Biodegradation of polyhydroxyalkanoates by soil microbial communities of different structures and detection of PHA degrading microorganisms

Biodegradation of microbial linear polymers of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) by soil microbial communities of different structures has been studied during two field seasons in different weather conditions. This process was shown to be influenced by the polymer chemical composition, temperature, humidity, and the microbial soil component. The PHA degradation was accompanied by a decrease in the polymer molecular weight and an increase in the degree of crystallinity, indicating the preferential destruction of the amorphous phase compared to the crystalline one. The quantity of the true PHA destructors developing at the surface of the polymer samples was lower than the quantity of accompanying bacteria. The dominant PHA degrading microorganisms under the test conditions were identified as bacteria of the genera Variovorax, Stenotrophomonas, Acinetobacter, Pseudomonas, Bacillus, and Xanthomonas and as micromycetes from Penicillium, Paecilomyces, Acremonium, Verticillium, and Zygosporium.     

Microbial polymers as a degradable carrier for pesticide delivery

The possibility of use of polyhydroxyalkanoates (PHAs), biodegradable microbial polyesters, as a carrier for pesticides (α-hexachlorcyclohexane and lindane) for targeted and controlled delivery of these compounds to soil was investigated. The kinetics of polymer degradation and the dynamics of pesticide release from the extended-release formulations was studied. It is shown that pesticides embedded in a degradable polymer (PHA) carrier are released gradually and slowly, without surges, as the polymer is degraded by the soil micro-flora. The microbial soil component actively responded to the addition of the polymer as an additional nutrient substrate: the latter was degraded and then utilized. The rate of the pesticide release to the soil can be regulated by varying the polymer-pesticide ratio.     

Microbial degradation of polyhydroxyalkanoates in tropical soils

The integrated study addressing biodegradation of microbial linear polyesters of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) in tropical conditions by microbial communities of Vietnamese soils was performed in locations close to Hanoi and Nha Trang, which differed in their weather conditions and microbial communities. It shows that PHA degradation in tropical soils is influenced by polymer chemical composition, specimen shape, and microbial characteristics. The homopolymer of 3-hydroxybutyric acid is degraded at higher rates than the copolymer of 3-hydroxybutyric and 3-hydroxyvaleric acids. The average rates of mass loss were 0.04–0.33% per day for films and 0.02–0.18% for compact pellets. PHA degradation was accompanied by a decrease in the polymer molecular mass and, usually, an increase in the degree of crystallinity, suggesting preferential degradation of the amorphous phase. Under the study conditions, representatives of the bacterial genera Burkholderia, Bacillus, Cupriavidus, Mycobacterium, and Nocardiopsis and such micromycetes as Acremonium, Gongronella, Paecilomyces, and Penicillium, Trichoderma have been identified as major PHA degraders.     

海拔对辽东栎林地土壤微生物群落的影响

以北京东灵山辽东栎林地土壤为对象,运用氯仿熏蒸-浸提法及磷脂脂肪酸分析(PLFA)法,研究林木生长季节土壤微生物群落随海拔梯度的变化特征.结果表明:随着海拔升高,辽东栎林土壤微生物生物量碳、氮,以及微生物各类群含量均有差异但不显著;土壤细菌/真菌升高,而革兰氏阳性菌(G+)/革兰氏阴性菌(G-)降低.土壤微生物生物量碳、氮以及细菌、真菌、G+细菌、G-细菌的含量与土壤含水量、有机碳、全氮呈显著正相关,土壤真菌含量与土壤碳氮比值呈正相关.土壤微生物群落组成结构(细菌/真菌和G+细菌/G-细菌)的变化主要受土壤温度和土壤含水量的显著影响,说明土壤微生物群落结构对环境条件的变化敏感.随着全球变暖的加剧,暖温带辽东栎林地土壤真菌和G+细菌的比例有升高的趋势.     

毛乌素沙地根际土壤微生物数量与植物盖度相关性的初步研究

对毛乌素沙地不同生境下根际土壤微生物细菌、真菌、放线菌的数量进行了研究,结果表明：根际不同菌种菌落总量表现为细菌最多,放线菌次之,真菌最少;地貌部位、植物类型等不同生境条件下,微生物菌落总量和垂直分布上均存在明显差异;固定沙丘迎风坡及荒草地的生物多样性指数高于其他地方,各微生物类群间分布比例较均匀。植物盖度与微生物的数量并不存在直接的相关性。因此,在改善毛乌素沙地生态环境时,应关注不同生境条件下植被演替的递续性和植物类型的配置。     

不同种植年限菜田土壤微生物区系的研究

采用稀释平板涂抹法对山东聊城周边地区不同种植年限的菜田土壤微生物数量、组成与养分状况及其相互关系进行了研究。结果表明：①该地区土壤微生物以细菌占绝对优势,其次为放线菌;②随着种菜年限的增加,土壤细菌和微生物总数表现出“低高低”的变化趋势,而真菌和放线菌则随种菜年限的增加而增加;③菜田土壤各类微生物主要集中在0～20cm的土层中,随着土层加深,其数量迅速减少;④菜田土壤放线菌组成复杂,共分离到了6个属的放线菌,但仍以链霉菌为主,其次为小单胞菌属和马杜拉放线菌属。链霉菌可分为9个类群,白孢类群占优势。种植1—2a的菜田土壤放线菌组成复杂,而链霉茼组成较简单;⑤蔬菜种类不同时,土壤微生物亦不同;⑥土壤养分含量与真菌和放线菌呈负相关,与细菌和微生物总数呈正相关。     

Determination of the extraction kinetics for the quantification of polyhydroxyalkanoate monomers in mixed microbial systems

For the first time, a systematic approach was conducted to determine the key factors influencing the kinetics of hydroxyalkanote (HA) extraction in biological systems. Six mixed microbial systems where polyhydroxyalkanoate (PHA) is produced were evaluated. Experiments were carried out for full-scale and lab-scale activated sludge systems using different configurations (containing floccular or granular sludge), as well as specific PHA accumulating cultures that contain high or low intracellular PHA fractions. The overall reaction was limited by the kinetics of the PHA hydrolysis in floccular cultures, whereas in granular cultures, it was limited by the cell lysis step. The monomeric composition of the polymer also had an impact on the HA extraction rate: higher acid concentration and a longer digestion time should be employed when cells accumulate monomers with more substituents, such as hydroxy-2-methylbutyrate (H2MB) and hydroxy-2-methylvalerate (H2MV). This study optimised the method for HA extraction, which impacts the assessment of the quantity and quality of PHA biopolymers.     

石羊河下游退耕地土壤微生物变化及土壤酶活性

采用时空替代法,对石羊河下游不同年限(1,2,3,4,5,8,15,24、31 a)退耕地土壤微生物(细菌、真菌和放线菌)数量、生物量(碳、氮和磷)及土壤酶(过氧化氢酶、蔗糖酶、脲酶和磷酸酶)活性变化及三者的相关性进行了测定和分析。结果表明,在退耕1—31 a的9个样地样方中土壤三大类微生物数量以细菌最高,放线菌次之,真菌最低。总体来看,三大土壤微生物数量的加权平均值最大值均在退耕后的前8 a。土壤微生物生物量碳在退耕初期随着退耕年限的增加而减小,退耕4 a后逐渐增大,退耕24 a期间达到了加权平均值的最大,最后趋于稳定;土壤微生物生物量氮在退耕初期随着退耕年限的增加而增加,退耕4 a加权平均值的最大值出现,随后逐渐减小的趋势,并且不同退耕年限土壤微生物生物量氮差异显著;土壤微生物生物量磷在退耕初期随着退耕年限的增加而增加,退耕8 a前后加权平均值达到最大值,随后逐渐减小,最终趋于稳定。土壤酶活性总趋势随着退耕地自然演替时间的增加呈波动式下降。不同土壤层次(0—10 cm,10—20 cm,20—30 cm及30—40 cm),土壤微生物数量、生物量及土壤酶活性随土层深度显著降低,并且表层土壤微生物生物量及土壤酶活性占有较大比例。土壤微生物及土壤酶活性的变化是一个极其缓慢的互动过程,存在着互相回馈的响应,特别是真菌与放线菌、微生物量氮及蔗糖酶,放线菌与过氧化氢酶、蔗糖酶,微生物量碳与磷酸酶,微生物量氮与脲酶,微生物量磷与蔗糖酶均存在极显著的相关性。总体来看在退耕年限4—5 a前,有利于土壤发育,退耕后期土壤肥力呈下降的趋势。     

典型黑土区侵蚀-沉积对土壤微生物数量空间分布的影响

研究土壤微生物群落对土壤侵蚀-沉积的响应可为农业生态系统功能提升提供重要指导.选择黑龙江省典型薄层黑土区宾县宾州河流域为研究区,分析土壤微生物数量的空间分布特征,并结合137Cs示踪方法估算土壤侵蚀模数,进一步分析坡面和流域尺度土壤侵蚀-沉积对土壤微生物数量的影响.结果表明:季节变化对土壤微生物数量的影响非常明显,夏季...     

青藏高原东缘高山森林-苔原交错带土壤微生物生物量碳、氮和可培养微生物数量的季节动态

为了了解青藏高原东缘高山森林-苔原交错带土壤微生物的特征和季节变化, 研究了米亚罗鹧鸪山原始针叶林、林线、树线、密灌丛、疏灌丛和高山草甸土壤微生物生物量碳(MBC)、氮(MBN)和可培养微生物数量的季节动态。结果表明, 植被类型和季节动态对MBC、MBN和微生物数量都有显著影响。不同时期的微生物在各植被类型间分布有差异, 植物生长季初期和生长季中期, 树线以上群落的MBC高于树线下的群落, 而到生长季末期恰恰相反, 暗针叶林、林线和树线的MBC显著升高, 各植被之间MBC的差异减小; 微生物数量基本上也是以树线为界, 树线以下群落土壤微生物数量显著低于树线以上群落, 其中密灌丛的细菌数量最高; 可培养微生物数量为生长季末期>生长季初期>生长季中期。生长季末期真菌数量显著增加, 且MBC/MBN最高。统计分析表明, MBN与细菌、真菌、放线菌数量存在显著的相关关系, 而MBC仅与真菌数量存在显著相关关系( p < 0.05)。植物生长季末期大量的凋落物输入和雪被覆盖可能是微生物季节变异的外在因素, 而土壤微生物和高山植物对有效氮的竞争可能是微生物季节变异的内在因素。植物生长季初期对氮的吸收和土壤微生物在植物生长季末期对氮的固定加强了高山生态系统对氮的利用。气候变暖可能会延长高山植物的生长季, 增加高山土壤微生物生物量, 加速土壤有机质的分解, 进而改变高山土壤碳的固存速率。     

桂林会仙喀斯特湿地芦苇群落土壤微生物数量动态分析

以广西桂林会仙喀斯特湿地典型芦苇植物群落为研究对象,于春、夏、秋、冬四个季节分别采集0~10cm,10~20 cm和20~30 cm不同层次的土壤样品,分析根际微生物与非根际微生物的数量特征及季节动态变化特点,探讨微生物数量对水热季节变化的响应规律。结果表明:不同季节的根际微生物与非根际微生物组成,均以细菌占绝对优势;微生物数量分布大小顺序为细菌放线菌真菌,细菌最高比例为96.62%,放线菌最高比例为35.38%,真菌的比例较低,最高仅为0.30%。细菌,真菌和放线菌的垂直变化明显,均随着土层的增加而呈现递减的趋势。不同土壤层次根际微生物与非根际微生物的季节变化一致,细菌数量表现为夏季秋季春季冬季,真菌数量表现为秋季夏季春季冬季,放线菌数量表现为秋季春季夏季冬季;细菌、放线菌、真菌的最大值分别为2.70×10~7、1.92×10~6、3.35×10~4cfu·g~-1,土壤微生物数量与土壤有机碳、全氮、全磷、全钾、速效氮、速效磷、速效钾等呈显著正相关。芦苇植物群落根际土壤微生物呈现出一定的根际效应,并与微生物数量、土壤深度、月平均降雨量和月平均气温变化等有关,而在冬季的根际效应则表现不显著。土壤养分含量是调节会仙喀斯特湿地土壤微生物数量变化的一个主要因素。     

黄萎病不同发生程度棉田中土壤微生物多样性

作物根际土壤微生物群落对土壤生态及作物健康至关重要。以棉花黄萎病不同发生程度棉田的土壤为研究对象,采用理化分析、微生物纯培养及高通量测序技术对土壤理化性质及微生物数量、细菌丰度多样性进行综合分析。结果表明:在纯培养条件下,大丽轮枝菌无菌发酵滤液对细菌生长有明显的抑制作用;棉田接种大丽轮枝菌对土壤中细菌、真菌、放线菌的数量及细菌菌群丰度多样性未产生明显影响,不同采样时间的土壤中细菌菌群结构差异更大。土壤中大丽轮枝菌微菌核数量与棉花黄萎病的发生程度呈显著正相关。土壤肥力对土壤中微生物数量起主导作用,而水稻-棉花轮作能够使棉田有效降盐、减病、改善土壤肥力。通过生物防治、作物轮作、深翻等调控措施增加土壤中有益菌群数量、改善土壤生态、降低棉田土壤中大丽轮枝菌菌源数量是减轻棉花黄萎病危害的基础。     

嫁接对铜胁迫下黄瓜根际土壤微生物特性和酶活性的影响

采用盆栽试验方法,研究了嫁接（以黑籽南瓜为砧木）对铜胁迫下黄瓜根际土壤微生物生物量、微生物种群数量和土壤酶活性的影响.结果表明： 铜胁迫下黄瓜根际土壤微生物生物量碳（MBC）和微生物生物量氮（MBN）含量显著下降,基础呼吸和代谢熵显著上升,但嫁接黄瓜根际土壤MBC和MBN含量显著高于自根黄瓜,而基础呼吸和代谢熵则显著低于自根黄瓜.铜胁迫下,根际土壤放线菌和自生固氮菌的数量显著下降,真菌数量显著上升,而细菌数量变化不显著;嫁接黄瓜根系土壤细菌、放线菌、自生固氮菌的数量显著高于自根黄瓜,而真菌数量显著低于自根黄瓜.嫁接黄瓜根际土壤脲酶、磷酸酶、蔗糖酶、过氧化氢酶活性在铜胁迫下显著高于自根黄瓜.试验结果证明嫁接使铜胁迫下黄瓜根际土壤微生物环境和酶活性得到了改善和提高,从而提高了黄瓜植株对铜胁迫的抵抗力.     

漓江河岸带不同水文环境土壤微生物与土壤养分的耦合关系

基于漓江河岸带典型地段内砾石滩、草地、灌草地和疏林地等4类水文环境梯度上的土壤样品分析,探讨了漓江河岸带土壤养分、微生物数量及微生物量等的变化特征及耦合关系。结果表明:土壤养分含量、土壤微生物数量及微生物量等在不同水文环境下均有显著差异。随着水淹时间的减少,土壤含水量、pH值、有效氮、有效磷和有效钾均呈先增后减的趋势,最大值多出现在灌草地,最小值多出现在砾石滩,而土壤微生物总量、细菌、放线菌数量和微生物量也为先增后减趋势,其数量或含量在灌草地最多,砾石滩最少。简单相关分析表明漓江河岸带各微生物指标与土壤有效氮、有效磷及速效钾相关性较强,与土壤全氮、全磷和全钾的相关性较弱。冗余分析表明土壤有效氮、有效磷和含水量是影响微生物总数、细菌数量、微生物量碳、微生物量氮和微生物量磷的重要因素,而有机质和全氮是影响放线菌数量的重要因素。研究结果说明适当的水文干扰有利于河岸带土壤养分的积累,对土壤微生物数量及其生物量也有显著促进作用;土壤有机质和有效养分含量(有效氮、有效磷等)与微生物数量及微生物量关系密切,其含量很大程度影响着微生物数量及其活性。在漓江河岸带生态保护和评估等研究过程中应充分考虑不同水文环境下的土壤微生物和养分的变化特征及耦合关系。     

Polyhydroxyalkanoate synthesis using different carbon sources by two enhanced biological phosphorus removal microbial communities

Polyhydroxyalkanoate (PHA) is a biodegradable plastic synthesised by bacteria as energy and carbon storage material. PHA production is mostly based on pure cultures operated under sterile conditions, which increase the costs of this biopolymer. The use of inexpensive mixed culture biomass, such as activated sludge, to produce biodegradable plastics from renewable waste streams has been proposed as an alternative.The effect of carbon sources (acetate, propionate, butyrate and glucose) on the type and quantity of PHA synthesis obtained with different enhanced biological phosphorus removal (EBPR) microbial communities enriched with acetate and propionate are reported in this work. Two sequencing batch reactors (SBRs) were seeded with biomass withdrawn from a non-EBPR wastewater treatment plant (WWTP). The same operational conditions were kept, but using acetate or propionate as the sole carbon source of each reactor. These conditions produced two microbial communities with different P-removal capacity. The results presented in this study show the effect of the carbon source on the PHA composition (amount of polyhydroxybutyrate (PHB), polyhydroxyvalerate (PHV) and polyhydroxy-2-methylvalerate (PH2MV)), which differed not just between substrates but also between the two EBPR communities used. In addition, some monomers not always analysed contribute significantly to the total amount of PHA, especially when using butyrate, showing that if they are not considered this can lead to erroneous calculated yields.     

辉腾锡勒草原干涸湖泊中氨氧化微生物群落结构分析

【目的】以内蒙古辉腾锡勒草原九十九泉湿地为对象,研究湖泊干涸过程中氨氧化微生物的群落结构及其变化。【方法】通过MPN-PCR定量测定氨氧化古菌(AOA)和氨氧化细菌(AOB)的数量;构建amoA基因克隆文库,进行系统发育分析;结合土壤环境因子,探讨湿地退化过程中影响氨氧化微生物的潜在因素。【结果】依湖泊湿地退水梯度的不同样点中,有75%的样点AOB的数量高于AOA,AOB与AOA的数量比率为0.3-18.1。从湖心到湖岸草原带,AOA和AOB的数量有明显增加,但生物多样性呈降低趋势,二者没有呈现正相关。研究发现,AOB的数量与土壤中NH 4+-N的变化存在良好响应。系统发育分析显示,退化湖泊湿地AOA克隆序列均来自于泉古菌门(Crenarchaeota);AOB的amoA基因的克隆序列大部分与亚硝化单胞菌属(Nitrosomonas)有一定同源性,较少部分与亚硝化螺菌属(Nitrosospira)有一定同源性。【结论】湖泊退水过程增加了湿地土壤氨氧化微生物的数量,而氨氧化微生物的种群丰度有所降低。AOA和AOB群落对湖泊湿地的退化过程做出了响应,其中AOB的响应较为明显,氧化条件和土壤铵浓度的改变可能是促成这种响应的重要原因。     

极端干旱区防护林地土壤微生物多样性

土壤微生物在森林土壤中有重要功能.采用传统培养、脂肪酸鉴定和PCR-DGGE 3种方法分别研究了塔里木沙漠公路防护林地土壤微生物数量、脂肪酸种类和细菌DNA片段的多样性,从微生物学角度为塔里木沙漠公路防护林的管理提供理论依据.结果表明,塔里木沙漠公路防护林的建设促进了风沙土土壤微生物的发育,随着防护林定植年限的增加,土壤微生物数量、脂肪酸和细菌DNA片段的多样性指数明显增大;土壤微生物区系组成中,细菌是优势类群,占微生物总数80%以上,而真菌很少,不到微生物总数1%,但在不同土层间有所差异;传统培养法与现代生物标记和分子生物学方法对塔里木沙漠公路防护林地土壤微生物多样性的研究结果基本一致,说明塔里木沙漠公路防护林的建设使林地土壤生物活性有所增强,有利于林地土壤养分循环与利用.     

土壤微生物学特性对土壤健康的指示作用

土壤健康是陆地生态系统可持续发展的基础。作者通过概述土壤微生物学特性(土壤微生物群落结构、土壤微生物生物量、土壤酶活性)与土壤质量的关系, 阐明了土壤微生物对土壤健康的生物指示功能。研究表明: 土壤中细菌、真菌和放线菌的组成及其所占比率在一定程度上反映了土壤的肥力水平: 在土壤性质和肥水条件较好的土壤中, 细菌所占比率较高。土壤微生物生物量与土壤有机质含量密切相关, 而且土壤微生物生物量碳与土壤有机碳的比值(C_mic : C_org)和土壤微生物代谢熵(qCO2)的变化在一定程度上反映了土壤有机碳的利用效率。一般情况下, 土壤酶活性高的土壤中, 土壤微生物生物量碳、氮含量也高。因此, 土壤微生物学特性可以反映土壤质量的变化, 并可用作评价土壤健康的生物指标。     

Plant species richness,identity and productivity differentially influence key groups of microbes in grassland soils of contrasting fertility

The abundance of microbes in soil is thought to be strongly influenced by plant productivity rather than by plant species richness per se. However, whether this holds true for different microbial groups and under different soil conditions is unresolved. We tested how plant species richness, identity and biomass influence the abundances of arbuscular mycorrhizal fungi (AMF), saprophytic bacteria and fungi, and actinomycetes, in model plant communities in soil of low and high fertility using phospholipid fatty acid analysis. Abundances of saprophytic fungi and bacteria were driven by larger plant biomass in high diversity treatments. In contrast, increased AMF abundance with larger plant species richness was not explained by plant biomass, but responded to plant species identity and was stimulated by Anthoxantum odoratum. Our results indicate that the abundance of saprophytic soil microbes is influenced more by resource quantity, as driven by plant production, while AMF respond more strongly to resource composition, driven by variation in plant species richness and identity. This suggests that AMF abundance in soil is more sensitive to changes in plant species diversity per se and plant species composition than are abundances of saprophytic microbes.