铜胁迫下4种外生菌根真菌的耐受性比较

外生菌根对重金属具有较强的耐受性,通过生物固定对重金属污染土壤有一定修复能力。本试验选择4种外生菌根真菌红绒盖牛肝菌(Xerocomus chrysenteron)、双色蜡蘑(Laccaria bicolor)、铆钉菇(Gomphidius viscidus)和彩色豆马勃(Pisolithus tinctorius)进行离体培养,通过分析不同浓度铜(0、10、25、35、50和90 mg/L)胁迫下外生菌根真菌生长、铜积累和分泌耐热蛋白的变化,以探索4种外生菌根真菌的重金属耐受性及其机制。研究结果发现,铜胁迫处理组X.chrysenteron、P.tinctorius的菌丝干重比对照组分别仅减少7.8%~33.6%、0.2%~11.5%,而单位菌丝铜积累量显著低于L.bicolor和G.viscidus(P0.05),在生长方面表现出对过量铜胁迫的较强耐受性;从单位菌丝耐热蛋白分泌量来看,G.visciduss(X.chrysenteron,P.tinctorius)L.bicolor,但由于X.chrysenteron、P.tinctorius生物量干重较大,耐热蛋白分泌总量足够多,从而降低培养环境中的Cu2+,X.chrysenteron、P.tinctorius通过分泌耐热蛋白耐受铜胁迫能力更强;通过SDS-PAGE图谱发现铜胁迫诱导X.chrysenteron、G.viscidus和L.bicolor分泌出分子量在31~43 k D之间的多种新耐热蛋白成分。     

过量铜、锌对外生菌根菌牛乳牛肝菌生物量、呼吸和糖酵解酶活性的影响

在过量铜或锌胁迫下,独立培养的外生菌根菌牛乳牛肝菌（Suillus     

外生菌根真菌同立枯丝核菌重寄生关系的研究

研究了18株外生菌根菌对Rhizoctonia solant Kuehn的重寄生作用。在光学显徽镜下观察到8株茵根真菌同R．Solani具有重寄生关系,通常表现为吸附生长,缠绕,侵入,消解等4种作用形式。首次将扫描电镜和能谱仪(EDAX)应用到真菌间相互作用关系的研究上,对3株菌根菌Gomphidius roseus、Lacearia bicolor、Lactarius deliciosus 进行了扫描电镜观察和X-射线微区分析。发现了在吸附生长,缠绕和侵入等3种作用形式中,R. solani和菌根菌菌丝之间有物质流的存在,物质流主要岔有P、S、K、Ca等元素物质,流向是由R. solani菌丝到菌根菌菌丝。而在消解作用中则没有发现这种物质流。     

宿主植物栽培密度对AM真菌生长发育的影响*

温室盆栽条件下宿主植物高粱(SorghumvulgarePers.)的栽培密度对丛枝菌根(Arbuscularmycorrhizae,AM)真菌Glomusmosseae(Nicol.&Gerd.)Gerdemann&Trappe生长发育的影响试验结果表明:60株/盆密度处理的根外菌丝量及孢子数均高于其它处理。在一定栽培密度下(20~60株/盆),植株根系可溶性糖浓度与根外菌丝量呈显著负相关,与菌根侵染率呈显著正相关。植株根中磷浓度与根外菌丝量、根外菌丝量与孢子数均呈显著正相关。植株根中磷浓度与菌根侵染率呈显著负相关。结果说明:适当密植虽对植株生长有一定影响,但却促进了真菌的生长,此时菌根共生体有可能由互惠共生开始向偏利共生或弱寄生转化。密植作为一种调控手段,在菌剂生产中能获得较大数量的侵染根段、菌丝及孢子等繁殖体。     

过量钠盐胁迫对3种外生菌根真菌生长和钠元素积累的影响*

通过测定3种外生菌根真菌-美味牛肝菌(Boletus edulis)、红绒盖牛肝(Xerocomus chtysenterort)和铆钉菇(Gomphidius viscidus)在Na2SO4和NaCl处理下的菌斑增长特性、生物量积累和菌丝中钠元素积累量显示,2种盐都没有影响菌丝的生长模式。但在0．1mol／L浓度下,NaCl显抑制了X．chrysertterort和G．viscidus的干重增长;在Na2SO4处理下,X．chrysenteron生物增长量比对照显增加,G．viscidus则降低,而2种盐处理对B．edtdis的生物量积累没有显影响。3供试菌种在2种盐处理下,其对钠元素的吸收和积累量差异显。NaCl处理下,X．chtysenteron菌丝积累的钠量最多,而在Na2SO4条件下,B．edulis菌丝中钠积累量显高于其他2菌种。     

过量铜、锌对外生菌根菌牛乳牛肝菌生物量、呼吸和糖酵解酶活性的影响

在过量铜或锌胁迫下,独立培养的外生菌根菌牛乳牛肝菌（Suillus bovinus)的生长和生理活动都受到影响。在添加不同浓度水平的铜或锌的培养基中,菌丝斑直径增长与对照相比显著减小。4周后,菌丝生物量与铜、锌处理浓度呈负指数关系,对菌丝的半致死浓度分别为铜6.5mg/L和锌120mg/L。取半致死浓度和4倍半致死浓度的铜,1/4半致死浓度和半致死浓度的锌处理菌丝,其呼吸强度表现为下降趋势,且已糖激酶和磷酸果糖激酶活性随营养液中铜或锌的增加而降低,丙酮酸激酶活性则随铜或锌浓度的增加而增加,生长在高处理浓度的铜、锌营养液中,已糖激酶和磷酸果糖激酶两种关键酶活性受到抑制,使菌丝的生长和呼吸都受到显著抑制,然而,在相对较低浓度锌50mg/L处理下,生物量、呼吸强度和糖酵解关键酶活性变化不显著,说明S.bovinus的菌丝对锌不敏感。     

黑松菌根共生体中真菌液泡形态构架及其活力

 为探讨菌根共生体中菌根菌液泡结构及其活力变化对宿主生长的影响, 对3种外生菌根菌黄色须腹菌(Rhizopogen luteous, 简称Rl)、彩色豆马勃(Pisolithus tinctorius, 简称 Pt₂)、美味牛肝菌 (Boletus edulis, 简称Be)离体菌丝及与黑松(Pinus thunbergii)形成菌根共生体的冰冻切片进行6-羧基二乙酸荧光素(6-CFDA)染色, 观察了真菌液泡形态构架及其活力的变化。结果表明, 在Pt₂与Rl离体菌丝中既可见单个分散的大液泡, 又可见相邻液泡间由细管状连接而成的管状液泡系统, Be离体菌丝的液泡则呈细小颗粒状, 密集分布于菌丝体的各个部分; Be离体菌丝的活性强于Pt₂和Rl。上述3种菌根菌与黑松形成菌根共生体后, Pt₂与Rl对黑松的促生效果好于Be, 菌根共生体中菌套和哈氏网菌丝的活性明显强于Be, 且仍可见数个液泡连接成管状系统; 而在Be菌根中菌丝液泡系统不形成管状, 活性较低。结果暗示, 菌根共生体中真菌的管状液泡系统及其活力与对黑松的促生作用似存在一定的相关性。该研究对于进一步了解菌根的促生作用机理及菌根真菌与宿主植物的协同进化具有一定的意义。     

辽东栎幼苗的外生菌根合成

 在温室花盆中播种辽东栎（Quercus liaotungensis）种子获得辽东栎幼苗,并对幼苗接种外生菌根真菌进行菌根合成试验。所用的外生菌根真菌有：铆钉菇（Gomphidius viscidus）、臭红菇（Russula foetens）、厚环乳牛肝菌（Suillus grevillei）、褐环乳牛肝菌（S. luteus）、彩色豆马勃（Pisolithus tinctorius）、美味牛肝菌（Boletus edulis）、赭丝膜菌（Cortinarius russus）、土生空团菌（Cenococ     

应用Coriolus vericolor菌丝球脱色染料及印染废水的研究

对白腐真菌(Coriolus vericolor)产生漆酶进行〖JP2〗了研究。发现该菌产漆酶的最适初始pH值为45。提高微量元素浓度或添加藜芦醇都可使C.versicolor的产酶能力增加,添加Tween80会有一定的抑制作用。采用C.versicolor菌丝球进行重复分批产酶试验,结果表明菌丝球的稳定性很好,同一批菌丝球可连续利用14次,平均每批酶活力可保持在672U/mL,产酶能力优于聚氨酯泡沫固定化菌丝。将粗酶液用于染料的脱色降解试验,在酶活力为3.3IU/mL〖JP〗,酸性橙浓度为500mg/L条件下,经过24h反应,脱色率达到98.5%;对含弱酸大红和卡布龙红的印染废水进行脱色试验,脱色率也达到了93%。     

我国西藏担子菌类数新种

西藏的东南隅是真菌区系极为丰富和复杂的一个地区。作者在此地区采到的910号真菌标本,经鉴定有担子菌类5个新种,即：牛肝菌科(Boletaceae)的木生小牛肝菌(Boletitzusligicolus Zang sp．nov),巨孢牛肝菌(Bolctus magasporus Zang sp. Nov.),喇叭菌科(can-tharellaceae)的疣孢鸡油菌(Cantharellus tuberculosporus Zang sp. Nov),灰包科(马勃科)(Lycoperdaceae)的纺锤形灰包(Lycoperdon fusiforme Zang sp. Nov.)和鸟巢菌科(Nidulariaceae)的盘状黑蛋巢(Cyathus dis~oideus Zang sp. Nov.)。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

放牧对牧草光合作用、呼吸作用和氮、碳吸收与转运的影响

研究放牧对草地植物生理活动的影响,对于揭示草地放牧演替的生理机制有重要意义.大量研究表明,家畜放牧对牧草光合作用、呼吸作用以及C和N吸收与转运的影响,可以分为生理伤害和生理恢复2个阶段.放牧通过改变草地冠层结构影响牧草光合作用,净光合作用速率短期内迅速下降,随着叶面积指数增加又逐渐上升,呼吸作用有相似的变化趋势.牧草放牧后再生长所需的C和N最初主要来自根系和留茬中的贮藏物质,此后随着牧草生长恢复逐渐由同化作用供给,C代谢与土壤N水平负相关.放牧后牧草生理活动变化与牧草遗传特性、种间竞争、家畜放牧特征、非生物环境等因素密切相关.     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Synthesis of hapten and conjugates of coumestrol and development of immunoassay

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。