Electrophoretic Patterns Of Soluble Proteins Research Articles

English

The present study was conducted to quantify the response of chlorophyll, protein concentration and electrophoretic patterns of leaf and root soluble proteins to increasing salinity in the rooting medium of three varieties (Payam, Kimia and Jambo) of sorghum (Sorghum bicolor L.) and to evaluate the usefulness of these parameters in identifying salt-resistant varieties. This work was carried out with five concentrations of salinity (0, 50, 100, 150 and 200 mM NaCl) and two sampling time points (sampling after 15 and 30 days salt treatment) using a split-split plot design with randomized complete block layout. With increasing salinity, the K+ concentration was found to decrease and the amount of Na+ and Na+/K+ increase significantly (P≤0.05) in roots and shoots. Chlorophyll a, b, total chlorophyll concentration and leaf and root soluble protein contents decreased with increasing salinity. The electrophoretic pattern of soluble proteins of cv. Jambo showed that after 15 days NaCl (200 mM) and 30 days NaCl (100 and 150 mM) treatment, a new polypeptide of molecular weight 50 kDa was expressed. The expression of this polypeptide might have been due to the plant adapting to NaCl via expression of a stress-resistant gene. This polypeptide was synthesized under salt stress and is suggested as a marker protein for salt adaptation.   Keywords: Chlorophyll, electrophoretic patterns, soluble proteins, salinity, sorghum.

Evaluation of protein content, lysine and sulfur-containing amino acids content and electrophoretic patterns of soluble proteins for gamma-irradiated semolina before and after milling of durum wheat

Influenced of gamma irradiation (0, 0.25, 1, 2.5, 5 and 10 kGy) on total nitrogen, lysine and sulfur-containing amino acids content and electrophoretic patterns of soluble proteins of semolina was studied. The effect of irradiation before and after milling on previous parameters was also investigated. Protein content of semolina was not affected with gamma irradiation before and after milling. Up to 10 kGy dose, cystine and methionine were not significantly changed, although they increased slightly with increasing irradiation dose. Lysine content decreased significantly ( P≤0.05) at irradiation dose higher than 5 kGy. At 10 kGy dose, lysine decreased 5% and 14% for irradiated semolina and that obtained from irradiated wheat grains, respectively. The bands number and intensity of soluble proteins decreased with increasing irradiation dose higher than 5 kGy, as shown on SDS-PAGE electrophoresis. Irradiated semolina and semolina obtained from irradiated wheat grains at 10 kGy showed 13 and 15 bands, respectively. Unirradiated sample showed 19 bands.

Effect of storage on biochemical and microbiological parameters of edible truffle species

The effects of different storage treatments on the most common edible truffle species, such as Tuber magnatum and Tuber borchii (white truffles), Tuber melanosporum and Tuber aestivum (black truffles), were analysed. Biochemical and microbiological profiles were monitored, in order to evaluate possible alterations during truffle preservation. After harvesting, some fresh samples were kept at 4°C for 30days, other samples were frozen at −20°C for one month, thawed and preserved at 4°C; the remainder were autoclaved.The biochemical parameters studied were sugar and protein content, the activity of some enzymes involved in the central metabolism of the fungi and the electrophoretic pattern of soluble proteins. Total mesophilic bacteria were also counted. The results obtained showed that the storage at 4°C is the treatment that best preserves the biochemical and microbiological characteristics of fresh truffles. Black truffles were more resistant to biochemical spoilage than the white ones, while T. magnatum was the most resistant to microbial spoilage.

Comparison of electrophoretic patterns of soluble proteins and isozymes of the red rot disease fungus Pythium sp. isolated from Porphyra yezoensis from Korea and Japan

SUMMARY:Pythium sp., the causative organism of red rot disease, isolated from Porphyra yezoensis from Wando in Korea was compared with those from Miyagi, Aichi, and Fukuoka Prefectures by means of electrophoretic patterns of soluble proteins and isozymes. Native and sodium dodecylsulfate–polyacrylamide gel electrophoretic patterns of soluble proteins revealed small but significant differences among the four isolates. Among the 14 detected enzymes, catalase (CAT), carboxylesterase (CE), glucose-6-phosphate dehydrogenase (G6PD), glucosephosphate isomerase (GPI), isocitrate dehydrogenase (IDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and xanthine dehydrogenase (XDH) possessed isozymes. The dissimilarity value of isozyme banding patterns for four isolates by unweighted pair-group method using arithmetic averages (UPGMA) cluster analysis was highest (0.51) between the isolate from Fukuoka and those from Miyagi and Aichi, and lowest (0.19) between the isolate from Miyagi and that from Aichi. Cluster analysis of data from isozyme banding patterns grouped the isolates from Miyagi and Aichi. These results indicate that the isolates from Miyagi and Aichi are similar, but each isolate from Wando and Fukuoka differed slightly from those of Miyagi and Aichi.

Phosphate Deficiency in Maize. VI. Changes in the Two-Dimensional Electrophoretic Patterns of Soluble Proteins from Second Leaf Blades Associated with Induced Senescence

Phosphate Deficiency in Maize. VI. Changes in the Two-Dimensional Electrophoretic Patterns of Soluble Proteins from Second Leaf Blades Associated with Induced Senescence

An asexual variant of Phytophthora insolita

All tested isolates of Phytophthora insolita and an unknown asexual Phytophthora species found in soil, ditch water, and diseased plant tissues in Taiwan produced ovoid, nonpapillate, nondeciduous sporangia on sporangiophores proliferating through an empty sporangium or in a nesting fashion, and formed irregular as well as spherical hyphal swellings. All tested Phytophthora isolates grew at an unusually high temperature of 39 °C, displayed similar or identical electrophoretic patterns of soluble proteins, and produced α1 hormone. The ability of one isolate of P. insolita to produce oospores was decreased and that of another was lost completely during storage. The results suggest that all isolates of the asexual Phytophthora sp. tested were Phytophthora insolita and were unable to produce oospores owing to a defect in the physiological process of sexual reproduction.Key words: asexual Phytophthora, Phytophthora insolita, high-temperature Phytophthora, sexual reproduction.

Antioxidant system in Boea hygroscopica: Changes in response to desiccation and rehydration

In leaves of Boea hygroscopica subjected to either rapid or slow dehydration and rehydration, the response to H 2O 2 production was studied by monitoring the changes in the amounts of ascorbic and dehydroascorbic acids as well as the amounts of reduced and oxidized glutathione and related enzyme activities. Boea hygroscopica is a resurrection plant in which drought tolerance depends on the rate of desiccation. For this reason, excised leaves of B. hygroscopica were subjected either to rapid or slow dehydration, by a 144-hr exposure to 0% or 80% relative humidity, respectively. Slowly dried leaves, rehydrated for 36 hr, were able to resume their activities completely, while rapidly dried leaves were not able to revive. After dehydration, H 2O 2 production decreased both in slowly and in rapidly dried leaves, whereas the levels of total ascorbate and glutathione became two and 50 times as high, respectively, as in the control. During slow drying, reduced glutathione was oxidized, since its content in slowly dried leaves was 50% lower than in rapidly dried leaves. The levels of soluble proteins were found to be, respectively, five and six times as high in rapidly and slowly dried leaves as in the control and rehydrated leaves. The electrophoretic patterns of soluble proteins were similar in both dehydrated leaves, but different from control and rehydrated leaves, which also presented similar patterns. The specific activities of dehydroascorbate reductase (EC 1.8.5.1), glutathione reductase (EC 1.6.4.2) and ascorbate peroxidase (EC 1.11.1.11) also had the same values in both slowly and rapidly dried leaves. Following rehydration, oxidative processes were intensified: H 2O 2 levels and the oxidation of ascorbate and glutathione mostly increased during this period in which the functioning of the ascorbate/glutathione cycle was induced.

Accumulation of High-Molecular-Weight Proteins in Response to Cold Hardening and Abscisic Acid Treatment in two Winter Wheat Varieties with Different Frost Tolerance

Summary Changes in two-dimensional gel electrophoretic patterns of soluble proteins from two winter wheat varieties ( Triticum aestivum L. cv. Roughrider and cv. Cappelle) differing in frost tolerance were studied. The proteins were obtained from the shoots of dark-grown unhardened seedlings, dark-grown cold hardened seedlings and dark-grown unhardened seedlings after abscisic acid (ABA) treatment. Cold hardening (at 2°C) or treatment with ABA 10 −4 M increased frost tolerance in both varieties. Two-dimensional gel electrophoretic studies revealed that cold hardening or ABA treatment caused the appearance of a group of up to seven high-molecular-weight proteins (150–176 kDa), which were not present on gels from unhardened or ABA-untreated seedlings. Their induction by cold hardening or ABA treatment was more pronounced in the cultivar Roughrider (more frost tolerant) than in the cultivar Cappelle (less frost tolerant). Kinetic studies revealed that in cv. Roughrider the high-molecular-weight protein pattern was completely expressed 48 h after the begin of cold hardening, long before maximal frost tolerance was developed. After ABA treatment the first high-molecular-weight protein spot appeared within 6 h on gels from cv. Roughrider, whereas the ABA induced increase in frost tolerance was slower (no change after 6 h, about 1°C after 24 h, about 2°C after 48 h). It is considered that the high-molecular-weight proteins are involved in the processes leading to increased frost tolerance.

Root rot of cabbage caused by Phytophthora drechsleri

A root rot of cabbage is reported from South Africa. The causal pathogen was identified as Phytophthora drechsleri, based on morphological characters and electrophoretic patterns of soluble proteins.

Etude comparative des protéases digestives de cinq espèces de crevettes pénéides

The activities of the digestive proteases, trypsin, chymotrypsin, leucine aminopeptidase and carboxypeptidases A and B were investigated in the hepatopancreas of Penaeus japonicus, P. stylirostris, P. vannamei, P. merguiensis and P. monodon. Using synthetic substrates, no difference is found for trypsin, leucine aminopeptidase and carboxypeptidases A and B from the five species. Total protease activity is lower in P monodon and chymotrypsin activity is variable depending on the substrate used. Optimal pH, established for total proteases, trypsin, leucine aminopeptidase and carboxypeptidases A and B is alkaline for the five species. No proteolytic activity is found below pH 6. The electrophoretic patterns of soluble proteins, trypsin and leucine aminopeptidase are different for each species. For carboxypeptidase A no differences are observed except for P. japonicus for which three phenotypes are present. As shown by a dot immunobinding assay, the digestive extracts of the five species interact with a P. japonicus trypsin antiserum. Using rocket immunoelectrophoresis, no immunoprecipitation was found between the hepatopancreas extract of P. monodon and the antiserum.

Biochemical transformation of bulbar conjunctiva into corneal epithelium: An electrophoretic analysis

When the entire corneal epithelium of the rabbit is mechanically removed, the denuded corneal stroma is completely resurfaced in 5–10 days with cells of conjunctival origin. The conjunctival cells differ from the corneal epithelial cells both morphologically and biochemically: however, within approximately 6 weeks, the conjunctival cells are converted into functional corneal epithelial cells. Using high-resolution agarose-gel electrophoresis, we compared the soluble proteins from functional conjunctiva (CON), functional corneal epithelium (EPI), and ‘regenerating’ corneal epithelium at eight intervals ranging from 24 hr to 6 weeks post-scraping. Comparison of CON and EPI patterns shows nine major mobility classes of proteins that fall into three subgroups: A = anodal, B = intermediate, and C = cathodal. The nine major classes have marked similarity in distribution and mobility, which may account for the apparent ease with which CON can transform into EPI during regeneration. Electrophoretic patterns of soluble proteins from regenerating epithelium suggest that the regeneration process occurs in three distinct phases: 24–72 hr post-scraping is a period of de-differentiation, 72 hr to 1 week is a period of reorganization, and 1–6 weeks is a period of differentiation. Specific protein groups are identified that represent fast-transforming, slow-transforming, conjunctival-specific, and corneal-specific proteins. Certain of these protein groups can be associated with particular stages in the regenerative process.

Difference in the pattern of soluble proteins from rat brain regions.

The electrophoretic pattern of soluble proteins from seven rat brain regions (amygdala, cerebellum, corpus striatum, cortex, hypothalamus, medulla, and midbrain) was examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Although the number of protein bands (36) was identical in all brain regions studied, there were differences in their relative densities, the greatest variation occurring in the low-molecular-weight region of the electrophoretogram. The bulk of the soluble proteins had molecular weights between 23,000 and 90,000 daltons. The medulla and amygdala showed the greatest range of protein band concentration. A large number of protein bands in the midbrain and corpus striatum showed a greater concentration of protein compared to the same bands in the other regions. A protein band that migrated with the same characteristic as albumin was found. It was consistently high in all regions, the midbrains showing a 1.5-fold greater concentration compared to other regions. Linear regression analysis of wet weight of regional brain tissue against protein concentration yielded a regression coefficient (r2) of 0.77. Midbrain and corpus striatum showed a relatively higher protein concentration: weight ratio than other regions.

A comparative study of avianbordetella‐like strains,bordetella bronchiseptica, alcaligenes faecalisand other related nonfermentable bacteria1

Cultural and biochemical characteristics, electrophoretic patterns of soluble proteins, and pathogenicity in turkey poults, of 30 avian gram-negative nonfermentable bacterial strains acquired from various geographical regions, of which 26 had been isolated from turkeys and one each from a chicken, duck, goose and sharp-tailed munia, were compared to those of Bordetella bronchiseptica, Alcaligenes faecalis, Alcaligenes denitrificans, Achromobacter xylosoxidans and Bordetella parapertussis. With the exception of two turkey isolates, the avian nonfermentable strains constituted a very homogeneous taxon with characteristics of the turkey coryza agent, which has been tentatively designated as a Bordetella-like bacterium. On the basis of the features studied it could be clearly differentiated from B. bronchiseptica, A. faecalis and the other bacteria, suggesting that the Bordetella-like bacterium occupies a distinct taxonomic position. No relevant differences were found between the strains of turkey coryza isolated in Germany and those from USA. The remaining two bacterial strains from turkeys showed the typical characteristics of B. bronchiseptica and A. faecalis respectively.

Soluble proteins and multiple forms of esterases in leaf tissue at first and flag leaf stages of spring barley plants in relation to their resistance to powdery mildew (Erysiphe graminis f.sp. hordei)

Electrophoretic patterns of soluble proteins and multiple forms of esterases were studied in non-infected first and flag leaves of six barley cultivars showing various levels of resistance to powdery mildew. Although the pattern of soluble acidic proteins was similar for all the cultivars tested, some low molecular weight proteins observed in the first leaves were absent in the flag leaves. The appearance of some multiple forms of esterases was genotype-dependent, irrespective of the stage of plant development, whereas the esterase pattern of the first and flag leaves showed characteristic differences, irrespective of the barley genotype. With regard to the different multiple forms of esterases, the barley cultivars tested were classified into three groups which also differed with regard to mildew resistance: (i) Mari S and Peruvian (highly susceptible to powdery mildew), (ii) Villa, Asse (adult plant resistant) and Stamm 4171 (moderately resistant) and (iii) Rupee (highly resistant). The possible significance of this correlation was discussed.

Electrophoretic patterns of soluble proteins of Hendersonia spp.

Electrophoretic patterns of soluble proteins of Hendersonia spp.

Changes in electrophoretic pattern of soluble proteins of potato tuber in early period of infection by phytophthora infestans.

Changes in electrophoretic pattern of soluble proteins of potato tuber in early period of infection by phytophthora infestans.

Electrophoretic Changes in Soluble Proteins during Vegetative and Floral Development of Male and Female Papaya Plants

Summary Electrophoretic patterns of soluble proteins were investigated at vegetative and reproductive growth stages of dioecious papaya. Different protein patterns were observed in the leaf extracts of negetative seedlings and male and female plants. Male and female flowers also possessed a specific pattern of proteins. As the floral ontogeny of sex type proceeded, new bands were observed, while others gradually disappeared. Implications of these data on sex differentiation are discussed.

Electrophoretic Patterns of Soluble Proteins and Isoenzymes of Gaeumannomyces graminis

The wheat and oat take-all fungi show a close serological relationship with Gaeumannomyces graminis var. graminis isolated from kikuyu grass, supporting the conclusions of Walker, who recognized the three fungal groups as varieties of G. graminis based on morphological characters. Three techniques for studying proteins (disc and gradient gel electrophoresis and isoelectric focusing) were assessed as possible aids in identifying and classifying isolates of G. graminis. Protein patterns were affected by the age of the mycelium and the composition of the growth medium used. Patterns prepared by disc electrophoresis were helpful for identifying isolates of G. graminis, but not for separating the varieties. Gradient gel electrophoresis patterns were sufficient for distin- guishing the kikuyu grass isolate from take-all fungi isolated from wheat or oats, but the differences between the patterns of the wheat and oat take-all fungi were too slight to aid in identifying these varieties. Esterase and peroxidase isoenzyme patterns of the isolates from the three varieties were too variable to assist in identifying isolates of G. graminis at either the species or the variety level. Isoelectric focusing patterns of the proteins for each variety were useful for distinguishing the kikuyu grass isolate from the wheat and oat take-all fungi. The protein patterns illustrate a closer relationship between the wheat and oat take-all fungi than either has with the variety of G. graminis from kikuyu grass. None of the methods gave a convenient means of separating the wheat and oat take-all fungi.

Changes in soluble protein and isoenzymes in normal and opaque-2 Zea mays endosperm during grain development

Electrophoretic patterns of soluble proteins, pH 5 enzyme fraction, peroxidase, glutamic dehydrogenase, leucine aminopeptidase in developing endosperm of normal and opaque-2 were studied. Multiple forms were found for all the enzymes studied. The GDH pattern showed considerable differences in normal and opaque-2 maize; the soluble protein pattern also differed, both qualitatively and quantitatively. The leucine-amino-peptidase pattern was identical and the peroxidase pattern showed slight differences.

Pineal proteins and free amino acids during ontogenesis in rats.

An attempt has been made to detect variations inlevels of pineal metabolic activity of rats of three ages: 31 days, 10 weeks, and 1 year. Weight, total protein content, electrophoretic pattern of the soluble proteins, and the free amino acids’ pool of the gland were examined. Absolute weights and total protein content of pineals showed an increase with age but the concentration of protein per mg pineal remained the same at all three ages studied. The electrophoretic pattern of soluble proteins in the pineal is similar at all ages studied, with only quantitative variations occurring between the various bands at different ages. The fast-moving proteins, comprising about 20 % of all soluble proteins, are present in considerably higher proportions in the immature rats; the reverse situation exists with the very slow-moving fraction, representing about 50% of all proteins measured. Certain amino acids were found to increase in their proportions during aging. Others are at their peak in immature rats. The data presented give some basic information on metabolic factors operating at several stages of life and do not appear to reflect a status of lower pineal function in aged animals.