玉米茎腐病菌产生的细胞壁降解酶的致病作用

 本文通过酶活分析和超微结构观察证明了玉米茎腐病菌能产生一系列细胞壁降解酶(CWDE),并能明显浸解胚根组织。浸解的组织发生质壁分离及原生质外流。随细胞壁降解酶浓度增加,酶浸解能力增强,两者呈显著的正相关。相比较而言,玉米赤霉病Fusarium graminearum在病株内产生的细胞壁降解酶的浸解能力比玉米腐霉茎腐病Pythium aphanidermatum强。2种病菌活体外产生的果胶酶与纤维素酶(Cx)存在明显的协同作用。毒素不能提高细胞壁降解酶的浸解能力,反而有一定的抑制作用。     

Maceration by Erwinia aroideae

Calcium which activates enzymes that split chains of pectic substances also increases the activity of macerating enzymes produced in culture byErwinia aroideae. EDTA suppresses both types of activity. Separation on carboxymethylcellulose gave a number of fractions in each of which there was reasonably close correspondence between macerating, chain-splitting and polygalacturonic acid transeliminase activities. One of the more active fractions released substances with an absorption peak at 235 mμ from purified potato cell walls as it did from polygalacturonate.Macerating solutions degraded neither cellulose nor a selection of hemicelluloses.The sum of this and related evidence suggests that a polygalacturonic acid transeliminase is important in maceration of potato parenchyma.     

Digalacturonates from pectin degradation induce tissue responses against potato soft rot

The pathogenicity of the bacteriumErwinia carotovorasubsp.atroseptica, which causes potato soft rot, is triggered by short oligogalacturonates released by enzymic degradation of plant cell wall pectin. In the first stage unsaturated digalacturonate (uDG), produced by the action of pectate lyases, is degraded by oligogalacturonide lyase (OGL) to keto-deoxyuronate (DKI). The OGL encoding gene fromE. carotovoraand the corresponding recombinant enzyme were characterized. Measuring the changes in plant cell viability and tissue maceration during soft rot pathogenesis in tissue slices of sprouting potato tubers, it was observed that exposure to uDG and DKI, produced by recombinant OGL, killed up to 30% of the plant cells over a period of 16h. This protected the tissue against maceration byE. carotovorasubsp.atroseptica. Endogenous OGL activity was detected in extracts from sprouting tubers where it may be involved in the conversion of uDG into cell toxic compounds. The results indicate that an additional function of small, diffusable digalacturonates is to induce plant cell death during the rotting process, thus contributing to defence reactions againstE. carotovorasubsp.atroseptica.     

Microcalorimetric studies of tissue cells and bacteria

Analytical microcalorimetric studies of cultures of Enterobacter aerogenes and Clostridium pasteurianum revealed a complex pattern of heat production. While both organisms gave thermograms which followed the exponential growth curves in the early stages, the later events in the heat effect appeared to reflect deficiencies in the growth medium and induction of “secondary” nutrients. Synchronously dividing Schizosaccharomyces pombe is known to give maxima in oxygen uptake during the cell cycle. These peaks are enhanced by the uncoupling agent, 2-oxomalononitrile m-chlorophenylhydrazone (CCCP). The rate of heat production in control cultures pumped through the LKB microcalorimeter increased at a linear rate, doubling over each complete cell cycle. Introduction of CCCP to cultures gave an altered thermogram in which the maxima of heat evolution which occurred corresponded in time to the peaks in oxygen uptake. When suspensions of muscle fibroblasts from 9 day old chick embryos were pumped through the aeration vessel, they produced heat at a rate which was proportional to cell concentration. Inhibitors of respiration decreased heat evolution by similar percentages to their effect on oxygen uptake. Cells of the same type in monolayer gave slightly lower levels of heat production. The thermogram was characterised by maxima over 4 h, which may be a reflection of temporal changes in intracellular ATP levels. When muscle fibroblasts were mixed with excess (zero order kinetics) ATP and pumped through the gold vessel of the microcalorimeter, the rate of heat production was proportional to cell concentration, the latter representing the hydrolysing enzyme. 4,4′-Dithiodi(nicotinic acid), a sulphydryl inhibitor which does not enter cells, depressed the calorimetric response vs cell concentration curve by 78%, indicating that a sulphydryl enzyme was present at the cell surface. Batch calorimetric experiments to determine the heat of interaction of antibodies with cells gave thermograms which differed in form and value, the latter changing in sign and magnitude. The results illustrate the need for careful model experiments to disect this complex system. The present results underline the value of microcalorimetry in giving analytical data on growth and respiration in cells and organisms and illustrates the potential of the technique in studies of surface-localised reactions.     

Selective alterations of membrane properties of cultured human liver cells caused by the insecticide DDT

A variety of membrane-specific parameters was examined in both intact cells and isolated plasma membranes following exposure of cultured human liver cells to the insecticide 1,1-(2,2,2-trichloroethylidene)bis(4-chloro)benzene (DDT). Uptake of DDT was at equilibrium within 6 hr. In contrast, a decrease in the number of β-adrenergic hormone receptors first became significant after 48 hr of cell exposure. Whereas the uptake was largely reversible, the loss in the number of β receptors did not recover after DDT-exposed cells were cultured in fresh medium lacking the insecticide. Experiments in vitro substantiated the time lag of the biological effect. The decrease in receptor proteins was persistent in membranes with increased phospholipid unsaturation. Temperature-activity profiles (“Arrhenius plots”) of $\text{Na}{}^{\mathrm{+}}\text{K}{}^{\mathrm{+}}\text{-ATPase}$ and 5′-nucleotidase were unchanged. Endogenous tryptophan fluorescence of membrane proteins was lower in membranes from DDT-exposed cells. These selective alterations in membrane parameters suggest a specific interaction of DDT with membrane proteins; interference with cellular protein synthesis is possible. The results indicate that membrane lipid “fluidization” does not play a physiologically important role in the mechanism of DDT action in biomembranes.     

Immunohistochemical investigation of cotton carpel tissue exposed to xylanolytic hydrolases of Aspergillus flavus

Cotton carpel tissue (35–45 days post-anthesis) that had been treated with a mixture of xylanolytic hydrolases derived from Aspergillus flavus was subjected to immunocytochemical analysis. Microscopic examination of treated tissues revealed severe degradation of the secondary wall structure. Control tissue cells revealed the presence of high concentrations of xylans/arabinoxylans throughout the cell wall, as well as significant concentrations of arabinogalactan proteins in secondary wall structure. Carpel cells treated with a mixture of A. flavus-produced xylanolytic hydrolases showed a much reduced presence of labeling by xylan-specific antibodies on the inner wall surface, suggesting a severe loss of these plant polysaccharides in the secondary wall structure. Carpel exposure to a purified 14 kD endoxylanase from A. flavus also resulted in a severe reduction of xylans from secondary wall structure, although penetration of the tissue was not as dramatic. Arabinogalactan proteins were not as severely affected by the xylanolytic hydrolases. Comparison of control tissue with hydrolase-treated tissue stained with toluidine blue revealed an apparent reduction in wall thickness, supporting the conclusion of secondary wall structure degradation. Interestingly, the pectins could only be detected in the samples treated with xylanolytic enzymes, indicating that the pectins were being masked by xylans. These results are consistent with the conclusion that the xylanolytic hydrolase complex of A. flavus is a critical factor for host cell wall maceration and may represent another important fungal virulence factor, in addition to pectolytic hydrolase activities.     

己唑醇及其对映体对人体乳腺癌细胞的选择毒性及氧化损伤研究

在对映体水平上研究己唑醇对人体乳腺癌细胞(MCF-7)的选择毒性及氧化损伤.以MCF-7细胞作为受试对象,采用Cell Counting Kit-8(CCK-8)试剂盒测定经己唑醇对映体处理后的细胞毒性;采用氧化损伤相关试剂盒测定经己唑醇对映体处理后,细胞内乳酸脱氢酶(lactate dehydrogenase,LDH...     

Effects of temperature,water regime and irrigation system on the release of ascospores of Mycosphaerella nawae,causal agent of circular leaf spot of persimmon

Experiments were conducted under controlled conditions to quantify the effects of temperature, water regime and irrigation system on the release of Mycosphaerella nawae ascospores from leaf litter in Spanish persimmon orchards. The effect of temperature on ascospore release was best described by a Gompertz model. The end of the lag phase of ascospore release occurred at 9·75°C, and the end of the exponential phase at 15·75°C. Few ascospores were discharged from dry leaves wetted with 0·1 or 0·5 mm water, but significant amounts were recovered with 1–50 mm water. About half of the total ascospores were released after three wetting and drying cycles, but 32 cycles were necessary for a complete discharge. No significant difference in ascospore release was detected when the leaf litter was wetted by flood and drip irrigation. However, considering the proportion of soil area wetted in both systems, inoculum release was significantly reduced by drip irrigation. The potential of drip irrigation as a cultural control measure should be investigated.     

空气湿度对灰霉菌侵染番茄叶片组织结构分析及抗氧化系统的影响

空气湿度对番茄灰霉病的发生有显著性影响。为了探明空气湿度对灰霉菌侵染番茄叶片的过程与机理,本研究以‘金棚14-6’番茄为材料,观察分析了高空气相对湿度(80%～95%)和低空气相对湿度(65%～80%)对灰霉菌侵染番茄叶片表型变化、细胞学差异、形态结构变化、活性氧含量和抗氧化酶的影响。结果表明：高湿接种60 h大量芽管伸长出现在叶片下表皮细胞并分化菌丝,叶肉细胞间隙分布了大量的菌丝并伴随病斑出现,灰霉菌在60 h完成侵染;低湿接种108 h芽管伸长出现在叶片下表皮并分化菌丝,叶肉细胞间隙有少量菌丝分布,没有明显病斑出现。随着灰霉菌的侵染,低湿与高湿相比栅栏组织和海绵组织结构从整齐紧密变为排列疏松的时间滞后;高湿和低湿处理的叶片厚度、栅栏组织和海绵组织厚度均呈先上升后下降的变化趋势,侵染后期低湿处理的叶片组织结构厚度显著高于高湿。随着接菌时间延长,高湿和低湿的活性氧含量和抗氧化酶活性处于相对活跃的调整、适应的变化过程,大致呈先上升后下降趋势,而对照和接菌相比无显著差异,变化趋势维持在基本的振幅上。研究显示灰霉菌发生的环境条件：湿度80%～95%和侵染完成时间60 h,即控制高空气湿度的持...     

Effects of the fungus Crinipellis perniciosa, causal agent of witches' broom disease, on cell and tissue cultures of cocoa (Theobroma cacao L.)

The symptoms of witches' broom disease in cocoa, caused by the Basidiomycete fungus Crinipellis perniciosa , are pronounced swelling of the terminal and axillary buds followed in the long term by necrosis of this tissue. The direct effect of C. perniciosa on cocoa cells was examined under controlled conditions by growing primary and secondary phase cultures of the fungus separately and also with callus cultures and with cell suspensions. Both primary and secondary phase mycelium reduced growth of callus cultures by about 47% after one week compared with the controls. However, cell suspensions containing primary phase mycelium showed initial growth double that of the uninfected controls after 5 days, but then growth was reduced below that of the control and particularly when the primary phase became secondary phase mycelium. This change in fungal development coincided with the time that the cell culture reached the stationary growth stage. Cell cultures inoculated with stationary phase mycelium showed the same growth as the control after 5 days but then growth was reduced to 50% of the control after 19 days incubation and remained at this low level subsequently. The inhibitory effect of secondary phase mycelium was examined by incubating callus and cell suspensions with culture filtrate from liquid cultures of the secondary phase. Inclusion of 50% by volume of culture filtrate from the secondary phase in the growth medium for callus and cell suspensions, respectively, resulted in a reduction in growth of the plant tissue cultures. Addition of fungal culture filtrates also led to loss in potassium and loss of viability of cell suspensions and of isolated cells as represented by protoplasts. The necrotrophic mode of the secondary phase may be achieved through the production of phytotoxins acting on the host cell membrane.     

The bleaching induced by the phenylpyridazinone herbicide Metflurazon in Chlorella is a metabolic and reversible process

The treatment of synchronized cells of the green alga Chlorella fusca under photoautotrophic conditions with metflurazon (SAN H 6706; 4-chloro-5-dimethylamino-2-(α,α,α-trifluoro-m-tolyl-3(2H)-pyridazinone)) induces a bleaching process and results in white-appearing cells. This process of bleaching was followed by quantitative analysis of cell growth and cell division, photosynthetic oxygen evolution, respiratory oxygen consumption, and of pigment pattern at 0, 6, 12, 18, 24, 48, and 72 hr after incubation with different concentrations of metflurazon. Increasing concentrations of metflurazon gradually affected cell growth of Chlorella measured as increase in cell diameter. Cell division was inhibited completely with 1, 10, and 100 μM metflurazon. Photosynthetic oxygen evolution and respiratory oxygen consumption were not inhibited by 1 μM metflurazon during the first 6 hr; after this time a gradually increased inhibition was observed. Both parameters were inhibited by 100 μM metflurazon immediately after herbicide addition. A detailed analysis of the pigment content during the bleaching process revealed that: (a) The bleaching of Chlorella cells by metflurazon is not a simple photochemical process like the photobleaching of boiled cells, but is directed by the active metabolism of Chlorella itself. (b) The bleaching process is characterized by two phases: an accumulation of pigments followed by their degradation. The accumulation phase extends to 6 hr after herbicide addition. (c) During the accumulation phase, chlorophyll is accumulated to 380 and 106% in cells treated with 1 and 100 μM metflurazon, respectively, compared to the initial pigment content. The breakdown of chlorophyll, however, during the degradation phase is 5 times faster in the 1 μM treatment than in the 100 μM treatment. This difference resulted in the faster appearance of white cells with the low metflurazon concentration. (d) During the accumulation phase in the 1 μM treatment, the biosynthesis of chlorophylls, xanthophylls, and carotenes is inhibited by 56, 74, and 78%, respectively, when compared to a nontreated control. When related to the initial amounts, chlorophylls, xanthophylls, and carotenes are accumulated to 380, 230, and 153%, respectively. However, the synthesis of violaxanthin is specifically inhibited, followed by α-carotene. During the degradation phase, violaxanthin and α-carotene again, are the most rapidly disappearing pigments. Continuous culturing of white Chlorella cells resulted in a regeneration to green cells after 96, 240, 384 hr for 1, 10, and 100 μM metflurazone, respectively. The bleaching of Chlorella by metflurazon is evidently dependent on a functioning metabolism and is itself a regulated disassembly of the photosynthetic apparatus, which is reversible and not lethal.     

Maceration of plant tissue by fungi is inhibited by recombinant antipectinase antibodies

Polyclonal antiserum from mice immunized with extracellular proteins from Rhizoctonia solani inhibited pectinase and cellulase activities in cell free culture supernatants of Rhizoctonia solani. Spleen mRNA from these mice was used to construct a cDNA library from which antipectinase ScFv antibodies were isolated using phage display techniques. Soluble ScFv antibodies produced by individual clones in Escherichia coli inhibited polygalacturonase in the culture supernatants of a range of fungal pathogens, including ascomycetes, basidiomycetes, and oomycetes. The soluble antibodies also inhibited maceration of potato tissue by these pathogens.     

Mitochondrial involvement in the mode of action of acifluorfen

The herbicidal action of acifluorfen {5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitrobenzoic acid} was studied with greened and expanded discs from cotyledons of cucumber (Cucumis sativus L.). Discs were floated on various treatment solutions for 20 hr in darkness before exposure to 400 μE m^?2 sec^?1 of white light. Herbicide damage, as measured by electrolyte leakage, began in the light after a 1- to 2-hr lag period. Cytochemical methods at the ultrastructural level indicated that acifluorfen caused marked increases in production of superoxide radical and hydrogen peroxide in the mitochondrion, but not in the plastid. The mitochondrial inhibitors antimycin A, rotenone, CCCP, and DNP antagonized the action of acifluorfen, lengthening the lag period and reducing the rate of electrolyte leakage. Ethanol, α-tocopherol, N-[2-(2-oxo-1-imidazolidinyl)ethyl]-N′-phenylurea, and copper-penicillin also lengthened the lag phase and slowed the rate of damage, indicating that acifluorfen damage involves toxic oxygen species. PS II-inhibiting levels of DCMU, atrazine, or bentazon did not affect acifluorfen-induced ion leakage. Yellow tissue produced by treatment with tentoxin was supersensitive to acifluorfen, but white tissue produced by treatment with norflurazon was relatively insensitive. These data indicate that, after an initial carotenoid-acifluorfen interaction, the mitochondrion is involved in production of toxic oxygen species and that this process is closely tied to the mechanism of action of this herbicide.     

芦笋茎枯病病菌鉴别寄主的构建

正芦笋(Asparagus officinalis Linn)又称石刁柏,百合科天门冬属,是世界十大名菜之一,在国际市场上被称为"蔬菜之王"。由于其营养价值高,并能润肺、镇咳、祛痰,具有抑制肿瘤生长的药理功能,深受人们的喜爱[1]。近年来,随着芦笋市场与栽培面积的扩大,病害的发生也逐年加重,尤其茎枯病的严重发生已影响芦笋的产量与质量,轻者生长发育不良,降低产量与品质,重者病株提前枯死,全田毁灭,严重影响芦笋生产与出口创汇[2]。目前该     

Polygalacturonase Isozymes Produced by Phomopsis cucurbitae in Relation to Postharvest Decay of Cantaloupe Fruit

ABSTRACT Production of polygalacturonase (PG), a cell wall-degrading enzyme, by Phomopsis cucurbitae (latent infection fungus) was studied in relation to different carbon sources and various stages of cantaloupe fruit development. P. cucurbitae produced multiple PG isozymes both in vitro and in vivo. The fungus produced the highest PG activity and the greatest number of isozymes on pectin compared with those produced on glucose, galactose, and sucrose. Eight P. cucurbitae PG isozymes (pIs 3.7, 4.2, 6.6, 7.0, 7.3, 7.5, 7.8, and 8.6) were detected in extract from inoculated mature fruit (40 days after anthesis) by isoelectric focusing. Isozyme bands with pIs of 4.2, 7.3, and 7.8 were the most prominent. A similar set of PG isozymes was produced by P. cucurbitae in autoclaved mature fruit tissue (mesocarp). When tissue discs taken from 20-, 30-, 40-, and 50-day postanthesis fruit were inoculated with P. cucurbitae, PG activity and the number of PG isozymes extracted from the macerated fruit tissue discs increased with the degree of fruit maturity and ripening. Increases in PG activity and PG isozymes were also correlated with reactivation of latent infections and the beginning of tissue maceration. An anionic PG isozyme (pI 4.2) was only visualized on decayed 50-day-old fruit exocarp, as well as 40- and 50-day-old fruit mesocarp. The experimental results support the hypotheses that P. cucurbitae PG isozymes play an important role in fruit decay once latent infection becomes active following harvest.     

Factors that influence the persistence of TCA in soil

The persistence of TCA in soil was examined using the pyridine-alkali colorimetric procedure. Loss of TCA was essenlially similar when determined by colorimetry, chloride release or bioassay. Persistence of TCA when incubated with soil at 25°C or sprayed on the soil surface in the field was slightly influenced by soil type. Degradation of TCA occurs largely by microbial action after a lag phase. Soils treated with TCA acquire the ability to degrade further additions of ihe compound without a lag phase. The three soils examined still possessed this ability to differing extents 32 months after they had been sprayed once with TCA at the rate of 22.4 kg/ha. In one experiment the persistence of TCA was shortened appreciably in a plot that had twice previously been sprayed with TCA.     

Performance of Bangladesh indigenous rice in a weed infested field and separation of allelopathy from resource competition

This study aimed to identify the potential allelopathic indigenous rice (Oryza sativa L. ssp. indica) varieties from Bangladesh using a performance study in a weed‐infested field and to assess the extent of allelopathic interference relative to resource competition in a glasshouse experiment. Six varieties – namely, “Boterswar,” “Goria,” “Biron” and “Kartiksail” as the most allelopathic, “Hashikolmi” as weakly allelopathic and “Holoi” as nonallelopathic – were raised following a nonweed control method. The infestation levels of weed species were calculated using Simpson's Diversity Index (SDI), which ranged from 0.2 to 0.56. However, a significant correlation coefficient (0.87, P < 0.001) was obtained from these field data compared with the root inhibition percentage from the laboratory bioassay, and the “Boterswar” variety was the most allelopathic. The interactions between the allelopathic variety “Boterswar,” weakly allelopathic variety “Hashikolmi” and Echinochloa oryzicola via a target (rice)‐adjacent (E. oryzicola) cogrowth culture were determined in a hydroponic arrangement. The relative competitive intensity (RCI) and the relative neighbor effect (RNE) values showed that the crop–weed interaction was facilitation for “Boterswar” and competition for “Hashikolmi” and E. oryzicola in rice/E. oryzicola cogrowth cultures. The allelopathic effects of “Boterswar” were much higher than the resource competition in rice/E. oryzicola cogrowth cultures. The converse was observed for “Hashikolmi.” Moreover, the mineral content of E. oryzicola was severely affected by “Boterswar”/E. oryzicola cogrowth cultures’ exudate solution. Therefore, the allelopathic potential of “Boterswar” variety might be useful for developing the weed‐suppressing capacity of rice, which will likely have a significant influence on paddy weed control.     

Microbial N-dealkylation of atrazine: Effect of exogeneous organic substrates and behaviour of the soil microflora

It has been demonstrated that atrazine side-chain mineralisation could be substantially stimulated by addition of carbon-containing substrates such as cellulose, green manure, straw or sawdust in the presence of NH₄⁺ nitrogen but poorly affected by amendments with glucose. Cellulose has the most beneficial effect. For that substrate it has been shown that (i) simultaneous application of the organic amendment and atrazine results in kinetics for side-chain dealkylation showing a lag phase which is reduced or even eliminated by preliminary incubation with the amendment, (ii) rate and extent of mineralisation of the ethylamino side chain are significantly accelerated by decreasing the C/N ratio of the amendment. By comparison, mineralisation of the isopropylamino side chain is not appreciably affected by a change in the value of the C/N ratio as far as atrazine is applied within a two- to three-week period following the organic treatment after which a small deficit in N supply has a definite beneficial effect on mineralisation. Cellulose and, to a lesser extent, straw induce a biphasic change in bacterial number with more numerous and/or active atrazine degraders being predominantly found in the later-developing bacterial community. The fungal microflora is relatively unaffected by all types of carbon substrates but glucose and, unexpectedly, by atrazine at high ratio of application. Activation of atrazine mineralisation seems to be a co-metabolic process which is kinetically controlled by the rate of release from polymerised C substrates of easily available and readily metabolisable low-molecular-weight co-substrates. Transient production of glucose as an end-product of cellulose depolymerisation might induce catabolic repression of dealkylation enzyme systems and be responsible for a lag in atrazine side-chain mineralisation. © 1998 Society of Chemical Industry