Esterase Isoenzyme Patterns Research Articles

Biochemical changes of the cereal cyst nematode, Heterodera avenae, at low temperatures

Cereal cyst nematode (Heterodera avenae) diapause is induced by high temperatures and is broken by low temperatures. In this study, metabolic responses were monitored in diapause and non-diapause H. avenae during exposure to 4°C for 10 weeks. The results showed that there was no difference in total carbohydrate content. The content of glycogen and glycerol at 0 week was relatively high but decreased with increased storage time at 4°C. The content of trehalose of the nematode at 10 weeks was significant lower than that at 5 weeks at 4°C. Protein content increased significantly after incubation for 5 and 10 weeks. Esterase and trehalase activity increased with the increasing period at 4°C and showed a significant difference between treatments for esterase activity but there was no significant difference between 5 and 10 weeks for trehalase activity. The SDS-PAGE pattern indicated that a 15.5 kDa protein was absent at 10 weeks and present at 0 and 5 weeks. Esterase isoenzyme patterns of H. avenae showed that at 10 weeks there were four bands: EST 0.21, EST 0.24, EST 0.30 and EST 0.34 (Rf values). EST 0.24 was the common band in the three treatments. Biochemical tests were conducted to correlate with hatching experiments using the same treatments. 2-DE patterns of H. avenae showed that diapause and non-diapause nematodes had 409 and 412 protein spots, respectively, and 19 protein spots were unique: 11 distinct proteins in non-diapause and eight distinct proteins in diapause. This information could be helpful in understanding the diapause mechanism of the cereal cyst nematode.

Physiological, biochemical, and molecular characterization of a new female sterile mutant in turnip

A female sterile plant with aborted pistils was previously obtained in an advanced self-pollinated turnip inbred line. Cross progeny with normal wild-type (WT) plants successfully characterized the mechanism of pistil abortion, and the mutant was designated ‘tpa’ (turnip pistil abortion). Further morphological investigations showed that the tpa mutant included two phenotypes, pistil abortion partially (PAP) and pistil abortion completely (PAC), and our recent result showed that the tpa trait was controlled by two recessive genes (Wu et al. 2011). In this study, comparisons of the floral organs indicated that corolla size and petal, sepal, filament, and anther length measurements did not significantly differ between the tpa and WT plants. Subsequently, the activities of enzymes (catalase, ascorbate peroxidase, and guaiacol peroxidase) related to the metabolism of reactive oxygen, the O2− production rate, as well as the contents of hydrogen peroxide and malondialdehyde were tested and found to be generally higher in tpa floral organs than in the WT. The reverse was true for the flower stems, however. Quantitative differences in peroxidase, cytochrome oxidase, and esterase isoenzyme patterns were also detected among these different organs. These findings suggest that pistil abortion is probably correlated with the unbalanced homeostasis of ROS antioxidant enzymes. Moreover, a specific band was recognized exclusively in the tpa plants using random amplified polymorphic DNA analysis. Sequencing and blasting results indicated that the 878-bp DNA fragment was located in the D1/D2/D3 region, near the 5′ end of the 26S rDNA. This study valuably adds to the current knowledge of and will help further elucidate the biological mechanism of female sterility induction.

<strong>Oribatid mites of China: a review of progress, with a checklist</strong>

The present paper gives a review of the taxonomic study on the suborder Oribatida (the cohort Astigmata excepted) in China including Hong Kong and Taiwan, with a checklist of 599 species and subspecies in 275 genera, representing 97 families. One junior synonymes is proposed, i.e., Peloribates praeoccupatus Subías, 2004 is regarded as a junior synonym of Peloribates tsengi Wen, Wang & Chen, 2003. The taxonomic study of oribatid mites in China was started by A. P. Jacot in 1922, and was very productive during the period from 1980 to the end of last century with the efforts of several Chinese and foreign oribatologists. In addition to taxonomic research, publications on biodiversity, seasonal dynamics, biological monitoring for soil pollution, and esterase isoenzyme patterns of oribatid mites have been published by Chinese scholars.

Esterase isoenzymes and insecticide resistance in Frankliniella occidentalis populations from the south‐east region of Spain

Frankliniella occidentalis (Pergande) is among the most important crop pests in the south-east region of Spain; its increasing resistance to insecticides constitutes a serious problem, and understanding the mechanisms involved is therefore of great interest. To this end, F. occidentalis populations, collected from the field at different locations in south-east Spain, were studied in terms of total esterase activity and esterase isoenzyme pattern. Individual thrips extracts were analysed by native polyacrylamide gel electrophoresis (PAGE) and stained for esterase activity with the model substrate alpha-naphthyl acetate. Significant correlations were found between resistance to the insecticides acrinathrin and methiocarb and the presence of a group of three intensely stained bands, named Triplet A. For each individual thrips extract, total esterase activity towards the substrates alpha-naphthyl acetate and alpha-naphthyl butyrate was also measured in a microplate reader. Insects possessing Triplet A showed a significantly higher alpha-naphthyl acetate specific activity and alpha-naphthyl acetate/alpha-naphthyl butyrate activity ratio. This observation allowed a reliable classification of susceptible or resistant insects either by PAGE analysis or by total esterase activity determination. The PAGE and microplate assays described can be used as a monitoring technique for detecting acrinathrin- and methiocarb-resistant individuals among F. occidentalis field populations.

First record of infection of Papaya trees with root-knot nematode (Meloidogyne javanica) in Australia

The root-knot nematodeMeloidogyne javanica was identified in papaya roots in Western Australia. Species identification was based on esterase isoenzyme patterns and PCR diagnostics.

First record of infection of Papaya trees with root-knot nematode (Meloidogyne javanica) in Australia

The root-knot nematode Meloidogyne javanica was identified in papaya roots in Western Australia. Species identification was based on esterase isoenzyme patterns and PCR diagnostics.

Superoxide dismutase activity as a function of culture aging of B-16 mouse melanoma cells

The C3 clone of B-16 mouse melanoma was cultured for 1, 6 and 9 days and analyzed. The changes which are not directly linked to melanogenesis in the B-16 / C3 cultures during their maturation were characterized. Early (1 day) confluent (6 days) and old (9 days) cell cultures are distinguished by their leucine aminopeptidase (LAP) and ?-naphthyl acetate esterase (ANAE) isoenzyme patterns. Both quantitative and qualitative changes in LAP and ANAE isoenzyme can be observed during culture maturation. There is an increase in the activity of the enzyme copper, zinc-containing superoxide-dismutase (CuZn SOD). The increaase in the CuZn SOD enzyme activity might be related to B-16/C3 cell melanogenesis and / or to differentiation.

Isoenzymes of peroxidase and esterase related to morphogenesis in Mammillaria gracillisPfeiff. tissue culture

In vitro propagated plants of the cactus Mammillaria gracillis Pfeiff. (Cactaceae) spontaneously produced callus. The habituated callus regenerated normal and hyperhydric shoots without the addition of grown regulators. Tumours were obtained by infecting cactus explants with Agrobacterium tumefaciens; the wild strain B6S3 (tumour TW) or with the rooty mutant GV3101 (tumour TR). Both tumour lines grew vigorously, never expressing any morphogenic potential. In this study, cactus shoots, callus, normal and hyperhydric regenerants and TW and TR tumours were compared with regard to peroxidase (EC 1.11.1.7) and esterase activity, and isoenzyme patterns. Guaiacol peroxidase activity was the lowest in the cactus shoots and in the normal regenerants. Callus, hyperhydric regenerants and tumours had peroxidase activity of 6 to 7 times higher. Esterase activity was measured with 1- and 2-naphthylacetate as broad-spectrum substrates. The highest esterase activity was determined in tumours with both substrates. All tissues, except the TR tumour, had higher esterase activity for 2-compared to 1-naphtylacetate. Peroxidase and esterase isoenzyme patterns were not completely identical among the investigated tissues.

Enhanced indole alkaloid production in suspension compact callus clusters of Catharanthus roseus: impacts of plant growth regulators and sucrose

A special culture system, compact callus clusters, was developed from Catharanthus roseus stem explants in a modified Murashige and Skoog (MS) liquid medium containing 5.37 µM α-naphthaleneacetic acid and 4.65 µM kinetin. Morphological and anatomical studies showed that the globular compact callus cluster cultures consisted of many cohesive callus aggregates displaying some level of cellular/tissue differentiation, which was also in agreement with the results from peroxidase and esterase isoenzyme pattern analysis. The compact callus cluster cultures could synthesise about 2-fold more indole alkaloids than the dispersed cell cultures, and this was postulated to be associated with their differential status. Plant growth regulators and sucrose concentration, as well as shaking speed significantly affected properties of the compact callus clusters. In detail, 2,4-dichlorophenoxyacetic acid destroyed the compact structure and reduced alkaloid production of the compact callus cluster cultures; but a high concentration of cytokinins was necessary to maintain the compact structure and high alkaloid production of the special cultures. The optimum sucrose (5–6%) gave the greatest alkaloid and biomass production, as well as the greatest degree of compaction of the compact callus clusters.

Somatic hybrids between Brassica juncea (L). Czern. and Diplotaxis harra (Forsk.) Boiss and the generation of backcross progenies

An attempt to transfer genes from droughttolerant Diplotaxis harra, a wild relative of Brassica species, to an elite oil-yielding cultivar, B-85, of mustard (Brassica juncea) was made through protoplast fusion, as the two plant systems are sexually incompatible. By following the standard protocol for PEG-mediated protoplast fusion followed by high pH, high Ca(++), DMSO treatment and appropriate cell-culture technique, 16 presumptive somatic hybrid plants could be regenerated. Chromosomal analysis of four such somatic hybrids revealed that three of them were asymmetric. Analysis of morphological characters, meiotic chromosomes, and esterase isoenzyme pattern revealed that all the somatic hybrids were different from each other. Furthermore four chromosomes of each genome could undergo homoeologous pairing at meiosis indicating the possibilities for genetic recombination and chromosomal rearrangements. Irregular distribution of chromosomes at anaphase-II at meiosis has been a consistent feature of these plants. Eventually, pollen of all the somatic hybrids showed complete infertility preventing the recovery of any selfed seed. Nevertheless, ovule fertility of one somatic hybrid was not totally impaired as it had set some seeds upon backcrossing with the B. juncea parent. The esterase isoenzyme banding pattern of 24 individual progeny plants of this backcross provided evidence for their recombinant nature. It was thus confirmed that a transfer of genetic traits from Diplotaxis harra to B. juncea had indeed taken place. Furthermore, it was conceptualised that a transfer of alien genes through the protoplast-fusion technique is primarily possible in situations where meiotic pairing of the chromosomes of the two participating genomes generates recombinant gametocytes which can pass through subsequent filial generations.

First Report of B‐Biotype Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Australia

ABSTRACTB‐type Bemisia tabaci was detected in Australia for the first time in October 1994. the insects were identified by esterase isoenzyme patterns. the initial identifications were made in B. tabaci collected from a rockmelon crop in Darwin (N.T.) and in a Tamworth (N.S.W.) plant nursery.

Esterase Isoenzyme Profiles in Acute and Chronic Leukemias

Using isoelectric focusing (IEF) a number of carboxylic esterase isoenzymes (EC 3.1.1.1) with isoelectric points between pH 4.5-8.0 can be separated. One particular isoenzyme with an isoelectric point at about pH 6.0, the Mono-band, can be selectively and completely inhibited by sodium fluoride; this isoenzyme comprises a number of closely related subcomponents and may appear in more than one band on the gel. We analyzed the expression of typical esterase isoenzyme patterns in cells from a large panel of leukemias which were tested under identical conditions by IEF on horizontal thin-layer polyacrylamide gels with an ampholyte of pH 2-11. The 442 cases of acute and chronic myeloid and lymphoid leukemia (AML/AMMoL, CML/CMML, ALL, CLL) were classified according to clinical, morpho-cytochemical and immunophenotyping criteria. While bands between pH 4.5-5.5 appeared not to be specific for lineage or stage of differentiation, isoenzymes between pH 6.6-7.7 provided information on the type of leukemia involved. Seven typical isoenzyme patterns termed Mono1/Mono2 (fo monocyte-associated), My1/My2 (myeloid), Lym1/Lym2 (lymphoid) and Und (undifferentiated) could be discerned. Lym and Und patterns are characterized by fewer bands with a weaker staining intensity than Mono and My patterns. Nearly all cases of lymphoid leukemias (acute and chronic) expressed only Lym or Und esterase isoenzyme patterns, but no Mono or My patterns. Cases of acute or chronic myeloid and (myelo)monocytic leukemia showed strong isoenzyme staining displaying predominantly Mono or My isoenzyme patterns. The isoenzyme patterns found in CML in lymphoid or myeloid blast crisis corresponded to those seen in the respective acute leukemias, ALL or AML. The Mono-band was found in most cases of leukemias with monocytic elements (AMMoL 80%, CML 44%, CMML 100%), in the occasional case of CML-myeloid blast crisis or AML, but in none of the cases of ALL or CLL. This isoenzyme is a distinctive, specific marker for leukemias of monocytic origin and is of discriminatory value for the differentiation of monocytic from non-monocytic leukemia variants. Esterase isoenzyme profiles can give additional evidence on the origin and stage of differentiation of leukemic cells.

Protein complex and esterase isoenzyme patterns ofAllium sativum L. cultivars and clones-regenerants

Protein complex patterns of cloves and esterase isoenzyme patterns of apical buds of cloves were studied with Czechoslovak virus-free cultivars ofAllium sativum L. and the wild speciesA. longicuspis Regel, Similarly, four clones-regenerants obtained using explant culture techniques from A.sativum L. cv. Bzenecký paličák (two somaclones and two clones derived from plants regenerated from meristem cultures treatedin vitro with colchicine) differing in their ploidy, morphology, and yields were studied. Immunophoreograms of protein complexes of theA. sativum L. cultivars under investigation differed from one another in the number and mobility of protein fractions in both the cathodic and anodic regions and thus these cultivars can be distinguished. On the basis of esterase isoenzyme patterns, the Czechoslovak cultivars of A. sativum L. can be arranged into three groups - bolting winter cultivars with broad leaves, non-bolting winter cultivars with broad leaves, and non-bolting spring cultivars with narrow leaves. All the clones-regenerants showed the same protein complex and esterase isoenzyme patterns as their original cultivar.A. longicuspis Regel markedly differed in its protein complex and esterase isoenzyme patterns from all the other genotypes studied.Received May 17, 1989: accepted January 5,1990

Isoenzyme Electrofocusing as a Biochemical Marker System of Embryogenesis and Organogenesis in Callus Tissues of Hordeum vulgare L.

Summary Esterase, peroxidase, and acid phosphatase isoenzyme patterns are different in mature and in immature barley tissue extracts. A clear distinction can also be made between fully developed plant organs and different stages of embryo development. Each isoenzyme system, however, has some particular advantages for one of these applications. We therefore propose the combined use of all three isoenzyme profiles as a rapid and sensitive marker system capable of indicating differentiation trends in barley callus tissues, even before visual morphological changes take place.

Esterases in inbred strains of mice with differential cholesterolemic responses to a high-cholesterol diet

Specific esterase isoenzyme patterns in plasma may be associated with responsiveness of serum cholesterol to dietary cholesterol. In rabbits and rats the presence and absence of a high-mobility, anodal esterase band on electrophoresis have been shown to be associated with hypo- and hyperresponsiveness, respectively. We fed for 28 days male mice of 7 inbred strains either a low-cholesterol, commercial diet or a diet containing 2% (w/w) cholesterol, 0.5% cholic acid and 5% olive oil. Feeding the high-cholesterol diet revealed marked inter-strain differences in the responses of plasma and liver cholesterol; the increases ranged from 21 to 129% and from 10 to 80-fold, respectively. There was no association between esterase isoenzyme patterns in plasma and the sensitivity to the high-cholesterol diet. The mean baseline plasma total esterase activity tended to be positively associated with the absolute response of plasma cholesterol to the high-cholesterol diet ( r = 0.56; n = 7), but the positive relationship between the baseline concentration of the ES-1 component in plasma and the cholesterolemic response was stronger ( r = 0.84; n = 7; P < 0.05). The high-cholesterol diet caused a significant increase in plasma total esterase activities in 6 out of the 7 strains. Evidence is presented that the increase in plasma total esterase activity, which was associated with an increase in the activity and concentration of the so-called ES-2 isoenzyme, is the result of an enhanced release of esterases from the intestine, rather than from the liver. A significant, positive correlation was found between the baseline intestinal esterase activity and the cholesterolemic response after cholesterol feeding ( r = 0.83; n = 7; P < 0.05).

Conversion of acute undifferentiated leukemia phenotypes: analysis of clonal development

The cellular origin of acute undifferentiated leukemia (AUL) is still a matter of controversy. We report on two cases in which the diagnosis of AUL was established according to restricted criteria. Blast cells of both patients showed phenotypic conversion during the course of disease. In one case, within 24 days from starting treatment, the leukemic phenotype changed from AUL to acute myelomonocytic leukemia (FAB Ll, TdT+ to FAB M4, TdT-). The initial phenotype of this acute leukemia was characterized by the co-expression of both B-lymphoid and myeloid markers on the same cell. Moreover, analysis of esterase isoenzyme pattern showed the whole spectrum of isoenzymes typically seen in myelomonocytic leukemias already at diagnosis, yet blast cells additionally contained all three isoenzymes of β-hexosaminidase typically seen in AUL. However, examination of immunoglobulin (Ig) heavy chain gene rearrangement initially and after conversion revealed an identical monoclonal configuration of Ig heavy chain sequences in both samples. The second AUL patient relapsed after allogeneic bone marrow transplantation with common ALL-antigen (CALLA) positive acute leukemia. Subsequent Southern blot analysis showed a novel rearranged Ig fragment compared to the analysis before transplantation indicating that the leukemic clones prior to and after transplantation were not identical. No chromosomal abnormalities were observed in both cases. These data (1) support the view that AUL cells originate from a pluripotent stem cell that is capable to differentiate in the myelomonocytic lineage (patient 1), and (2) confirm the value of Ig gene analysis as marker for cellular clonality.

Identification of Kentucky Bluegrass Cultivars with Esterase and Phosphoglucomutase Isoenzyme Markers1

Starch gel electrophoresis was used to further extend the identification of Kentucky bluegrass cultivars. A total of 24 cultivars were examined for isoenzyme variation in esterase (EST), phosphoglucomutase (PGM), phosphoglucoisomerase (PGI), and glutamate‐oxaloacetate transaminase (GOT) using starch gel‐electrophoresis. Esterase and phosphoglucomutase patterns were varible among the 24 cultivars. ‘Baron’, ‘Fylking’, ‘Merion’, and ‘Newport’ were examined for isoenzyme pattern differences between seed and seedling materials. Differences of esterase isoenzyme patterns were found between the two kinds of tissues. Seeds of ‘Adelphi’ and ‘Glade’ harvested from different fields and years were examined for isoenzyme consistency and within seed lot variation. Variation within seed lots differed from seed lot to seed lot. If seedlings from a seed lot were mixed, a constant pattern of both EST and PGM isoenzyme systems was obtained for different seed lots of the same cultivar. Therefore the process of mixing seedlings allows the finger‐printing characterization for Kentucky bluegrass cultivars even if there is occasional segregation from sexual reproduction.

Esterase isoenzyme variation in the genus Saprolegnia, with particular reference to the fish-pathogenic S. diclina-parasitica complex.

Esterase isoenzyme patterns determined by slab gel electrophoresis were compared for nearly 60 Saprolegnia isolates, particularly from the S. diclina-parasitica complex. Consistent differences were found between S. diclina and S. parasitica isolates, with the latter being characterized by having between one and five very fast moving esterase bands that were generally absent from the former. A range of asexual (unidentifiable) isolates, taken from the vicinity of fish hatcheries or from fish lesions, were also examined and their esterase isoenzymes shown to be similar to those of S. parasitica, although usually showing fewer bands. The use of esterase isoenzymes for screening potential fish pathogenic isolates of Saprolegnia is briefly compared with results obtained using oogonium morphology or cyst ornamentation, and the relative merits of each method are discussed. It is proposed that, on the basis of their distinctive cyst coat ornamentation (bundles of long 'boathooks') and esterase isoenzymes, fish lesion isolates can be distinguished from saprophytic isolates, and that analysis of isoenzymes should provide a useful method for the screening of potential pathogenic isolates.

Soluble proteins and isoenzymes from seeds of diverse male steriles of sorghum, (Sorghum bicolor (L.) Moench)

A comparison of soluble protein, esterase, GDH and ADH isoenzyme patterns in seeds of different steriles, maintainers and restorer lines exhibited similarities as well as differences. Soluble protein patterns from sterile and maintainer lines differed both qualitatively and quantitatively. Based on the esterase patterns, male steriles with different cytoplasms could be separated into three groups (i) Ck 60A and B; Nagpur A and B, (ii) M 35-1A and 1 B, M 31-2A and 2B, (iii) G1A and B, VZM2A and 2B. Each group could further be differentiated on the basis of minor differences in esterase isoenzyme patterns within each group. ADH and GDH patterns in general were similar in both sterile and maintainer lines.

Isoenzyme studies in human leukemia — II. Carboxylic esterase (E.C. 3.1.1.1.)

Isoelectric focusing analysis of esterase activity (E.C. 3.1.1.1.) has been carried out on polyacrylamide gel slabs of a variety of immunologically defined acute leukemia subtypes. Distinct isoenzyme bands and isoenzyme groups have been identified and characterized biochemically with regard to their features for substrate specificity, pH optimum and inhibition experiments. The esterase isoenzyme patterns permitted the distinction between acute myeloid and non-myeloid leukemias. Acute lymphocytic leukemias localized by their immunological profile along the T-cell axis exhibited a readily recognizable isoenzyme group (T group). Reproducible differences in isoenzyme patterns were observed in childhood common ALL which indicate that the immunologically determined entity cALL displays a clear heterogeneity with respect to biochemical profiles of enzyme markers. These results suggest that further combined biochemical and immunological characterization may significantly contribute to a better understanding of lymphopoietic and leukemic cell differentiation.