Isoenzyme Polymorphism Research Articles

Analysis of the Spring Barley (<i>Hordeum vulgare</i> L.) Isoenzyme Polymorphism Connection with Its Tolerance to the Cadmium Influence

In was the laboratory experiment carried on with the spring barley variants, which shown a contrasting reactions to the cadmium influence. The topic of this work was a searching of the connection of the barley variants response to the toxic stress with the isoenzyme polymorphism of some ferments, which are determined a plants tolerance to the environmental stress. It was taken 14 spring barley variants with the different geographic origin (7 – tolerant to Cd2+ and 7 – sensitive) for this task. Such variants were selected on the base of the morphometric criteria in our previous work. The seeds of these variants were germinated and then a protein extracts were prepared from the seedlings. The extracts were separated by electrophoresis in the polyacrilamide gel. The gel blocks after this process were stained for discovering of the enzyme activity zones. The list of the ferments used was follows: superoxidedismutase, peroxidase, glutamatedehydrohenase, alcoholdehydrohenase, malatedehydrohenase, glutationedehydrohenase, glucose-6-phosphatedehydrohenase, catalase. The frequencies of all enzyme activity zones were counted, and thus such frequencies were compared for the groups of the Cd2+ tolerant and sensitive barley variants. Consequently, it was discovered the specific alleles, which are found in the tolerant or sensitive variants with the much probability. Therefore, some conclusion can be stated: the reaction of the selected barley variant to the Cd2+ influence is connected with some specific isozyme variants. The data, collected in this work, can be used for forecasting of the tolerance of the selected barley variants to the Cd2+.

Monitoring Voriconazole Pharmacogenomics and Plasma Concentrations in the Treatment and Prevention of Invasive Fungal Disease for Chinese Hematological Patients

Background Voriconazole has been widly used in treatment and prevention invasive fungal disease for immunodeficiency hematological patients. And the voriconazole plasma drug levels were associated with its efficacy and toxicity. The hepatic cytochrome P450 isoenzyme 2C19 plays a important role in voriconazole metabolism. However if CYP2C19 genetic polymorphism can result in voriconazole metabolism and drug plasma level in setting of Asian population especially in hematologic patients is unknown.ObjectiveTo evaluate the effect of CYP2C19 polymorphism on the voriconazole (VCZ) plasma concentration of patients with hematological disease and the value of serial monitoring voriconazole plasma concentrations in the treatment and prevention of invasive fungal disease(IFD).Methods Between January to August 2016, 76 hematological patients who received voriconazole for the treatment or prevention of invasive fungal disease were enrolled in this study. The population CYP2C19 polymorphism of voriconazole were performed using PCR-Pyrosequencing. The trough plasma concentrations of vriconazole (Ctrough) was determined using high-performance liquid chromatography (HPLC).Results1. Genotyping for CYP2C19 polymorphic isoenzyme variations showed that 32 subjects (43.42%) for the CYP2C19 wild-type, 43 (56.58%) for the CYP2C19 no-wild-type. 2. 45 of 76 patients received voriconazole intravenous administration, Based on the genotype analysis, 45 subjects were identified as extensive metabolizers' group for EMs (CYP2C19*1/*1), poor metabolizers' group for IMs+PMs (CYP2C19 * *1/*2,*1/*3,*2/*3,*2/*2,*3/*3), and there was a significant difference between Ctrough values in the two groups (1.66±1.86ug/ml vs 3.30±2.35ug/ml, p=0.00). 3. The Ctrough of the 45 patients were detected for 119 times totally. The medium of the Ctrough 45 hematological patients were described. Lack of response to therapy was more frequent in patients with voriconazole levels <1.5mg/L (23.5%) than in those with voriconazole levels >1.5mg/L (21.4%])(p=0.87). And the risk of adverse events was more frequent in patients with voriconazole levels >5.5mg/L (40.0%) than in those with voriconazole levels <5.5mg/L (25.0%) (p=0.49). Furthermore, the Ctrough values of patients with adverse events is higher than the others(3.21±2.46ug/ml vs 2.17±2.14 ug/ml, p=0.042).Conclusions The single-center study showed that the mutation of CYP2C19 was quite common in Chinese hematological patients. Patients with CYP2C19 wild-type phenotype are extensive metabolizers, their Ctrough of voriconazole are significantly lower than patients with CYP2C19 non-wild-type phenotype (poor metabolizers). Appropriate concentrations of vriconazole can improve the effiency of therapy and safety outcome. DisclosuresNo relevant conflicts of interest to declare.

ВИЗНАЧЕННЯ ГЕНЕТИЧНОГО ПОЛІМОРФІЗМУ CYP2C9 НА ТЛІ ПРИЗНАЧЕННЯ ВАРФАРИНУ У ПАЦІЄНТІВ НА ІШЕМІЧНУ ХВОРОБУ СЕРЦЯ, УСКЛАДНЕНУ ПОСТІЙНОЮ ФОРМОЮ ФІБРИЛЯЦІЇ ПЕРЕДСЕРДЬ

&#x0D; &#x0D; &#x0D; &#x0D; This article describes the results of a genetic study of the spread of CYP2C9 polymorphic variants in patients in the Podilskyi region of Ukraine. In different ethnic groups, the frequency of cytochrome polymorphic isoenzymes can vary significantly. The presence of genetic mutations of this cytochrome (Arg144Cys, Ile359Leu) is associated with an increased concentration of warfarin in the blood, which can lead to excessive hypocoagulation and risk of bleeding. Particular attention should be paid to patients with a detected mutation in homozygous form, as both alleles of the gene are mutant (their carriers are "slow metabolizers" and therefore require more careful dose selection). Patients are advised to undergo individual genotyping, which will help to predict the risk of each individual patient.&#x0D; &#x0D; &#x0D; &#x0D;

Surface mycobiota of home-made dry cured sausages from the main producing regions of Argentina and morphological and biochemical characterization of Penicillium nalgiovense populations

The characteristics and quality of home-made dry cured sausages can be recognized and associated with the region of origin. The characteristics of this type of sausages result from the superficial mycobiota that spontaneously colonizes the products. The aim of this study was to identify the house mycobiota associated with home-made dry cured sausages from different localities of Argentina and characterize the populations of Penicillium nalgiovense present by morphological and biochemical markers. To this end, 79 samples were collected from 10 localities of three main producing regions (Buenos Aires, Córdoba and La Pampa provinces). A total of 196 isolates belonging to six genera and 17 species were obtained. The predominant genus was Penicillium (134 of the isolates) and the predominant species was P. nalgiovense (108 isolates). The isoenzyme patterns of α-esterase (α-EST; EC 3.1.1.1) and Malate dehydrogenase NADP+(MDHP; EC 1.1.1.40) were characterized in 48 of these isolates (ten from Colonia Caroya, ten from Oncativo, ten from Tandil, nine from Mercedes and nine from La Pampa). A total of 26 bands were observed: 17 for α-EST and 9 for MDHP. α-EST was the most polymorphic isoenzyme, whereas MDPH presented no polymorphism. The results were subjected to numerical analysis. Cluster analysis revealed the formation of two groups: Group I formed by 24 isolates from Córdoba and Buenos Aires provinces and Group II with 24 isolates from La Pampa and Buenos Aires province. These data suggest the existence of morphological and biochemical variations among P. nalgiovense populations with different geographical origin.

Isoenzyme and molecular approach for authenticating and monitoring of animal cell lines.

Authentication of cell lines is of paramount importance to validate the results from their use in biomedical research. Although isoenzyme polymorphism is the standard method, molecular methods based on mitochondrial DNA (mtDNA) have been developed to replace it. The aim of this study was the improvement of our isoenzyme electrophoretic analysis and the validation of one molecular technique targeted at mtDNA for the authentication of our animal cell lines. The combined method of cellular lysing through osmotic shock, followed by freezing-thawing in N2 to obtain isoenzyme extracts, and with 42 × 106 cells maintained the best efficiency. The superior electrophoretic conditions were PAGE run at 200 V. All cell lines had isoenzymatic mobility corresponding to their species to lactate dehydrogenase, malate-dehydrogenase, and glucose-6-phosphate dehydrogenase isoenzymes, and could be distinguished from each other. Two molecular techniques based on mtDNA were tested, one on the cytochrome b gene and other on cytochrome c oxidase I subunit gene. Due to difficulties in distinguishing all cell lines using only one these techniques, we merged the primers of two methods in such a way that there was a sufficient differentiation of all DNA fragments. The sequencing of these PCR products was also performed to validate these data.

Gene pool state and degree of infestation by bark beetle (Ips tipographus L.) of Norway spruce (Picea abies L. Karst.) natural populations and planted stands in Moscow region

A comparative analysis of the gene pool state in natural populations and planted stands of Norway spruce and the degree of their infestation by the bark beetle in the Moscow region was conducted taking into account the dynamic state of communities (4 populations, 148 samples, 24 isoenzyme loci). The degree of infestation by the bark beetle of conditionally native communities is 0%; for planted stands, it is 90–100%; and for a short-term community, it is 15–20%. The comparison of “healthy” populations and those infested with bark beetle by average values of observed heterozygosity (H O) detected no significant differences. However, the test on allelic frequency heterogeneity demonstrated the difference of planted stands from conditionally native populations both by three loci (Fe-2, Idh-1, Mdh-3) and by the totality of 18 polymorphic isoenzyme loci; the short-term population differs from conditionally native population only by two loci. The value of the inbreeding coefficient by the Idh-1 locus is significantly higher in both populations infested with the bark beetle than in “healthy” populations. The results of conducted studies demonstrate the necessity of continuation of the study on the gene pool state in Norway spruce populations owing to the degree of their infestation by the bark beetle along with the study on the dynamic state of the communities; this can provide a key to solving the problem of the forest preservation from pests.

The role of CYP3A5 polymorphism and dose adjustments following conversion of twice-daily to once-daily tacrolimus in renal transplant recipients.

BackgroundTacrolimus is available as twice-daily Prograf® (Tac-BID) and the once-daily formulation, Advagraf® (Tac-OD). Although therapeutically equivalent, some transplant recipients require dose adjustments to achieve similar tacrolimus trough concentrations [Tac C0] after conversion between formulations. Tacrolimus is primarily metabolized by cytochrome P450 3A5 (CYP3A5). We sought to determine whether genetic polymorphisms in the CYP3A5 enzyme; CYP3A5 *1/*1 and CYP3A5 *1/*3 (expressers) compared to CYP3A5 *3/*3 (non-expressers) could account for discrepancies in dose requirements following conversion from Tac-BID to Tac-OD.MethodsA cohort of 60 renal transplant recipients (RTR) from our larger conversion study of 496 patients underwent additional testing for CY3A5 genetic polymorphisms. Analysis included demographics, tac dosing and [Tac C0] pre- and post-conversion and dosing changes relative to CYP3A5 genotypes. CYP3A5 genetic polymorphisms were identified through analysis of genomic DNA.ResultsConversion from tac bid to tac OD in this cohort required a mean (SD) dose increase from 3.1 (1.0) mg/day to 3.8 (1.3) mg/day (p = 0.007), to achieve similar [Tac C0]. The *1/*3 expresser group required a greater percentage dose adjustment (56.7 %) in converting from Tac-BID to Tac-OD as compared to the *3/*3 non-expresser group (26.6 %). Similar findings were observed with the both expresser groups combined (*1/*1 &*1/*3). The expressers were significantly more highly represented in the East Asian cohort.ConclusionsThe CYP3A5 expresser polymorphism necessitates an increase in dosing upon conversion from Tac-BID to Tac-OD, with the expresser genotypes contributing significantly to this finding. Given the variability in frequency of CYP3A5 genotypes in various ethnic groups, future studies should account for both isoenzyme polymorphism and ethnicity in optimizing dosing requirements.Trial registrationClinical trials.gov identifier: NCT01884480

Glutathione S-Transferase M1 and T1 Gene Polymorphisms in Patients with Chronic Plaque-Type Psoriasis: A Case-Control Study

Objective: To determine the role of glutathione S-transferase (GST) isoenzyme polymorphisms as susceptibility factors in patients with psoriasis in a Turkish cohort. Subjects and Methods: In this case-control study, 105 patients with plaque-type psoriasis and 102 healthy controls were recruited from the dermatology outpatient clinics of two university hospitals. Genomic DNA was extracted from whole blood using a DZ DNA isolation kit. Multiplex PCR was used to determine GSTM1 and GSTT1 polymorphisms in the isolated DNAs. Results: Of the 150 patients with psoriasis, 83 (79%) were identified with the GSTT1 genotype and 22 (21%) with the null genotype. Of the 102 patients in the control group, 69 (67.6%) subjects were identified with the GSTT1 genotype and 33 (32.4%) with the null genotype. There was no significant difference between the patient and control groups (p = 0.063). Regarding the GSTM1 polymorphism, 54 (51.4%) patients were identified with this genotype and 51 (48.6%) with the null genotype; in the control group, 50 (49%) were identified with this genotype and 52 (51%) with the null genotype. Again there was no statistically significant difference between the groups (p = 0.957). Conclusion: In this Turkish cohort of patients with psoriasis, neither GSTT1 nor GSTM1 polymorphisms were associated with disease susceptibility. Larger studies with a wider range of GST isoenzyme are needed.

Prevalence of CYP2B6 polymorphisms in Argentinians: the role of genetic testing.

CYP2B6 is a highly polymorphic isoenzyme involved in the metabolism of many drugs including cyclophosphamide, bupropion, and efavirenz. A single nucleotide polymorphism (SNP) in CYP2B6 (516G>T) resulted in decreased expression and function associated with the CYP2B6*6 haplotype. Among the clinical implications of this phenotype, decreased activation of cyclophosphamide and increased plasma levels of efavirenz associated with increased central nervous system toxicity have been reported. The frequency of the CYP2B6 (516G>T) SNP has been studied in several different populations, but there is no data regarding distribution among Argentinians. In this study, 102 DNA samples from healthy volunteers were analyzed using a polymerase chain reaction-restriction fragment length polymorphism reaction specific for the CYP2B6 (516G>T) SNP. Our results showed a prevalence of 71.08% for the G allele and 28.92% for the T allele. This was distributed as 52.9% for the GG genotype (reduced dosage required), 36.6% for the GT genotype (normal dosage range), and 10.8% for the TT genotype (high drug toxicity). There was no preferential gender distribution observed. The relatively high prevalence of the TT genotype in our population supports the clinical use of genotyping as an additional tool in personalized medicine.

Attempt at conserving the genetic resources of Hu sheep by fibroblast line cryopreservation

Hu sheep is one of the most distinctive local sheep breeds in China, while investigations concerning its cell biology and molecular genetics are still scarce. To preserve the valuable genetic resources of the Hu sheep, we established a Hu sheep fibroblast line from 35 sheeps' ear marginal tissues using explant culture and cryopreservation techniques, and its biological characteristics were identified, including cell morphology, growth curve, karyotype, isoenzyme polymorphism, and expression of exogenous fluorescent genes. The results showed that the viability of the fibroblasts before freezing was 96.54 ± 3.22% and 92.36 ± 2.77% after thawing, and there was no microbial contamination and cross-contamination; the growth curve of Hu sheep ear marginal fibroblasts displayed an obvious “S” shape; all somatic chromosomes were acrocentric autosomes, and only the two sex chromosomes (XY) were submetacentric; the normal number of chromosomes (2n = 54) was observed in 96.56 ± 2.89% of the cells; and the transfer efficiencies of four fluorescent proteins could achieve 37.4% with an optimized ratio of plasmid and lipofectamine. It could be concluded that this study has not only provided the biological characteristics of Hu sheep at cellular level, but also makes a valuable contribution to the preservation of the genetic resources of the Hu sheep.

Combined assessment of genetic variability of Coregonus albula (L.) populations in Latvia based on allozymes and RAPD markers

The vendace (Coregonus albula L.) inhabits several Latvian lakes. In Latvia this species is included into the list of specially protected species with limited use. Taking into consideration the high variability of vendace and the fact that it belongs to valuable and marketable fish, there arises a scientific interest to reveal possible reasons influencing its variability. There is no precise scientific data on the local vendace populations and its biology and this hampers the rational usage of this fish and possibilities of its reproduction in Latvian lakes. In the present work the genetic polymorphism and the structure of vendace populations in Rāznas, Nirzas and Sventes lakes in Latvia have been defined based on the analysis of isoenzyme systems and RAPD markers. The genetic variability and population structure both within and among vendace populations have been studied. The activity of eight isoenzyme systems was studied and 16 loci were selected for genetic analysis. Eight of them were polymorphic and used for the statistical analysis. Also, 60 decanucleotide (RAPD) primers were tested. For the analysis, eight decanucleotide primers were used. The average level of isoenzyme polymorphism in vendace populations from three lakes was 41.67%. The diversity of alleles based on allozyme data in the studied populations was approximately the same. Data of RAPD analysis showed that Coregonus albula populations from the studied lakes were highly polymorphic. The results of this study indicate that the estimated intraspecific variation may be more pronounced with RAPD markers than with allozymes when the two approaches were applied to the same populations.

Biological characteristic of an embryonic fibroblast line from Xiaoshan chicken for genetic conservation

A fibroblast line from embryos of Xiaoshan chicken was established successfully by direct culture of explants and cryopreservation techniques. The cells were morphologically consistent with fibroblasts, and the population doubling time (PDT) was about 46.72 h. According to karyotyping and G-banding, the diplontic cells with 78 chromosomes accounted for 98.58±1.27% of the total cells. The cells were tested for microbial contamination and they were free of infections from bacteria, fungi, viruses and mycoplasmas. There were no cross-contamination from other cell lines as revealed by isoenzyme polymorphism analysis of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH). Three fluorescent protein genes were transfected into the Xiaoshan chicken embryonic fibroblasts and the transfection efficiencies of these genes were between 12.72 and 35.89%. All the tests showed that the quality of the cell line conformed to the quality criteria of the American type culture collection (ATCC). This work had not only preserved the precious genetic resources of Xiaoshan chicken, but also explored a new protocol to preserve the endangered animal breeds. Key words: Xiaoshan chicken, fibroblasts, genetic conservation, biological characteristics.

Genetic variation of isolated and peripheral populations of Pinus sylvestris (L.) from glacial refugia

Analysis of variation at 10 polymorphic isoenzyme loci in 51 European populations of Pinus sylvestris both from its main continuous range and from isolated, marginal populations out of the continuous range, revealed differences in genetic structure. The highest genetic richness, measured as mean number of alleles per locus, was recorded for populations from the Iberian Peninsula, followed by those from Scotland and the Balkans. Marginal, isolated populations were characterized by much greater interpopulation variation than populations from the continuous range of the species. The highest mean observed heterozygosity was recorded for the Iberian populations. The highest gene flow was observed among populations in the continuous range of the species. The populations could be classified into five groups based on genetic similarities. Populations from the continuous range formed one group, those from Spain two groups, pines from Scotland one group and those from the Balkans one group. One population from Scotland was found to be most distinct from the other Scottish pines and was found to be grouped with the Balkan populations. The occurrence and frequencies of rare alleles in the populations showed a characteristic pattern, suggesting that populations from the Iberian Peninsula probably did not participate in the colonization of Europe by Pinus sylvestris after the last glaciation. In contrast, the migration of populations from the Balkan refugium into Central and Western Europe had a significant effect on the contemporary gene pools of populations of Pinus sylvestris in its continuous range.

Pharmacogenetics Variations in Anesthesia

Pharmacogenetics is an emerging discipline that attempted to understand the hereditary basis for differences in responsiveness or inter-individual variation to therapeutic agents. It is the study of variability in drug response as a result of heredity factors. The importance for pharmacogenetics for the clinician is to enable optimum therapeutic efficacy, to avoid toxicity of those drugs whose metabolism is catalyzed by polymorphic isoenzymes, and to contribute to the rational design of new drugs.

Dehydrogenase isoenzyme polymorphism in genus Prunus, subgenus Cerasus

Dehydrogenase polymorphism was studied in 36 sour cherry (Prunus cerasus L.), sweet cherry (Prunus avuim L.), mahaleb (Prunus mahaleb L.), ground cherry (Prunus fruticosa Pall.), duke cherry (Prunus gondounii Redh.), Japanese flowering cherry (Prunus serrulata Lindl.) and four iterspecific hybrids (standard cherry rootstocks ?Gisela 5?, ?Gisela 6?, ?Max Ma? and ?Colt?). Inner bark of one-year-old shoots, in dormant stage, was used for enzyme extraction. Vertical PAGE was used for isoenzyme analysis: alcohol dehydrogenase (ADH), formate dehydrogenase (FDH), glutamate dehydrogenase (GDH), isocitrate dehydrogenaze (IDH), malate dehydrogenase (MDH), phosphogluconate dehydrogenase (PGD), and shikimate dehydrogenase (SDH). All studied systems were polymorphic at 10 loci: Adh -1 (3 genotypes) and Adh-2 (5 genotypes), Fdh-1 (2 genotypes), Gdh-1 (3 genotypes), Idh-1 (4 genotypes) i Idh -2 (5 genotypes), Mdh-1 (3 genotypes), Pgd-1 (4 genotypes), Sdh-1 (1 genotype) i Sdh-2 (3 genotypes). Cluster analysis was used to construct dendrogram on which four groups of similar genotypes were separated. Obtained results indicate that studied enzyme systems can be used for determination of genus Prunus, subgenus Cerasus. Among studied enzyme systems ADH, IDH and SDH were the most polymorphic and most useful to identify genetic variability. Polymorphism of FDH and GDH in genus Prunus, subgenus Cerasus was described first time in this work. First results for dehydrogenase variability of Oblacinska indicate that polymorphism of loci Idh-2 and Sdh-2 can be useful for discrimination of different clones.

Establishment and biological characteristics of a Jingning chicken embryonic fibroblast bank

Using tissue explantation and cryopreservation biotechniques, a Jingning chicken embryonic fibroblast bank was successfully established, which includes 43 embryo samples and a stock of 178 cryovials, each one containing 3.0×106 cells. Most of the cells were apparently fibroblasts in their morphology, and the population doubling time (PDT) was about 48 h. The total chromosome number of a diploid cell was 78. According to karyotyping and G-banding, the diploid rate in the cell bank was 97.62±2.12%. The cells were tested for microbial contamination and found free of infections from bacteria, fungi, viruses and mycoplasms. There was no cross-contamination from other cell lines as revealed by lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) isoenzyme polymorphisms. Six fluorescent proteins were transfected into the Jingning chicken embryonic fibroblasts, and the transfection efficiencies of these genes were between 10.1 and 41.9%. All the tests showed that the quality of the cell line conforms to the quality criteria of the ATCC (American type culture collection). This work succeeded not only in preserving the genetic resources of Jingning chicken, but it also established a new protocol to preserve endangered animal breeds.

Establishment and biological characteristics of Qingyuan partridge chicken fibroblast line

The aim of this work was to seek a different approach to conserving important domestic animals in imminent danger. A Qingyuan partridge chicken embryonic fibroblast line, containing 336 cryovials with 8x10(6) cells each, was successfully established from 60 Qingyuan partridge chicken embryos using explant culture and cryopreservation techniques. The cells were morphologically consistent with fibroblasts. The growth curve was a typical "S" shape having a detention phase, a logarithmic phase, and a plateau phase. The population doubling time was approximately 72 h. Tests for bacteria, fungi, viruses, and Mycoplasma were all negative. Isoenzyme polymorphism indicated that the genetic characteristics of the cell line were stable in vitro. Karyotyping analysis suggested that the chromosome number of a normal cell was 2n=78 and 93.4% of the entire population was diploid. The transfection efficiencies of 6 fluorescent proteins (pEGFP-C1, pEGFP-N3, pDsRed-N1, pEYFP-N1, pECFP-N1, and pECFP-mito) optimal at 48 h were from 15.1 to 39.8%. The cell line met all of the criteria from the American Type Culture Collection. Not only has the germline of this important chicken breed been preserved at the cell level, but also valuable material has been provided for genome, postgenome, and somacloning research. Moreover, the establishment of this technical platform may provide both technical and theoretical support for storing the genetic resources of other animals and poultry at the cell level.

Isoenzyme polymorphism of almond genotypes selected in the region of northern Serbia

Isoenzyme polymorphism was studied in 20 almond (Prunus dulcis [Mill.] D.A. Webb) genotypes selected from seedling populations of unknown almond cultivars in the region of northern Serbia (Vojvodina). Fourteen enzyme systems were studied using the method of vertical polyacrylamide gel electrophoresis. Ten systems were polymorphic in twelve loci. This polymorphism allowed unique identification of all studied genotypes. The most useful enzyme for analysis of almond genetic variability was menadione reductase. Polymorphism identified for alkaline phosphatase, formate dehydrogenase, glutamate dehydrogenase, malic enzyme, and menadione reductase was reported for the first time in almond. Cluster analysis was used to construct a dendrogram on which five clusters with different number of genotypes could be identified.

ANALYSIS OF DONGBI LONGAN WITH ISOENZYME AND RAPD TECHNOLOGIES

Longan (Dimocarpus longan Lour.) is one of the most important subtropical fruit trees in Fujian province. Due to little genetic background, the development of longan resources study was restricted. Random amplified polymorphic DNA (RAPD) and POD isoenzyme techniques were used to study the genetic polymorphism and relationship among strains of Dongbi longan. Twenty two arbitrary 10mer primers were used in RAPD analysis of 13 Dongbi longan strains. A total of 124 bands were amplified from 13 samples, each primer pair produces 3 (S299) to 9 (S60) bands, and the average bands were 5.64. The RAPD results showed that the genetic comparability was significantly high and there was high genetic stability among 13 strains, but genes varied with the time and environment. Compared to RAPD results, there was no significant difference in POD isoenzyme among 13 samples from POD isoenzyme gel. According to the results, we concluded that RAPD technology is more suitable than POD isoenzyme technology for longan germplasm resources analysis.

Isoenzymatic polymorphism in the leaf-cutting ant Atta capiguara Gonçalves (hymenoptera: formicidae)

This study was carried out to analyze the genetic population structure of Atta capiguara from 12 nests collected in Tapejara in the state of Paraná, Brazil, using isoenzyme polymorphisms. The analyzed isoenzymes were esterases (EST-EC 3.1.1.1), acid phosphatase (ACP-EC 3.1.3.2) and carbonic anhydrase (CA-EC 4.2.1.1). Ten loci were found in A.capiguara and four polymorphic loci were detected. The observed heterozigosity (0.0296) was low when compared to the expected heterozigosity (0.1461). The high value of F(IS) (0.7954) shows an excess of homozygous genotypes probably caused by inbreeding.