Genetic linkage relationships between two enzyme loci,Est-2 and acph,in the mosquito Aedes (Finlaya) togoi

An esterase locus (Est-2), coding for carboxylesterase, and an acid phosphatase locus (Acph) were genetically studied by agar gel electrophoresis in the mosquito Aedes (Finlaya) togoi. The Est-2 and Acph variants occur as a monomer and a dimer, respectively. Both enzyme loci are linked to the sex locus (M) and s (straw-colored larva); the gene arrangement and recombination distances were Est-2—12.6%—s—31.7%—M—2.9%—Acph—3.2%—Est-3. The Est-3 locus was previously shown to code for carboxylesterase.This work was supported by Grant AI 16983-02 from the National Institutes of Health, Bethesda, Md.     

Differential staining of polytene chromosome bands in Chironomus by Giemsa banding methods

Two Giemsa banding methods (C banding and RB banding) are described which selectively stain the centromere bands of polytene salivary gland chromosomes in a number of Chironomus species. — By the C banding method the polytene chromosome appearance is changed grossly. Chromosome bands, as far as they are identifiable, are stained pale with the exception of the centromere bands and in some cases telomeres, which then are intensely stained reddish blue. — By the RB method the centromere bands are stained bright blue, whereas the remainder of the polytene bands stain red to red-violet. — Contrary to all other species examined, in Chironomus th. thummi numerous interstitial polytene chromosome bands, in addition to the centromere regions, are positively C banded and blue stained by RB banding. In the hybrid of Ch. th. thummi x Ch. th. piger only those interstitial thummi bands which are known to have a greater DNA content than their homologous piger bands are C banding positive and blue stained by the RB method whereas the homologous piger bands are C banding negative and red stained by RB banding. Ch. thummi and piger bands with an equal amount of DNA both show no C banding and stain red by RB banding. — It seems that the Giemsa banding methods used are capable of demonstrating, in addition to centromeric heterochromatin, heterochromatin in those interstitial polytene chromosome bands whose DNA content has been increased during chromosome evolution.     

Linkage group IV of fish of the genus Xiphophorus (Poeciliidae): Assignment of loci coding for pyruvate kinase-1, glucosephosphate isomerase-1, and isocitrate dehydrogenase-1

Electrophoretic variation ascribable to three enzyme loci, coding for a pyruvate kinase (PK1), a glucose phosphate isomerase (GPI1), and an isocitrate dehydrogenase (IDH1), was observed in three species of fish of the genus Xiphophorus. Electrophoretic patterns in F₁ hybrid heterozygotes confirmed the dimeric structures of GPI and IDH, and indicated a multimeric structure for pyruvate kinase. Variant alleles at the three loci exhibited normal Mendelian segregation in backcross hybrids. Linkage analyses indicate a gene order and estimated recombination of PK1—10%—GPI1—41%—IDH1. No significant interference or sex- or population-specific recombination difference was detected. This group (designated linkage group IV) was shown to assort independently from the nine loci comprising linkage groups I, II, and III and from 23 other informative markers, within the limits of the data. No conclusions with respect to homology of linkage relationships could be reached, due to the presence of presumably duplicated loci in these fish coding for isozymes whose homology with enzymes in other vertebrate species is as yet unestablished.This work was supported in part by Public Health Service Research Grant CA-28909.     

Genetics of lucilin,a storage protein from the sheep blowfly,Lucilia cuprina (Calliphoridae)

Lucilin, the main storage protein of larval fat body and hemolymph in the sheep blowfly, Lucilia cuprina, has been isolated as a series of trimers composed of subunits of 83,000±5% daltons. Extensive electrophoretically detectable polymorphism of lucilin subunit patterns occurs in wild and laboratory populations of Lucilia; from four to nine bands are seen in any one individual. Evidence from genetic, electrophoretic, immunological, and structural studies suggests the existence of a series of 12 or more closely related structural loci (designated Luc-1 to Luc-12) which may have arisen through gene duplication. Codominant allelic variation has been found at several of these loci. Luc-1 and Luc-3, and probably the other structural loci of the series, are located on chromosome 2.Financial support for this work was largely obtained through the Australian Research Grants Committee (Grant D65/15167). J. A. T. held an Honorary Fellowship at the Australian National University during 1972–1973.     

The semidwarf gene,sd-1, of rice (Oryza sativa L.). II. Molecular mapping and marker-assisted selection

To establish the location of the semidwarf gene, sd-1, the anthocyanin activator (A), purple node (Pn), purple auricle (Pau), and the isozyme locus, EstI-2, in relation to DNA markers on the molecular linkage map of rice, 20 RFLP markers, previously mapped to the central region of chromosome 1 (McCouch et al. 1988), were mapped onto an F₂ population derived from the cross Taichung 65 (A,Pn,Pau)/Taichung 65 (sd-1). sd-1 and EstI-2 were determined to be linked most tightly to RFLP markers RG 109 and RG 220, which cosegregated with each other. The distance between these RFLP markers and sd-1 was estimated to be 0.8 cM, based on an observed recombination value of 0.8%. The order of genes and markers in this region of chromosome 1 was determined to be sd-1 — (EstI-2 — RG220 — RG109) — RG381 — A — Pn — Pau. To test the efficacy of selection for sd-1 based on these linked markers, 50-day-old F₂ seedlings derived from another cross, Milyang 23/Gihobyeo, were analyzed for marker genotype. At this age, the semidwarf character could not be clearly detected based on phenotype. In addition, plant height was normally distributed in this population, making it difficult to unambiguously identify plants carrying sd-1. Thirteen seedlings homozygous for the sd-1-associated allele at EstI-2, RG220 and RG109, and 13 seedlings homozygous for the Sd-1-associated allele at all three marker loci were selected for further genetic analysis. At 20 days after heading, the culm lengths of these 26 plants were measured and the expected phenotype was confirmed in every case. These 26 plants were then selfed for four generations and F₆ lines were again evaluated to determine whether any recombination among the three molecular markers, or between these markers and the sd-1 gene, could be detected. No recombinants were identified, confirming the tight linkage of these loci and the usefulness of genotypic selection for this recessive semidwarf character prior to the time when it can be evaluated based on phenotype.     

RFLP-based genetic map of rye (Secale cereale L.) chromosome 1R

Summary A map of chromosome 1R of rye was constructed using 16 molecular and biochemical loci. From long arm to short arm, known-function loci were placed in the order: XAdh — XLee — Glu-R1[Sec-3] — XPpdk-1R — XEm-1R-1 — XEm-1R-2 — Centromere — XNor-R1 —Gpi-R1 — XGli-R1 [Sec-1a] along with six anonymous genomic and cDNA clones from wheat. The map, which spans 106 cM with 12 loci clustered in a 15-cM region around the centromere, shows reasonably good agreement with previously published maps for the centromeric region, whereas the XNor-R1 — Gpi-R1 region gives a much larger distance than previously reported.     

Linkage of lactate dehydrogenase-2, Ldh-2, in the mouse

An electrophoretically detectable variant of lactate dehydrogenase-2 in Mus musculus has been found and used to locate the structural gene, Ldh-2, on chromosome 6. Gene order and recombination frequencies are estimated as Sig—36.0±4.8—Lc 21.0±4.1—Mi ^wh—20.0±4.0—Ldh-2.     

Giemsa banded karyotypes,systematics, and evolution inAnacyclus (Asteraceae-Anthemideae)

Giemsa C-banding allows the differentiation of six, otherwise very similar karyotypes from the small genusAnacyclus. Banding style—with stable centromeric and nucleolar bands, and diverse specific banding patterns in distal chromosome segments—contributes significantly to generic demarcation and systematic grouping. The amount of banding corresponds to heterochromatic chromocentres and increases from perennials to annuals. Relationships with other nucleotype parameters and evolutionary mechanisms are discussed.First contribution of a series on Giemsa Banded Karyotypes, Systematics, and Evolution inAnthemideae (Asteraceae).     

The human collagenase-3 (CLG3) gene is located on chromosome 11q22.3 clustered to other members of the matrix metalloproteinase gene family

The gene coding for human collagenase-3 (CLG3), a recently described matrix metalloproteinase produced by breast carcinomas, has been localized by fluorescence in situ hybridization on chromosome 11q22.3. Physical mapping of an isolated YAC clone containing CLG3 has revealed that this gene is tightly linked to those encoding other matrix metalloproteinases, including fibroblast collagenase (CLG1), stromelysin-1 (STMY1), and stromelysin-2 (STMY2). Further mapping of this region using pulsed-field gel electrophoresis has shown that the CLG3 gene is localized to the telomeric side of the matrix metalloproteinase cluster, the relative order of the loci being centromere—STMY2—CLG1—STMY1—CLG3—telomere.     

Physical mapping of the Esterase-6 locus of Drosophila melanogaster

The Esterase-6 gene locus of Drosophila melanogaster although well-characterized, has not been definitly mapped by in situ hybridization. In this paper, a high resolution in situ hybridization protocol using an avidin/biotinylated-horseradish peroxidase/diaminobenzidine system was adopted to refine the physical map position of the Esterase-6 locus. Clarity of signal, detail of banding pattern and absence of background allowed the assignment of a 1.8 kb cDNA encoding Esterase-6 to three bands within subsections 69 A1–A3 on the left arm of polytene chromosome 3. These data refine earlier deletion mapping and low resolution in situ hybridization results, which assigned Esterase-6 to 69 A1–A5. The potential use of this high resolution in situ hybridization technique in the analysis of the physical organization of the Esterase-6 gene duplication and surrounding region is discussed.     

Linkage studies on an esterase locus in the mosquito Aedes togoi

An electrophoretic survey of esterases in 7 wild-type and 10 mutant strains of the mosquito Aedes (Finlaya) togoi was undertaken using thin-layer agar gels. Three esterases (designated the Est-1, Est-2, and Est-3 loci in decreasing order of electrophoretic mobility) could be detected from fourth-instar larvae, pupae, and 2- to 5-day-old adults. Homogenates of the larvae gave the most intensely stained bands in the gels, especially for Est-3. The three esterases were designated carboxylesterases based on their response to the two esterase inhibitors, eserine and paraoxon (diethyl-p-nitrophenyl phosphate). The Est-3 locus was found to have five alleles including at least one null. The linkage results of six backcrosses suggest that Est-3 is located only 5–8 map units from the sex allele (m) and the gene arrangement is Est-3-m-s (straw-colored larva) in linkage group I.This work was supported by National Institutes of Health Grant AI 16983-01.     

Trisomic analysis of isozymic loci in tomato species: Segregation and dosage effects

Five isozymic loci were localized in the tomato (Lycopersicon esculentum) genome by trisomic analysis. Results revealed the following locations: Aps-1 on chromosome 6, Est-1 and Prx-2 on chromosome 2, Prx-4 on chromosome 10, and Prx-7 on chromosome 3. Three genes—Aps-1, Prx-2, and Prx-4—showed an arithmetic increase in allozyme concentration in direct proportion to the increase of gene dosage in respective primary trisomics. In contrast, no increase in relative Est-1 isozyme concentration was observed for any primary trisomic type. The phenotypes of the Aps-1, Prx-2, and Est-1 genes showed a pattern of banding intensity proportional to the allelic ratio (+/+/a vs. + /a/a) in primary trisomics; zymotypes of these differential trisomic heterozygotes appeared as converse images of each other.This research was performed under the auspices of NSF Grants BMS75-03024 and DEB77-02248 to C. M. Rick.     

Electrophoretic variation in esterases and peroxidases of native greek diploidAegilops species(Ae. caudata andAe. comosa,Poaceae)

The esterase and peroxidase patterns in five varieties ofAegilops caudata (genome type C) andAe. comosa (genome type M) were studied in order to elucidate the phylogenetic relationships within and between the two groups. The electrostarch gel electrophoresis technique was applied to extracts of shoot and root of 4-day-old seedlings, and the electropherograms were evaluated by gel densitometer traces. Inspite of considerable isozyme polymorphism, closer relationships in the banding patterns were found between different varieties of a single species than between varieties of the two different species. Esterase and peroxidase patterns of the twoAe. caudata varieties (caudata andpolyathera) are very similar and prove their close phylogenetic relationship. The isozyme affinities withinAe. comosa varieties are illustrated by the seriessubventricosa—biaristata—thessalica. The latter endemic variety has quite a number of characteristic bands and is relatively isolated. Altogether, the electrophoretic data agree well with morphological and cytological similarities (Zhukovsky 1928,Eig 1929,Karataglis 1973, 1975b).     

Genetics of alkaline phosphatase of the small intestine of the house mouse (Mus musculus)

Four inbred strains of mice exhibited either slow (PL/J), intermediate (DBA/2J, LP/J), or fast (SWR/J) rates of migration of duodenal alkaline phosphatase on cellulose acetate electrophoresis. Hybrids of these strains also had intermediate rates of migration regardless of the combination of strains used as parents. Strain differences were present in all regions of the small but not the large intestine. Crosses of the PL/J strain to hybrids between this strain and the other three strains gave a 1:1 segregation of the slow and intermediate patterns. The symbol Akp-3 is proposed for the locus responsible for the slower migration of the enzyme in this strain. Data from the LP/J × PL/J hybrid crossed with the PL/J strain showed linkage with two loci on chromosome 1 as follows: centromere—Idh-1–13.8±3.1 cM—Akp-3–8.9±2.6 cM—Pep-3. The available data do not reveal the genetic basis for the faster migration rate of the enzyme from the SWR/J strain, but a different response to neuraminidase and apparent nonlinkage to the Pep-3 locus suggest that a locus other than Akp-3 is responsible.This work was supported by a grant from the University Research Committee, Indiana State University.     

C banding in polytene chromosomes of Simulium ornatipes and S. melatum (Diptera: Simuliidae)

Polytene and mitotic chromosomes of Simulium ornatipes and S. melatum were subjected to C banding procedures. In both species polytene chromosomes consistently show C banding of centromere regions, telomeres, nucleolar organiser and, unexpectedly, numerous interstitial sites. The interstitial C banding sites correspond to morphologically single polytene bands. Their response is graded and independent of band size. Interstitial C bands in S. ornatipes are scattered throughout the complement, whereas in S. melatum they are clustered. Supernumerary heterochromatic segments in S. ornatipes also exhibit strong C banding and inverted segments can differ from standard in C banding pattern. — Mitotic chromosomes of both species show a single centric C band with indications of two weak interstitial bands in S. ornatipes, suggesting that many C band regions, detectable in polytene chromosomes, are not resolved by present techniques in mitotic chromosomes. — Contrary to current opinion that C banding is diagnostic for constitutive heterochromatin, the interstitial C band sites of polytene chromosomes are regarded as euchromatic. Conversely, the heterochromatic pericentric regions of S. ornatipes are not C banded. — It appears that polytene chromosomes offer a promising system for the elucidation of C banding mechanisms.     

Genetics of esterases in species of Lycopersicon

Summary Improvements in plant culture and electrophoretic technique permit detection and genetic analysis of seven esterase loci in Lycopersicon esculentum and related species with homosequential chromosomes. At all of these loci except one, each allele codes for a single anodal band, and the electrophoretic variants are inherited in monogenic fashion. For the exceptional Est-4, allozymes are 1–3 banded in various combinations at four positions, and rare recombinants in one cross appeared at a frequency of 0.0005, suggesting the existence of several very tightly linked genes. Est-2 segregated solely for intensity differences in dominant/recessive fashion; Est-3 and Est-4 behave as monomers; the remaining Est-l, 5, 6, and 7 — coding for contiguous bands in the region closest to the origin — are dimeric. The latter group are tightly linked inter se in the proximal portion of 2L (long arm of chromosome 2), the total map distance of the complex being approximately 1.5 cM; Est-2 is situated on 9L between ah and marm; Est-3 on 1L between inv and dgt; Est-4 has not yet been located. Even in the interspecific hybrids, map distances are similar to the standard values for L. esculentum. Tandem duplication is hypothesized for the origin of the Est-l, 5–7 complex, which adds another example to the growing list of linked mimic genes in the tomato genome. In respect to the position of their bands and tight inter se linkage, this series exactly parallels the EA, EB, EC esterase series in Hordeum vulgare — a fact which suggests great antiquity for this block of genes.     

Cytogenetic localization by variation in electrophoretic allozyme phenotype: Drosophila Odh

A gene-dosage effect is characteristic of eukaryotic structural genes and is therefore useful in gene mapping. However, attributing quantitative variations in enzyme activity to a gene-dosage effect or other putative regulatory loci can be suspect when the locus in question may be inducible by variations in culture conditions. The problem of controlling for allele-specific variations in activity and regulation can be circumvented in Drosophila melanogaster by the use of synthetic duplications and deficiencies in conjunction with enzyme polymorphism. A method for constructing segmental aneupliods heterozygous for electrophoretic variants of octanol dehydrogenase (Odh) is presented which permitted variations in allozyme phenotype and enzyme activity—which show a strict dosage dependency—to be produced simultaneously. The structural gene region for Odh was identified using T(Y;A) stocks and the deficiency M(3)S31 was used to assign the locus to polytene band region 86D1–4. With this method a segmental aneuploid survey of Drosophila for purposes of gene localization can be accomplished in one generation with appropriate stocks.This work was supported by National Institutes of Health Research Grant GM18678 awarded to E. Novitski.     

Genetic control and linkage relations of additional isozyme markers in chick-pea

Summary Allozyme polymorphisms of nine enzymes — aspartate aminotransferase (AAT), diaphorase (DIA), esterase (EST), formate dehydrogenase (FDH), -galactosidase (GAL), -glucosidase (GLU), malate dehydrogenase (MDH), malic enzyme (ME), and peroxidase (PRX) — were described in chick-pea (Cicer L.). Thirteen isozyme loci, Aat-c, Dia-4, Est-2, Est-4, Est-10, Fdh, Gal-2, Gal-3, Gal-4, Glu-3, Mdh-2, Me-2, and Prx-2, were genetically defined. Alleles of each of these isozyme loci expressed codominantly in heterozygotes and exhibited a codominant, single-locus segregation ratio in F₂. The loci Est-2, Mdh-2, and Me-1 were expressed only in flower. Linkage relations were determined for these 13 and several previously defined isozyme loci. The following new genetic linkages were identified: Pgm-p (locus for plastid phosphoglucomutase) — Est-10; Ald-p1 (one of the duplicate loci for plastid aldolase) — Glu-3 — Gal-2 — Est-2,3; Gal-3 — Aco-m (locus for mitochondrial aconitase) — Prx-2,3; Gpi-c (locus for cytosolic glucosephosphate isomerase) — Fdh; and Est-4 — Me-1. This study provides further confirmation on the existence of several conserved linkage groups among Cicer, Pisum, and Lens.     

Giemsa C-banded karyotypes and taxonomic relationships of some North AmericanAllium species and their relationship to Old World species (Liliaceae)

The somatic karyotypes of six North AmericanAllium species and the EuropeanA. scorzonerifolium have been investigated using a Giemsa C-banding technique. All species have a chromosome number of 2n = 14. InA. scorzonerifolium and the three North American speciesA. dichlamydeum, A. fibrillum andA. unifolium C-bands are restricted to two pairs of nucleolar chromosomes. Each chromosome has a band proximal to the nucleolar constriction and a positively banded satellite. InA. acuminatum, in addition to the bands associated with the nucleolar constrictions, all chromosomes also have pericentromeric bands.A. cernuum exhibits a distinctive banding style: two chromosome pairs with bands adjacent to the nucleolar constrictions and four pairs with telomeric bands on their short arms. In the karyotype ofA. geyeri neither C-bands nor nucleolar chromosomes were found.—A comparison of the banding styles together with other cytological and morphological characters of these species with old world members ofAllium reveals:A. cernuum closely resembles species within subgenusRhizirideum, whereas the other species studies exhibit many similarities with subgenusMolium. Their sectional grouping and their relationships with Old World species are discussed.     

A polymorphism detected in a variable number of tandem repeats (VNTR) of the seminal vesicle secretion (SVS) IV gene in inbred rats

The second intron of the rat SVS IV gene contains a tandem repeat region of 20-bp sequences. This region was amplified using the polymerase chain reaction to detect variations. Three alleles, characterized by amplified fragments of 750, 490, and 390 bp, respectively, were found in 24 strains examined. This variation segregated in F₁ and backcross progeny in an autosomal codominant manner. We tentatively designated this locusSvs-4. Analysis of linkages between theSvs-4 locus and other loci revealed that it was closely linked to theSvp-1 (<2.9%) and the a (10.0±6.7%) loci, which belong to rat linkage group IV. TheSvp-1 andSvs-4 loci, however, were differently distributed among the inbred rat strains.This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.