rates of gene flow.     

Chromosome labeling with transposable elements in maize

Transposable elements randomly insert into a targeted locus at a frequency of 10^-6 to 10^-5. The En element has been shown in previous studies to transpose more frequently into closely linked sites. Thus, it is appropriate to place an En element onto each of the 20 chromosome arms in maize to maximize tagging efficiency. This is called chromosome labeling for tagging purposes with transposons. After a chromosome arm has been labeled with a transposon, genes residing in that arm will have a greater chance to be tagged by the transposon. To date, all of the maize chromosome arms have been labeled with at least one of five Encontaining alleles. The elements were linked to the arms using reciprocal translocations. The usage of these arm-labeled lines is discussed in the context of gene tagging.Journal Paper No. 15224 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa; Project No. 3176     

REGIONAL,LOCAL AND MICROGEOGRAPHIC ALLELE FREQUENCY VARIATION BETWEEN APPLE AND HAWTHORN POPULATIONS OF RHAGOLETIS POMONELLA IN WESTERN MICHIGAN

In the preceding study (Feder et al., 1990), we report that paired apple and hawthorn infesting populations of Rhagoletis pomonella are genetically differentiated for six allozymes. Here, we show that patterns of intra- and inter-host allele frequency variation seen for these six loci across the eastern United States are consistent on a more fine grained spatial scale in western Michigan. Malic enzyme, Aconitase-2, Mannose phosphate isomerase, and Hydroxyacid dehydrogenase all displayed significant linear relationships with latitude among five “regional” hawthorn populations sampled along a north-south transect between the cities of Cadillac and Portage, Michigan. Clines were not as evident among “regional” apple populations in western Michigan, although allele frequencies for Malic enzyme¹⁰⁰, Mannose phosphate isomerase¹⁰⁰ and Aconitase-2⁹⁵ varied with latitude among six “local” apple populations within a 60 km² area near the town of Grant. Significant allele frequency differences were observed between hawthorn and apple populations at all “regional” and “local” collecting sites analyzed in the study (a total of 20 different apple and hawthorn populations). As was the case in the geographic survey of the eastern United States, the magnitude and pattern of inter-host frequency differences at “regional” and “local” sites were a function of latitude. Host related genetic differentiation was consistent on a “microgeographic” scale as well. Allele frequencies for Malic enzyme¹⁰⁰ and Aconitase-2⁹⁵ were significantly higher over a four-year period (1984 to 1987) for flies sampled from individual hawthorn trees (N = 6) than apple trees (N = 7) within an old field (0.09-km² area) located near Grant. The fine level of genetic subdivision between hawthorn and apple populations of R. pomonella in western Michigan substantiates the existence of host associated polymorphism in the fly and supports a sympatric mode of divergence for the “apple race”.     

Variation in polychlorinated biphenyl levels in common carp from a primarily agricultural drainage

Concentrations of polychlorinated biphenyls (PCBs) in common carp, Cyprinus carpio, from the Des Moines River, Iowa, were assessed for variability related to sampling location, sampling period, fish age, and fat content. Concentrations were highest at a location near the City of Des Moines; they were substantially lower in 1981 than in 1980. Age and fat content had little influence on PCB concentrations in carp. Overall concentrations were some of the lowest recorded in the United States and Canada in recent times.The Unit is jointly supported by Iowa State University, the Iowa State Conservation Commission, and the U.S. Fish and Wildlife Service.Journal Paper No. 10754 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 2465. Financed by a grant from the U.S. Department of Defense Army Corps of Engineers and made available through the Engineering Research Institute, Iowa State University.     

Polymorphism of hordein-coding loci in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) populations of Iran and Central Asian countries

Starch gel electrophoresis was performed to study polymorphism of hordeins encoded by the Hrd A, Hrd B, and Hrd F loci in 366 local old barley accessions from Iran and Central Asian countries, including Turkmenistan, Uzbekistan, Tajikistan (Mountain Badahsan), and Kirgizia. In total, 60 alleles with frequencies of 0.0003–0.2818 were observed for the Hrd A locus, 106 alleles with frequencies of 0.0003–0.1603 were observed for the Hrd B locus, and five alleles with frequencies of 0.0164–0.4131 were observed for the Hrd F locus. The alleles and allele frequencies displayed irregular distributions in barley populations of the above countries. Cluster analysis of the matrix of allele frequencies in populations from known collection sites revealed a cluster structure of local barley populations within each country. Local populations formed five differently sized clusters in Iran, six in Turkmenistan, three in Uzbekistan, and three in Kirgizia. The variation and allele frequency distribution of the hordein-coding loci in Iran and Central Asian countries were assumed to result from the introduction and spreading of barley forms via migrations of husbandmen.     

Genetic structure of natural populations of the red-spotted newt,Notophthalmus viridescens

Genetic variation is described at 15 loci in 2 neotenic and 12 nonneotenic populations of red-spotted newts. Though high levels of genetic similarity (I=0.990) were found among all populations, allele frequencies at six of the eight most polymorphic loci show significant heterogeneity across populations. Change in allele frequencies at two of these loci (Pep-2 and Ldh-1) is significantly correlated with latitude. Interspecific homologies are established for newt peptidases based on substrate specificities and lactate dehydrogenases based on tissue distribution, thermal stability, and kinetic properties. Nonneotenic populations are highly variable (H=0.157) and neotenic populations are only slightly, but significantly, less variable (H=0.120). The high levels of heterozygosity detected in nonneotenic populations may result from large effective population size and/or environmental heterogeneity. The unexpectedly high heterozygosity values obtained for the neotenic populations may indicate adult dispersal or the presence of some previously undetected red efts at these localities. In any case, a major change in life history has apparently had little effect on the genetic structure of these populations.This research was supported by grants from the Blakeslee Fund of Smith College.     

Change in ribosomal DNA spacer-length composition in maize recurrent selection populations. 2. Analysis of BS10, BS11, RBS10, and RSSSC

Four maize (Zea mays L.) populations selected for grain yield (BS10, Iowa Two-ear Synthetic; BS11, formerly Pioneer Two-ear Composite; RBS10, Illinois strain of BS10; and RSSSC, Illinois strain of Iowa Stiff Stalk Synthetic) were assayed for molecular variation in the ribosomal DNA (rDNA) intergenic spacer (IGS) at initial and advanced cycles of selection. RSSSC and RBS10 underwent reciprocal recurrent selection with an inbred tester in a high-yield environment, whereas BS10 and BS11 were subjected to full-sib reciprocal recurrent selection. Maize rDNA, which encodes the ribosomal RNA genes, is highly repetitive and shows IGS length variation within and among individuals. Five different ribosomal spacer-length variants (rslvs) and a polymorphic SstI restriction site in the IGS were detected in the four populations. The five rslvs and the polymorphic restriction fragment were observed in 20 different combinations or hybridization fragment patterns (HP). RSSSC, RBS10, and BS11 showed significant changes in the overall rslv and HP frequencies between cycle 0 and the advanced cycle of selection, whereas BS10 did not. In general, two specific HPs were more frequent in the majority of the advanced cycles of the four populations. The frequency changes between initial and advanced cycles were more dramatic for HPs than rslvs. These results are consistent with earlier findings and further support the hypothesis that certain rDNA HPs and/or linked loci may be responding to selection for grain yield and may be associated with a selective advantage in US Corn Belt environments.     

Tagging of a maize gene involved in kernel development by an activated Uq transposable element

Summary A quiescent Uq transposable element has been activated in a maize plant treated with 5-aza-2-deoxycyti-dine. This activated Uq cosegregates with a heritable dominant miniature (Mn) kernel phenotype, indicating its physical association with a maize miniature locus (Mn:: Uq). The Mn:: Uq mutant is dominant in producing a miniature seed phenotype of variable size and in reducing seedling vigor in the early growth stage. Genetic experiments indicate that the Mn:: Uq mutant also affects the activity of the male gametophyte, whereby pollen germination is inhibited, thus lacking pollen tube growth resulting in the male nontransmissibility of this mutant. Proof for the Uq element in this mutant is derived by its ability to transactivate the standard a-ruq reporter allele to yield spotted aleurone tissue. However, the Mn:: Uq mutant does not transactivate a normally Uq-responsive c-ruq allele, suggesting a structural difference between the two ruq receptors at the A1 and C1 loci. It is anticipated that cloning of the Uq transposable element would facilitate the molecular cloning and characterization of the maize miniature gene.Journal Paper No. J-13425 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011, USA, Project No. 2850