梯棱羊肚菌Morchella importuna对重金属离子的耐受性研究

以加入6种不同浓度的重金属离子的培养基对梯棱羊肚菌Morchella importuna进行培养的方法,研究其对重金属离子的耐受性。以平板进行培养,测量和计算菌丝平均生长速率和生长促进率,绘制生长曲线获取菌丝生长最佳耐受浓度;采用定性方法对气生菌丝浓密、菌核大小和数量进行描述,获得菌核生成最佳耐受浓度。结果显示:在试验浓度内梯棱羊肚菌对Cr~(6+)、Mn~(2+)、Pb~(2+)、Mo~(6+)、Cu~(2+)具有较强的耐受性,这些重金属离子促进了羊肚菌菌丝生长,而对Cd~(2+)耐受性差,低浓度抑制菌丝生长。促进菌丝生长最适浓度分别为Cr~(6+)100.0mg/L、Mn~(2+)100.0mg/L、Cd~(2+)5.0mg/L、Pb~(2+)50.0mg/L、Mo~(6+)20.0mg/L、Cu~(2+)100.0mg/L,促进菌核生成最适浓度分别为Cr~(6+)50.0mg/L、Mn~(2+)100.0mg/L、Cd~(2+)5.0mg/L、Pb~(2+)20.0mg/L、Mo~(6+)10.0mg/L、Cu~(2+)100.0mg/L。Cd~(2+)抑制菌丝生长临界浓度为介于20.0mg/L与50.0mg/L之间。     

土壤中木霉的分离及其对毒死蜱降解特性研究

为挖掘保护地土壤中的能降解毒死蜱的木霉资源,从长期污染的土壤中共分离6个不同木霉菌株,通过形态学特征和ITS r DNA序列对各菌株进行鉴定,6个菌株分别为哈茨木霉(Trichoderma harzianum)、绿色木霉(T.viride)、康宁木霉(T.koningii)、拟康宁木霉(T.koningiopsis)、长枝木霉(T.longibrachiatum)和短密木霉(T.brevicompactum)。选择降解活性最高的长枝木霉TC5菌株,测定各种条件下对毒死蜱的降解特性。结果表明,木霉TC5在中性或偏碱性条件下对毒死蜱具有更好的降解活性;添加碳源或提高木霉的接种浓度,能提高对毒死蜱的降解活性;在50-300 mg/L范围内,随毒死蜱浓度升高,木霉的降解活性也明显提高。盆栽试验中木霉TC5对毒死蜱保持较强的降解活性,但在自然土壤中降解活性显著低于在灭菌土壤中。分离的木霉菌株在土壤农药污染修复方面具有应用开发潜力。     

细菌还原酪酸盐

酪酸盐是电镀业和其它工业生产过程的一种有毒副产物,存在于这些行业的废水里,利用荧光假单胞菌(Preudomonas Fluorescens)的一个菌株能够去除酪酸盐。荧光假单孢菌耐酪酸盐,在厌氧或好气条件下能够使溶液中的Cr~(6+)还原成为Cr~3。细菌也使污染生活废水的酪酸盐降低,因     

Desulfovibrio desulfuricans G20生理特性及处理含铬(Ⅵ)硫酸盐废水的研究

选取一株硫酸盐还原菌Desulfovibrio desulfuricans G20,探讨其生理特性及其对含重金属硫酸盐废水的处理效果。结果显示,D.desulfuricans G20在2~18 h进入对数生长期,18~26.5 h进入稳定期。该菌株最适宜温度为37℃,最佳初始p H为8.2;最佳生长碳源为乳酸钠和蔗糖;最适硫源为亚硫酸钠。D.desulfuricans G20对Cr~(6+)最大耐受度为150 mg/L。随着Cr~(6+)初始浓度的减少,SO2-4去除率逐渐增加,最高达75.67%。Cr~(6+)质量浓度低于120 mg/L的去除率接近100%。可见,D.desulfuricans G20有潜力应用于处理含重金属Cr~(6+)的硫酸盐废水。     

木霉菌合成银纳米粒子条件的优化及其对甜瓜尖孢镰刀菌抑制作用

随着绿色环保观念的普及,生物合成金属纳米粒子的方法备受青睐。纳米银(Silver nanoparticles,AgNPs)由于其抗菌活性强且不易产生抗药性等特点在农业病害防治中越来越受到关注。文中利用橘绿木霉Trichoderma citrinoviride和毛簇木霉Trichoderma velutinous研究了AgNPs的最适合成条件和AgNPs对尖孢镰刀菌抑菌活性。结果表明,所有合成的AgNPs均在400–500 nm处有吸收峰,两种木霉生物合成AgNPs的最适合成条件为CL法(菌丝滤液)静置光照培养,底物AgNO_3浓度为2.0mmol/L,pH值为7,反应温度为45℃。橘绿木霉和毛簇木霉合成的AgNPs均对尖孢镰刀菌有抑制作用,抑菌效果随浓度的增加而增大,AgNPs在浓度为200 mg/L时,抑菌率分别达到33.745%和36.083%。     

Cr(VI)还原菌的筛选、鉴定及其还原物质分析

【背景】铬污染土壤是我国土壤污染修复的重点治理对象,在众多修复技术中,微生物法因具有简单、经济、无二次污染等特性已成为研究热点,而微生物法中筛选出既能适应污染场地环境又能高效还原Cr(VI)的菌株尤为重要。【目的】筛选适应西北寒旱区高效还原Cr(VI)的菌株,丰富铬还原菌资源库,为铬污染土壤修复奠定基础。【方法】采用富集驯化、分离纯化法进行筛菌;通过形态学和分子生物学相结合的方法对目的菌株进行鉴定;采用傅里叶变换红外光谱法对还原机理进行研究。【结果】菌株G-13有较强的Cr(VI)还原能力,pH 9.0、温度为30°C条件下,60 h对Cr(VI)(100 mg/L)的还原率达到82.8%。经形态学和分子生物学鉴定,菌株G-13为Micrococcus luteus。反应中Cr(VI)的降低伴随着Cr(III)的增加,说明以还原反应为主,并且还原能力与细菌生长呈依赖型关系。对细胞各组分及变性研究表明,胞外酶在还原反应中占主要作用。除Pd~(2+)、Cd~(2+)外,其余金属离子对酶活性无明显抑制作用。通过傅里叶变换红外光谱分析,发现G-13与Cr(VI)结合位点主要为羟基、羰基、羧基、–CH、酰胺基等。【结论】菌株G-13有较强的Cr(VI)还原能力,能为西北寒旱区铬污染土壤修复丰富菌种资源。     

不同耐铬性青菜中Cr~(6+)的亚细胞分布及其耐性机理

为探明青菜对Cr6+的吸收及其耐性机理,选取2个不同耐铬(Cr)性青菜品种‘美加华’(Cr耐性品种)和‘三月慢’(Cr敏感品种)进行水培试验,比较0、10、50和100μmol·L~(-1) Cr~(6+)处理下青菜地上部和根系Cr含量、Cr的亚细胞分布、非蛋白巯基(NPT)、植物螯合肽(PCs)和抗氧化酶活性的差异。结果表明:(1)2个青菜品种地上部Cr含量显著小于根系的含量,且‘美加华’根系Cr含量、Cr的根系滞留率、地上部和根系细胞壁中Cr含量与总Cr含量的比值均显著大于‘三月慢’,而其可溶性Cr含量与总Cr含量的比值显著小于‘三月慢’。(2)Cr~(6+)处理显著增加了青菜地上部NPT和PCs的含量,且‘美加华’增加的幅度显著高于‘三月慢’。(3)低浓度的Cr~(6+)(10μmol·L~(-1))能够提高青菜SOD、POD和CAT的活性,且‘美加华’的增幅大于‘三月慢’;而高浓度的Cr~(6+)(100μmol·L~(-1))显著降低了SOD、POD和CAT的活性,且‘美加华’的降幅小于‘三月慢’。研究认为,在Cr~(6+)胁迫条件下,与青菜品种‘三月慢’相比品种‘美加华’能将更多的Cr分布于根和细胞壁中,减少了跨膜进入细胞内的Cr,并且具有较高的抗氧化酶活性和PCs含量,从而表现出更强的耐Cr~(6+)性。     

棘孢木霉菌对钠胁迫的生理响应机制

【背景】棘孢木霉菌制剂被广泛应用于生物防治和次生盐渍化土壤的微生物修复,但是关于棘孢木霉在盐渍化胁迫条件下生长的耐盐机理及其富集盐离子的能力尚缺乏深入研究。【目的】揭示一株耐盐棘孢木霉菌(Trichodermaasperellum)CTCCSJ-W-SBW10264(T264)对钠胁迫的生理响应机制及其对钠离子的吸附和累积特性。【方法】设计梯度浓度钠胁迫培养实验,采集不同培养时期的菌丝样本,测定细胞氧化损伤相关指标H_2O_2和丙二醛(Malondialdehyde,MDA)的含量及细胞抗氧化相关酶的活性变化。【结果】钠胁迫实验表明,棘孢木霉T264能够在1.22mol/L的钠胁迫环境中生存,在低于0.25 mol/L的钠胁迫下其生长不会被明显抑制。细胞氧化损伤及氧化损伤响应相关指标的研究结果表明,培养液中钠离子浓度越高,棘孢木霉的膜系统氧化水平(MDA含量)越高,而且随着细胞中MDA和H_2O_2的累积,细胞抵御氧化损伤相关酶的活性也有明显提高,在钠盐处理24 h后,0.5、1.0和1.22 mol/L的钠离子胁迫分别使过氧化物酶(Peroxidase,POD)、超氧化物歧化酶(SuperoxideDismutase,SOD)和过氧化氢酶(Catalase,CAT)活性达到峰值,依次为36.66、3.34和233.3 U/mg。钠离子吸附和累积特性实验结果表明,棘孢木霉T264的菌丝对钠离子有强的吸附能力。在0.05 mol/L的钠离子环境中培养72 h后,菌丝表面钠离子吸附量为1 347.6 mg/g,菌丝内部钠离子累积量为218.6 mg/g,木霉菌菌丝通过菌丝表面吸附和菌丝内部累积对培养液中钠离子的去除率达到32%。【结论】T264的抗氧化损伤相关酶在其耐受钠离子胁迫过程中发挥重要作用,菌株T264对高浓度钠离子有强适应性,而且对环境中钠离子有高效的吸附和累积作用。     

Cr(Ⅵ)还原菌Microbacterium sp.BD6的分离鉴定及还原特性

【目的】从电镀厂下水道的淤泥中分离筛选Cr(Ⅵ)高效还原菌,并对其生长和还原特性进行研究,以期为Cr(Ⅵ)污染的生物修复提供优质的菌种资源和应用参考。【方法】采用富集培养法从淤泥中分离、筛选出Cr(Ⅵ)还原菌,通过生理生化及16S rRNA基因序列分析进行初步鉴定。采用单因素实验确定菌株的最佳培养条件和抵抗胁迫环境的能力,利用外加电子供体改善菌株的Cr(Ⅵ)还原能力,筛选出最佳电子供体研究对菌株还原的影响。【结果】经分离筛选得到1株Cr(Ⅵ)耐受还原菌,初步鉴定为微杆菌属(Microbacterium sp.),命名为BD6。菌株BD6适宜在中温、偏碱性的环境条件下生长,能耐受50.0 g/L NaCl的高盐环境。Mn~(2+)对菌种的生长表现出较高的抑制,Ni~(2+)、Zn~(2+)、Cd~(2+)的抑制作用较小,Cu~(2+)产生了一定的促进作用。Cr(Ⅵ)对BD6的最低抑菌浓度为1700 mg/L。添加甘油、果糖、乳糖、葡萄糖、丙酮酸钠作为电子供体促进了菌株对Cr(Ⅵ)的还原。选择甘油作为菌株还原Cr(Ⅵ)的最佳电子供体,无电子供体添加时菌株96 h内对100 mg/L Cr(Ⅵ)的还原率仅为69.63%,添加2 g/L的甘油菌株在36 h内的还原率达到了100%。通过加大甘油的添加量可以促进菌株对初始浓度较高Cr(Ⅵ)的还原,但要受到Cr(Ⅵ)的毒性限制。菌株的最适还原条件和最适生长条件吻合,在50.0 g/L NaCl的高盐条件和50 mg/L Cd~(2+)的毒性环境中,添加2 g/L的甘油,菌株对100 mg/L Cr(Ⅵ)的还原率分别为72 h 96.79%、54 h 99.86%。【结论】分离筛选得到的Microbacterium sp. BD6是一株潜在的可用于Cr(Ⅵ)污染生物还原修复的候选菌株。     

我国农作物根际土壤木霉菌资源分布调查

通过对我国18个省市地区的农作物根际土壤进行采样,使用木霉菌选择性培养基(TSM)和马铃薯葡萄糖培养基(PDA),对采样土壤中的木霉菌(Trichoderma spp.)进行分离和纯化,再与同一地区的多种植物病原菌进行平板对峙测试,筛选和保存了1 300余株有拮抗功能的木霉菌株。经形态学和分子生物学(ITSTef1-α)鉴定,这些菌株分属9个木霉菌种。其中哈茨木霉(T.harzianum)和棘孢木霉(T.asperellum)为优势菌种,分别占29.07%和29.91%(总数:393和382株);其次是绿色木霉(T.viride),占筛选总数的14%(总数:185株);而渐绿木霉(T.viridescens)和康宁木霉(T.koningii)分别占7.53%和7.00%(总数:99和92株);深绿木霉(T.atroviride)和钩状木霉(T.harmatum)分别只占5%以上(总数:69和67株);长枝木霉(T.longibrachiatum)和橘绿木霉(T.citrinoviride)的群落在我国农作物根际土壤中的分布最少,分别只占1.5%和0.5%以上(总数:21和7株)。同时,为了解不同地区的木霉菌株对上海地区的草莓灰霉菌(Botrytis cinerea)的拮抗作用的差异性,选用14个不同地区的木霉菌株,通过对峙培养拮抗测定,筛选出6个木霉菌株,进一步做了抑制草莓灰霉病的室内生测试验。结果显示,来源于华东地区的木霉菌株TR78和TR85,在平板对峙试验中对草莓灰霉菌生长的抑菌率分别为85.49%和82.90%,室内生测对灰霉病的防治效果分别为80.19%和80.02%,显著高于(P0.05)其他菌株对灰霉病的防治效果。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

人血管能抑素基因的克隆、表达及其表达产物的纯化和生物活性测定

以人胎盘脐带组织为材料,提取组织总RNA,用netRTPCR方法合成人血管能抑素cDNA基因,将该cDNA克隆进pSP72载体获得重组质粒pSP72C, DNA序列分析结果与预期序列一致。用BamHⅠ和NdeⅠ双酶切,切下pSP72C上的血管能抑素cDNA,插入pET3c载体的相应位点获得重组表达质粒pETC, 转化E. coli BL21(DE3), SDSPAGE分析显示：在IPTG诱导下,血管能抑素基因获得了高效表达,表达量约占菌体总蛋白的 27.9 %,主要以包涵体形式存在。包涵体经过洗涤、裂解、蛋白复性以及Sephadex G75凝胶过滤层析等步骤后,获得了纯度达91.4 %的人血管能抑素。CAM实验证明10 μg纯化蛋白就能显著抑制鸡胚新生血管生成。     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Vascular volumes and hematology in male and female runners and cyclists

To examine the hypothesis that foot-strike hemolysis alters vascular volumes and selected hematological properties is trained athletes, we have measured total blood volume (TBV), red cell volume (RCV) and plasma volume (PV) in cyclists (n = 21) and runners (n = 17) and compared them to those of untrained controls (n = 20). TBV (ml x kg(-1)) was calculated as the sum of RCV (ml x kg(-1)) and PV (ml x kg(-1)) obtained using 51Cr and 125I-labelled albumin, respectively. Hematological assessment was carried out using a Coulter counter. Peak aerobic power (VO2peak) was measured during progressive exercise to fatigue using both cycle and treadmill ergometry. RCV was 15% higher (P < 0.05) in male cyclists [35.4 (1.0), mean (SE); n = 12] and runners [35.3 (0.98); n = 9] compared to the controls [30.7 (0.92); n = 12]. Similar differences existed between the female cyclists [28.2 (2.1); n = 9] and runners [28.4 (1.0); n = 8] compared to the untrained controls [24.9 (1.4); n = 8]. For the male athletes, PV was between 19% (cyclists) and 28% (runners) higher (P < 0.05) in the trained athletes compared to the untrained controls. The differences in PV between the female groups were not significant. Although the males had a higher (P < 0.05) TBV, RCV and PV than the females, no differences between cyclists and runners were found for either gender. Mean cell volume was not different between the athletic groups. VO2peak (ml x kg(-1) x min(-1)) was higher (P < 0.05) in both male [68.4 (1.5)] and female [54.8 (2.1)] runners when compared to the untrained males [47.1 (1.0)] and females [40.5 (2.1)]. Although differences existed between the genders in VO2peak for both cyclists and runners, no differences were found between the athletic groups within a gender. Since the vascular volumes were not different between cyclists and runners for either the males or females, foot-strike hemolysis would not appear to have an effect on that parameter. The significant correlations (P < 0.05) found between VO2peak and RCV (r = 0.64 and 0.64) and TBV (r = 0.82 and 0.63) for the males and females, respectively, suggests a role for the vascular system in realizing a high aerobic power.