Microsatellite markers for northern red oak (Fagaceae: Quercus rubra)

We provide primer sequences for 14 (GA) _n microsatellite loci developed from northern red oak, an important timber species. We screened loci using two sets of samples. A parent–offspring set included DNA from seven acorns collected from one mother tree along with maternal DNA, to determine that all progeny carried a maternal allele at each locus. The other set was comprised of 10 adult trees sampled from Indiana old‐growth forest, providing a measure of diversity revealed by each locus.     

The highly polymorphic microsatellite markers for the greater long‐tailed hamster (Tscherskia triton)

We isolated 25 dinucleotide microsatellite loci from the greater long‐tailed hamster (Tscherskia triton) populations in North China. We developed the amplification conditions of polymerase chain reaction for producing high‐resolution genetic markers for each locus. We found 10 microsatellite loci were highly polymorphic in 90 individual hamsters from three areas of North China, and the number of alleles in each locus varied from three to 11. These markers are potential tools for studying the genetic variation of the natural populations of this species.     

Characterization of seven polymorphic microsatellites for the study of two Ranunculaceae: Ranunculus nodiflorus L., a rare endangered species and Ranunculus flammula L., a common closely related species

Ranunculus nodiflorus and Ranunculus flammula are two closely related Ranunculaceae. In France, the first one is rare and endangered, whereas the second is common. To compare their population genetic structure, seven polymorphic microsatellite markers were characterized. We found no polymorphism in R. nodiflorus, whereas R. flammula was polymorphic at each locus. The total number of alleles per locus varied from three to eight. Observed heterozygosities ranged from 0 to 0.261. This set of microsatellites offers an efficient tool to study population genetic structure and to compare the results between closely related endangered and common species.     

Isolation and characterization of six microsatellite loci in the drywood termite Cryptotermes secundus (Kalotermitidae)

The Kalotermitidae is a family of lower termites and its species are ideal to study the evolution and maintenance of eusociality within the Isoptera. Co‐dominant genetic markers are an essential tool to elucidate the putatively complex colony and population structure. Six polymorphic microsatellite loci were identified in the drywood termite Cryptotermes secundus (Kalotermitidae). To test the variability of the markers, 18 colonies (n = 18) from Darwin (Australia) were assayed. We found two to eight alleles per locus with the level of observed heterozygosity at each locus ranging from 0.00 to 0.53. The significantly positive F_IT value suggests that some kind of inbreeding does occur in this population.     

Involvement of transposition in dispersion of tandem repeat sequences (TrsA) in rice genomes

We describe a method to identify and characterize DNA fragments containing the junction of AA genome-specific tandem repeat sequences (here called TrsA) with adjacent chromosomal sequences of rice by the polymerase chain reaction (PCR) using a pair of primers that hybridize with TrsAs and a flanking non-TrsA sequence. With this method, we obtained results suggesting that TrsA sequences present at two loci (here called trsA1 and trsA2) are flanked by direct repeats of chromosomal sequences of 172 by and about 440 by in length, respectively. These results support the idea that the TrsA sequences have been inserted into each locus by transposition, resulting in duplication of the chromosomal sequence used as target. We also describe a method to identify and characterize TrsA sequences repeated in only a few copies in the rice genome by PCR, using a pair of primers that hybridize with two different portions in the TrsA sequence, and demonstrate that TrsA sequences are present not only in rice strains with the AA genome, but also in those with non-AA genomes. The TrsA sequences were present at the trsA1 locus in all the rice strains examined, indicating that TrsA was inserted and amplified at the locus before the divergence of the various species of rice in the Oryza genus. TrsA sequences were present at the trsA2 locus, however, only in an O. sativa IR36 strain, indicating that TrsA was inserted and amplified at this locus during divergence of rice strains with the AA genome.     

Eight polymorphic microsatellite markers isolated from the widespread avian louse Colpocephalum turbinatum (Phthiraptera: Amblycera: Menoponidae)

We report eight novel microsatellite loci for Colpocephalum turbinatum, a parasitic louse of the endangered Galápagos hawk (Buteo galapagoensis). Two island populations of C. turbinatum (N = 30) were genotyped for each locus. We found between two and 12 alleles per locus, polymorphic information content from 0.268 to 0.798, observed heterozygosity from 0.067 to 0.667 and no linkage disequilibrium was detected between loci. These markers will be useful in understanding contemporary gene flow of C. turbinatum among islands in the Galápagos and in understanding transmission dynamics between B. galapagoensis hosts, within and between social groups. Because this louse is unusually widespread among avian host taxa, parasitizing at least 53 bird species in the Falconiformes, Strigiformes and Columbiformes, these markers are likely to be useful outside the context of the Galápagos Islands.     

The dynamics of interlocus associations in the three-locus hitchhiking model

The hitchhiking effects of a selected locus upon the dynamics of the pairwise association,D _nn between two neutral loci is examined analytically for the special case where at least one of the neutral loci is in linkage equilibrium with the selected locus. The results apply approximately whenever the product of the pairwise associations between the selected locus and each neutral locus is negligible with respect to the three-way linkage disequilibrium. It is shown that precisely four broad classes of trajectories are possible, whether the selected locus is between (nsn) or to one side (snn) of the neutral loci, and whatever the mode of selection operating.D _nn may: (1) decay rapidly to zero, at a rate faster in each generation than that expected for two isolated neutral loci; (2) monotonically decay to zero at a rate which is slower in every generation than under the usual neutral regime; (3) increase initially and/or in intermediate periods before eventually slowly decaying to zero; or (4) exhibit type 1 behavior in the first segment of the trajectory and either type 2 or 3 behavior in the subsequent generations, with the transition marked by a change in sign. The nature of a given trajectory is largely determined by the direction of gene frequency change at the selected locus, and the initial signs of bothD _nn and the three-way linkage disequilibrium.The single most important consequence of these results is that there is no simple relation between the amount of pairwise association between two neutral markers and the recombination fraction between them. Several factors influencing the magnitude of the hitchhiking effect are also examined. It is shown that, all else being equal, the greater the three-way linkage disequilibrium, the greater the departure ofD _nn from the expected neutral dynamic. Increased recombination among the loci reduces the hitchhiking effect onD _nn . The dependence of the behavior upon the exact position of the selected locus is also determined both within and betweennsn andsnn chromosomal systems. An interesting discovery is that given equivalentnsn andsnn systems, with each having the same recombination between their two neutral loci,D _nn will deviate more from the standard neutral dynamic in thesnn system if its selected locus is sufficiently tightly linked to the neutral loci.     

Isolation and characterization of microsatellite loci in the beaver (Castor canadensis)

We have isolated and characterized 10 microsatellite loci in the beaver (Castor canadensis). Sixty individuals from southern and central Illinois were screened at each locus. All loci exhibited moderate levels of polymorphism, ranging from five to 13 alleles per locus with average heterozygosity ranging from 0.317 to 0.867. Locus Cca5 deviated significantly from HWE (P < 0.001). The locus pair Cca4/Cca5 was shown to be in linkage disequilibrium in southern Illinois, but not in the central Illinois population. The remaining eight loci will be useful in investigations of mating and kinship patterns in beaver populations in Illinois.     

Microsatellite isolation from the gall midge Spurgia capitigena (Diptera: Cecidomyiidae), a biological control agent of leafy spurge

Spurgia capitigena is a gall midge that has been released as a biological control agent of leafy spurge (Euphorbia esula), a noxious rangeland weed in North America. We isolated 15 microsatellite loci from S. capitigena with from two to 27 alleles at each locus. These markers will allow us to examine the structure and levels of neutral genetic variation in native and introduced populations.     

Linkage relationships between regulatory and structural gene loci involved in zein synthesis in maize

Summary The synthesis of at least 15 zein polypeptides is under the control of regulatory gene loci. One of these, Opaque-2 (chromosome 7, position 16) strongly reduces the zein accumulation without modifying the zein molecular components. The linkage relationship between the regulatory gene 02 and the 5 structural loci (Zp1, Zp2, Zp3, Zp6, Zp12) segregating with sample Mendelian ratios have been studied. Zp1, Zp2, Zp3 are closely linked to each other; moreover this gene cluster is located on chromosome 7 at 5.5 cM from the Opaque-2 locus. The structural loci Zp6 and Zp12 are not linked with each other, with the 02 locus or with Zp1, Zp2, Zp3. From our data it follows that the zein structural genes are located in at least three positions on the maize genome. The scattering in the genome of the genes controlled by the Opaque-2 locus suggests a transacting role for this regulatory element.     

Isolation and characterization of microsatellite markers in the solitary hunting wasp Ancistrocerus adiabatus (Vespidae: Eumeninae) and their cross utility in other species of Ancistrocerus

We have developed seven microsatellite primers for the solitary wasp Ancistrocerus adiabatus. The number of alleles per locus ranged from three to 14. The markers were developed to study population structure, inbreeding and sex determination of A. adiabatus. For each locus, we genotyped 30 female wasps collected from Southwest Michigan. The observed heterozygosity ranges from 0.067 to 0.933. We have also determined that primers for these microsatellite loci amplify in three other species in the genus: Ancistrocerus antilope, Ancistrocerus catskill, and Ancistrocerus campesterus.     

Fine genotyping of a highly polymorphic ASTRINGENCY-linked locus reveals variable hexasomic inheritance in persimmon (Diospyros kaki Thunb.) cultivars

Persimmon (Diospyros kaki Thunb.) is one of the major tree crops in East Asia and is generally hexaploid. A single ASTRINGENCY (AST) locus controls the astringency/non-astringency (A/NA) trait of persimmon fruit, one of the most important traits for consumption, on each of the six corresponding chromosomes. Although several molecular approaches are in progress to elucidate the molecular mechanisms of astringency trait in persimmon, the distinct polysomic behavior of the AST locus remains to be solved. The aim of this study was to perform fine genotyping of a highly polymorphic marker locus linked to the AST locus, detect the allele pairing in ten segregated F₁ lines derived from hybridization of A-type × NA-type cultivars, and identify the basis of hexaploid inheritance at the AST locus in persimmon. The results showed that persimmon cultivars frequently produce aneuploid offspring bearing an extra chromosome with the AST locus, with the incidence of aneuploidy varying among the cultivars. On the examination of hexasomic behavior in persimmon cultivars, the ratios of individuals bearing each allele pair segregated from A-type parents showed a good fit to the expected ratios in an autohexaploid inheritance model, except for cvs. Luo-tian-tian-shi and Sa-gok-shi which fitted to an autoallohexaploid inheritance model. These results suggest variable hexasomic behavior among persimmon cultivars.     

Biochemical composition of maize (Zea mays L.) pollen

Summary Pollen grains containing either the Wx, wx, Su ₁, su ₁, Sh ₂ or sh ₂ alleles were stored at 0, 1, 2, 3, 4 and 5 days at 2 °C. After each storage period, a portion of pollen from each genotype was analyzed for free amino acid content. Over all genotypes, storage significantly altered the content of all 16 amino acids measured. With increasing storage, a relatively consistent increase in aspartic acid, isoleucine, leucine, phenylalanine, ethanolanine, aminobutyric acid, NH₃ and lysine was found. A relatively consistent decrease in glutamic acid, proline, glycine and alanine occurred with increasing storage. No consistent response to storage was obtained with threonine-serine, valine, histidine and the unknown. Apparently, storage or stage of viability loss has a pronounced effect on amino acid metabolism in maize pollen grains. The experiment was designed so that comparisons free of genetic background effects could be made between alleles at each locus. Significant allele X storage interactions at each locus were found as follows: at the waxy locus, aspartic acid, glycine, alanine and ethanolanine; at the sugary locus, aspartic acid, alanine, ethanolanine and aminobutyric acid; and at the shrunken locus, aspartic acid, alanine, valine, leucine and ethanolanine. Amino acid metabolism is apparently influenced by the action of the alleles at these loci. The differences between the loci in the amino acids affected indicate the different areas of amino acid metabolism are influenced by each locus.Journal Series Paper No. 4425, Florida Agricultural Experiment Station.     

Identification of two loci involved in phytochrome expression in Nicotiana plumbaginifolia and lethality of the corresponding double mutant

Four Nicotiana plumbaginifolia mutants exhibiting long hypocotyls and chlorotic cotyledons under white light, have been isolated from M2 seeds following mutagenesis with ethyl methane sulphonate. In each of these mutants, this partly etiolated in white light (pew) phenotype is due to a recessive nuclear mutation at a single locus. Complementation analysis indicates that three mutants, dap5, ems28 and ems3-6-34, belong to a single complementation group called pew1, while dap1 defines the pew2 locus. The mutants at pew1 contain normal levels of immunochemically detectable apoprotein of the phytochrome that is relatively abundant in etiolated seedlings, but are deficient in spectrophotometrically detectable phytochrome, whether seedlings are grown in darkness or light. Moreover, biliverdin, a precursor of the phytochrome chromophore, restores light-regulated responses in pew1 mutants and increases their level of photoreversible phytochrome when grown in darkness. These results indicate that the pew1 locus may be involved in chromophore biosynthesis. The mutant at the pew2 locus displays no photoreversible phytochrome in etiolated seedlings, but does contain normal levels of photoreversible phytochrome when grown in the light. Biliverdin had little effect on light-regulated responses in this mutant. In addition, biliverdin did not alter the level of phytochrome in etiolated seedlings. These observations lead us to propose that this mutant could be affected in the phyA gene itself. We have also obtained the homozygous double mutant at the pew1 and pew2 loci. This double mutant is lethal at an early stage of development, consistent with a critical role for phytochrome in early development of higher plants.     

The <Emphasis Type="Italic">Tyr</Emphasis> (<Emphasis Type="Italic">albino</Emphasis>) locus of the laboratory mouse

The albino mouse was already known in ancient times and was apparently selectively bred in Egypt, China, and Japan. Thus, it is not surprising that the c or albino locus (now the Tyr locus) was among the first used to demonstrate Mendelian inheritance in mammals at the dawn of the past century. This locus is now known to encode tyrosinase, the rate-limiting enzyme in the production of melanin pigment, and the molecular basis of the albino (Tyr ^c ) mutation is known. Here we describe the congenic series of Tyr-locus alleles, from wild type to null (albino). We compare eye and skin pigmentation phenotypes and the genetic lesions that cause each. We suggest that this panel of congenic mutants contains rich, untapped resources for the study of many questions of basic cell biological interest.     

Positive selection on the mitochondrial ATP synthase 6 and the NADH dehydrogenase 2 genes across 22 hare species (genus Lepus)

Amino acid changes in mitochondrial (mt) oxidative phosphorylation (OXPHOS) genes have been suggested as a key adaptation to environmental variation. Here, we analyzed 416 sequences of ATPase synthase 6 (MT‐ATP6) and NADH dehydrogenase 2 (MT‐ND2) in 22 different hare (Lepus) species from across a wide range of habitats and climates. We used site‐ and branch‐based methods to test for positive selection on specific codons and lineages. We found four codons in MT‐ATP6 and five in MT‐ND2 under positive selection, affecting several species lineages. We investigated the association of protein variants at each locus with climate zone, using multinomial generalized linear models (glm), including species, regions, historical introgression events, and the co‐occurring protein variant at the other locus as additional explanatory variables. A significant climate effect as based on the “Köppen climate classification” was observed for MT‐ND2 protein variants as translated from our nucleotide sequences. Moreover, MT‐ND2 protein variants were significantly affected by the co‐occurring MT‐ATP6 protein variant in the same mtDNA molecule. Contrary to the expectation for non‐recombining mitochondrial DNA molecules, the presence of an evolutionarily relatively ancestral protein variant at one locus was associated with a relatively derived protein at the other locus in the same mitochondrial molecule, respectively. The relative evolutionary status of a protein variant was evaluated according to its positions relative to the respective out‐group protein variant in a network analysis of nucleotide sequences. All our results suggest a complex effect of various climatic parameters acting on multiple mtOXPHOS genes in a co‐adaptive way, favoring combinations of ancestral and derived variants.     

A gene's eye view of epistasis,selection and speciation

In this mini‐review, I discuss the effects of gene interaction or epistasis from a `gene's eye view.' By a `gene's eye view' of epistasis, I mean that I will consider a single, bi‐allelic locus, A , whose effects on fitness result only from its interactions with alleles of another, unknown locus, X . I will show how changes in the frequencies of alleles at the background locus affect the relationship of alleles at the A ‐locus to fitness. Changing the genetic background changes the fundamental characteristics of the A ‐locus, such as the magnitude and sign of allelic effects on fitness, and, consequently, it changes the strength and pattern of selection. I consider each of the four kinds of pair–wise interactions between two loci and show that some kinds of epistasis are more sensitive than others to population genetic subdivision. Lastly, I show that some kinds of epistasis are more likely than others to affect the process of speciation and contribute to or be responsible for general genetic features of interspecific hybrids, such as Haldane's rule.     

Characterization of 14 tetranucleotide microsatellite loci derived from Pacific herring

Spawning aggregations of Pacific herring (Clupea pallasi) often exhibit significant interannual variation in allele frequencies of neutral gene markers. We isolated 14 tetranucleotide microsatellites to examine hypothetical processes that may produce this unique genetic signal. We developed and tested primer pairs for each locus and then estimated locus variability in samples (n = 60) from two populations. The number of alleles per locus ranged from five to 49. The expected heterozygosity across loci and populations ranged from 0.20 to 0.96. These microsatellites will be useful for estimating genetic variation in herring on a fine geographical scale.     

Mapping and characterization of the DQ subregion of the ovine MHC

A map of the ovine MHC class II DQ subregion has been constructed from overlapping cosmid clones. This region consists of two loci linked on a linear tract of 130 kb DNA. Each locus consists of a DQA and a DQB gene in a tail-to-tail orientation. The genes in each locus are transcribed but only those designated DQ1 express class II molecules at the surface of mouse L cells following DNA-mediated gene transfection. The DQA1 and DQB1 genes are separated by 11kb while the DQA2 and B2 genes are 25 kb apart. The loci are separated by 22 kb.