Inheritance of enzymes and blood proteins in the leopard frog,Rana pipiens: Three linkage groups established

Individuals from natural populations of the leopard frog, Rana pipiens, were analyzed for electrophoretic differences in blood proteins and enzymes from an amputated digit. The proteins examined represent products of 72 loci. Presumptive heterozygotes at multiple loci were selected for experimental crosses. Mendelian inheritance of 18 protein variations were demonstrated in the offspring. Tests for linkage or independent assortment were performed for 75 locus pairs. Three linkage groups were established. Linkage group 1 contains two loci, aconitase-1 (Acon1) and serum albumin (Alb), with a 19% recombination frequency between them. Linkage group 2 contains four loci, glyoxalase (Gly), acid phosphatase-1 (Ap1), acid phosphatase-2 (AP2), and esterase-5 (Est5). The data show the relationships Gly-21.1%-AP1-0%-AP2-6.3%-Est5, and Gly-25.6%-Est5. Linkage group 3 consists of four closely linked esterase loci. The data, Est1-5.1%-Est6, Est6-1.8%-Est10-1.9%-Est4 and Est6-3.0%-Est4, do not establish a complete order but suggest that Est10 is between Est4 and Est6. These results, with data demonstrating apparent independent assortment of 67 other locus pairs, provide a foundation for establishing the frog genetic map.The project was supported by Grant No. RR-00572 from the Division of Research Resources, National Institutes of Health. This paper is contribution No. C-87 from the Amphibian Facility, George W. Nace, Director.     

Worldwide patterns of genetic variation among four esterase loci in Barley (Hordeum vulgare L.)

Summary Electrophoretic assays of 1506 accessions of domestic (Hordeum vulgare L.) and wild (H. spontaneum Koch.) barley, maintained in the USDA World Barley Collection, led to the following conclusions: (1) worldwide the four esterase loci, Est 1, Est 2, Est 3, and Est 4, have a minimum of 7, 12, 6, and 7 alleles, respectively; (2) little or no genetic differentation has developed between H. vulgare and H. spontaneum at these four esterase loci; (3) substantial genetic polymorphism and heterozygosity occur within many of the accessions despite the heavy inbreeding which results from the mating system of predominant self fertilization and from genetic drift associated with maintenance in small populations; (4) patterns of geographical distribution of alleles at these four loci are not at random over both small and large geographical areas, including differences on a continental scale; (5) four among 16 four-locus combinations of alleles are found in excess and all other combinations occur in deficiency on a worldwide basis.This work was supported in part by National Science Foundation Grant DEB 78-02046     

Es-6, a further polymorphic esterase in the rat

A further polymorphic rat esterase with broad tissue expression and restricted substrate specificity is described and tentatively called Es-6. Inbred rat strains have either fixed allele Es-6^F or fixed allele Es-6^S. Es-6 is not linked to the established esterase cluster consisting of the eight esterase loci Es-1, Es-2, Es-3M, Es-4M, Es-4W, Es-5 (=Es-3W), Es-7, and Es-8 in LG V of the rat or to RT1, Gc, c, a, and h. Esterases with apparently identical biochemical and genetical characteristics are Es-17 of the mouse and Es-A4 of humans.Supported by the Deutsche Forschungsgemeinschaft (Be 352/13 and Gu 105).     

Geographic distribution of alleles at the Ga2 locus for segregation distortion in barley

Summary A distorted segregation of esterase alleles at the complex loci, Est1, Est2 and Est4, was found in an F₂ population. This distortion is typical for cross combinations between the Ga2Ga2 and ga2ga2 genotypes responsible for segregation distortion, since the Ga2 locus is linked with the complex loci encoding the esterase isozymes. The segregation of esterase isozyme patterns in F₂ populations between 473 varieties of barley and a tester of ga2ga2 genotype was examined, and the genotypes inducing segregation distortion were detected. Varieties with a ga2ga2 genotype are widely distributed throughout the world, whereas Ga2Ga2 varieties are found only in eastern and southern regions of Asia, from Japan to North India, with a low frequency. In varieties collected from these regions, some associations were detected between alleles at the Ga2 locus and esterase isozyme patterns. Additionally, most of the Ga2 barley varieties are naked and possess a BtBtbt2bt2 genotype for a non-brittle rachis.     

Chromosomal location of esterase,peroxidase and phosphoglucomutase isozyme structural genes in cultivated rye (Secale cereale L.)

Summary Isoelectric focusing of esterase (EST), peroxidase (PRX), and phosphoglucomutase (PGM) isozymes in Chinese Spring wheat, Imperial rye and several Chinese Spring/Imperial and Holdfast/King II addition, translocation and substitution lines revealed the chromosomal location of nine Est loci previously described and of one Prx and Pgm locus. Loci Est1, Est2, Est3, Est5, Est6 and Est7 were found on chromosome arm 5RL, Est8 and Est9 on chromosome 6R in Imperial rye, and the Est10 locus on chromosome arm 4RL in Imperial rye and King II rye. A discrepancy was found between the chromosomal location of the Prx locus in Imperial where chromosome 2R was responsible for the expression of the peroxidase enzyme, and King II with chromosome 1R carrying the Prx gene. As a possible explanation, the occurrence of translocation events during the production of wheat/rye aneuploid lines is discussed. The rye Pgm locus could be associated with chromosome 4RS in Imperial and King II rye. Except for the location of Est loci on chromosome 5RL, the results reported in this paper lend further evidence for the assumed homoeology relationships between the chromosomes of Triticinae and for the conservation of gene synteny groups during the evolution of the Triticeae tribe.     

Analysis of Linkage Between Biochemical and Morphological Markers of Rye Chromosomes 1R, 2R,and 5R and Mutations of Self-Fertility at the Main Incompatibility Loci

In F₂ hybrids between self-sterile plants of the Volkhova cultivar and self-fertile lines with established self-fertility mutations (sf mutations) at the major incompatibility loci S (1R), Z (2R), and T (5R), the effect of sf mutations on the inheritance of secalin-encoding, isozyme, and morphological markers located on the same chromosomes was investigated. Linkage between loci Prx7 and Sand locus Sec3 coding for high-molecular-weight secalins on chromosome 1R was shown for the first time. The frequency of recombination between Prx7andSec3and between S and Sec3was 29.1 ± 4.8% and 30.9 ± 7.0%, respectively. Independent inheritance of locus Z and isozyme markers of chromosome 2R, Est3/5 and -Glu, from locus Sec2 encoding 75-kDa -secalins was shown; in hybrids, the recombination frequency between Est3/5 and locus Z varied from 19.2 ± 8.1 to 50%. Independent inheritance of morphological (Ddw and Hs) and isozyme markers (Est4, Est6/9,and Aco2) of chromosome 5R from locus Tlocated on the same chromosome was demonstrated.     

Structural and biochemical characterization of a metagenome‐derived esterase with a long N‐terminal extension

The genes encoding six novel esterolytic/lipolytic enzymes, termed LC‐Est1～6, were isolated from a fosmid library of a leaf‐branch compost metagenome by functional screening using tributyrin agar plates. These enzymes greatly vary in size and amino acid sequence. The highest identity between the amino acid sequence of each enzyme and that available from the database varies from 44 to 73%. Of these metagenome‐derived enzymes, LC‐Est1 is characterized by the presence of a long N‐terminal extension (LNTE, residues 26–283) between a putative signal peptide (residues 1–25) and a C‐terminal esterase domain (residues 284–510). A putative esterase from Candidatus Solibacter usitatus (CSu‐Est) is the only protein, which shows the significant amino acid sequence identity (46%) to the entire region of LC‐Est1. To examine whether LC‐Est1 exhibits activity and its LNTE is important for activity and stability of the esterase domain, LC‐Est1 (residues 26–510), LC‐Est1C (residues 284–510), and LC‐Est1C* (residues 304–510) were overproduced in E. coli, purified, and characterized. LC‐Est1C* was only used for structural analysis. The crystal structure of LC‐Est1C* highly resembles that of the catalytic domain of Thermotoga maritima esterase, suggesting that LNTE is not required for folding of the esterase domain. The enzymatic activity of LC‐Est1C was lower than that of LC‐Est1 by 60%, although its substrate specificity was similar to that of LC‐Est1. LC‐Est1C was less stable than LC‐Est1 by 3.3°C. These results suggest that LNTE of LC‐Est1 rather exists as an independent domain but is required for maximal activity and stability of the esterase domain.     

ESTERASE VARIATION AND GENETIC STRUCTURE IN THREE GEOGRAPHIC POPULATIONS OF SITODIPLOSIS MOSELLANA (GEHIN) (DIPTERA: CECIDOMYIIDAE) IN WESTERN CHINA*

Abstract This paper investigates the esterase variation and genetic structure in three geographic populations of Sitodiplosis mosellana (Géhin) in western China by PAGE. The localities surveyed are Gaolan (36.3°N, 103.9°E) and Wuwei (37.9°N, 102.6°E) in spring wheat region and Chang'an (34. 1°N, 108.9° E) in winter wheat region. The results suggest that the esterase is coded by two loci: Est‐1 and Est‐2. Est‐1 is coded by a plastogene producing only one band that is the fastest on the gel among all bands. The Est‐2 is duplicated loci with 8 alleles, namely, a, b, c, d, e, f, g, h, which produce altogether 8 bands in all the populations and 1–4 bands in individual samples. There are 19 zymogram types observed in the three geographic populations. Seventeen zymogram types emerge in Chang'an population, but 5 and 4 zymogram types are found respectively in Gaolan and Wuwei populations. II₂ zymogram type is the commonest in all the populations. The alleles that had the highest frequencies in all the populations are d, e, g. All 8 alleles at the Est2 were observed in Chang'an population, but only total 3 alleles‐d, e, g at the Est‐2 appeared in Gaolan and Wuwei populations. The analysis of genetic identity and cluster (UPGMA) on the alloenzyme indicates that the relationship between the two populations of spring wheat region seems to be closer, as compared with the relationship between spring wheat population and winter wheat population. It is evident that there exists some infraspecific variation caused mainly by genetic drift in S. mosellana and the gene flow among the populations possibly took place to some extent.     

Studies of esterase 6 in Drosophila melanogaster XII. Evidence for temperature selection of Est 6 and Adh alleles

Changes in allele frequencies at the esterase 6 (Est 6) and alcohol dehydrogenase (Adh) enzyme loci of Drosophila melanogaster and simulans are examined in natural populations and artificial populations maintained at two temperatures. Results from cage populations at 18 °C and 25 °C provide evidence for temperature selection at both loci. Seasonal population samples show no significant change in gene frequencies for either locus, a reasonable outcome given the small selection coefficients found in cage populations. The temperature effect for the Adh locus appears to be direct: natural selection of the fast allele in cool environs and of the slow allele in warm environs. The temperature effect for Est 6 is weaker and complicated by sex differences and deviations from Hardy-Weinberg expectation. This evidence and different Est 6 frequencies found for melanogaster and simulans, in conjunction with evidence of the male reproductive function of this enzyme, suggest that Est 6 polymorphisms are maintained in natural populations by a complex form of sexual selection.     

Identification and characterization of a novel salt‐tolerant esterase from a Tibetan glacier metagenomic library

A salt‐tolerant esterase, designated H9Est, was identified from a metagenomic library of the Karuola glacier. H9Est gene comprised 1071 bp and encoded a polypeptide of 357 amino acids with a molecular mass of 40 kDa. Sequence analysis revealed that H9Est belonged to the family IV of bacterial lypolitic enzyme. H9Est was overexpressed in Escherichia coli and the purified enzyme showed hydrolytic activity towards p‐nitrophenyl esters with carbon chain from 2 to 8. The optimal esterase activity was at 40°C and pH 8.0 and the enzyme retained its activity towards some miscible organic solvents such as polyethylene glycol. A three‐dimensional model of H9Est revealed that S200, D294, and H324 formed the H9Est catalytic triad. Circular Dichroism spectra and molecular dynamic simulation indicated that the esterase had a wide denaturation temperature range and flexible loops that would be beneficial for H9Est performance at low temperatures while retaining heat‐resistant features. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:890–899, 2015     

Latitudinal clines for nucleotide polymorphisms in the Esterase 6 gene of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

Previous studies have found non-neutral patterns of nucleotide polymorphism in the promoter and coding regions of Est6 in D. melanogaster. Coding region polymorphism peaks around two closely linked replacement differences associated with the EST6-F/EST6-S allozyme polymorphism. The promoter contains two common, highly diverged haplotype groups, P1 and P7, that differentially affect Est6 expression. Allozyme studies have also revealed latitudinal clines in EST6-F and EST6-S frequencies that recur across continents. Here we analyse nucleotide polymorphisms across the promoter and the region of peak coding sequence polymorphism in 10 Australian populations along a 25° latitudinal gradient in order to examine the basis for the allozyme clines. As with the earlier studies, we find an excess of intermediate to high frequency variants in both the P1/P7 region and around the two EST6-F/EST6-S replacements in some populations. The two EST6-F/EST6-S replacement polymorphisms show latitudinal clines whereas the P1 and P7 groups of promoter haplotypes do not. However the strongest clines are for three co-segregating silent site polymorphisms in a 4 bp stretch at the 3′ end of the sequenced region. Monte Carlo simulations show that the clines for those three sites can explain all others in the data but none of the others can explain those three. Thus the allozyme clines may not reflect selection on either the P1/P7 polymorphism or the two replacements previously associated with the EST6-F/EST-S difference.     

Genetics of three esterase loci in Anopheles stephensi liston

A survey of laboratory strains of Anopheles stephensi for nonspecific esterases by polyacrylamide gel electrophoresis revealed 10 zones of esterase activity. In 3 of the 10 zones, three electromorphs were observed. Genetic analysis revealed that these three zones are controlled by three loci, viz., Est-3, Est-4, and Est-5, and that the electromorphs are codominant alleles at each locus. The three esterase loci were found linked to each other and to an autosomal marker colorless-eye. The esterase loci have tentatively been placed in linkage group II. The probable gene sequence on chromosome 2 is either c-Est-3-Est-4-Est-5 or c-Est-4-Est-3-Est-5.     

Contrasting population genetic structures using allozymes and the inversion polymorphism in Drosophila buzzatii

Second chromosome inversion and genotypic frequencies at seven allozyme loci, differentially associated with inversions, were determined in seven natural populations of Drosophila buzzatii. The patterns of variation of allozymes and the inversion polymorphisms were significantly different, indicating the role of adaptive differentiation for the latter. Moreover, the patterns of population structure varied among allozyme loci, suggesting the operation of diversifying selection for certain loci. Differentiation was negligible for Leucyl‐amino peptidase (Lap) and Peptidase‐2 (Pep‐2), low to moderate for Aldehyde oxidase (Aldox), Peptidase‐1 (Pep‐1) and Esterase‐1 (Est‐1) and high for Esterase‐2 (Est‐2) and Xanthine dehydrogenase (Xdh). Significant linkage disequilibria were detected between inversions and Aldox, Est‐1, Est‐2 and Xdh. Multiple regression analyses of inversion and allele frequencies on environmental variables revealed the existence of clines for inversions, Est‐1, Est‐2, Xdh and Aldox along altitudinal, latitudinal and/or climatic gradients. Tests using conditional allele frequencies showed that Est‐1 and Aldox clines could be accounted for by hitchhiking with inversions, whereas natural selection should be invoked to explain the clines observed for Est‐2 and Xdh.     

Linkage of Genes Controlling Morphological Traits with Isozyme Markers of Rye Chromosomes

Data on linkage of 12 rye genes controlling morphological traits (el, Vs, ln, w, np, ct2, Hs, Ddw, cb, mn, vi1, mp) with one or several isozyme markers of individual rye chromosomes (2R–7R) are presented. Linkage of the following gene pairs was established: chromosome 2R: Est3/5–el, el–-Glu, Sod2–el, Sod2–Vs; chromosome 3R: ln–Got4; chromosome 4R: w–Got1, np–Got1; chromosome 5R: Est4–ct2, Est6/9–ct2, ct2–Est2, ct2–Aco2, Est2–Hs, Aco2–Hs, Est2–Ddw, Aco2–Ddw; chromosome 6R:Lap2–cb, cb–Aco1, Est10–mn; chromosome 7R: Acph2/3–vi1, Got2–vi1, mp–Acph2/3. The reasons for mapping a very small number of genes in rye in spite of high intraspecific variability of this species are discussed. An approach is suggested to improve this situation by simultaneous identification and mapping of all diverse spontaneous mutations maintained in heterozygous state in various rye cultivars.     

Interval mapping of genes for quantitative resistance of maize to Setosphaeria turcica,cause of northern leaf blight,in a tropical environment

Quantitative trait loci (QTL) involved in the resistance of maize to Setosphaeria turcica, the causal agent of northern leaf blight, were located by interval mapping analysis of 121 F_2:3 lines derived from a cross between Mo17 (moderately resistant) and B52 (susceptible). A linkage map spanning 112 RFLP loci with 15 cM mean interval length was constructed, based on marker data recorded in a previous study. Field tests with artificial inoculation were conducted at three sites in tropical mid- to high-altitude regions of Kenya, East Africa. Host-plant response was measured in terms of incubation period, disease severity (five scoring dates), and the area under the disease progress curve (AUDPC). Heritability of all traits was high (around 0.75). QTL associated with the incubation period were located on chromosomes 2S and 8L. For disease severity and AUDPC, significant QTL were detected in the putative centromeric region of chromosome 1 and on 2S, 3L, 5S, 6L, 7L, 8L and 9S. On 2S the same marker interval which carried a gene enhancing latent period was also associated with reduced disease severity of juvenile plants. QTL on chromosomes 3L, 5S, 7L and 8L were significant across environments but all other QTL were affected by a large genotype x environment interaction. Partially dominant gene action for resistance as well as for susceptibility was prevailing. Single QTL explained 10 to 38% of the phenotypic variation of the traits. All but the QTL on chromosomes 1, 6 and 9 were contributed by the resistant parent Mo17. On chromosome 8L a QTL mapped to the same region as the major race-specific gene Ht2, supporting the hypothesis that some qualitative and quantitative resistance genes may be allelic.Abbreviations AUDPC area under the disease progress curve - CIMMYT International Maize and Wheat Improvement Center - KARI Kenya Agricultural Research Institute - NCLB northern corn leaf blight - QTL quantitative trait locus/loci     

Genetic analysis of disease resistance to all strains of BaYMV in a Chinese barley landrace, Mokusekko 3

 A Chinese landrace of barley, Mokusekko 3, is unique in being completely resistant against all strains of barley yellow mosaic virus (BaYMV). The present investigation revealed that the resistance of Mokusekko 3 is governed by two recessive genes. As one of the resistance genes was known to be tightly linked with alleles at the Est complex locus, consisting of the Est1, Est2 and Est4 loci for esterase isozymes, each of the resistance genes could be separated by means of marker-assisted selection using an isozyme allelic combination as a marker. One of the resistance genes, ym1, is linked to K (hooded lemma) and gl3 (glossy leaf 3) with recombination values of 25.3% and 9.7% respectively, and these three genes are located in the order K-gl3-ym1 on chromosome 4. Another newly designated resistance gene, ym5, is linked to alleles at the Est complex locus and cu2 (curly growth 2), with recombination values of 1.9% and 19.5% respectively, in the order cu2-Est-ym5 from proximal to distal on the long arm of chromosome 3. The complete resistance of Mokusekko 3 is caused by combining two resistance genes, ym1 and ym5. However, almost all the “resistant” cultivars derived from crosses with Mokusekko 3 are susceptible to the recently detected strain BaYMV-III in Japan, since they contain only one resistance gene, ym5. Marker-assisted selection to combine resistance genes into a cultivar is discussed for the breeding of stabilizing resistance to BaYMV. Received: 23 September 1996 / Accepted: 8 November 1996     

The genetics and expression of an esterase locus inAnopheles gambiae

The main polymorphic system of esterase isoenzymes in adults of the G3 laboratory strain ofAnopheles gambiae consists of two to five major bands of activity per individual. The bands are designated 5S, 5F, 13, 14, and 15. In genetic crosses, the genes which coded for the bands assorted as three codominant alleles, Est A, Est B, and Est C, at a single autosomal locus. Homozygotes for the Est C allele were significantly underrepresented among backcross progeny. The developmental pattern of esterase expression was examined. Esterase gene expression in embryos was first detectable between 2 and 12 hr after oviposition. The initiation or termination of expression of some of the bands corresponded to boundaries between developmental stages. Most of the esterase fractions were not specifically localized within the tissues tested, with the exception of a series of bands which were restricted largely to adult male testes.     

Mapping strategy for resistance genes in tomato based on RFLPs between cultivars: Cf9 (resistance to Cladosporium fulvum) on chromosome 1

Summary The contribution of introgressed regions derived from wild species to the genetic variation within the species of Lycopersicon esculentum was investigated by comparing the RFLP patterns of 2 introgression-free, obsolete cultivars (Moneymaker and Premier) and a modern cultivar (Sonatine) that carries at least 5 introgressed resistance genes. In this analysis 195 mapped nuclear markers were used in combination with 6 restriction enzymes. Among the 1170 probe-enzyme combinations tested, only 3 showed a polymorphism between the 2 introgression-free cultivars. On the other hand 24 probe-enzyme combinations were found to exhibit polymorphisms between Moneymaker and Sonatine. These represented ten polymorphic loci distributed among 5 linkage groups on chromosomes 1, 3, 4, 6, and 9.On the assumption that most of the polymorphic loci corresponded to introgressed chromosome segments of wild species carrying resistance genes, linkages between these loci and the component resistance genes were examined by RFLP analysis of pairs of near-isogenic lines differing only for one particular resistance gene, and a variety of commercial cultivars having different resistance gene compositions. Two of the polymorphic linkage groups could thus be ascribed to resistance genes whose map positions were already known: Cf2 on chromosome 6 and Tm2a on chromosome 9, whereas another marker, TG301 on chromosome 1, could be assigned to the Cladosporium fulvum resistance gene Cf9 with a hitherto disputable map position. By linkage analysis of a segregating F₂ population the genetic distance between the Cf9 gene and the marker TG301 was estimated at 5.5 ± 2.3 cM.     

Genetics of an esterase in Aedes aegypti

Zymograms of single individuals of Aedes aegypti were obtained by means of starch gel electrophoresis, using alpha-naphthyl acetate as substrate. Inbred lines gave consistently homogeneous patterns; earlier results from random-breeding laboratory strains had shown considerable variability. Six distinct bands were observed. The furthest moving band, designated Esterase 6, showed differential migration in two inbred lines. Reciprocal crosses between these lines gave F₁ progeny showing both bands. Backcrosses of F₁ to either parental line gave a 1:1 segregation. These results are consistent with the hypothesis that the two forms of Esterase 6 are controlled by a single pair of codominant alleles at a single gene locus (Est 6 ^a and Est 6 ^b). Linkage tests with marker genes have demonstrated that Est 6 is on linkage group 2, with the following alignment: spot-abdomen (9.0±1.0) yellow-larva (17.4±1.3) Est 6. Crosses with another inbred line demonstrated a third band with intermediate mobility, designated Est 6 ^c. An additional electrophoretic variant which seems to have a simple Mendelian basis was found in esterase band 1.This work was supported by NIH Research Grant No. A1-02753.     

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.