外源磺胺二甲嘧啶对施用粪肥的菜地土壤中微生物群落多样性的影响

【目的】明确磺胺二甲嘧啶(SM2)随粪便施入土壤对土壤微生物群落造成的影响,为合理进行养殖场周围土壤环境质量评估和施肥管理提供参考依据。【方法】收集养殖场附近未经抗生素污染的粪便和土壤,构建粪便–土壤模型,设置不同SM2添加剂量的处理,分别为对照组(0)、低剂量组(5μg·kg~(-1))、中剂量组(500μg·kg~(-1))和高剂量组(5 000μg·kg~(-1)),并分别于粪便和SM2施入1、7、14和50 d后采集土壤样品,采用Biolog技术以及磷脂脂肪酸(Phospholipid fatty acid,PLFA)标记法对比分析不同处理组土壤微生物群落的碳源利用能力、菌群结构和功能多样性变化。【结果】SM2的施入提高了土壤微生物群落的总体碳源利用能力,中、高剂量组在施入50 d时对除酯类以外的其他碳源利用能力均强于对照组。低剂量组在施入7 d内对各类碳源利用能力增强,中、高剂量SM2的施入可能会影响土壤群落某些优势物种的繁殖和数量分配。随施入时间的延长,土壤微生物群落对不同剂量SM2反应不同,革兰阳性菌、革兰阴性菌以及真菌数量变化显著,放线菌变化不明显。【结论】SM2的施入将长期影响土壤微生物群落的结构与功能多样性。养殖场应及时处理动物排泄物以消除抗生素残留,减少粪肥对土壤环境造成的生态破坏。     

循环水养鳗系统生物过滤器中微生物群落的代谢特性

为了研究鳗鱼循环水养殖系统不同水处理单元的微生物群落碳代谢特征,实验采用Biolog Eco技术,分析了流化床两个槽和滴流式生物过滤器上、中、下三层的生物膜微生物群落功能多样性。结果显示,流化床两个槽和滴流式生物过滤器中、下层微生物多样性指数（Shannon-Wiener指数、Simpson指数和Pielou指数）无显著差异（P>0.05）,但均显著高于滴流式生物过滤器上层（P<0.05）。平均色度变化（AWCD）与主成分分析（PCA）均证明滴流式生物过滤器上层与其它采样点微生物代谢差异较大。各采样点微生物未对ECO板某一大类碳源表现出偏好,但对衣康酸、D-半乳糖醛酸、L-精氨酸、L-天门冬酰胺、L-丝氨酸、D-甘露醇、D-木糖、N-乙酰-D-葡萄糖氨、吐温40、吐温80、苯乙胺等单一碳源利用较好;而对γ-羟丁酸和α-丁酮酸以及D,L-α-磷酸甘油和1-磷酸葡萄糖利用较差。某些碳源种类如D-葡糖胺酸、α-D-乳糖、2-羟基苯甲酸仅能被部分采样点的微生物利用。本实验利用Biolog EcoPlateTM技术研究中试规模循环水处理单元微生物群落代谢特征,研究结果为生物过滤器的调控提供了一种新的思路即可以通过碳源调节,来促进生物膜微生物群落结构的改变以此提高水处理效率。     

典型血吸虫病疫区河滩地五氯酚污染土壤微生物特征——以松滋老陈镇庙河为例

采用现场采样及室内分析方法,对长期施用五氯酚(PCP)作为钉螺杀灭剂的典型血吸虫病流行疫区河滩地的土壤微生物特征进行了初步调查研究.结果表明,长期大量施用PCP对河滩裸地土壤微生物种群造成了一定程度的伤害,降低了微生物的总体活性;而种植杨树有利于恢复土壤微生物生物量碳,对修复PCP污染土壤有一定的促进作用.Biolog结果显示土壤微生物利用单一碳源能力的大小顺序为杨树林地>对照>河滩裸地,表明有机污染在一定程度上抑制了土壤微生物的生长.主成分分析显示对照、杨树林与河滩裸地的碳源利用能力差异显著,表明PCP污染对微生物的碳源利用造成了显著影响.土壤微生物对不同种类碳源利用计算结果表明,种植杨树导致林地土壤环境条件发生变化,杨树林地土壤微生物对碳源的利用也发生了明显变化.     

3株斑羚、岩羊化脓隐秘杆菌的鉴定及系统发育分析

本研究利用Biolog快速鉴定系统、16S rRNA序列分析及传统细菌鉴定方法对3株分离自北京动物园斑羚、岩羊化脓隐秘杆菌进行了鉴定和系统发育分析。结果表明,3株菌株均为化脓隐秘杆菌;系统发育分析结果表明,3株化脓隐秘杆菌16S rRNA序列与NCTC5224模式株同源性均为100%,且位于系统发育树的同一分支。     

土壤微生物多样性的研究方法

为了更加全面有效地研究土壤微生物多样性,笔者通过对于当前国内外研究土壤微生物多样性的研究方法,包括微生物稀释平板记数法、Biolog微平板法、PLFA分析法、分子生物学方法进行了阐述,说明了几种方法操作性的优缺点,重点说明了在实验过程中应该选取的方法,为研究者选取研究方法时提供基本信息及建议。     

Pollution-induced tolerance of soil bacterial communities in meadow and forest ecosystems polluted with heavy metals

Pollution-induced community tolerance (PICT) allows finding a cause–effect relationship between pollution and adverse changes in a community. In our previous study we found that functional diversity of bacterial communities decreased significantly with increasing metal concentration, in both forest humus and meadow topsoil. Thus, the aim of the present study was to test whether tolerance of soil bacterial communities had increased as an effect of long-term metal pollution. Bacterial tolerance was tested with the use of the Biolog^® ECO plates in soils originating from the most polluted and the least polluted sites from three forest and five meadow transects located near smelters in Avonmouth (England), Clydach (Wales), and Głogów and Olkusz (Poland). We found that tolerance of bacterial communities was significantly increased in polluted meadow soils when compared to control meadow soils. On the contrary, no increase in tolerance was detected in polluted forest humus.     

Do water regimes affect iron‐plaque formation and microbial communities in the rhizosphere of paddy rice?

Pot experiments were conducted to investigate the effect of soil water regimes on the formation of iron (Fe) plaque on the root surface of rice seedlings (Oryza sativa L.) and on the microbial functional diversity in a paddy soil. The rice seedlings were subjected to three moisture regimes (submergence, 100%, and 60% water‐holding capacity [WHC]), and were grown for 5 and 11 weeks. Aerobic lithotrophic Fe(II)‐oxidizing (FeOB) and acetate‐utilizing Fe(III)‐reducing bacteria (FeRB) in the rhizosphere and non‐rhizosphere soil were determined at 5 weeks using the most probable number (MPN) method. The carbon substrate use patterns of the microbial communities in the rhizosphere and non‐rhizosphere soil samples were determined at 11 weeks using Biolog‐GN2 plates. The amount of Fe plaque (per unit dry root weight) was much higher under submerged conditions than at lower soil moisture contents and decreased with plant age. There was a positive correlation between the amount of Fe plaque and phosphorus accumulated in the Fe plaque at both sampling times (r = 0.98 and 0.92, respectively, n = 12). Numbers of FeOB and FeRB in the submerged soil were lower than in aerobic soil, but by two orders of magnitude higher in the rhizosphere than in the bulk soil. On the other hand, the functional diversity of the rhizosphere microbial communities was much higher than that of the non‐rhizosphere soil, irrespective of soil water regimes. We conclude that soil flooding results in a decreased number and diversity of Fe‐oxidizing/reducing bacteria, while increasing the Fe‐plaque formation.     

Persistence of Cry1Ac Protein from Transgenic Bt Cotton Cultivation and Residue Returning in Fields and Its Effect on Functional Diversity of Soil Microbial Communities

The persistence of Cry1Ac protein in the soil and its effect on soil microbial communities are a core issue in assessing the ecological risk of transgenic Bacillus thuringiensis (Bt) cotton. In this study a field experiment was conducted on the cultivation of transgenic Bt cotton (Jin 26 and BtJi 668) with the immediate returning of residues to the fields, in order to quantify the Cry1Ac protein content in the fields and investigate its effects on the functional diversity of soil microbial communities. Cry1Ac protein in the residue-soil mixture was gradually degraded in the transgenic Bt cotton fields. After transgenic Bt cotton straw was returned to the fields for 30 d, 63.73% and 58.33% of the initial amounts of Cry1Ac protein were degraded in the Jin 26 and BtJi 668 fields, respectively. Before the crops were sown in the following year (180 d after returning the straw), no Cry1Ac protein was detected in the fields. After returning the cotton straw to the fields for 30 d, the Shannon-Wiener and McIntosh indices of soil microbial communities in the transgenic Bt cotton fields were significantly higher than those in the non-transgenic cotton fields. Meanwhile, the utilization of carbon sources including amino acids, amines, and carbohydrates by the soil microbial communities significantly increased. Both the McIntosh index and the utilization of carbohydrates increased until 180 d. Principal component analysis revealed that amino acids, amides, and carbohydrates were the main carbon sources distinguishing the two principal component factors. These findings indicated that Cry1Ac protein did not accumulate in the fields after transgenic Bt cotton was planted for one year and the residues were immediately returned to the fields; however, the original functional diversity of soil microbial communities was affected continuously.     

鲢鳙放养对水体微生物碳源利用影响的围隔试验

利用Biolog技术就滆湖原位围隔内鲢鳙放养密度和比例对水体微生物碳源利用的影响进行了研究。结果显示,在试验阶段后期不同围隔内水体微生物对碳源总量的利用相对于试验中期有较明显的变化。未放养鲢鳙的对照组(A0)在试验后期对于碳源总量的利用低于各实验组(A_1:鱼密度40 g/m~3、鲢鳙比7∶3;A_2∶40 g/m~3、3∶7、A_3∶80 g/m~3、7∶3和A_4∶80 g/m~3、3∶7)。试验中期和后期,利用比例较高的碳源类型均为聚合物。主成分分析表明,试验后期不同围隔水体微生物群落碳代谢方式具有显著差异。其中,与主成分1显著相关的碳源主要有16种,分别属于聚合物、糖类、羧酸、氨基酸。实验中期和后期围隔水体微生物碳代谢多样性指数Shannon物种多样性指数、丰富度指数和Pielou均匀度指数均存在显著差异,代谢多样性指数最高的是实验组A1,实验组A4碳代谢多样性指数相对较低。根据鳙的绝对生物量、多样性指标以及对碳源的总量利用,可以得出以鲢鳙主导的微型生态系统中,鳙的绝对生物量对于水体微生物影响更加显著。     

沙漠微生物群落功能多样性分析

以宁夏回族自治区沙坡头地区的沙漠微生物群落为研究对象,采用Biolog方法研究了已固定沙丘结皮(J1)及结皮下层(J2),半流动沙丘上层(L1)和下层(L2)中微生物群落的功能多样性。结果表明,J1的微生物群落活性最高,J2的微生物群落次之,再次为L2的微生物群落,最后为L1的微生物群落;J1,J2,L1和L2的微生物群落对羧酸类、聚合物类和氨基酸类碳源的利用表现一致,对糖类、胺/氨类和双亲化合物类碳源的利用表现不同;利用ECO板上31种碳源进行主成分分析得出,PC1,PC2和PC3特征值的贡献率为分别为70.16%,19.23%,10.61%。研究结果表明:(1)经过固沙后的微生物群落活性明显高于半流动沙漠微生物群落的活性;(2)微生物群落结构上的差异致使其对6类碳源的利用方式不同;(3)主成分分析表明沙漠微生物群落对羧酸类、糖类和氨基酸类碳源的利用最高,主要利用碳源有22种。