营养物质及金属离子对DLL-E4菌降解对硝基苯酚的影响

研究了酵母膏、葡萄糖、蛋白胨、土壤浸液及Ca2 、Mg2 、Fe2 、Fe3 、Al3 、Mn2 、Co2 、Zn2 、Li 、Cu 、Cu2 、Ba2 、Ni2 等 13种金属离子对甲基对硫磷降解菌DLL E4降解对硝基苯酚的影响。结果表明 :适量添加酵母膏、葡萄糖和蛋白胨都能有效提高菌株对对硝基苯酚的降解 ,土壤浸液没有影响 ;金属离子中 ,除Li 和 0 1mmolL-1Fe3 外均对DLL E4降解对硝基苯酚的性状有一定的影响 ,其中Ca2 、Mg2 、Mn2 影响不大 ,Fe3 、Fe2 、Al3 、Ba2 、Zn2 高浓度时影响较大 ,Co2 、Cu2 、Cu 、Ni2 对DLL E4降解对硝基苯酚有较大的影响     

基于构建微生物传感器的甲基对硫磷降解菌的分离鉴定及其降解特性研究

有机磷农药是目前环境中残留量最多的农药之一,对其残留量的检测及降解机制的研究对于环境污染及生态修复具有重要意义。微生物传感器由生物学元件与换能器构成,因具有成本低廉、易于微型化及选择性高等特点而被广泛应用于各种生化物质的分析和检测。本文从长期受农药污染的土壤中分离出4株能以甲基对硫磷为碳源生长的菌株,根据形态特征和16S r RNA基因序列同源性分析,对4株降解菌进行鉴定,利用高效液相色谱测定降解率,选取降解率最高的1株菌进行降解机制研究,以期将其应用于测定环境中甲基对硫磷残留的电位型微生物传感器的构建。结果表明,在甲基对硫磷初始浓度50 mg·L-1、30℃、p H 7.0的培养条件下培养7 d,4株菌对甲基对硫磷的降解率均在78%以上,其中1株菌的降解效率可达100%。16S r RNA基因序列测定表明,该菌株属于克雷伯氏菌属,命名为Klebsiella sp.MP-6。利用液相色谱-质谱联用对其降解产物的研究表明,菌株MP-6水解甲基对硫磷主要产生二甲基硫代磷酸(dimethyl thiophosphoric acid,DMTP)和对硝基苯酚(p-nitrophenol,PNP),极少部分PNP通过产生4-硝基邻苯二酚(4-nitrocatechol,4-NC)和1,2,4-苯三酚(1,2,4-BT)进一步代谢。结果表明,基于测定中间产物对硝基苯酚(p-nitrophenol,PNP)的电位响应信号,该菌株适用于构建测定海水及土壤等环境中有机磷农药的微生物传感器。     

节杆菌属甲基对硫磷的降解菌株L-W的分离及降解特性

从生产甲基对硫磷的华阳农药厂污水曝气池中,分离到1株能以甲基对硫磷及其降解中间产物对硝基苯酚为唯一碳源生长,且能够将其彻底降解为CO2和H2O的细菌L-W,经鉴定,为节杆菌属(Arthrobacter sp)。用气相色谱法和分光光度计法深入研究了L-W的降解能力,其7h内对50mg/L甲基对硫磷的降解率为85%,对50mg/L对硝基苯酚的降解率为99%。     

蜡状芽孢杆菌HY-1降解甲基对硫磷和毒死蜱的影响因素研究

采用富集驯化培养和紫外分光光度计定量的方法,从农药生产企业的废水处理系统中分离筛选出1株能够降解甲基对硫磷和毒死蜱的蜡状芽孢杆菌（Bacilluscereus）HY-1,并系统研究了影响其降解甲基对硫磷和毒死蜱的主要因素。研究表明,菌株HY-1能够利用甲基对硫磷和毒死蜱为唯一磷源降解农药。HY-1降解甲基对硫磷的适宜条件为：培养温度30～35℃,pH为6～8,甲基对硫磷初始浓度为10～50mg·L^-1,接种量20%（体积比,菌体密度：稀释到菌悬母液（OD600=3.0）的0.8～1倍）,添加葡萄糖不能促进菌株对甲基对硫磷的降解。HY-1降解毒死蜱的适宜条件为：葡萄糖浓度6g·L^-1,培养温度30～35℃,pH为7.0,毒死蜱初始浓度80～200mg·L^-1,接种量20%（体积比,菌体密度：稀释到菌悬母液（OD600=3.0）的0.8～1倍）。结果表明,HY-1菌株降解甲基对硫磷和毒死蜱的适宜条件相类似,只是降解所需的最适葡萄糖浓度和底物浓度不同。     

一株新的多菌灵高效降解茵的筛选与降解特性分析

从长期施用多菌灵农药的土壤中,通过富集筛选,获得1株新的多菌灵高效降解菌株。通过生理生化实验和16SrDNA序列同源性分析鉴定该菌株,应用高效液相色谱法对纯培养条件下菌株的降解特性和粗酶提取液的降解性能进行了分析。结果表明,筛选所获得的菌株与Raoultella菌属的亲缘关系最近,将其命名为Raoultellasp．MBC,该菌株能在以多菌灵为唯-碳源的无机盐培养基中生长;25℃、pH7．0、200r·min。的最适生长条件下避光振荡培养72h,多菌灵的降解率达到100％;在最适培养条件下外加氮源和碳源在培养后期均可以提高多菌灵的降解率,外加氮源对多菌灵的降解效果优于外加碳源;该菌体的粗酶提取液具有降解多菌灵活性,且多菌灵降解酶为诱导酶。研究结果为多菌灵污染土壤的生物修复和酶修复提供了材料和理论依据。     

高效纤维素优势分解菌的筛选和鉴定

本试验基于获得高效纤维素优势分解菌的目的,通过分离纯化初步得到30株菌株,利用刚果红染色法初筛共得到14株纤维素分解菌,并通过滤纸条崩解实验进一步进行筛选得到5株效果较好的纤维素分解菌,通过发酵产酶利用DNS显色法测定CMC酶活力和FPA酶活力最终确定了4株优势纤维素分解菌,通过测定4株菌株的羧甲基纤维素酶(CMCase)、滤纸酶(FPA)以及β-葡萄糖苷酶(β-Gase)活,验证4株纤维素优势分解菌的产酶能力,并分别命名为X-1、X-6、X-7和X-11,并将该4株优势纤维素分解菌应用于秸秆的液态发酵,其对秸秆的降解率较自然降解相比,降解率分别提高了31.92%、40.15%、35.29%和39.98%。对4株优势菌株进行了分子鉴定,根据16S r DNA序列比对结果表明,菌株X-1、X-7和X-11均为粪产碱杆菌;菌株X-6属于解糖假苍白杆菌。     

降解玉米秸秆真菌复合菌系的构建及其降解效果评价

  【目的】  秸秆的木质纤维素含量丰富、结构复杂，在自然界中降解较慢，增加秸秆降解菌剂中菌株的多样性有利于提升还田秸秆的降解效果。探究菌株多样性水平和组成影响复合菌系秸秆降解的效果及原因，为复合菌系在秸秆降解中的应用提供理论支撑。  【方法】  通过富集驯化培养，从玉米秸秆还田土壤中筛选具有秸秆降解能力的真菌，从中挑选5株高效秸秆降解真菌进行基因间隔区序列 (ITS) 测定和物种鉴定，明确其分类地位。通过全组合构建菌株多样性为1～5的复合菌系，分别检测复合菌系的秸秆相对降解率及其滤纸酶、纤维素内切酶和木聚糖酶活性，利用方差分析和相关性分析等方法研究菌株多样性和组成对复合菌系玉米秸秆降解效果及其纤维素酶活性的影响。  【结果】  共筛选获得了15株具有秸秆降解能力的真菌，其中5株真菌的秸秆降解效果好、纤维素水解能力强。经ITS序列鉴定和系统发育分析，发现5株降解真菌的遗传差异较大，Z7-6、F7-5、F4-3、L1-1和J2-5分别与草酸青霉 (Z7-6: Penicillium oxalicum)、烟曲霉 (F7-5: Aspergillus fumigatus)、哈茨木霉 (F4-3: Trichoderma harzianum)、白囊耙齿菌 (L1-1: Irpex lacteus) 和木贼镰刀菌 (J2-5: Fusarium equiseti) 的ITS序列相似度均超过99.95%。全组合复配结果表明，复合菌系的秸秆降解能力和纤维素酶活力均高于各单一菌株，且随着菌株多样性水平的增加而提高。滤纸酶、纤维素内切酶和木聚糖酶的活力越强，复合菌系对玉米秸秆的降解效果越好，而其秸秆相对降解率主要取决于滤纸酶和纤维素内切酶的活性。抽样效应分析发现，不同菌株对复合菌系的秸秆降解效果、滤纸酶和纤维素内切酶活性的影响不同。不含菌株F7-5的复合菌系降解效果显著优于含有该菌株的组合，以Z7-6 (P. oxalicum)、F4-3 (T. harzianum)、L1-1 (I. lacteus) 和J2-5 (F. equiseti) 组合F1的玉米秸秆降解效果最佳、酶活性最高。  【结论】  秸秆降解复合菌系的构建过程需要同时考虑多样性效应和抽样效应，增加降解菌的多样性有助于增强秸秆的降解效果。本研究筛选获得的复合菌系F1在玉米秸秆降解中具有潜在的应用前景。     

沼灌土壤17β-雌二醇专性降解菌的分离筛选及降解特性

集约化畜禽养殖场产生的沼液通常就地回用,在循环利用有机物的同时也会带来类固醇雌激素（Steroid Estrogens,SEs）的累积及污染。为降低沼灌后SEs对水土环境的污染风险,该研究采用富集和纯化培养法,对西南地区某奶牛养殖场沼灌区土壤中雌激素降解菌进行分离及筛选,获得一株利用17β-雌二醇（17β-E2）为唯一碳源生长繁殖的降解菌。通过16S rDNA 基因序列进行同源性比对以确定种属,并研究其降解特性。分别研究了菌株在不同温度、pH值、底物浓度三种单因素条件下的降解特性,然后利用三因素三水平正交试验继续优化菌株最适降解条件。结果表明：分离出的优势菌为生丝微菌属（Hyphomicrobium sp.）,命名为Hyphomicrobium sp.SS-1,该菌株在10～40 ℃、pH值为5～9、底物浓度为1～10 mg/L的条件下,均能不同程度降解17β-E2。其中菌株在温度为30 ℃、pH值为7、底物浓度5 mg/L的条件下,培养7 d对17β-E2的降解率可达71%,并伴随毒性低于E2的降解产物E1和E3生成,总雌激素去除率为56.8%。正交试验结果显示,各因素对菌株降解能力的影响顺序从小到大为：底物浓度、温度、pH值,且都为显著影响（P<0.05）;菌株最适降解条件为温度35 ℃、pH值为7、底物浓度5 mg/L,该条件下培养7 d,菌株对17β-E2的降解率可达97.09%。研究结果可为复杂基质环境中微生物降解SEs提供优质菌种资源,并为沼液灌溉区土壤的雌激素污染修复提供有效途径。     

盐渍化土壤中土著菌的石油烃降解潜力研究

自盐渍化地区（黄河三角洲）采集4种不同石油污染程度的土壤样品,从中筛选出高效降解石油烃的4个菌系和8个单菌株。分别以原油、柴油、烷烃和多环芳烃（PAHs）为底物进行培养,测定降解菌的生物量和降解率,研究其对不同底物的耐受浓度和降解潜力。结果表明,获得的石油烃降解菌为轻度嗜盐菌;不同菌株对不同底物的耐受浓度不同,混合菌系对不同底物的降解能力强于单菌株,对单一组分底物的降解优于复杂组分的底物;单菌株I-2、3、5、7能较好地降解PAHs并且对原油的降解能力高于柴油,单菌株I-1、4、6、8能够利用烷烃且对柴油的降解能力要比原油高;降解菌对柴油和原油的最高降解率分别可达78．4％和70．7％,对正十六烷和菲的生物降解率分别高达87．7％和88．1％,表现出较强的降解能力。研究结果表明黄河三角洲盐渍化土壤中土著菌对石油烃污染土壤具有较强的牛物修复潜力。     

油泥高效降解菌的筛选及其性能研究

从胜利油田及河南油田分离筛选出6株除油菌,对单一菌株和XT-4、F-2、C-2、A-1和AB-1菌组成的混合菌进行了性能评价,并对混合菌降解石油烃的能力进行了研究。通过研究表明,混合菌在耐温、耐酸碱和耐矿化度方面均比单一菌种优越,混合菌具有更好的协同作用和对环境的适应性,其对含油量高和含盐量高的油泥具有很好的降解效果,当油泥含油量高达126g kg-1时,经过混合菌120d的降解,降解率可以达到61.35%。     

氯苯、对硝基酚对土壤生物活性的影响

本文通过对土壤呼吸强度，各微生物生理群的数理以及土壤酶活性的测定，研究了氯苯，对硝基酚对土壤生物活性的影响。实验表明氯苯，对硝基酚对土壤活性的影响是复杂的。氯苯在２００ｍｇ／ｋｇ以下对土壤呼吸强度，细菌放线菌及真菌的数量都有一定的刺激作用；而对氨化细菌硝化细菌以及反硝化细菌则表现出抑制作用；     

Phosphatase activity and phosphorus fractions in Karri (Eucalyptus diversicolor F. Muell.) forest soils

Summary Karri forest soils contain negligible concentrations of labile-P, low concentrations of total P and more P in organic forms than inorganic. The ratio of organic P to inorganic P was lowest (1:2) in recently burnt surface soils and greatest (7:1) at depth in soil that had been undisturbed for long periods of time. Phosphomonoesterase and phosphodiesterase activities (to 10 cm depth, phosphomonoesterase 700–1300; phosphodiesterase 2000–2400 g nitrophenol released h^-1 g^-1 fresh weight) were comparable to those in other, organically rich forest soils. The optimum pH for phosphatase activities were within 1–2 units of soil pH (6) and little reduction in activity was observed over the pH range 4–8. Phosphatase activity was reduced by air-drying (up to 20-fold reduction) and was almost entirely absent in soils that were heat-affected as a result of logging/burning operations. Neither phosphomonoesterase nor phosphodiesterase were directly related to soil P fractions or total P. A reduction in P demand is postulated as the cause of reduced phosphatase activity and the increased concentration of organic P with increasing soil depth.     

Salinity and the persistence of parathion in flooded soils

The persistence of parathion in five coastal saline soils of varying electrical conductivity and in one nonsaline soil sample was studied under flooded conditions. Parathion was decomposed faster in nonsaline soil than in saline soils and its stability increased with increasing electrical conductivity. The addition of salts to the nonsaline soil at 4, 8 and 16 dS^?1 increased the persistence of parathion. Nitro-group reduction, and not hydrolysis, was the major route of parathion degradation in saline and nonsaline soils. The accumulation of aminoparathion was less pronounced in saline soils than in nonsaline soil concomitant with slow degradation of parathion in saline soils. The inhibition of nitro-group reduction in saline soils was related to low microbial activities as reflected in decreased dehydrogenase activity and slow iron reduction.     

Biochar impact on the estimation of the colorimetric‐based enzymatic assays of soil

This study was carried out in order to assess the influence of biochar applications on the estimation of colorimetric‐based enzymatic assays and to verify the effectiveness of the most common methods. Since most methods used to determine enzymatic activities in the soil are based on colorimetry, biochar may absorb substrates and/or coloured products thereby distorting the analytical result. Biochar was added to two soils, with different textures and cation exchangeable capacities, at a rate of 2% (w/w), and seven enzyme activities were determined following standard methods. The biochar amendment lowered the spectrophotometer reading of the activity of FDAase and dehydrogenase in the sandy soil. In the three enzymatic activities based on p‐nitrophenol production (β‐glucosidase, phosphatase and arylsulphatase), the addition of biochar did not change the enzyme assays. The biochar led to an overestimation in terms of the protease and urease activities in the sandy soil. In the clay loamy soil, biochar did not change the response of any of the enzyme activities tested. A biochar dose of up to 2% only guarantees the effectiveness of the most common spectrophotometric methods for not excessively sandy soils.     

Comparative phosphatase activities in four species during germination in Nacl media.

In a first experiment, phosphatase activities of seedlings, at the first day of radicle emergence, and of roots of young plants, at the fifth day of radicle emergence, have been studied in three species differing in their NaCl tolerance threshold: tomato NaCl‐sensitive, red cabbage NaCl‐tolerant and radish semi‐tolerant. The Pase activities (p‐nitrophenol phosphatase, ATPase, Phytase) of red cabbage, at the both germinative stage, are the less modified by NaCl salinity. In a second experiment, Pase activities of excised and attached cotyledons of pea have been compared: Pase activities are dependent upon the presence of the axis; the ATPase activity is the main enzyme controlled by the axis. The pnpase activity of seedlings in NaCl free media appears a biochemical marker of NaCl tolerance degree; comparing ATPase and phytase or pnpase activities, also was discussed the availability of energy as a criterion of seed vigour and adaptation to Nacl media.     

Acid phosphatase activity and vesicular‐arbuscular mycorrhizal infection associated with roots of four wheat cultivars

Acid phosphatase activity and vesicular‐arbuscular mycorrhizal (VAM) infection associated with four spring wheat genotype roots were investigated. Plants were grown in a typic low P volcanic soil with and without P‐fertilizer addition and harvested at 21, 42, 63, 84 and 96 days. Results show that phosphatase activity, expresed as ug p‐nitrophenol released per g dry root, decreased from 21 to 96 days in all cultivars. Conversely, trends of VAM infection were similar in all genotypes being higher in P added plants at 63 days after sown. This opposite effects may be viewed as alternative and/or complementary adaptations for P‐uptake by plants growing in low nutrient situations. It was concluded that one of the ways of P‐alleviation in wheat growing in our volcanic soils might be the search of genotypes having high biochemical and/or biological root activities.     

低温生防菌株的诱变选育及生防效果

为了提高生防菌株wswshg-10低温生长速度和抑菌活性,采用常压室温等离子体诱变技术(ARTP)对其进行诱变选育。结果表明,最佳照射时间为120 s,此时菌株致死率为95.4%,单菌落变大的突变率为24.5%。经低温平板培养和抑菌活性测定,从19个大菌落的菌株中筛选出2株低温生长速度和抑菌活性明显高于出发菌株的突变菌株WM4和WM7,其低温(15℃)生长速度分别是出发菌株的1.94倍和2.09倍,平板抑菌活性分别较出发菌株高26.2%和29.8%。传代培养10代,2株突变菌株在低温生长速度和抑菌活性方面均表现出良好的遗传稳定性。突变菌株WM7对玉米茎基腐病的温室防效为70.8%,比出发菌株高15.1%。本研究结果为适于低温地区生物农药的研发提供了菌株资源。