泡囊丛枝(VA)菌根对玉米根际磷酸酶活性的影响

以玉米为材料,利用三室隔网培养方法,研究了缺P土壤上施用植酸和卵磷脂时接种几种菌根真菌(Glomus mosseae, Glmous versiformea, Gigaspora margarita)对根际土壤酸性磷酸酶和碱性磷酸酶活性的影响.玉米生长70d后,收获测定距根表不同距离土壤中的磷酸酶活性.结果表明,接种菌根真菌增加了根际土壤酸性和碱性磷酸酶活性,Gigaspora margarita菌根菌的作用大于其它2个菌根菌.不同P源对磷酸酶活性有明显影响.     

不同磷源对红三叶草根际和菌根际磷酸酶活性的影响

以红三叶草为研究对象,利用三室培养系统,在接种菌根真菌(Glomus mosseae)的条件下研究了不同磷源对根际和菌根际磷酸酶活性的影响,植株生长8周后收获并测定根室、菌丝室的土壤磷酸酶活性、植株干重及含磷量．结果表明,根室酸性磷酸酶活性比碱性磷酸酶活性更裔,接种条件下二者都稍有增加,特别是在供给有机磷(植酸钠)的条件下明显增加了菌丝室土壤磷酸酶活性．接种菌根真菌显著增加了植株干重、磷含量和总磷吸收．施用磷酸二氢钾(KH2PO4)时菌丝吸磷量占吸磷总量的43．1%,而施用植酸钠(Na-phytate)时菌丝吸磷量占吸磷总量的60．8%。     

植酸钠对菌根真菌根内菌丝碱性磷酸酶活性及根外菌丝生长的影响

采用三室隔网培养装置,以玉米为宿主植物,接种丛枝菌根真菌(AM)(Glomus intraradices),研究了不同用量的植酸钠对AM真菌生长和代谢活性的影响．研究发现,接种AM真菌的植株地上部和根系的P浓度和吸P量,比非菌根植物的提高了1～2倍．外源植酸钠的存在,显著降低了AM真菌根内菌丝的碱性磷酸酶活性,增加了AM真菌在土壤中的菌丝密度．结果表明,外源植酸钠对根内AM真菌碱性磷酸酶活性和真菌根外菌丝的生长具有调控(增减)作用,并且AM真菌提高了植物对土壤固有养分和外源植酸钠中P的吸收和利用．     

不同pH水平下两种菌根真菌对紫云英生长的影响及其相互作用

设计在不同pH水平（4．3、5．1、5．8、6．8）下两种VA菌根真菌Glomus mosseae和Gigaspora margarita对紫云英Astragalus sinicus进行单接种、混合接种及无接种对照的盆栽实验。对紫云英地上和地下部分生物量、根部侵染率、SDH和ALP酶活进行了检测。实验结果表明：紫云英的生长效应与VA菌根真菌的侵染率及两种酶活成明显相关性。土壤pH升高,单接种Glomus mosseae和混合接种的侵染率也随之升高,而单接种Gigaspora margarita的侵染率呈现出先上升后下降的趋势。本实验设计了特异性扩增Glomus mosseae和Gigaspora margarita的引物gml和gigl,在混合接种实验中,nested PCR扩增结果显示：在低pH水平下（4．3-5．1）大多数根段为Gigaspora margarita所侵染,在高pH水平下（5．8—6．8）Glomus mosseae表现出较强的竞争力,但并没有检测到两种VA真菌存在于同一条侵染根段;对比单接种实验,在低pH水平下,Glomus mosseae显著抑制了Gigaspora margarita的侵染,而在高pH水平下Gigaspora margarita明显促进Glomus mosseae的侵染。     

不同pH水平下两种菌根真菌对紫云英生长的影响及其相互作用

设计在不同pH水平(4.3、5.1、5.8、6.8)下两种VA菌根真菌Glomus mosseae和Gigaspora margarita对紫云英Astragalus sinicus进行单接种、混合接种及无接种对照的盆栽实验.对紫云英地上和地下部分生物量、根部侵染率、SDH和ALP酶活进行了检测.实验结果表明:紫云英的生长效应与VA菌根真菌的侵染率及两种酶活成明显相关性.土壤pH升高,单接种Glomus mosseae和混合接种的侵染率也随之升高,而单接种Gigaspora margarita的侵染率呈现出先上升后下降的趋势.本实验设计了特异性扩增Glomus mosseae和Gigaspora margarita的引物gml和gigl,在混合接种实验中,nested PCR扩增结果显示:在低pH水平下(4.3-5.1)大多数根段为Gigaspora margarita所侵染,在高pH水平下(5.8-6.8)Glomusmosseae表现出较强的竞争力,但并没有检测到两种VA真菌存在于同一条侵染根段;对比单接种实验,在低pH水平下,Glomus mosseae显著抑制了Gigaspora margarita的侵染,而在高pH水平下Gigasporamargarita明显促进Glomus mosseae的侵染.     

不同pH水平下两种菌根真菌对紫云英生长的影响及其相互作用

设计在不同pH水平（4.3、5.1、5.8、6.8）下两种VA菌根真菌Glomus mosseae和Gigaspora margarita对紫云英Astragalus sinicus进行单接种、混合接种及无接种对照的盆栽实验。对紫云英地上和地下部分生物量、根部侵染率、SDH和ALP酶活进行了检测。实验结果表明：紫云英的生长效应与VA菌根真菌的侵染率及两种酶活成明显相关性。土壤pH升高,单接种Glomus mosseae和混合接种的侵染率也随之升高,而单接种Gigaspora margarita的侵染率呈现     

丛枝菌根真菌影响宿主植物蒺藜苜蓿根系酸性磷酸酶活性的跨世代效应

丛枝菌根真菌(AMF)能够通过增强宿主植物根系分泌酸性磷酸酶帮助其适应低磷环境,但这种可塑性改变能否跨世代传递并影响后代适应低磷环境,仍不清楚。本研究通过亲代实验(实验1)和子代实验(实验2),研究AMF影响宿主植物蒺藜苜蓿(Medicago truncatula)根系酸性磷酸酶分泌的跨世代效应。实验1表明,土壤低磷水平下接种AMF的亲代宿主植物根系酸性磷酸酶活性显著提高,且根际土中有更高的酸性磷酸酶活性和有效磷含量。而高磷水平下,接种AMF处理的宿主植物酸性磷酸酶活性与不接种AMF的处理无显著差异。实验2表明,在当代低磷环境下,来自亲代低磷接种AMF的后代,其根系和根系分泌的酸性磷酸酶活性显著高于来自亲代低磷不接种AMF的后代,而亲代高磷处理的后代(有AMF和无AMF)之间酸性磷酸酶活性无显著差异。在当代高磷环境下,来自亲代不同处理的后代根系和根系分泌的酸性磷酸酶活性均无显著差异。本研究表明,AMF对宿主植物根系分泌酸性磷酸酶的生理可塑性能够跨世代传递,且该跨世代效应受到亲代磷水平的影响。     

接种AM菌对西部黄土区采煤沉陷地柠条生长和土壤的修复效应

以采煤沉陷区柠条为宿主植物,研究接种丛枝菌根真菌(arbuscular mycorrhizal fungi,简称AM菌)对柠条生长和根际土壤的改良效应。结果表明:8月份接种AM菌比不接菌柠条的株高、冠幅和地径显著增加了29.11%,29.83%和14.81%,9月份接菌区柠条的根长、平均直径、根表面积和根体积分别比对照区增加了151.0%,34.2%,116.0%和129.3%。接种AM菌增强柠条的抗逆性,接菌区的柠条叶片可溶性糖含量和过氧化氢酶活性分别比对照区增加了13.4%和111.1%。8月份接种AM菌改善了土壤的生物理化性质,接菌区有机质、碱解氮、速效磷和速效钾比对照区分别增加7.06g/kg,140.0 mg/kg,1.82 mg/kg和16.72mg/kg,接种AM菌显著增加了根际土壤中真菌、放线菌、细菌数量和酸性磷酸酶活性。总之,接种AM菌促进采煤沉陷区柠条的生长和土壤的改良。     

西藏高原天然长芒草地丛枝菌根真菌接种效应

采用草地均匀打孔方法,就草地土壤未消毒条件下接种丛枝菌根（AM）真菌对长芒草(Stipa bungeana)的侵染效应以及对植物生长、吸磷效率、土壤微生物区系等的影响进行研究.结果表明,1)接种处理、不接种处理的菌根效应存在着明显的差异,多数接种处理根围土壤AM真菌孢子密度、菌根侵染率和侵染强度显著提高,但对丛枝丰度的影响相对较低.2)接种后AM真菌孢子密度对菌根侵染率具有极显著影响(r=0.7679**);随菌根侵染率的增加,植株总干物重和吸磷总量均呈极显著提高,r值分别为0.7556**、0.8018**.3)与植株地上部相比,接种AM真菌对提高根系干物重、根系吸磷量和含磷量的促进作用相对较大.4)多数接种处理根际土壤酸性磷酸酶、碱性磷酸酶活性均呈一定程度的提高,根际土壤细菌数量显著增加,真菌、放线菌的数量变化则不甚明显.5)各接种处理对寄主植物的综合侵染效应在总体上呈Glomus mosseae+G. intraradices+Scutellospora calospora＞G. mosseae+G. aggregatum＞Glomus sp.＞G. mosseae＞G. mosseae+ G. etunicatum+G. intraradices+S. erythropa＞G. geosporum的趋势.     

大豆毛状根-VA菌根真菌双重培养体系的建立

以大豆毛状根为宿主,接种VA菌根真菌珠状巨孢囊霉（Gigaspora margarita）,经过3.5个月的双重培养,观察到VA菌根真菌珠状巨孢囊霉对大豆毛状根的侵染,辅助细胞形成,并获得VA菌根真菌成熟孢子,在无菌条件下建立了大豆毛状根-VA菌根真菌双重培养体系,为研究菌根真菌侵染大豆根部形成共生体系及相关分子机制提供了一种有效的研究方法。     

Activity of acid and alkaline phosphatases (intracellular and extracellular) in rhyzosphere fungi from Arachis hypogaea (Papiloneaceae)

The potential role of the fungi, isolated from the peanut rhizosphere, in the production of extracellular and intracellular acid and alkaline phosphatase, was evaluated in vitro. Acid and alkaline extracellular phosphatases showed the highest activities, and the Penicillium species were the most efficient in their production. The correlation analysis showed that extracellular acid and intracellular acid phosphatase produced by Aspergillus niger A. terreus, Penicillium sp. y P. brevicompactum were negatively correlated; while the extracellular and intracellular phosphatase activities, were positively correlated. The extracellular acid phosphatase activities produced in vitro by majority of fungi assayed, were not correlated with the acid phosphatase activity present in the peanut soil rhizosphere. Nevertheless, the extracellular alkaline phosphatase activities produced in vitro, were negatively correlated with the extracellular alkaline phosphatase activities present in the rhizosphere. The ability of phosphatase production by fungi isolated from peanut rhizosphere suggests they have great potential to contribute to the P mineralization in this zone.     

Nucleotide sequence of the gene for alkaline phosphatase of Thermus caldophilus GK24 and characteristics of the deduced primary structure of the enzyme

The gene encoding Thermus caldophilus GK24 (Tca) alkaline phosphatase was cloned into Escherichia coli. The primary structure of Tca alkaline phosphatase was deduced from its nucleotide sequence. The Tca alkaline phosphatase precursor, including the signal peptide sequence, was comprised of 501 amino acid residues. Its molecular mass was determined to be 54? omitted?760 Da. On the alignment of the amino acid sequence, Tca alkaline phosphatase showed sequence homology with the microbial alkaline phosphatases, 20% identity with E. coli alkaline phosphatase and 22% Bacillus subtilis (Bsu) alkaline phosphatases. High sequence identity was observed in the regions containing the Ser-102 residue of the active site, the zinc and magnesium binding sites of E. coli alkaline phosphatase. Comparison of Tca alkaline phosphatase and E. coli alkaline phosphatase structures suggests that the reduced activity of the Tca alkaline phosphatase, in the presence of zinc, is directly involved in some of the different metal binding sites. Heat-stable Tca alkaline phosphatase activity was detected in E. coli YK537, harboring pJRAP.     

Boophilus microplus: characterization of larval phosphomonoesterases and isolation of subcellular fractions with high phosphatase activity.

Phosphomonoesterase activity was determined for a 115,000g pellet and soluble fractions resulting from a subcellular fractioning of a homogenate of larval Boophilus microplus. Both fractions showed maximum phosphatase activity at pH 5.5 and 10. Acid phosphatase (EC 3.1.3.2) activity was found to be greatest in the soluble fraction. When the reaction rate was plotted against homogenate concentration, the soluble acid phosphatase deviated from the linear relationship. For both fractions different thermostability patterns were obtained, inactlvation beginning for the alkaline phosphatase (EC 3.1.3.1) at 45–55 C. When the effect of substrate concentration on activity was studied, deviations from the typical hyperbolic behavior were observed. Homogenization of larvae with 5 mm EDTA buffer failed to yield a low-speed pellet with high alkaline phosphatase activity, as it is expected if absorptive structures sediment. Moreover, total alkaline phosphatase activity recovered by this method is significantly lower than activity recovered when homogenization is carried out without EDTA. Alternately, homogenization with 10 mM Tris buffer and 0.25 M sucrose gave 27,000g and 115,000g fractions with high phosphatase activity when fractioned by centrifugation. Alkaline treatment of the 115,000g fraction with 10 mM Tris buffer, pH 7.8, failed to separate endoplasmic reticulum contaminants without loss of phosphatase activity. When the 115,000g fraction was centrifuged in a sucrose density gradient, two activity peaks, coincident for both acid and alkaline phosphatases, were obtained. Antigenic analysis showed the existence of similar antigenic determinants in both peaks “immunologically” presented in different ways.     

Regional and Subcellular Distribution in Mammalian Brain of the Enzymes Producing Adenosine

The activities of 5'-nucleotidase, 2'-nucleotidase, alkaline phosphatase, and acid phosphatase were measured in rat and autopsied human brains. The four phosphatases in the rat brain showed little change in activity after death. The activities of adenosine-producing enzymes were compared in various parts of rat and human brains. When phosphatase activity was measured at pH 7.5, 5'-nucleotidase showed the highest activity in the most parts of the brain. The activity of 2'-nucleotidase and that of nonspecific phosphatase were almost the same at pH 7.5. However, higher phosphatase activity was observed in all parts of the brain when nonspecific phosphatase activity was measured at pH 10.0 or 5.5. High specific activity of 5'-nucleotidase in the brain was detected in the membranous components, especially in the synaptic membranes. The activity of 2'-nucleotidase was distributed in the soluble and synaptosomal fractions. The highest activity of both alkaline and acid phosphatases was recovered in the crude mitochondrial fraction, with the highest specific activity in the microsomal fraction. Phosphatase activity was distributed widely in the rat brain. The activity of 5'-nucleotidase was high in the medulla oblongata, thalamus, and hippocampus, but low in the peripheral nerve, spinal cord, and occipital lobe. The activity of 2'-nucleotidase was high in the vermis and frontal lobe. The highest acid and alkaline phosphatase activities were detected in the frontal lobe and in the olfactory bulb, respectively. The distribution of the four phosphatases in the autopsied human brain was similar to that in the rat brain. The highest 5'-nucleotidase activity was observed in the temporal lobe and thalamus.(ABSTRACT TRUNCATED AT 250 WORDS)     

A light- and electron-microscopic study of enzymes in the embryonic shell-forming tissue of the freshwater snail, Biomphalaria glabrata

Abstract. The mode of formation of the molluscan exoskeleton is still poorly understood, but studies on adult snails indicate that enzymes involved in vertebrate bone formation also participate in mollusc shell formation. The enzymes peroxidase, alkaline phosphatase, and acid phosphatase are expressed in a constant pattern and help to identify the different zones of the adult shell-forming tissue. The present study evaluates whether the expression of these enzymes is also a tool for the identification of the developing zones of the embryonic shell-forming tissue. Thus, we analyzed the temporal and spatial activity of the above-mentioned enzymes and of tartrate-resistant acid phosphatase in the shell forming tissues in Biomphalaria glabrata. Embryos of different age groups and adults were studied; alkaline phosphatase activity was seen in very young embryos in the shell field invagination prior to the secretion of any shell material, while peroxidase activity was present from the start of the periostracum production. Acid phosphatase, found in considerable amounts in yolk granules and albumen cells, appeared in the embryonic shell-forming tissue in relatively few Golgi stacks. Tartrate-resistant phosphatase was not present in embryos, but was found in adults in the same zone of the mantle edge as acid phosphatase. Using the enzymes as cell markers, the differentiation of the embryonic shell-forming tissue to the different zones of the adult mantle edge could clearly be followed.     

Phosphatase activity in <Emphasis Type="Italic">Amoeba proteus</Emphasis> at pH 9.0

In the free-living ameba Amoeba proteus (strain B), after PAAG disk-electrophoresis of the homogenate supernatant, at using 1-naphthyl phosphate as substrate and pH 9.0, three forms of phosphatase activity were revealed; they were arbitrarily called “fast,” “intermediate,” and “slow” phosphatases. The fast phosphatase has been established to be a fraction of lysosomal acid phosphatase that preserves some low activity at alkaline pH values. The question as to which particular class the intermediate phosphatase belongs to has remained unanswered: it can be either acid phosphatase, or protein tyrosine phosphatase. Based on data of inhibitor analysis, broad substrate specificity, results of experiments with reactivation by Zn ions after inactivation with EDTA, and another localization in the ameba cell than of the fast and intermediate phosphatases, it is concluded that only the slow phosphatase can be classified as alkaline phosphatase (EC 3.1.3.1).     

施用胶质芽胞杆菌菌剂对黑麦草根际土壤脲酶、磷酸酶及过氧化氢酶活性的影响

土壤酶活性是反映土壤肥力最为重要的生物学指标之一。采用稀释涂布平板法研究了从玉米（K02）、草地早熟禾（K05）、披碱草（K09）、多年生黑麦草（K11）、匍匐翦股颖（K12）根际土壤中分离到的5株胶质芽胞杆菌菌株对黑麦草根际土壤脲酶、磷酸酶及过氧化氢酶活性的影响。结果表明：在苗期、中期、收获期各处理黑麦草根际土壤脲酶、磷酸酶和过氧化氢酶活性均高于对照（P〈0.05）。总体来看,各处理黑麦草根际土壤脲酶和过氧化氢酶活性呈先增加后降低的变化趋势;土壤磷酸酶活性与对照（CK）相比除苗期处理K12外,其他处理均呈上升趋势,以处理K05磷酸酶活性最大。研究表明,施入胶质芽胞杆菌菌剂对黑麦草根际土壤脲酶、磷酸酶及过氧化氢酶活性有一定的积极作用,其结果可为生物钾肥的研制提供必要的数据。     

Inhibition of alkaline phosphatase by thioureido derivatives of methylenebisphosphonic acid

A series of thioureido derivatives of methylenebisphosphonic acid were synthesized by the reaction of aminomethylenebisphosphonic acid with the corresponding isothiocyanates, and their effect on the activity of alkaline phosphatases from bovine small intestine mucosa (BSIM) and human placenta was studied. It was found that (3-phenylthioureido)methylenebisphosphonate is approximately one order of magnitude more effective in inhibiting the activity of alkaline phosphatase from BSIM than the alkyl derivatives of thioureidomethylenebisphosphonic acid with methyl, ethyl, tert-butyl, or cyclohexyl substituents. The introduction of substituents into the benzene ring of (3-phenylthioureido)methylenebisphosphonate decreased the effect of the inhibitor on the activity of the enzyme. The affinity of (3-phenylureido)methylenebisphosphonate to the alkaline phosphatase of BSIM was also weaker as compared with the corresponding thioureidomethylenebisphosphonate. The insertion of thioureidobisphosphonates into the active site of alkaline phosphatase of human placenta by the method of molecular docking indicated that the methylenebisphosphonate residue and the substituted amino groups of the inhibitor are involved in the mechanisms of complex formation with the enzyme. It is supposed that the improvement of the inhibitory activity of (3-phenylthioureido)methylenebisphosphonate toward alkaline phosphatase of BSIM is due to the additional fixation of the phenyl substituent in the active site of the enzyme.     

Dual inoculation with Aspergillus fumigatus and Glomus mosseae enhances biomass production and nutrient uptake in wheat (Triticum aestivum L.) supplied with organic phosphorus as Na-phytate

In a pot experiment, wheat was grown for 50 days in two heat-sterilized low-phosphorus (P) soils supplied with organic P as Na-phytate. Seed inoculation with the phosphatase-producing fungus (PPF) Aspergillus fumigatus or soil inoculation with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae increased shoot and root dry weight and root length, phosphatase activity in the rhizosphere and shoot concentrations of P and to a lesser extent of K and Mg. As a rule, the greatest effects on those parameters were most in the combined inoculation treatment (PPF + VAM). Shoot concentrations of Cu and Zn were only enhanced by VAM, not by PPF. At harvest, depletion of organic P in the rhizosphere soil increased in the order of: sterilized soil < PPF < VAM < PPF + VAM which corresponded with the enhanced P concentrations in the plants. The results demonstrate that organic P in form of Na-Phytate is efficiently used by VAM and that use of organic P can be increased by simultaneous inoculation with phosphatase-producing fungi.