Esterase Isozyme Patterns Research Articles

Somatic hybridization between Dianthus chinensis and D. barbatus through protoplast fusion

Protoplasts isolated from leaf mesophyll cells of Dianthus chinensis and D. barbatus were fused by polyethylene glycol (PEG). Calli exhibiting vigorous growth were selected from the PEG-treated protoplasts and shoots were regenerated from one of these calli after 5 months of culture. These shoots readily rooted and continuously produced flowers in the in-vitro condition. The data on flower color, chromosome number, and esterase isozyme patterns indicated that this plantlet was an interspecific somatic hybrid. The hybridity of the plantlet was also confirmed by nuclear rDNA analysis. This report provides the possibility of applying the somatic hybridization technique for the genetic improvement of the genus Dianthus.

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

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.

Production of sodium-chloride-tolerant Brassica juncea plants by in vitro selection at the somatic embryo level

Somatic embryos, developed from hypocotyl segments of light-grown seedlings of Brassica juncea cv RLM198, were subjected to selection at varying concentrations of sodium chloride (NaCl). Plants were developed from proliferated somatic embryos selected on NaCl-containing medium. The selections were characterized for salt tolerance, esterase isozyme pattern, and proline accumulation. It has been found that: (i) selected tolerant lines showed better root growth, shoot growth, and fresh weight accumulation on salt-containing medium when compared to the control; (ii) salt tolerance was transmitted to the next generation in seed progeny of tolerant plants grown in the absence of exposure to salt; (iii) both the starting material and the tolerant selections accumulated proline, even when grown in salt-free medium. On salt-containing medium, however, the differences in accumulated proline between the control and tolerant lines became more pronounced, and (iv) the patterns of esterase isozymes of two tolerant selections were similar but distinctly different from that of the parental control.

Isozyme profiles associated with the hypersensitive response of Chenopodium foetidum to plum pox virus infection

Isozyme profiles of esterases (E.C. 3.1.1.1), glutamate oxaloacetate transaminase (E.C. 2.6.1.1) and peroxidases (E.C. 1.11.1.7) have been determined in healthy tissues of Chenopodium foetidum as well as their modifications during leaf development. The effect of plum pox virus infection on the isozyme profiles has also been studied. Virus‐induced necrotic lesions displayed a peroxidase (POX) pattern that has not been found in any other tissue of the plant so far analyzed. The pattern was similar to that of old yellow leaves, except that POX‐B, which was detected in the necrotic lesions, has not been detected in any developmental stage of healthy leaves. Changes in the peroxidase profile seem to begin early during infection, even before necrosis is visible. We suggest that senescence is established at necrotic lesions extending from there to the rest of the infected leaf affecting the peroxidase isozyme pattern. However, other changes, which induce POX‐B, must also take place at necrotic lesions. These do not extend to the rest of the infected leaves. Plum pox virus infection has less effect on the glutamate oxaloacetate transaminase and esterase isozyme patterns, inducing an almost normal senescence pattern.

Variation in isozyme patterns of esterase and poly phenol oxidase among isolates ofArmillaria ostoyaefrom British Columbia

Variation in isozyme patterns of esterase and poly phenol oxidase among isolates of<i>Armillaria ostoyae</i>from British Columbia

Isozyme and general protein patterns of Armillaria spp. collected from the boreal mixedwood forest of Ontario

Isolates of four North American biological species of Armillaria, I (A. ostoyae), III, V, and VII (A. lutea), that occur in the boreal mixedwood forest of Ontario were differentiated by their esterase isozyme patterns and total protein profiles. Within biological species I, there are two types of esterase patterns that correspond to the genotypically distinct clones observed in isolates from the same geographic area. In addition, isolates of biological species I from different geographic areas showed differences in their esterase patterns. The general protein patterns of four biological species of Armillaria isolated from maple also showed the distinct pattern of each intersterility group.

Sexual difference in enzyme activities in the mouse.

Electrophoretic analyses of esterases and superoxide dismutase activities of serum, kidney and liver were carried out in ddY mice and six strains of inbred house mice, BALB/c, C3H/He, DBA/2, B10A, B10, C57 BL/6. Sexual dimorphism of esterase activities was observed in serum and kidney but not in the liver, whereas superoxide dismutase (SOD) activities in serum, kidney and liver were not apparently sex-dependent. Serum esterase isozyme patterns among the six strains of inbred house mice showed four different types of banding, namely, BALB/c and C3H/He; B10A and B10; and DBA/2 and C57 BL/6. The patterns of serum esterase in B10A and B10 mice were converted from male type to female type for 2 weeks and 4 weeks following castration, however the male phenotype was restored by administration of testosterone but not of saline.The manifestation of trypsin like protease activities were distinct in submaxillary glands of B10A and B10 mice. In B10 male mice, protease activities were continuously reduced during a 4 week-period following castration, and were completely restored by the administration of testosterone for 2 weeks. In B10A male mice, however, the protease activities were reduced by castration within 2 weeks and remained unchanged for another 2 weeks. Moreover protease activities were not significantly increased after the administration of testosterone for 2 weeks. Since B10 and B10A mice are congenic strains, the different responses of these mouse strains suggest that the H-2 locus region on chromosome 17 may be involved in the hormonal regulation of these enzyme activities.

Interspecific and Intraspecific Differentiation Within the Genus Cronartium by Isozyme and Protein Pattern Analysis

Isozyme analyses were made on aeciospore collections from several species of the genus Cronartium and from several formae speciales of C. quercuum. Aeciospores from closely related Endocronartium harknessii also were tested. Esterase isozyme patterns provided the best differentiation. Patterns for Cronartium spp. differed markedly from those for E. harknessii. C.q.f.sp. banksianae, C.q.f.sp. virginianae, C.q.f.sp. echinatae, and C.q.f.sp. fusiforme were easily distinguished

Sperm competition ofMacrocheles muscaedomesticae (Scopoli) (Acarina: Mesostigmata: Macrochelidae), with special reference to precopulatory mate guarding behavior

The adult males of the manure-inhabiting predatory mite,Macrocheles muscaedomesticae, mounted on the backs of pharate female deutonymphs and guarded them from other males for several hours until female emergence and ensuing copulation. I assumed that an adaptive significance of such characteristic precopulatory mate guarding behavior was closely related to inter-male sperm competition. An existence of 2 sexually compatible strains having different esterase isozyme patterns were found through polyacrylamidegel electrophoresis. A double copulation experiment utilizing these patterns as genetic markers was conducted. In 410 F1 progeny from doubly copulated mothers examined, 409 daughters were fathered by the first males, and only 1 was fathered by the second male (F1 males were eliminated because of the arrhenotokous sex determination system). These results seem to indicate that males ofM. muscaedomesticae guard the immature females in order to secure virgin individuals to mate with.

Electrophoretic and zymogram analysis of the cytoplasmic soluble protein fraction of Spiroplasma citri

The biological function of proteins in the soluble cytoplasmic fraction of two isolates of Spiroplasma citri was investigated by zymogram analysis. There were quantitative and qualitative differences in the isozyme patterns of esterase and RNase between the two isolates of S. citri. Esterase activity in S. citri, reported here for the first time, was higher in one isolate, while RNase activity was higher in the other. No activity was detected in either isolate for acid phosphatase, alkaline phosphatase, or peroxidase. Spiralin, detected in the the cytoplasmic fraction using both native and denaturing polyacrylamide gel electrophoresis, did not appear to be an isozyme of any of the enzymes tested.

Comparison of the Gene frequency between sympatric populations of ninespine sticklebacks, genusPungitius, in Hokkaido, Japan

Gene frequencies, estimated by electrophoretical analysis, were compared between sympatric populations of ninespine sticklebacks,Pungitius pungitius, P. tymensis andP. sinensis, in Hokkaido, Japan. The loci examined wereLdh- E,Sod, Pgm andMp. Consequently, significant differences were detected betweenP. tymensis and other species in all rivers examined. This result strongly suggests thatP. tymensis is reproductively isolated from the other species even when they coexist, although a few natural hybrids betweenP. pungitius andP. tymensis were found through its heterozygosity in esterase isozyme patterns and inPgm. On the other hand, no significant difference was detected betweenP. pungitius andP. sinensis populations in the Biwase River of eastern Hokkaido. Therefore, it is suggested that they possibly belong to a single interbreeding population.

Morphological and Isozyme Survey of Chalkbrood Disease of the Alfalfa Leafcutting Bee in the Western United States

Morphological characteristics of reproductive organs in selected Ascosphaerales were examined utilizing light and scanning electron microscopy. These methods did not establish criteria for distinguishing Ascosphaera aggregata from all other chalkbrood fungi. Esterase isozyme patterns of 50 of 52 geographic isolates of chalkbrood fungi collected from western North America were identical to those of A. aggregata.

Morphological and Isozyme Survey of Chalkbrood Disease of the Alfalfa Leafcutting Bee in the Western United States

Morphological characteristics of reproductive organs in selected Ascosphaerales were examined utilizing light and scanning electron microscopy. These methods did not establish criteria for distinguishing Ascosphaera aggregata from all other chalkbrood fungi. Esterase isozyme patterns of 50 of 52 geographic isolates of chalkbrood fungi collected from western North America were identical to those of A. aggregata.

Difference of Esterase Isozymes between Non-diapausing and Diapausing Strains of the Citrus Red Mite, Panonychus citri (McGREGOR) : Acarina : Tetranychidae

The relationship between the diapause attribute and the non-specific esterase isozyme patterns was investigated in several populations of the citrus red mite, Panonychus citri (McGREGOR), collected from various localities in Japan. The diapause attribute of each population was determined by their developmental success on citrus leaves. The esterase isozyme patterns were detected by agarose thin layer electrophoresis, and classified to Types I, II, III and IV. Each population of the non-diapausing strain showed 1 or 2 patterns among Types I, II and III. On the other hand, Type IV alone was detected from the populations of diapausing strain. Even if the populations included both strains, the diapause attribute of individuals could be clearly determined by their esterase isozyme patterns.

Fingerprinting Apple Cultivars by Electrophoretic Isozyme Banding Patterns

Anionic electrophoretic isozyme patterns of peroxidase, esterase and acid phosphatase and cationic peroxidase isozyme patterns from shoot bark protein extracts were used to identify clonal apple scion cultivars. Each of the 21 cultivars included in this study developed a unique combination of isozyme patterns which allowed it to be distinguished from the others. Sports within cultivars exhibited identical patterns of enzymes, with the exception of ‘Wijcik’, a natural compact mutant of ‘McIntosh’ which could be distinguished from the latter, although it was indistinguishable from the cultivar ‘Spartan’. Isozyme patterns remained constant when samples were taken from wood of different ages, at several times of the year and with trees growing in different locations and on different rootstocks.

Variations of morphological characters and isozyme patterns in Japanese cultivars of Colocasia esculenta Schott and C. gigantea Hook.

Morphological characters related to the vegetative organs, and peroxidase and esterase isozyme patterns were investigated for cluster analyses in 38 strains of taro and three of C.gigantea in Japan. Chromosome numbers of the strains were also determined. There were large variations in the morphological characters and the isozyme patterns among the taro strains. In contrast, few morphological variations and no zymographic variations were observed among the strains of C.gigantea. The small clusters based on the isozyme patterns corresponded more closely to the cultivar groups defined by KUMAZAWA et al. (1956) than those based on the morphological characters. Three strains were markedly distant from the clusters of their groups and they could have been included in the other cultivar groups. Diploid (2n=28) and triploid forms (2n=42) were found in taro, whereas only diploid ones (2n=28) in C.gigantea. These two forms could not be clearly separated by the method mentioned above. There are numerous discrepancies in the classification of the Japanese taro cultivars. KUMAZAWA et al. (1956) classified the largest number of strains based on the largest number of characters related to the morphology of the vegetative and flowering organs, on the texture of corms, and the chromosome number. In the present study, the classification by cluster analysis based on the isozyme patterns agreed well with that of KUMAZAWA's cultivar groups, unlike the classification based on the morphological characters. This finding suggests that KUMAZAWA's classification system for the cultivar groups is supported chemotaxonomically, and that cluster analysis based on two isozyme patterns is a suitable method for assigning the strains into the cultivar groups.

Intraspecific protoplast fusion of Pleurotus ostreatus using auxotrophic mutants.

Hybrid strains of Pleurotus ostreatus were produced by intraspecific protoplast fusion crf auxotrophic mutants derived from monokaryotic strains. Auxotrophic mutants were obtained by treating fragmented mycelia by N-methyl-N'-nitro-N-nitrosoguanidine. One of them obtained from 241 ss 8 strain required uracil for growth on minimal medium, and another one obtained from 235 pSA strain required methionine. After fusion of mixed protoplasts of these two auxotrophic mutants by polyethylene glycol, many colonies were formed on a plate containing minimal agar medium. By isolating those colonies, several dikaryotic strains were obtained. They were cultured in sawdust-rice bran medium at 25C. After transfering into a condition alternating in temperature and light condition in cycle, fruit bodies were formed. Color of the fruit body was gray which was the similar color to one of the parent FMC 241, while the temperature for primordia formation of those fruit bodies was similar to the other parent FMC 235. Thus, the hybrid strain had the characteristics of both parents. Analysis of esterase isozyme pattern also showed the hybrid nature.

Culture of plant somatic hybrids following electrical fusion

Electrically-induced protoplast fusion has been used to produce somatic hybrids between Nicotiana plumbaginifolia and Nicotiana tabacum. Following fusion of suspension culture protoplasts (N. plumbaginifolia) with mesophyll protoplasts (N. tabacum) heterokaryons were identified visually and their development was followed in culture. Because electrical fusion is a microtechnique, procedures were developed for culturing the heterokaryons in small numbers and at low density. The fusion and culture procedures described are rapid, uncomplicated and repeatable. Good cell viabilities indicate that the fusion procedure is not cytotoxic. Fused material was cultured 1-2 days at high density in modified K3 medium (Nagy and Maliga 1976). The heterokaryons were isolated manually and grown, at low density in conditioned media. Calli have been regenerated. Esterase isozyme patterns confirm the hybrid character of calli and clonally-derived plantlets recovered from these fusions.

Isozyme Characterization of Ascosphaerales Associated with Bees. I. Ascosphaera apis, Ascosphaera proliperda, and Ascosphaera aggregata

Acrylamide gel electrophoretic studies were conducted to characterize three chalkbrood fungi associated with bees. Of 28 enzyme assays, acrylamide gel electrophoresis detected 12 enzymes and 285 isozymes. Indices of genetic identity and genetic distance were computed. Proteins showed distinct isozyme profiles. Intraspecific genetic similarities were greater than interspecific ones, but some intraspecific variation was also apparent. Esterase isozyme patterns of chalkbrood fungi, isolated from the alfalfa leafcutting bee in the western United States, were identical to those of Ascosphaera aggregata. Isozyme patterns of A. apis, A. proliperda, and A. aggregata support the current concept of three separate species.

Isozyme variation in populations ofSphagnum recurvum var. mucronatumfrom Britain and Finland

Variations in (1) banding patterns of peroxidase, acid phosphatase and esterase isozymes and (2) morphological characters, of two British and two Finnish populations of Sphagnum recurvum var. mucronatum are described. No links were found between the presence of specific isozyme bands and individual morphological characters, nor were the individual populations typified by particular bands. The results do present some evidence for the linking of certain morphological characters with environmental conditions and suggest that there may be a relationship between the complexity of banding patterns and the stability of site conditions.