Genetic and physiological variation among bean lines resistant and susceptible to bean anthracnose

Summary Electrophoresis was used to determine genetic and or biochemical variation, if any, among bean lines resistant and susceptible to anthracnose. This was based on two enzyme systems: peroxisase and esterase. It was revealed that resistant and suceptible plants differed in their band patterns and intensities. Band intensity differences occurred mainly among monomorphic bands with higher intensities expressed by susceptible plants, while band pattern differences were expressed both by resistant and susceptible plants. These differences appeared only at certain stages of development. These stages were identified as 3 and 40 days after emergence and were considered as critical stages for screening purposes. The peroxidase isozyme A₅ and the esterase isozyme C₁ at 3 days, and the peroxidase band C₁ and esterase bands A₁ and A₂ at 40 days were important because these differences could be used as genetic/biochemical markers for screening the population for resistance. Thus, electrophoretic differences could be used as a screening aid and this could save time and effort in breeding programmes. Comparisons between inoculated and non-inoculated leaves of resistant and susceptible lines indicated that infection induced changes in both the amount and kind of peroxidases even before symptoms of the disease appeared. However, there were no specific differences between resistant and susceptible lines, indicating that resistant and susceptible lines responded to infection in the same manner.     

Activity and isoenzyme composition of peroxidase and esterase in fertile and male-sterile lines of tomatoes (Lycopersicon esculentum Mill.)

A comparative study of the isoenzyme patterns of esterase and peroxidase and overall peroxidase activity in stamens of male-sterile (MS) lines of Pearson ms-35 and P ms-35aa and of the respective male-fertile (MF) tomato plants have been conducted. The study has been made at two stages of stamens development — tetrad and pollen. Higher activities of the esterase isoenzymes in the MF stamens than that of MS in both ontogeny stages have been found. The slow moving esterase isoenzymes both of the MF and the MS stamens are the major isoenzymes in the early stage and are connected with tapetum development while the fast moving esterase isoenzymes are connected with pollen formation in the later ontogeny stage. Overall peroxidase levels in the MS stamens were higher than those of MF. The peroxidase patterns of the MS lines are also characterized by the greater number of isoenzymes and also the presence of specific isoenzymes, the contrast between the MF and the MS stamens being more strongly expressed at the later stage of development. A strong similitude between esterase and peroxidase patterns behaviour in both MS lines has been found.     

黄连木雌雄株叶水溶性酚类物质和两种氧化酶活性的比较

黄连木为雌雄异株植物。为探索鉴别黄连木幼苗性别的方法,应用比色法对其雌雄株不同发育阶段叶片中水溶性酚类物质的含量、过氧化物酶(POD)活性和多酚氧化酶(PPO)相对活性进行测定和比较分析,结果表明:不同性别的黄连木叶片水溶性酚类物质含量存在极显著的差异(p<0.01);POD、PPO相对活性在雌雄株间也存在显著差异,特别在老叶中则存在极显著差异,认为黄连木雌雄株老叶中的水溶性酚类物质含量差异,可以作为其性别鉴定的依据,为进一步研究黄连木植株早期性别鉴定提供参考。     

雌雄文昌鱼同工酶的表型差异

本文应用聚丙烯酰胺凝胶电泳结合生化染色方法分析了雌雄文昌鱼中苹果酸酶、苹果酸脱氢酶、酸性磷酸酶和酯酶四种同工酶的酶谱。首次发现苹果酸酶、苹果酸脱氢酶和酸性磷酸酶表型在文昌鱼雌性和雄性个体之间存在差异 ,而在同一性别不同个体之间无差异。酯酶表型较复杂 ,不但在不同性别个体之间而且在同一性别不同个体之间都出现一定差异     

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

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

Role of peroxidase and esterase activities during cotton fiber development

Cytoplasmic and salt-extracted wall peroxidase and nonspecific esterase activities along with growth analysis were investigated during the entire period of cotton fiber development. Both the peroxidase fractions, when assayed with chlorogenic and ferulic acids as substrates, recorded low levels during the fiber elongation phase, and a close relationship between cessation of elongation growth and increase in peroxidase activity was discernible. Nonspecific esterase activity in both cytoplasmic and salt-extracted fractions, on the other hand, showed higher activity during the elongation phase, whereas during the secondary thickening phase it decreased. The role of cytoplasmic peroxidase in IAA oxidation is discussed. It is suggested that esterases and peroxidases associated with wall fractions may well be involved in turnover of phenolic acids that are cross-linked to wall polysaccharides.     

罗汉果雌雄株同工酶性别鉴定研究

采用电泳技术结合同工酶染色,分析了罗汉果雌雄株叶片的过氧化物酶同工酶、酯酶同工酶、超氧化物歧化酶同工酶、多酚氧化酶同工酶和过氧化氢酶同工酶。结果表明:罗汉果雌雄叶片在同工酶谱上,存在着与性别性状相关的酶带;雌雄间的差异酶带在每一种同工酶中均有一条以上,可作为罗汉果雌雄株间的性别鉴定。此外,还比较了高产、低产、不结果雌株之间同工酶的酶带和活性差别。     

六种鼠组织中过氧化物酶和酯酶变异的比较

本文研究姬鼠属（Ａｐｏｄｅｍｕｓ）２种鼠和鼠属（Ｒａｔｕｓ）４种鼠的心组织过氧化物酶和肝组织酯酶同工酶等位基因的变异。结果表明,６种鼠心组织过氧化物酶皆为４条区带,其中社鼠和白腹巨鼠的酶带泳动速度略快于褐家鼠和黄毛鼠的酶带;鼠属４种鼠的酶带间距大致相同,而姬鼠属的酶带间距略大于鼠属,这种谱型的区别就所分析的６种鼠而言,可以认为是该同工酶表现的属间区别。肝组织酯酶区带较多,其酶带数和泳动速度的差异具有明显的种间特征;泳动最快的酶带在姬鼠中是２条,在鼠属中是１条,可以认为是属间特征的区别。从９项生化指标的异同作动物配对比较,表明褐家鼠与黄毛鼠亲缘上较近,社鼠与白腹巨鼠亲缘上也较近,而且白腹巨鼠在进化上可能要比其余３种鼠属动物更接近于姬鼠属     

几丁质酶基因转化亚麻、红豆草、骆驼刺的同工酶研究

以亚麻、红豆草、骆驼刺的下胚轴为外植体,用含几丁质酶ＲＣ２４基因的根瘤农杆菌ＬＢＡ４４０４进行转化。对３种植物转化植株叶片及对照植株叶片的愈伤组织的过氧化物酶及酯酶同工酶分析表明,转基因植株叶片愈伤组织的同工酶谱与对照系相比具有明显的差异,由此推测转化系愈伤组织中可能存在变化了的基因产物,并认为导入的外源几丁质酶基因可能引起了转化材料的遗传变异。     

Sexual differentiation in Asparagus officinalis L.

Summary Using an immunological method we assayed the levels of auxin, abscisic acid and three cytokinins (transzeatin riboside, dihydrozeatin riboside, isopentenyladenosine) in flowers of female and male plants of Asparagus officinalis L. at different stages of development. The largest differences between the sexes were found for auxin: auxin content was found to be about three times higher in young male flowers than in female flowers at a corresponding developmental stage. In order to identify some of the biochemical markers linked to sex differentiation, we also examined peroxidase isoenzyme patterns during flower development. We found five flower-specific peroxidase bands, three of which appear to be localized in the anthers. In young flowers still sexually undifferentiated in their morphology these bands are present in both sexes. They subsequently rapidly disappear in the female flower (approximately at the same time as when anther development is blocked), while they persist for a much longer time in the male. The temporary presence of these peroxidase isoenzymes in female young flowers together with the large difference in auxin content indicate that the stage of the young flower is a crucial moment in the process of sex determination.     

Characterization of oxidative enzyme changes in buffalograsses challenged by Blissus occiduus

This research investigated the role of oxidative enzymes in the defense response of buffalograss, Buchlo? dactyloides (Nuttall) Engelmann, to Blissus occiduus Barber. Changes in catalase and peroxidase activity were observed in both resistant and susceptible buffalograsses in response to chinch bug feeding. Susceptible plants were shown to have a lower level of catalase activity compared with their respective control plants. By contrast, catalase activities of resistant plants were similar between infested and control buffalograsses throughout the study. Resistant plants had higher levels of peroxidase activity compared with their control plants, whereas peroxidase activities for control and infested susceptible plants remained at similar levels or were slightly lower for infested plants. These findings suggest that chinch bug feeding leads to a loss in catalase activity in susceptible buffalograsses. In contrast, resistant buffalograsses may be able to tolerate chinch bug feeding by increasing their peroxidase activity. Polyphenol oxidase activities were similar between control and infested plants for the buffalograsses evaluated. Among the enzymes examined, no differences in isozyme profiles for peroxidase and polyphenol oxidase were detected between control and infested 378, NE91-118, Cody, and Tatanka plants. Gels stained for catalase identified differences in the isozyme profiles of infested and uninfested 378 plants; however, infested and control NE91-118, Tatanka, and Cody plants has similar isozyme profiles. No differences in protein profiles were observed between chinch buginfested 378, NE91-118, Cody, and Tatanka plants and their respective uninfested controls.     

Peroxidase and IAA Oxidase Activity and Isoenzyme Patterns in Cucumber Plants, as Affected by Sex Expression and Ethephon

Peroxidase and IAA-oxidase activity, electrophoretic patterns of total proteins and isoenzymes and the effect of Ethephon (= Ethrel, 2-chloroethane phosphonic acid) on these patterns were compared in extracts of monoecious and gynoecious cucumber plants. The activity of both peroxidase and IAA oxidase was greater in gynoecious than in monoecious plants. Ethephon treatments given at the 2-leaf stage increased peroxidase activity, especially in monoecious plants, and decreased IAA-oxidase activity. Ethephon treatment did not affect total protein or isoenzyme patterns, but increased band intensity, especially that of one band in monoecious plant extracts. No differences between monoecious and gynoecious plants were found in total protein, peroxidase and IAA-oxidase isoenzyme patterns. Peroxidase and IAA-oxidase activity sites on the gel were similar.     

黄连木雌、雄株内源植物激素和POD同工酶的比较

为揭示黄连木（Pistacia chinensis Bunge）不同性别植株的生理差异,以雌株和雄株叶片为试材,对叶片赤霉素（GA3）、脱落酸（ABA）、吲哚-3-乙酸（IAA）、亚精胺（Spd）、精胺（Spm）和腐胺（Put）含量变化及过氧化物酶同工酶进行了测定。结果表明,黄连木雌、雄株叶片GA3含量在6至9月均达到极显著差异,含量均在10月8日达到最大值,分别为100.9μg/g FW和92.9μg/g FW;雄株叶片ABA含量均高于雌株,雌、雄株叶片ABA含量在5月21日和7月8日达到极显著差异,5月21日均为最小值,含量分别为222.0 ng/g FW和340.1 ng/g FW;雄株叶片IAA含量均高于雌株,雌、雄植株叶片中IAA含量在8月8日达到极显著差异,9月8日达到最大值,分别为984.8 ng/g FW和1000.6 ng/g FW;雌株GA3/ABA的比值在7月至9月极显著高于雄株,9月8日差异值最大,为42.8;雌、雄株叶片IAA/ABA在5月21日差异值最大,为0.54;雌、雄株叶片Spd含量存在显著或极显著差异,均在7月8日达到最大值,分别为47.9μg/g FW和42.8μg/g FW;雄株叶片Spm含量均大于雌株,雌、雄株叶片Spm含量在10月8日达到极显著差异,均在7月8日出现最大值,分别为24.3μg/g FW和34.5μg/g FW;雄株叶片Put含量在各个时期均低于雌株,雌、雄株叶片Put含量除5月21日外,存在显著或极显著差异,在9月8日均达到最大值,分别为156.0μg/g FW和135.1μg/g FW。雌、雄株叶片、叶柄过氧化物酶同工酶有显著差异,雌株叶片过氧化物酶同工酶有5条带,分别为POD-1、POD-2、POD-3、POD-4、POD-5;雄株叶片只有4条带,分别为POD-2、POD-3、POD-4、POD-5;雌株叶柄内的过氧化物酶同工酶有4条带,分别为POD-1’、POD-3’、POD-4’、POD-5’,雄株叶柄内的过氧化物酶同工酶有3条带,分别为POD-2’、POD-4’、POD-5’。POD-1为雌株叶片特有条带,POD-1’、POD-3’为雌株叶柄特有条带,POD-2’为雄株叶柄特有条带。通过试验得出,黄连木叶片中ABA、IAA、Spd、Spm和Put含量与性别有关,叶片和叶柄的POD同工酶条带也是鉴别黄连木性别的有效指标。     

Extracellular Enzyme Activities during Lignocellulose Degradation by Streptomyces spp.: A Comparative Study of Wild-Type and Genetically Manipulated Strains

The wild-type ligninolytic actinomycete Streptomyces viridosporus T7A and two genetically manipulated strains with enhanced abilities to produce a water-soluble lignin degradation intermediate, an acid-precipitable polymeric lignin (APPL), were grown on lignocellulose in solid-state fermentation cultures. Culture filtrates were periodically collected, analyzed for APPL, and assayed for extracellular lignocellulose-catabolizing enzyme activities. Isoenzymes were analyzed by polyacrylamide gel electrophoresis and activity staining on the gels. Two APPL-overproducing strains, UV irradiation mutant T7A-81 and protoplast fusion recombinant SR-10, had higher and longer persisting peroxidase, esterase, and endoglucanase activities than did the wild-type strain T7A. Results implicated one or more of these enzymes in lignin solubilization. Only mutant T7A-81 had higher xylanase activity than the wild type. The peroxidase was induced by both lignocellulose and APPL. This extracellular enzyme has some similarities to previously described ligninases in fungi. This is the first report of such an enzyme in Streptomyces spp. Four peroxidase isozymes were present, and all catalyzed the oxidation of 3,4-dihydroxyphenylalanine, while one also catalyzed hydrogen peroxide-dependent oxidation of homoprotocatechuic acid and caffeic acid. Three constitutive esterase isozymes were produced which differed in substrate specificity toward α-naphthyl acetate and α-naphthyl butyrate. Three endoglucanase bands, which also exhibited a low level of xylanase activity, were identified on polyacrylamide gels as was one xylanase-specific band. There were no major differences in the isoenzymes produced by the different strains. The probable role of each enzyme in lignocellulose degradation is discussed.     

根瘤菌属几种同工酶的研究

本文对根瘤菌属12株不同根瘤菌的过氧化物酶,谷氨酸草酰乙酸转氨酶及酯酶同工酶进行了测定,结果发现:所有菌株的过氧化物酶和谷氨酸草酰乙酸转氨酶均呈阴性。酯酶同工酶酶带数在快生型和慢生型根瘤菌之间存在明显的区别,在YMA培养基上,快生型根瘤菌仅有1—2条,迁移率在0.71—0.98之间变化;而慢生型根瘤菌却有4—6条酶带,迁移率在0.15—0.98之间变化,同一菌株在不同碳源上酯酶同工酶酶带数也发生显著变化。     

Effect of drought on proteins and isoenzymes in rice during germination

Two drought tolerant varieties TKM-1 and TKM-2 and two drought susceptible varieties Jaya and Improved Sabarmati of rice were studied for soluble protein pattern and isoenzymes of malate dehydrogenase, glutamate dehydrogenase, esterase and peroxidase during germination at different water stress. MDH, GDH and esterase patterns were not affected, but the soluble proteins were changed. Peroxidase isoenzyme pattern from drought tolerant and susceptible varieties showed characteristic differences. The intensity of bands with higher electrophoretic mobility decreased in Jaya and Improved Sabarmati while in TKM-1 and TKM-2 the intensity of these bands did not change much after 72 hr water stress. In shoots of Jaya and Improved Sabarmati, the activity of the peroxidase isoenzymes decreased more than in TKM-1 and TKM-2 shoots with increase in water stress.     

The Relationship Between Peroxidase Activity and the Resistance of Tomatoes (Lycopersicum esculentum) to Verticillium dahliae

The levels of peroxidase activity in roots of non-infected tomato plants resistant to Verticillium dahliae Kleb., were found to be considerably higher than those in the roots of susceptible plants. Similar differences were found in the leaves but were not significant as in the roots. Peroxidase activity increased in leaves, stems and roots of both susceptible and resistant plants after infection with V. dahliae. The ratio of the increase in leaves and roots of susceptible plants was greater than those of resistant plants. A positive correlation was found between the peroxidase activity in the roots of eleven cultivars/breeding lines and presence of Ve gene.     

Sex-specific physiological, allocation and growth responses to water availability in the subdioecious plant Honckenya peploides

The gender of dimorphic plant species is often affected by ecophysiological variables. Differences have been interpreted as a response of the sexes to meet specific resource demands associated with reproduction. This study investigated whether sex‐specific variations in ecophysiological traits in response to water availability determine the performance of each sex in different habitats, and therefore promote extreme spatial segregation of the sexes in the subdioecious plant, Honckenya peploides. Twenty‐seven plants of each sex were individually potted in dune sand and assigned randomly to one of three water treatments. Well‐watered plants were watered daily to field capacity, whereas plants in the moderate and high‐water stress treatments received 40% and 20%, respectively, of the water given to well‐watered plants. Photochemical efficiency, leaf spectral properties and components of relative growth rate (leaf area ratio and net assimilation rate) were measured. Photochemical efficiencies integrated over time were higher in male than in female plants. Water deficit decreased maximum quantum yield in female plants more rapidly than in male plants, but female plants (unlike male plants) had recovered to initial values by the end of the experiment. Maximum quantum yield in male plants was more affected by water stress than in female plants, indicating that male plants were more susceptible to photoinhibition. The two sexes did not differ in growth rate, but male plants invested a higher proportion of their biomass in leaves, had a higher leaf area per unit biomass and lower net assimilation rate relative to female plants. Female plants had a higher water content and succulence than male plants. Differences in stomatal density between the sexes depended on water availability. The results suggest that the two sexes of H. peploides have different strategies for coping with water stress. The study also provides evidence of sex differences in allocation traits. We conclude that between‐sex differences in ecophysiological and allocation traits may contribute to explain habitat‐related between‐sex differences in performance and, therefore, the spatial segregation of the sexes.     

Characterization of Common Carbohydrate Antigenic Determinants on Soya Bean Cell-Wall Enzymes

Three soya-bean (Glycine max) cell-wall enzymes (ß-glucosidase,pectin methyl esterase and phosphatase) have been found to beglycoproteins. The polyclonal antibodies raised against pectinmethyl esterase and ß-glucosidase lacked specificity,cross-reacted highly with native enzymes and also both reactedwith pure soya-bean phosphatase, horseradish peroxidase andhoneybee venom phospholipase A2. They did not react with eithernon-glycosylated bacterial phosphatase or deglycosylated cell-wallenzymes. The two antisera contained both non-specific anti-glycanantibodies and specific anti-polypeptide antibodies that werequantified. Antiglycan antibodies specific to 1–3 fucoseand ß1–2 xylose were detected in both antiseraand were separated and quantified. The occurrence of terminalfucose (and mannose) was confirmed with specific lectins. Theseresults indicate that most of the common glycan epitopes probablycorrespond to the asparagine-linked complex glycan previouslydetected in several glycoproteins of plants as well as in thoseof molluscs and insects. (Received March 10, 1993; Accepted November 5, 1993)     

Differential identification of mouse granulocyte (CFU-g) and macrophage (CFU-m) precursors in plasma clots

Because benzidine and its derivatives have possible carcinogenic activity, a safe method is needed to demonstrate endogenous peroxidase activity. Colonies derived from mouse bone marrow cells in plasma clot culture were classified as granulocyte (CFU-g) or macrophage (CFU-m) precursors by peroxidase and naphthol AS acetate (NASA) esterase staining, respectively. Endogenous peroxidase activity was measured using benzidine or p-phenylenediazine-pyrocatechol (PPD-PC). The effectiveness of peroxidase staining with both reagents was evaluated under several conditions, and the enzyme property was confirmed by inactivation with a variety of inhibitors. The level of peroxidase activity did not differ significantly between PPD-PC and benzidine. Colony number and number of cultured cells were strongly correlated (P greater than 0.983). We conclude that PPD-PC safely demonstrates peroxidase activity in cultured cells and is as accurate, reliable, and efficient as benzidine.