Esterase Isozyme Patterns Research Articles

Phylogenetic relationships in the family Resedaceae L.

A study was made on the phylogenetic relationships of species of the family Resedaceae, based on morphological features, chromosome meiotic behaviour, karyotype features, size and fertility of pollen grains, nucleotypic parameters, seed protein profiles and esterase isozyme patterns.

Isolation of pea nodule bacteroids utilizing silica sol (Percoll) density gradient centrifugation

Isolation of bacteroids from effective (Fix+) and ineffective (Fix−) pea nodules, inoculated withRhizobium leguminosarum K, were performed by a density gradient centrifugation method using silica sol (Percoll). Only one zone (ρ=1.064–1.072; n-zone) was recognized in the Fix+ nodule which contained typical Y-shaped bacteroids while two zones (n-zone and ρ=1.125–1.145; n'-zone) were obtained from the Fix− nodule. The cells in the n'-zone, which are long rods differed morphologically from free-living cells at any growth phase (ρ=1.108–1.125; f-zone and ρ=1.074–1.078; f'-zone), and differed from Y-shaped bacteroids by cell density. The esterase isozyme pattern of bacteroids in the n-and n'-zones also showed clear differences from that of f-and f'-zone of free-living cells.

Electrophoretic analysis of esterase isozymes in organophosphate-resistant mosquitoes ( Culex pipiens)

Esterase isozymes from individual mosquitoes, Culex pipiens, were analyzed by thin layer agar gel electrophoresis. Changes in esterase isozyme patterns during the developmental stages were studied. In most strains, only one band was found in eggs. During larval development many bands were gradually formed, and the most anodal bands were lost upon pupation. These larva-specific esterases may be controlled by three or more co-dominant alleles. In the adult, many bands were present that were probably controlled by at least seven loci. Est-2 may be controlled by co-dominant alleles; a recessive silent allele was also found. Esterase isozyme patterns of organophosphorus insecticide-resistant and susceptible strains were compared. Resistant strains had very active esterase bands but different patterns were found in malathion and temephos-resistant strains. The strong bands presumably hydrolyse the insecticides to which the strains are resistant, or their PO analogues. Their substrate specificity was studied with various esters.

The effects of freezing and thawing on lactate dehydrogenase and esterase isozyme patterns of the goldfish Carassius auratus, demonstrated by isoelectric focusing

AbstractCurrent literature suggests that the reliability of results obtained from electrophoretic studies using material which has been frozen is in doubt. Banding patterns of lactate dehydrogenase and non‐specific esterase produced by isoelectric focusing (IEF) of fresh tissues from the goldfish, Carassius auratus, are presented. Changes which occur in these patterns due to repeated freezing and thawing involve a reduction in both the intensity and the number of bands. Differences in the IEF profiles are extensive and consistent. Changes also occur in material which has been frozen for a ten‐week period and thawed once. Storage in liquid nitrogen, ‐180°C, as distinct from a conventional freezer at ‐20°C, produes no significant improvement. Interpretation of results from material that has been frozen must therefore be undertaken with extreme care, especially when sensitive separation techniques like IEF are used.

コバヤシミカンのキメラ性に関する研究-特に器官, 組織のアイソザイムパターンについて

The present experiment was undertaken to confirm the generally recognized assumption concerning the chimerism of Kobayashi-mikan. This cultivar is supposed to be a synthetic chimera which has arisen from the junction of satsuma mandarin (Citrus unshiu Marc.) scion and Natsudaidai (C. natsudaidai Hayata) stock. Peroxidase and esterase isozymes of foliage and fruit parts as well as immature leaves and rootlets of young seedlings were analyzed by starch gel electrophoresis. The results obtained from a zymographic comparison of Kobayashi-mikan, satsuma mandarin and Natsudaidai are summarized as follows.1. In the case of samples taken in July, peroxidase isozyme patterns of new and old leaves as well as albedo of Kobayashi-mikan distinctly resembled those of Natsudaidai. The pattern of juice sac of Kobayashi-mikan considerably resembled that of satsuma mandarin. Esterase isozyme patterns of flavedo and albedo of Kobayashi-mikan were completely the same as those of Natsudaidai.2. In the case of samples taken in October, peroxidase isozyme patterns of old leaves as well as flavedo of Kobayashi-mikan were nearly the same as those of Natsudaidai. The patterns of albedo and new leaves of Kobayashi-mikan resembled those of Natsudaidai. Esterase isozyme patterns of old leaves and seeds of Kobayashi -mikan were equal to those of Natsudaidai. The patterns of flavedo, alvedo and new leaves of Kobayashi-mikan were rather similar to those of Natsudaidai.3. Peroxidase and esterase isozyme patterns of immature leaves and rootlets of young seedlings grown from seeds of Kobayashi-mikan were similar to those of young seedlings grown from seeds of Natsudaidai. From these results, the hypothesis that Kobayashi-mikan is a graft-induced periclinal chimera between satsuma mandarin and Natsudaidai has been further confirmed.

数種殺虫剤に対するコガタイエカ幼虫の感受性について

A colony of the larvae of Culex tritaeniorhynchus which was obtained in Toyama Prefecture by laboratory rearing of eggs deposited by wild caught females collected at a pig house surrounded by rice paddies, was shown to be highly resistant to organophosphorus insecticides. The values of LC-50 against malathion, fenitrothion, fenthion, cidial, BPMC, and p, p' DDT were 34,13.5,9.6,8.8,2.16,and 0.078ppm, respectively. It was presumed that repeated applications of organophosphorus insecticides against rice plant pests have resulted in the development of such resistances in this mosquito species, which mainly breeds in rice paddies and is known to be acting as the principal vector of Japanese encephalitis. From analysis of dosage-mortality regression lines of the larvae against malathion and fenitrothion, it was shown that the colony composed of a mixture of different populations with various susceptibility levels against the insecticides. In the study of esterase isozyme patterns of individual female adult mosquitoes reared from larvae that survived after exposure to 5ppm fenitrothion and 10ppm malathion, it was also shown that they were composed of heterologous populations with different active band patterns.

B-Chromosomes and E-1 isozyme activity in mosaic bulbs of Scilla autumnalis (Liliaceae)

The electrophoretic patterns of esterase (E-1), alcohol dehydrogenase (ADH), and glutamate oxaloacetate transaminase (GOT) isozymes were studied in two Spanish populations of the lily Scilla autumnalis with B-chromosome carrying individuals. The E-1 isozyme activity appears only in those individuals with B-chromosomes. None of the bulbs free of B's show it. Five bulbs, mosaic for B-content, were identified. Electrophoretic analysis shows that these bulbs are characterised by mosaicism for E-1 isozyme activity. An analysis of individual roots by both electrophoretic and cytological methods shows that tissue mosaicism for B-content correlates with tissue mosaicism for E-1 isozyme activity. The electrophoretic analysis of different roots from bulbs heterozygous for the Est-1 locus indicates that the structural gene for E-1 is not located on the B-chromosome itself. Rather there is a derepressor effect of Bs on E-1 isozyme activity. Since ADH and GOT patterns are unaffected by the presence of B-chromosomes it is clear that they do not exhibit a generalised derepressor effect.

Variation among British isolates of Armillaria mellea

Twenty-one British isolates of Armillaria mellea could be assigned to one of three groups based on rhizomorph growth habit in soil. Each group of isolates showed differences in pathogenicity to conifer seedlings and in esterase isozyme patterns. The results suggest that the groups are three members of the A. mellea complex.

The genetic relationship between the human foetal acetylesterase ESA7 and the adult acetylesterase ESA5.

(1) There are very few clear examples among human enzymes of foetal isozymes which are the products of foetal specific gene loci. Earlier studies had pointed to the foetal brain esterase ESA7 as a probable example. (2) Detailed biochemical investigation of partially purified human adult brain ESA5 and the foetal esterase ESA7 has revealed a close resemblance in the biochemical properties of these two isozymes. In addition to similarities in substrate specificity and inhibition sensitivity the two esterases have the same molecular size (c. 57,000), are both relatively unstable at 37 degrees C and show decreased anodal electrophoretic mobility after storage at 20 degrees C. Furthermore there was suggestive evidence that ESA7 and ESA5 may be interconvertible. (3) A variant esterase isozyme pattern, which shows unusual features of both ESA7 and ESA5, was found in a survey of 120 foetal brains. This variant pattern is consistent with a monomeric structure for both esterases and points strongly to a common genetic determination.

Unterscheidung von Gerstenartbastarden mit Hilfe von Isoenzymmustern

Soluble proteins and isozy mes have been studied with the species Hordeum bulbosum (L.) and H. vulgare (L.) as well as with their hybrid plants and from the crossings of H. vulgare with the hybrids, characterized by typical bulbosum morphology (“modified barley„). The results showed tissue-specific patterns of esterase isozymes in caryopses, coleoptiles and leaves of different age, especially when slow and mediate fast migrating bands are regarded. Species-specific patterns were visible in the fast migrating band group. Three marked bands of esterases as well as three peroxidase bands in the Rf-range between 0.65 and 0.75 have been shown to be present in all tissues of Hordeum bulbosum. H. vulgare indicates only one esterase band at Rf 0.73. Hybrid plants and “modified barley„ reflected the typical patterns of their parents in a different way. Hybrids of the F1 (BBVV) genome developed the vulgare patterns as well as the typical bulbosum bands. In “modified barley” plants (VVB) bulbosum patterns have been observed in few cases only.

Origin and differentiation of Aegilops triuncialis L. as determined by esterase isozyme analysis

Banding patterns of esterase isozymes in Aegilops triuncialis (2n = 28, genome formula C(u)C(u)CC) and its putative parental species, Ae. umbellulata (2n = 14, C(u)C(u)) and Ae. caudata (2n = 14, CC), were studied by the gel isoelectric focusing method using pH 6-8 carrier ampholite. Zymogram phenotypes of both parents were quite uniform. Seven zymogram phenotypes (designated as phenotypes 1 to 7) were found among the 260 strains of Ae. triuncialis examined. Of these phenotypes, phenotype 1 was identical to the zymogram phenotype produced by the ancestral species, Ae. umbellulata, and bands considered to have been derived from Ae. caudata were absent in this phenotype. Phenotype 3 had all bands of both parents. The other phenotypes differed greatly from phenotype 3. Therefore, phenotype 3 was considered to be most primitive of the 7 types, and the Ae. triuncialis strains which showed phenotype 3 to be the most primitive of the strains examined. If Ae. triuncialis originated as a hybrid between Ae. umbellulata and Ae. caudata, the zymogram phenotype must have been phenotype 3, in which the isozymes of both parental species are present. Whether the phenotypes other than type 3 were due to introgressive hybridization could not be verified, but they were considered in this article to be a consequence of a rearrangement of chromosomes.

Culture of sheep X mouse hybridoma cells in vitro.

Hybridomas were made between NS1/1-Ag4-1 mouse myeloma cells and spleen and lymph node cells from a sheep immunized with sheep red cells (RBC). The hybrid colonies grew well in culture but there was a substantial loss of sheep chromosomes. No hemolytic or agglutinating antibodies were detected in the culture supernatants after the 17th day following fusion, but immunofluorescence tests indicated that a few of the cells may have been expressing sheep IgG. Cytogenetic comparison of cells grown with and without HAT medium provided evidence that the enzyme HGPRT is located on the X chromosome of sheep as it is in man and mouse. Hybridoma isozyme patterns of esterase, G6PD, 6PGD, NP, LDH and SOD tested between the 63rd and 71st day of culture were like those of NS1; NP and LDH also showed zones that probably came from the sheep component.

D genome doners for Aegilops cylindrica (CCDD) and Triticum aestivum (AABBDD) deduced from esterase isozyme analysis

Putative D genome donors for Aegilops cylindrica (2n = 28, CCDD) and Triticum aestivum (2n = 42, AABBDD) were studied with the isoelectric focusing patterns of esterase isozymes. 103 strains of Ae. cylindrica were uniform in their isozyme pattern. 30 strains of the putative parent, Ae. caudata, showed no zymogram variation, whereas the other parent, Ae. squarrosa, comprised 3 phenotypes. Natural Ae. cylindrica had an isozyme pattern which corresponded to a mixture of esterases from Ae. caudata and type 3 Ae. squarrosa. Therefore, it is concluded that the D genome donor of Ae. cylindrica is derived from type 3 Ae. squarrosa. These results suggest that Ae. cylindrica originated with a single amphiploidy event, and the C and D genomes have remained remarkably constant regarding esterase isozyme composition.On the other hand, T. aestivum comprised three zymogram phenotypes. These phenotypes contain bands which can be ascribed to the D genome of type 2 Ae. squarrosa. These results suggest that the D genome of Ae. cylindrica differs from that of T. aestivum. Evolution of the AB and D genomes of T. aestivum is indicated by the zymogram polymorphism. The origin of Ae. cylindrica is possibly more recent than that of T. aestivum.

The B chromosome system of Scilla autumnalis (Liliaceae): effects at the isozyme level

Supernumerary (B) chromosomes have been studied in a Spanish population of Scilla autumnalis L. (Liliaceae). Out of the 140 individuals analysed, seven had 2n=14+1B, one 2n=14+2B, one 2n=14+3B and one 2n=14+9B. An analysis of esterase isozyme patterns shows that all 130 individuals with a standard karyotype (2n=14) have two esterase loci, Est-2 and Est-3, whereas all 10 individuals with Bs have three, Est-1, Est-2 and Est-3, irrespective of the precise number of Bs present. The role that the Bs may have played in the appearance of this new locus (Est-1) is discussed in relation to their possible origin.

THE RELATION BETWEEN ISOZYMES AND PLOIDY LEVEL. ITS APPLICATION TO BIOGEOGRAPHICAL STUDIES OF MUSCARI ATLANTICUM (LILIACEAE)

SummaryIn this paper a correlation between polyploid level and esterase isozyme pattern in individuals from different Spanish populations of Muscari atlanticum has been established. We have taken advantage of this correlation to make a study of the distribution of the reported polyploid levels in 1,017 individuals from thirteen natural populations. The interest in and application of this method for bio geographical analysis are discussed.

Isozyme variations in Aegilops and Triticum. IV. The origin of the common wheats revealed from the study on esterase isozymes in synthesized hexaploid wheats.

Esterase isozymes in two kinds of synthesized hexaploid wheat and its parental strains were analyzed by the isoelectric focusing method using pH 6-8 carrier ampholite. In present-day hexaploid wheat, three similar patterns of esterase isozymes were found. The large number of them had nine esterase bands, which are controlled by the genes on homoeologous chromosomes 3. On the other hand, the parental species of the hexaploid, also, showed inter- as well as intraspecific variations. Seven different zymograms were observed in the tetraploid wheats, among which type 4 distributed most widely in tetraploid species. Moreover, most bands of this zymogram were commonly found in the zymograms of common wheat. Therefore, the donor of AB genome to common wheat seems to have had this zymogram. The Tetra-Canthatch (AABB) which the D genome was removed from Canthatch (AABBDD) had the same zymogram, the type 4, of present-day tetraploid species. The type 4 of the esterase zymogram was, also, posturated as AB genome donor from this evidence. Another parent, Ae. squarrosa, involved three different types of zymograms. Artificially synthesized hexaploid wheat as the hybrids between type 4 tetraploid and three different types of Ae. Squarrosa were produced. Combinations between type 4 tetraploid and type 2 Ae. Squarrosa showed the same zymogram as compared with present-day hexaploid wheat. Therefore, type 2 Ae. Squarrosa was expected as D genome donor. Concerning the geographical distribution of type 2 Ae. Squarrosa, this type was found in the Elbruz Mountains along the coast of the Caspian Sea and Transcaucasus. Yet type 2 was not found in both countries, Pakistan or Afghanistan. From this evidence, birthplace of hexaploid wheat seemed to originate in the region from the south west Caspian belt to Transcaucasus.

Molecular aspects of the environmental induction of heritable changes in flax

The conditions for the induction and stabilisation of environmentally induced changes in flax, and the characterisation of these changes for six characters have been reviewed. A model for the induction and expression of these changes has been proposed. The environmental conditions for the successful induction of heritable changes comprise at least two components; a specific inducing component, for example a particular fertiliser treatment, which is required to induce the change, and a general inducing component, providing for vigorous healthy growth, which is necessary for the induced changes to be inherited. Stable induced changes require appropriate environments for their maintenance, for example heated greenhouses for the first 5 weeks of seedling growth and adequate pot size. Under certain growth conditions the induced changes can themselves be modified. DNA differences induced in the different genotrophs can occur at a number of sites within the genome, including the ribosomal RNA cistrons. Environmental induction causes changes in the isozyme band patterns of peroxidase, acid phosphatase and esterase isozymes. The genotrophs behave as distinct genetic types in crosses, with a complex pattern of inheritance of the hairy septa character. The association between the different induced characters is examined. All the genotrophs can be distinguished from each other on at least one of the six characters. A model for the induction of the DNA changes, involving a type of IS-element has been proposed. The expression of the induced changes, in terms of altered numbers of copies of particular sequences at specific sites within the genome, has been considered.

Effects of acute and chronic dieldrin administration on liver and plasma esterases of the rat

The administration of a single oral dose of dieldrin, 40 mg/kg, or the feeding of a diet containing 50 ppm of dieldrin for up to 8 weeks to male and female rats produced changes in plasma and liver esterases that were dependent on the type of esterase, the duration of exposure, and the sex of the animal. Two hours after oral administration of dieldrin, plasma carboxylesterase activity declined in males but not in females; hepatic microsomal carboxylesterase activity was also slightly lower in dieldrin-treated males, but not females. Forty-eight and 120 hr after dieldrin administration, hepatic microsomal carboxylesterase activity was elevated in rats of both sexes. After 8 weeks of dieldrin feeding (50 ppm), both plasma and microsomal carboxylesterase activities were significantly elevated in males and females. In most instances, elevations in the activities of hepatic microsomal esterases were due to dieldrin-induced increases in microsomal protein content and not to increased esterase specific activity. Electrophoresis of solubilized microsomal membranes from control and dieldrin-treated animals revealed no sex differences or major dieldrin-induced alterations on esterase isozyme patterns. Plasma cholinesterase activity was significantly reduced in females fed 50 ppm of dieldrin in the diet for 8 weeks. Since the level of activity of this enzyme is regulated in part by estrogen, depression of cholinesterase activity may result from an increased rate of steroid metabolism in dieldrin-treated female rats. Plasma cholinesterase activity of male rats was unaffected by either acute or chronic dieldrin administration.

The effects of freeze-preservation on some pollen enzymes. 1. Freeze-thaw stresses

Summary Isozyme patterns of nonspecific esterase, leucine aminopeptidase, acid and alkaline phosphatases and peroxidase were studied to aid explanation of the nature of freeze-thaw injury to pollen. Fresh samples of Lilium longiflorum L. and Zea mays L. pollen were frozen at rates between 200 and 0.5°C/min and thawed at rates ranging between 218 and 2.5°C/min. Soluble enzymes were extracted from pollen and analyzed by disc electrophoresis on 7.5% polyacrylamide gels. The gels were stained for enzymes using substrate specific stains and the stained gels were scored for isozyme patterns, migration rate and relative activity. The data suggest that isozymic alterations of nonspecific esterase, acid and alkaline phosphatases, peroxidase and to a lesser extent leucine aminopeptidase are associated with freezing and thawing treatments. Generally, a decrease in enzyme stainability could be observed with freezing rates below 180 or 150°C/min. A diffused staining pattern of some enzymes may be a reflection of conformational changes. Increased acid phosphatase activity of frozen-thawed samples may indicate lysosomal damage. The isozymes of leucine aminopeptidase remained fairly stable in most treatments.

Esterase isozyme patterns in Glycine max exposed to gamma radiation

The esterase isozyme patterns for irradiated and non-irradiated soybean seed and seedlings were determined by starch gel electrophoresis. Patterns were determined for cotyledons, hypocotyl, epicotyl, first pair of leaves, and roots. Two different esterase systems, E1 and E2, were detected by using appropriate substrates. E1 produces three anodic bands with both α- and β-naphthyl acetate. E2 acts only on α-naphthyl acetate producing three cathodic bands. Radiation did not change the patterns. Results indicate that esterase isozymes are more abundant in the aerial than in the underground portion of developing seedlings.