Effects of lanthanum on the active oxygenscavenging enzyme activities and ultrastructure in Potamogeton crispus leaves

在菹草培养液中施用LaCl3·8 H2O培养菹草,研究了稀土镧对菹草活性氧清除酶活性及叶细胞超微结构的影响.结果表明,菹草在含适宜浓度La3+的培养液中长势均好于对照.较低浓度La3+处理时,菹草叶绿素含量、叶片自发荧光强度、硝酸还原酶(NRase)活性、过氧化物酶(POD)活性、过氧化氢酶(CAT)活性及可溶性蛋白质含量均升高,而超氧化物歧化酶(SOD)活性、细胞膜透性、O2-产生速率及丙二醛(MDA)含量则呈降低趋势,电镜观察结果为La3+对菹草生长的影响提供了细胞学证据.实验认为促进菹草生长的最适La3+浓度为5～10 mg·L-1.     

La(NO3)3浸种对镉胁迫下油菜种子萌发及幼苗生长的影响

通过水培实验研究了La(NO3)3浸种对镉胁迫下油菜(Brassicanapus)种子萌发及幼苗生长的影响。结果表明,当镉浓度≤10mg·L-1时,La(NO3)3浸种(0～50mg·L-1)能提高油菜种子的活力,促进油菜幼苗的生长,并提高脂肪酶的活性和幼苗根细胞有丝分裂指数。其中以10mg·L-1La(NO3)3浸种处理效果最佳,与对照相比,种子活力指数及油菜幼苗各生长指标显著提高,脂肪酶活性增加40.41%～65.09%,幼苗根细胞有丝分裂指数增加62.76%～77.02%。当镉浓度>10mg·L-1时,La(NO3)3浸种对镉胁迫的缓解效用明显减弱,与对照相比,仅脂肪酶活性显著提高,其它指标无明显变化。     

草酸青霉DFH对菌核病菌的抑制作用

菌核病菌(Sclerotinia sclerotiorum)所分泌的草酸是油菜菌核病菌重要的生化致病因子。通过在培养基中添加高浓度草酸,从土壤中分离出一株在预培养1 d的菌核病菌平皿中形成2.1 cm(菌块直径1 cm)抑菌圈的菌株,编号为DFH,经形态学与分子生物学鉴定为草酸青霉(Penicillium oxalicum),在油菜离体叶片试验中,DFH与对照组抗菌核病菌的效果差异不明显。初步证明DFH对菌核病菌有抑制作用,抗菌活性有待提高。     

镧对菹草活性氧清除酶活性及叶细胞超微结构的影响

在菹草培养液中施用LaCl3.8H2O培养菹草,研究了稀土镧对菹草活性氧清除酶活性及叶细胞超微结构的影响。结果表明,菹草在含适宜浓度La3+的培养液中长势均好于对照。较低浓度La3+处理时,菹草叶绿素含量、叶片自发荧光强度、硝酸还原酶(NRase)活性、过氧化物酶(POD)活性、过氧化氢酶(CAT)活性及可溶性蛋白质含量均升高,而超氧化物歧化酶(SOD)活性、细胞膜透性、O-2.产生速率及丙二醛(MDA)含量则呈降低趋势,电镜观察结果为La3+对菹草生长的影响提供了细胞学证据。实验认为促进菹草生长的最适La3+浓度为5～10mg.L-1。     

紫外光诱导的油菜菌核病菌抗扑海因菌株的特性研究

通过室内紫外光诱导,获得了油菜菌核病菌多株抗扑海因突变体。采用菌丝生长速率法.对诱变得到的抗扑海因菌株的抗药性进行了测定。结果显示,油菜菌核病菌抗扑海因菌株的EC_（50）值分布范围为0.1159-604.2200μg/mL,平均值为117.1363μg/mL,突变株UVIP＇SS-7、UVIP＇SS-8、UVIP＇SS-15、UVIP＇SS-16和UVIP＇SS-18的EC_（50）值均大于160μg/ML。油菜菌核病菌抗扑海因突变株在含扑海因质量浓度为1μg/mL时的菌核产生数量绝大多数都大于不含药PSA平板的菌核数量,而高浓度下菌核几乎不产生。     

硝酸镧浸种对NaCl胁迫下柳枝稷种子萌发及幼苗生理特性的影响

为探讨镧对NaCl胁迫下柳枝稷种子萌发及幼苗生理特性的影响,该研究以柳枝稷(Panicum virgatum L.)种子为研究材料,通过不同浓度(0、0.05、0.1、0.5和1.0mmol/L)La(NO3)3溶液浸种24h处理后,添加100mmol/L NaCl溶液,以蒸馏水为对照,在种子发芽箱中培养并观察柳枝稷种子萌发和幼苗生长,测定幼苗叶片相对电导率、丙二醛、脯氨酸和叶绿素的含量。结果显示:(1)0.05mmol/L La(NO3)3浸种处理可缓解100mmol/L NaCl对种子发芽的影响。(2)不同处理对幼苗苗长无显著影响,0.1mmol/L La(NO3)3浸种显著增加了幼苗的鲜重和叶面积,当浓度≥0.5mmol/L时根长、根冠比、鲜重和叶面积受到抑制。(3)幼苗叶片相对电导率和脯氨酸含量随La(NO3)3浓度的增加呈先降低后增加的趋势,其中0.05和0.1mmol/L La(NO3)3浸种处理下效果较好,丙二醛含量随La(NO3)3浓度的增加持续降低,但各浓度处理间差异不显著,叶绿素含量在0.05mmol/L La(NO3)3浸种处理下较NaCl处理显著增加,当浓度≥0.5mmol/L随着浸种浓度的增加呈下降趋势。(4)不同指标间发芽势与发芽率、根长和叶绿素含量间呈极显著正相关关系,与根冠比、电导率和脯氨酸含量均呈极显著负相关关系,而与苗长不相关。研究表明,低浓度(0.05mmol/L)La(NO3)3可缓解NaCl胁迫对柳枝稷种子萌发及幼苗生长的影响,而高浓度(1.0mmol/L)的La(NO3)3则会加重NaCl的胁迫危害。     

孜然种子提取物枯茗醛和枯茗酸抑菌作用研究

以番茄早疫病菌、棉花黄萎病菌、小麦纹枯病菌、烟草赤星病菌、小麦全蚀病菌、油菜菌核病菌、辣椒疫霉病菌、水稻纹枯病菌、小麦白粉病菌和番茄灰霉病菌等为供试菌种,采用离体与活体相结合的方法系统地测定了枯茗醛和枯茗酸的抑菌活性。离体抑菌活性测定结果表明,枯茗醛和枯茗酸对多种病原菌具有一定的抑制效果,其中对油菜菌核病菌菌丝生长的抑制效果高于其它供试病原菌,有效中浓(EC50)分别为2.1和7.3 mg/L;枯茗醛和枯茗酸对小麦白粉病的防治实验结果表明,供试浓度为1000 mg/L时,两种药剂的保护防效均高于50%;相同处理浓度下,枯茗酸对油菜菌核病的保护防效与速克灵处理相当,达到57.52%。     

抗植物病原真菌紫叶小檗内生真菌的筛选和鉴定

对紫叶小檗根、茎、叶和果实中分离纯化得到的28株内生真菌进行液体培养,培养物烘干研磨后用丙酮浸提,测定各提取物对5种供试植物病原真菌的抑菌活性,对抗菌谱较广的菌株进行分子鉴定,并分别测定其胞内、胞外代谢产物的抑菌活性.结果显示:(1)紫叶小檗25株内生真菌中有5株菌(R3、S4、L6、F2、F6)的抗菌谱较广,其中,茎中的内生菌S4抑菌活性最强,对5种植物病原真菌的抑菌活性均大于90％,对马铃薯干腐病菌的抑菌率最高达93.06％.(2)经鉴定S4为子囊菌门的Para phaeos phaeria属真菌.(3)S4菌丝体丙酮提取液对马铃薯干腐病菌的抑菌率随其浓度(加入体积)的增加而增大,呈正相关线性关系(y=15.334x+14.618,r=0.99),而S4发酵液对马铃薯干腐病菌的抑菌率随其浓度(加入体积)的增加而减小,呈负相关线性关系(y=-20.196x+64.984,r=0.99),即在较高浓度时促进病原菌生长.研究表明,紫叶小檗内生真菌S4菌株的抑菌活性成分主要为胞内代谢产物.     

铀尾沙对油菜幼苗生长和生理特征的影响

采用沙培盆栽试验,以铀尾沙所占比例分别为0%(CK)、25%(T1)、50%(T2)、75%(T3)、100%(T4)的培养基质,研究了铀尾沙对芥菜型油菜、甘蓝型油菜和白菜型油菜出苗率、幼苗生物量、叶绿素含量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和还原型谷胱甘肽(GSH)含量的影响.结果表明:不同比例的铀尾沙对油菜出苗率的影响差异不显著(P＞0.05).3种油菜的生物量均随铀尾沙处理量的增加而呈先增后降的现象,甘蓝型油菜和白菜型油菜在T2时生物量最大,芥菜型油菜在T3时生物量最大.MDA含量则呈先降低后升高,甘蓝型油菜和白菜型油菜在T2时MDA含量最低,芥菜型油菜在T3时的MDA含量最低,并与叶绿素含量呈显著的负相关.随铀尾沙处理量的增加,3种类型的油菜SOD、CAT、POD、APX和GR活性逐渐增加,GSH含量也持续升高.铀尾沙对油菜幼苗生长和抗氧化酶系统产生了一定的诱导作用,而这3种油菜也对铀尾沙也表现出较强的耐受能力.     

镉对花生幼苗生长及生理生态特性的影响

通过溶液培养,研究不同浓度镉(Cd2 ,0～1.00mmoL/L)时花生幼苗生长及生理生态特性的影响.结果表明,Cd2 影响花生种子的萌发和幼苗的生长.当Cd2 浓度高于0.50mmol/L时,显著抑制种子的发芽率,随Cd2 浓度的增加,苗高、根长和侧根数减少;叶绿素含量随Cd2 浓度的增加而下降,过氧化物酶(POD)活性随Cd2 浓度增加而增加;丙二醛(MDA)含量同样呈上升趋势;超氧化物歧化酶(SOD)与根系活力表现为低浓度下升高、超过0.5mmoL/L后下降的变化趋势.     

Lanthanum increases the rat thymocyte cytoplasmic free calcium concentration by enhancing calcium influx

In the present study, I have examined the effect of lanthanum (La3+) on cytoplasmic free calcium concentration in isolated rat thymocytes employing the quin2 technique. As with its effect on 15Ca accumulation in rat thymocytes (Segal, J. and Ingbar, S.H. (1984) Endocrinology, 115, 160-166), La3+ produced a concentration-related increase in thymocyte cytoplasmic free calcium concentration. This effect of La3+ was very prompt in onset, evident within about 30 s from the time of addition of La3+. The lowest effective concentration of La3+ was 6 microM (+22.7% above control), and it increased progressively to reach maximal values at 25 microM (+100% above control). La3+ added to quin2-loaded thymocytes suspended in a calcium-free medium was without effect. In addition, La3+ had no significant effect on 45Ca efflux, and La3+ did not inhibit calcium-ATPase activity in the rat thymocytes. These results demonstrate that in rat thymocytes La3+ increases cytoplasmic free calcium concentration by increasing the extracellular calcium influx into the cell rather than the release of calcium from an intracellular pool.     

Involvement of alternative oxidase in the regulation of growth, development, and resistance to oxidative stress of Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a cosmopolitan, filamentous, fungal pathogen that can cause serious disease in many kinds of crops. Alternative oxidase is the terminal oxidase of the alternative mitochondrial respiratory pathway in fungi and higher plants. We report the presence of this alternative pathway respiration and demonstrate its expression in two isolates of S. sclerotiorum under unstressed, normal culture conditions. Application of salicylhydroxamic acid, a specific inhibitor of alternative oxidase, severely inhibited the mycelial growth of S. sclerotiorum both on potato dextrose agar plates and in liquid culture media. Inhibition of alternative oxidase could influence the growth pattern of S. sclerotiorum, as salicylhydroxamic acid treatment induced obvious aerial mycelia growing on potato dextrose agar plates. Under the treatment with salicylhydroxamic acid, S. sclerotiorum formed sclerotia much more slowly than the control. Treatment with hydrogen peroxide in millimolar concentrations greatly decreased the growth rate of mycelia and delayed the formation of sclerotia in both tested S. sclerotiorum isolates. As well, this treatment obviously increased their alternative pathway respiration and the levels of both mRNA and protein of the alternative oxidase. These results indicate that alternative oxidase is involved in the regulation of growth, development, and resistance to oxidative stress of S. sclerotiorum.     

Indolyl-3-acetaldoxime dehydratase from the phytopathogenic fungus Sclerotinia sclerotiorum: purification, characterization, and substrate specificity

The purification and characterization of indolyl-3-acetaldoxime dehydratase produced by the plant fungal pathogen Sclerotinia sclerotiorum is described. The substrate specificity indicates that it is an indolyl-3-acetaldoxime dehydratase (IAD, EC 4.99.1.6), which catalyzes transformation of indolyl-3-acetaldoxime to indolyl-3-acetonitrile. The enzyme showed Michaelis-Menten kinetics and had an apparent molecular mass of 44 kDa. The amino acid sequence of IAD, determined using LC-ESI-MS/MS, identified it as the protein SS1G_01653 from S. sclerotiorum. IADSs was highly homologous (84% amino acid identity) to the hypothetical protein BC1G_14775 from Botryotinia fuckeliana B05.10. In addition, similarity to the phenylacetaldoxime dehydratases from Gibberella zeae (33% amino acid identity) and Bacillus sp. (20% amino acid identity) was noted. The specific activity of IADSs increased about 17-fold upon addition of Na(2)S(2)O(4) under anaerobic conditions, but in the absence of Na(2)S(2)O(4) no significant change was observed, whether aerobic or anaerobic conditions were used. As with other aldoxime dehydratases isolated from microbes, the role of IADSs in fungal plant pathogens is not clear, but given its substrate specificity, it appears unlikely that IADSs is a general xenobiotic detoxifying enzyme.     

Effect of oxygen concentration on nitrification and denitrification in single activated sludge flocs

Simultaneous nitrification and denitrification (SND) was investigated in the single aeration tank of a municipal wastewater treatment plant. Microelectrode measurements and batch experiments were performed to test for the presence of SND. Microelectrodes recorded the presence of O(2) concentration gradients in individual activated sludge flocs. When the O(2) concentration in the bulk liquid was <45 microM, anoxic zones were detected within flocs with a larger diameter (approximately 3000 microm). The O(2) penetration depth in the floc was found to be dependent on the O(2) concentration in the bulk liquid. Nitrification was restricted to the oxic zones, whereas denitrification occurred mainly in the anoxic zones. The nitrification rate of the activated sludge increased with increasing O(2) concentration in the bulk liquid, up to 40 microM, and remained constant thereafter. SND was observed in the aerated activated sludge when O(2) concentration was in the range of 10 to 35 microM.     

Effect of Sclerotial Damage of Sclerotinia sclerotiorum on the Mycoparasitic Activity of Trichoderma hamatum

Damaged sclerotia of Sclerotinia sclerotiorum buried in soil infested with Trichoderma hamatum isolate TMCS - 3 were degraded rapidly when the medulla of sclerotia was com pletely exposed by the feeding activity of larvae of the fungus gnat Bradysia coprophila. These heavily damaged sclerotia also enhanced , in vitro, the growth of TMCS - 3 . Growth of TMCS - 3 in liquid culture was studied using different carbon sources as substrates , including sclerotia of S. sclerotiorum. Significantly more biomass of TMCS - 3 was recovered using sclerotia as a substrate compared to other carbon sources tested . Exudates from sclerotia whose melanized rinds had been completely removed by feeding larvae accelerated the germination of conidia of TMCS - 3 . Concentrations of amino acids , carbohydrates and proteins in the sclerotial exudates were not increased as damage to sclerotia was increased . Exudation of electrolytes was higher in undamaged than damaged sclerotia . Glucanase activity of TMCS - 3 was slightly increased when the fungus was exposed to damaged sclerotia . However , chitinase activity was not increased by damaging the sclerotia . Larval damage altered the sclerotia not only physically but also chemically , thereby enhancing the activity of the fungus T. hamatum.     

Oxalic acid, a pathogenicity factor for Sclerotinia sclerotiorum, suppresses the oxidative burst of the host plant

Effective pathogenesis by the fungus Sclerotinia sclerotiorum requires the secretion of oxalic acid. Studies were conducted to determine whether oxalate aids pathogen compatibility by modulating the oxidative burst of the host plant. Inoculation of tobacco leaves with an oxalate-deficient nonpathogenic mutant of S. sclerotiorum induced measurable oxidant biosynthesis, but inoculation with an oxalate-secreting strain did not. Oxalate inhibited production of H(2)O(2) in tobacco and soybean cultured cell lines with a median inhibitory concentration of approximately 4 to 5 mM, a concentration less than that measured in preparations of the virulent fungus. Several observations also indicate that the inhibitory effects of oxalate are largely independent of both its acidity and its affinity for Ca(2)+. These and other data demonstrate that oxalate may inhibit a signaling step positioned upstream of oxidase assembly/activation but downstream of Ca(2)+ fluxes into the plant cell cytosol.     

Defense against Sclerotinia sclerotiorum in Arabidopsis is dependent on jasmonic acid, salicylic acid, and ethylene signaling

Genotypic differences in susceptibility of Arabidopsis thaliana to Sclerotinia sclerotiorum have not been reported due to the extreme susceptibility of this cruciferous plant. To overcome this limitation, we have established inoculation conditions that enable evaluation of differences in susceptibility to S. sclerotiorum among Arabidopsis mutants and ecotypes. Two coil mutant alleles conferred hypersusceptibility to S. sclerotiorum. The plant defensin gene PDF1.2 was no longer induced after challenging the coi1-2 mutant with S. sclerotiorum. Hypersusceptibility of the coi1-2 mutant to S. sclerotiorum was not correlated with oxalate sensitivity. The mutants npr1 and ein2 were also hypersusceptible to S. sclerotiorum. Induction of PDF1.2 and the pathogenesis-related gene PR1 was reduced in ein2 and npr1 mutants, respectively. Actigard, a commercial formulation of the systemic acquired resistance inducer benzothiadiazole, reduced susceptibility to S. sclerotiorum. Based on histochemical analysis of oxalate-deficient and wild-type strains of S. sclerotiorum, oxalate caused a decrease in hydrogen peroxide production but no detectable changes in plant superoxide production or gene expression.     

Study of lanthanum on seed germination and growth of rice

A study of lanthanum nitrate on the seed germination and the growth of rice was conducted. The results of this experiment indicate that soaking in a proper concentration of La(NO3)3 could accelerate the germination of the rice seeds, significantly increase seed vigor and chlorophyll contents, and improve root growth. Because during the soaking stage with La(NO3)3 the absorbed water and imbibition process of the rice seeds was quickened, the plasma membrane permeability of the seeds was increased, O2 and H2O were easier to get into the cell, and the respiratory rate was enhanced. During the germination stage, La(NO3)3 could enhance the activities of alpha-amylase, proteinase, lipase, and other hydrolytic enzymes and the contents of plant hormones, such as IAA, GAs, CTK, and so forth, except ABA contents, changed little.