Analysis of Amylase and Superoxide Dismutase Isozymes During the Germination Process of Emmenopterys henryi Oliv Seeds

[ Objective] The study was to understand the changes of amylase（AMY） and superoxide dismutase（SOD） isozymes during the ger- mination process of Emmenopterys henryi Oliv seeds. [ Metbod] By employing polyacrylamide gel electrophoresis method, the expressions of AMY and SOD isozymes during seed germination process were analyzed. ~ Result] The main AMY bands remained strong during the whole peri- od and a new band A2 appeared in the middle and late period of seed germination. Some new SOD bands occurred at the early stage, then be- came weak or disappeared in the middle period, and band S6 became intense in the late peried. [ Conclusion.] The expression of AMY and SOD isozyme gene has temporal difference during germination of E. henryi Oliv seeds.

Effects of Hg^(2+) on Isozymes of Peroxidase,Catalase and Superoxide Dismutase in Wheat Seedlings

[Objective] This work was aimed to explore the mechanism of Hg2+ toxicity on plants.[Method]Activities of peroxidase(POD),catalase(CAT)and superoxide dismutase(SOD)were investigated in wheat(Triticum aestivum L.)seedlings under Hg2+ stress at different concentrations.[Result]① There were no obvious effects on the growth of seedlings when the concentration of Hg2+ was lower than 0.10 mmol/L.However,toxic effects on the growth of seedling were observed when the concentration of Hg2+ was higher than 0.10 mmol/L.② Different tissues showed different resistant ability in response to Hg2+ stress.The leaves and roots of wheat seedlings were more insensitive to Hg2+ toxicity.③ CAT was more sensitive to Hg2+ stress compared to POD and SOD.[Conclusion]The toxic effect was related to the concentration of Hg2+(0.10 mmol/L).The higher concentration of Hg2+ could affect the expression of POD,CAT,and SOD isozymes in the leaves,roots of wheat seedlings and germinated seeds,which further affect the normal metabolism of membrane lipid and inhibit the growth of wheat seedlings at last.

Electrophoretic Analysis of Isozymes and Discussion about Species Differentiation in Three Species of Genus Gymnocypris

By using the method of electrophoresis,three isozymes (lactate dehydrogenase,malate dehydrogenase and esterase) of three species of genus Gymnocypris were described and analyzed from North Tibet in this paper. The results showed that all three isozymes presented interspecific difference and distinct differentiation among individuals in the same population,and there was no electrophorectic difference between males and females. Analysis of relationships among three naked carps indicated a high degree of similarity between G. selincuoensis and G. cuoensis ,whereas low degree between G. selincuoensis and G. namensis . Furthermore,three isozymes presented expression of null alleles,and the duplicate genes of LDH A 2,LDH B 2,s MDH A 2 and m MDH B 2 also expressed in some individuals. Compared to other tetraploid fishes,three naked carps retained more functional duplicate genes and null alleles. This suggests fishes of genus Gymnocypris are at the early stage of evolution after polyploidization than that of fishes of Catostomidae,it directly related to the later time of schizothoracine fishes originate as well as severe environment.

Comparative Study of Immunological Properties on Glutamine Synthetase Isozymes in Rice Plants

In rice (Oryza sativa L.) roots two glutamine synthetase (GS) isozymes, GSra and GSrb, were identified recently in the author's experiments, but the homology of both GSra and GSrb as well as their localization in the rice roots are unclear. In the present study, the purified GSra and GSrb from rice roots were used to immunize rabbits to obtain the respective antibodies. The immunodiffusion and immunoblotting experiments showed that the antibody against GSra or GSrb was specific for GS and its isozymes. The immunoprecipitation test indicated that the antibody of GSra or GSrb not only recognized its respective antigen, but also well recognized each other's antigen. GSra or GSrb antibody recognized also better cytosolic GS1 of rice leaves, but the recognization for chloroplast GS2 from rice or spinach (Spinacia oleracea Mill.) leaves was weaker. Our results indicate that GSra and GSrb from rice roots are quite similar in antigenicity and are extremely similar proteins and that both GSra and GSrb may also be a form of cytosolic GS just as the cytosolic GS1 of rice leaves.

Effect of biliary obstruction and internal biliary drainage on hepatic cytochrome P450 isozymes in rats

AIM: To investigate the total cytochrome P450 (CYP) content, microsomal mixed-function oxidase (MFO) activity, and expression of mRNAs for various CYP isozymes in a simple rat model of reversible obstructive jaundice. METHODS: Obstructive jaundice was created in male rats by causing bile duct obstruction with polyester tape. In another group of rats, bile duct obstruction was followed by internal biliary drainage after releasing the tape. The expression of various CYP isozyme mRNAs was semi-quantitatively assessed by competitive RT- PCR. RESULTS: The total CYP content and microsomal MFO activity showed a significant decrease after biliary obstruction, but returned to respective control levels after biliary drainage. A marked reduction in the expression of CYP1A2, 2B1/2, 2C11, 2E1, 3A1, and 3A2 mRNA was detected during biliary obstruction, while expression increased significantly toward the control level after biliary drainage. Although expression of CYP4A1 mRNA showed no reduction during biliary obstruction, it still increased significantly after biliary drainage. CONCLUSION: These results suggest that not only obstructive jaundice, but also the subsequent internal biliary drainage may affect regulatory medications of the synthesis of individual CYP isozymes differently.

6 Kinds of Isozymes after Long-term Subculture of Emmenopterys henryi Oliv.

[Objective] The aim was to study the changes of zymography in 6 kinds of isozymes after long-term subculture of Emmenopterys henryi Oliv.. [Method] Non-denaturing polyacrylamide gel electrophoresis was used to analyze isozyme patterns such as esterase (EST),acid phosphatase (ACP),ATP enzyme (ATPase),amylase (AMY),superoxide dismutase (SOD) and peroxidase (POD) in long-term subculture callus of Emmenopterys henryi Oliv. [Result] The research showed that there were differences among the 6 kinds of isozymes in embryogenic callus and non-embryogenic callus of Emmenopterys henryi Oliv.,and both levels could be taken as the basis for identification,the EST,ACP,and POD of non-embryogenic callus were significantly higher than embryogenic callus. The browning of non-embryogenic callus was non-level in the AMY,SOD and POD isozymes when was compared with normal non-embryogenic callus,while the EST,ACP and ATPase isozymes decreased; When the browning of embryogenic callus was contrasted with normal embryogenic callus,EST isozyme increased and the other 5 kinds of enzymes decreased. [Conclusion] The study provided theoretical basis for research morphological difference and browning of long-term tissue culture of Emmenopterys henryi Oliv..

ONTOGENIC STUDY OF ISOZYMES IN GIANT TIGER PRAWN,PENAEUS MONODON (CRUSTACEA:DECAPODA)

The patterns of isozyme variation throughout the life cycle of the giant tiger prawn, Penaeusmonodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH,PGD, and PROT). The zymograms showh little variation in the number of isozymic loci expressed duringlarval development compared with results reported for other penaeid prawns. There were specific ontogenicpatterns at some loci. As larvae developed beyond the embryonic stage, there was tendency for intensityof staining and complexity of banding patterns to increase.

The inhibitory effect of lead on the seedling growth and the induced formation of oxidase isozymes in higher plant

The inhibitory effect of lead on the seed germination and seedling growth was obvi-ously seen in barley,sorghum and wheat.Their differential reactions on the lead solution werefound under the lead stress during the seed germination and seedling growth stages.Wheat ismore sensitive to lead than the other three plants.This can be used to select new crops which canresist lead damage.The changes of the peroxidase and polyphenol oxidase isozymes have beenfound in individual plants which was injured by a lead solution at the seedling and root develop-ment stages.The authors suggest that these isozymes,which have been changed in patterns andnumbers,may be adaptive isozymes.Adaptive isozymes are produced in plant tissue when plantsare in harmful environmental conditions.The change in environmental conditions influences theexpression of the gene which encode peroxidase,polyphenol oxidase isozymes,and promote forma-tion of the adaptive isozymes.

REGULATING FUNCTION OF COENZYMIZATION AND DECOENZYMIZATION OF THE LACTATE DEHYDROGENASE ISOZYMES IN THE MOUSE TISSUES DURING HYPOXIA

Objective. To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.Methods. Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by the specific staining technique. LDH (1 -5) relative percentage enzymatic activity (RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner.Results. The RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymes LDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH,(H4), LDH2(H3M) increased, while RPEA of LDH5(M4) in various tissues decreased prominently except the cardiac muscle, and that of LDH4(HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypoxia treated cardiac muscle specimen was made, activity subbands originated regularly in the isozyme patterns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands), LDH4 (1-3 subbands), LDH5 (2-4 subbands). After adding appropriate amount of NAD+ to the hypoxia treated cardiac muscle specimen, PAGE showed the subbands of four isozymes (LDH2-LDH5) reduced or even totally disappeared in the isozyme patterns.Conclusions. The negative feedback regulation of coenzymization and decoenzymization of LDH isozymes is one of the mouse stress responses to slight hypoxia.

Electrophoretic Analysis of Isozymes and Proteins in F_1 Hybrids of Ctenopharyngodon idellus ♀ × Elopichthys bambusa ♂

[ Objective] This study aimed to investigate the biochemical and genetic characteristics of F1 hybrids of Ctenopharyngodon idellus ♀× Elopichthys bambusa ♂ - [ Method ] By using vertical polyacrylamide gel electrophoresis technology, three isozymes （LDH, EST, MDH） in nine tissues （heart, brain, eyes, liver, kidney, spleen, fin, muscle, plasma） of F1 hybrids of C. ideUus and parents were compared. [Result] Three isozymes in F1 hybrids of ♀× E. bambusa ♀ were analyzed; differences in isozyme and proteins between F1 hybrids idellus ♀× E. bambusa ♀ exhibited varying degrees of tissue-specificity; isozymes and proteins in F, hybrids were signiticandy different from the parents. [ Conclusion] These nine differences could be used as indicators to identify F1 hybrids and parents.

Isozymes Analysis of the Golden Cuttlefish Sepia esculenta (Cephalopoda: Sepiidae)

Thirty nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH, MDH, MEP, IDHP, PGDH, GRS, NP, AAT, CK, AK, EST, ALP, ACP, FBP, MPI, GPI and PGM) show strong activities and good convergence in zymogram. They are proved to be suitable genetic markers in Sepia esculenta. Among the tissues used, mantle muscle is the best for electrophoretic analysis of isozymes. Eye and liver are fairly good for some special enzymes, such as LDH, EST, MPI, etc. Twenty six loci are detected. The proportion of polymorphic loci is 0.115 in the Qingdao sample and 0.153 in the Rizhao sample ( P < 0.99 ). The mean values of the observed and expected heterozygosity per locus of Qingdao sample are 0.016 and 0.017 , while those of the Rizhao sample are 0.023 and 0.025 respectively.

Effects of 1 — 4℃ low temperature treatment on seedling growth and POD isozymes in higher plants

The effects of iow temperature treatment on seedling growth and POD isozymeswere studied using hyacinth bean (Dolichos tablab L. )and Soybean[Glycine max(L. )Merr. ]culti-vars as experimental materials. Their seedtings were exposured to 1 - 4℃ low temperature for 4 days to inhibit growth. The changes of POD isozymes in roots and seedlings were observed after seedlings were exposured to 1 - 4℃ for 4-8 days. There was a significant linear relation between inhibition of seedling growth and change of POD isozymes in these two kinds of plants.

Tissue Expression and Stock Variation of Isozymes of Stone Flounder(Kareius bicoloratus)

Tissue expression and stock variation of isozymes of stone flounder （Kareius bicoloratus） were analyzed with horizontal starch gel electrophoresis. For the fourteen enzymes assayed, 31 loci were recorded. The results indicated that all the isozymes examined were obviously tissue-specific. The expressions of SOD＊, GDH＊, G3PDH-2＊ and ADH-2＊ were detected only in liver, SDH-1＊, MDH-1＊ and ADH-1＊ only in muscle, and LDH-B＊ and LDH-C＊ only in eyes. In comparison, MDH-2＊, GPI-3＊ and SDH-2＊ were detected in all tissues examined. Other loci examined were detected in a variety of tissues. Muscle and liver were selected to detect the isozyme variation of the two geographic stocks of Qingdao and Weihai, Shandong Province, China. The percentages of polymorphic loci （P0.99） were 29.17% and 25.00%, the observed heterozygosities （H0） were 0.028 ±0.014 and 0.040 ± 0.019, and the expected heterozygosities （He） were 0.039±0.017 and 0.052±0.022 in Qingdao and Wethai stock, respectively. The coefficient of gene differentiation （Fst） and genetic distance （D） between the two stocks was 0.012 and 0.0011, respectively, indicating that the genetic differentiation is low between them. Compared with other species of Pleuronectiformes, both the percentage of polymorphic loci and the mean heterozygosity ofK. Bicoloratus were at a middle level.

Effect of Coconut Milk and Rare Earth on Proliferation, Rootage and Isozymes of Dendrobium in Tissue Culture

In tissue culture, effects of coconut milk, rare earth and medium on proliferation, rootage and isozymes of Dendrobium were studied. The result indicated that coconut milk and RE could promote proliferation and rootage, with their combined treatment the most effective. In proliferation culture, coconut milk enhanced peroxidase (POD) and esterase(EST) activity and types, RE decreased POD, but increased EST activity and types and combined treatment did not show the same influence on POD and EST too. In rootage culture, adding coconut milk decreased POD activity, but improved EST activity in 1/2MS medium and declined EST activity, expression type in WPM medium. when using RE, POD and EST activity both reduced, types of the former decreased, types of the latter changed slightly with the basic medium. And combined treatment would decreased POD activity and types.

The differenee in protein content,MDH,POD and COD isozymes during root different developmental stages in lowland rice and upland rice

The difference in protein content, MDH,POD and COD isozymes have been seenbetween lowland rice roots and upland rice roots during root growth and development. High pro-tein content has been examined in upland rice roots,but isozyme bands in lowland rice roots aremore than that in upland rice roots.Total area of POD isozyme bands at seedling and tilleringstages of upland rice is larger than that of lowland rice.The POD isozymes zymogram in the rootof upland rice is more stable compared with that of lowland rice.More COD isozyme numbershave been showed in roots of upland rice,but the relative activity of COD isozyme in roots of up-land rice at the seedling and tillering stages is lower than that of lowland rice.COD7 band can beshown in upland rice root at flowering stage,but it is absent in lowland rice.Therefore,COD7,band can be taken as an index for resistance of the upland rice to drought environment.

Comparison on Esterase Isozymes and Peroxidase Isozymes of Black Worm Tenebrio molitor Linnaeus at Different Developmental Stages

[ Objective ] To respectively compare esterase isozymes and peroxidase isozymes among black worm Tenebrio molitor Linnaeus （ Tenebno molitor L. ） at different developmental stages and thus provide a theoretical basis for species identification and large-scale farming. [ Method] The esterase isozymes and peroxidase isozymes of black worm Tenebno molitor L. at different developmental stages were respectively analyzed by polyacrylamide gel electrophoresis. [ Result] Both esterase isozymes and peroxidase isozymes were different among black worm Tenebrio molitor L. at developmental stages, larvae in different instars or between the male adults and the female adults. [ Conclusion] The esterase isozymes, peroxidase isozymes and physiological characteristics of black worm Tenebrio molitor L. varies with developmental stages.

Study on the isozymes in two species of prawns

This paper studies the activity and absolute content of six kinds of isozymes, such as lactate dehydrogenase,malate dehydrogenase, glucose-6-phosphate dehydrogenase, glutamate dehydrogenase, isocitrate dehydrogenase, alcoholdehydrogenase in Penaeus chinensis and P. japonicus. And by comparing, this study approaches the mechanism of high-er bought-endurance in P. japonicus.

Effect of copper on the development, protein and esterase isozymes of Drosophila melanogaster

Copper is an important ingredient in fungicides,which are used for many economic plants.However,there is concern about the side-effects of copper-based fungicides due to their potential to affect beneficial mites.The purpose of the present paper was to investigate the effects of copper on the development of the fruit fly,Drosophila melanogaster.It was found that higher doses of copper significantly prolonged the developmental time of the fruit flies,especially during the larval stages.When 320 mg L-1 copper-contaminated food was used,most of the larvae died when they were small and before the pupal stage.The protein contents and esterase isozymes extracted from the larvae changed according to the doses of copper.The small-molecular-weight protein bands gradually became weaker or were lost as the copper levels increased.However,low doses of copper stimulated a stronger expression of a few proteins.These results indicate that low doses of copper generally have no lethal effects on D.melanogaster because a specific group of genes,which encode specific proteins,are probably activated in order to withstand the onslaught of stressful conditions.At high doses of copper in food(e.g.320 mg L-1),fly development and viability are significantly affected.

Characteristics of Cellulases from Anoplophora glabripennis Motsch (Coleoptera: Cerambycidae)

The property of major cellulases from the guts of Anoplophora glabripennis larvae have been characterized. The optimal temperatures of both β 1,4 glucosidase (β glucosidase) and endo β 1,4 endoglucanase (endoglucanase, Cx) are 40℃. The β glucosidase was optimally active at pH 4\^8, while the optimal activity of the endoglucanase occurred at pH 4 4 5 6 The endoglucanase was active with a wide range of pH and temperature, the levels of activity from 25℃ to 50℃ were more than 80%, and the activity remained 60% between pH 3 2 and pH 7 2. The endoglucanase exhibited higher thermal stability than β glucosidase. Both enzymes lose their activities by heat treatment at 60℃. Two isozymes of endoglucanase were detected in sodium carboxymethylcellulose polyacrymide gels (CMC gel) by chemical colorization, and purified by elution from the gel slices. The molecular weights of the two isozymes were estimated as 26kD and 39kD respectively. Moreover molecular characteristics of the two isozymes are currently underway.

Antioxidase System of a Rice Mutant with Low Chlorophyll b

[Objective] The mitigative effect of antioxidase system of a rice mutant with low chlorophyll b on photooxidative damage was studied.[Method] A rice mutant with low chlorophyll b and its wild type were taken as experimental materials to comparatively research their peroxide (H2O2) contents, the activity and isozymes of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in chloroplast.[Result] Compared with the wild type, there were many kinds of SOD, POD and CAT isozymes in leaf cells and chloroplast cell of mutant, and the activity of SOD, POD and CAT isozymes in leaf cells and chloroplast cell of mutant was also correspondingly higher. Under intense light condition, the H2O2 content of chloroplast in mutant was less than that in the wild type. [Conclusion] The higher activity of scavenging active oxygen can relieve the photooxidative damage made by excessive light energy of intense light on photosynthetic membrane, which is an important reason for higher photosystem Ⅱ (PS II) stability of this mutant.