Short-term population dynamics of ammonia oxidizing bacteria in an agricultural soil

Ammonia oxidizing bacteria (AOB) control the rate limiting step of nitrification, the conversion of ammonia (NH₄⁺) to nitrite (NO₂⁻). The AOB therefore have an important role to play in regulating soil nitrogen cycling. Tillage aerates the soil, stimulating rapid changes in soil N cycling and microbial communities. Here we report results of a study of the short term responses of AOB and net nitrification to simulated tillage and NH₄⁺ addition to soil. The intensively farmed vegetable soils of the Salinas Valley, California, provide the context for this study. These soils are cultivated frequently, receive large N fertilizer inputs and there are regional concerns about groundwater N concentrations. An understanding of N dynamics in these systems is therefore important. AOB population sizes were quantified using a real-time PCR approach. In a 15 day experiment AOB populations, increased rapidly following tillage and NH₄⁺ addition and persisted after the depletion of soil NH₄⁺. AOB population sizes increased to a similar degree, over a 1.5-day period, irrespective of the amount of NH₄⁺ supplied. These data suggest selection of an AOB community in this intensively farmed and C-limited soil, that rapidly uses NH₄⁺ that becomes available. These data also suggest that mineralization may play an especially important role in regulating AOB populations where NH₄⁺ pool sizes are very low. Methodological considerations in the study of soil AOB communities are also discussed.     

Influence of water potential on nitrification and structure of nitrifying bacterial communities in semiarid soils

The objective of this study was to examine the relationship between soil water potential, nitrifier community structure and nitrification activity in semiarid soils. Soils were collected after a 5-month dry period (end of summer) and subsequently rewetted to specific water potentials and incubated for 7 days prior to analysis of nitrification activity and nitrifier community structure. The approach used in this study targeted a 491bp segment of the amoA gene which encodes the active site of the ammonia monooxygenase enzyme, which is the key enzyme for all aerobic ammonia oxidisers. amoA serves as a useful target for environmental studies since it is both specific and universal for all ammonia oxidisers and reflects the phylogeny of the ammonia oxidisers. Our results suggest that in semiarid soils water potential plays a key role in determining the structure of ammonia oxidising bacteria (AOB), and that additionally AOB community structure is correlated to potential nitrification rate in these soils.     

Population size and specific potential of P-mineralizing and -solubilizing bacteria under long-term P-deficiency fertilization in a sandy loam soil

The effects of organic manure, mineral fertilizer (NPK), and P-deficiency fertilization (NK) on the individual biomass of young wheat plants, arbuscular mycorrhizal (AM) colonization in wheat root systems, population sizes of soil organic phosphorus mineralizing bacteria (OPMB) and inorganic phosphate solubilizing bacteria (IPSB) as well as soil P-mineralization and -solubilization potential were investigated in a long-term (18-year) fertilizer experiment. The experiment included five treatments: organic manure, an equal mixture of organic manure and mineral fertilizer, fertilizer NPK, fertilizer NK, and the control (without fertilization). Plant biomass, population sizes of soil OPMB and IPSB were greatly increased (P<0.05) by the application of organic manure and slightly increased by the balanced application of mineral fertilizer, while undiminished AM colonization in wheat root system was only observed in the case of the NK treatment. Compared to balanced fertilization, P-deficiency fertilization resulted in a significant increase (P<0.05) of OPMB-specific mineralization potential (soil P-mineralization potential per OPMB cell) and highest IPSB-specific solubilization potential (soil P-solubilization potential per IPSB cell), suggesting that OPMB and IPSB are likely more metabolically active in P-deficiency fertilized soils after long-term fertilizer management, and mycorrhizal plants are more dependent on AM in P-poor soils than in P-fertilized soils. Our results also showed the different effects of mineral fertilizer versus organic manure on soil P-mineralization and -solubilization potentials, as well as specific potentials of OPMB and IPSB in arable soils.     

高产水稻土细菌多样性的培养法与非培养法比较研究

利用细菌的通用引物扩增江西余江县高产水稻土红壤细菌总DNA和平板培养细菌混合总DNA的16S rDNA基因片段,在此基础上分别建立两种16S rDNA文库(文库a和文库b)。从两个文库中各随机挑选100个克隆,扩增出阳性克隆中的插入片段后选用HhaⅠ和RsaⅠ两种四碱基酶进行ARDRA(amplified rDNA restriction analysis)分析。统计比较分析发现,文库a的Shannon-Wienner指数、Simpson指数、丰富度、均一度分别为4.432、0.987、18.885和0.973,均高于文库b中相应的多样性参数(分别为2.271、0.758、5.736和0.501),即平板培养方法所展现的细菌群落结构多样性低于土壤中原始的多样性。结果表明,传统培养方法存在着很大的局限性,必须结合新的分子生物学技术手段才能更全面完善地认识土壤微生物群落结构多样性,以期充分利用其中丰富的微生物资源。     

棉花黄萎病拮抗细菌HMB-1005鉴定及定殖

以大丽轮枝菌(Verticillium dahliae)为指示菌,对从棉田土壤中已经分离筛选到的细菌HMB-1005进行形态、19项生理生化特征及16S rDNA试验,结果表明,HMB-1005为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);通过灭菌土和非灭菌土进行盆栽试验,拮抗菌HMB-1005芽孢液浸种、土壤全部混菌和部分混菌3种处理,经过5个月的盆栽试验表明,拮抗细菌HMB-1005能够在灭菌土和非灭菌土中定殖,且定殖数量达106cfu/g土左右,并仍能保持较高的抑菌活性。通过对棉苗根内细菌的分离,证实拮抗细菌HMB-1005能在棉花根内定殖,数量达到103cfu/g。实验结果显示,灭菌土的拮抗菌数高于非灭菌土,并且3种处理方式差异不明显,在实际生产应用中,建议使用浸种方法。     

反硝化技术对模拟养殖池塘修复的研究

浙江奉化市池塘的底泥经过反复培养、驯化,从中筛选、分离出反硝化细菌,在模拟实验条件下,研究其对不同浓度的硝酸盐氮和亚硝酸盐氮的去除情况,讨论反硝化菌种的生长情况。结果表明,在初始浓度为1、25、50mg·L^-1的硝酸盐氮和亚硝酸盐氮模拟池塘中,随着实验的进行,对污染物的去除效果逐渐提高。其中在1mg·L^-1的浓度组中,3d内硝酸盐氮和亚硝酸盐氮去除率就分别达到了95．8％和90．2％;在25mg·L^-1的浓度组中,第6d硝酸盐氮和亚硝酸盐的去除率分别为93．8％和87．8％;在50mg·L^-1的浓度组中,第6d硝酸盐氮和亚硝酸盐的去除率分别为89．7％和78．7％。此外,反硝化菌对硝酸盐氮的去除效果略优于亚硝酸盐氮,而且硝酸盐氮和亚硝酸盐氮的浓度越低,对其去除效果越好,达到稳定状态的时间越短。在模拟池塘中,菌种的生长情况与硝酸盐氮和亚硝酸盐氮的浓度呈负相关,即污染物的浓度越高反硝化菌的生长情况越差。对反硝化菌的生态影响因子研究表明,其反硝化最适宜的pH值为6～7,温度为25～35℃;而且在同一pH值和温度条件下,硝酸盐氮和亚硝酸盐氮浓度越低,反硝化菌对其去除效果越好。     

蚯蚓对金霉素污染土壤酶活性和微生物群落的影响

为探究蚯蚓对抗生素污染土壤酶活性和微生物群落的影响,选取金霉素为目标污染物,威廉环毛蚓（内层种）和赤子爱胜蚓（表层种）为实验生物,通过高效液相色谱-串联质谱法、土壤酶试剂盒（微板法）、高通量测序技术和冗余分析对土壤中金霉素残留、酶活性和微生物群落进行了测定与分析。结果表明：经过28 d的培养,蚯蚓处理组中金霉素浓度（0.022 4~1.006 0 mg·kg^-1）显著低于对照组土壤（0.033 5~1.585 0 mg·kg^-1）。金霉素抑制了土壤脱氢酶、蔗糖酶、脲酶和碱性磷酸酶的活性,激活了过氧化氢酶的活性。两种蚯蚓均激活了土壤脱氢酶、过氧化氢酶、蔗糖酶和脲酶的活性,最大激活率分别为77.80%~88.50%、4.00%~6.16%、69.20%~72.60%、43.20%~48.80%;碱性磷酸酶的活性受到抑制,最大抑制率为23.8%~25.0%。高通量测序结果表明两种生态型蚯蚓均可改变土壤中细菌群落结构,不同蚯蚓类型对细菌群落结构的影响存在有差异。两种蚯蚓的加入均未改变土壤中优势菌门的组成,但都改变了其丰度占比。在属水平上,Flavobacterium、Aeromonas、Luteolibacter、Adhaeribacter和Pseudomonas等潜在金霉素降解菌被两种蚯蚓刺激丰度显著增加,加速了土壤中金霉素的降解。冗余分析结果表明,土壤中pH、有机质含量、金霉素浓度和细菌群落结构是影响土壤酶活性变化的重要因子。研究表明,威廉环毛蚓对脱氢酶、蔗糖酶和脲酶的促进作用优于赤子爱胜蚓。     

生物炭对烤烟成熟期土壤养分及根际细菌群落结构的影响

采用大田小区处理方式,分别设置常规施肥(CK)和750 kg/hm~2生物炭+常规施肥(T)2组试验,探究生物炭对植烟土壤微生物群落和土壤养分的变化规律及其对烟叶质量的影响。结果显示:生物炭可以显著提高土壤pH、速效磷、速效钾和有机碳含量,促进烟株生长发育;与CK相比,生物炭处理后绿弯菌门(Chloroflexi)、酸杆菌门(Acidobacteriota)和厚壁菌门(Firmicutes)丰度增加,分别提高了1.82%、12.36%和64.55%;放线菌门(Actinobacteriota)和变形菌门(Proteobacteria)丰度分别降低了2.02%和9.00%;聚类和主成分分析结果显示,烤烟根际土壤中优势细菌与土壤pH、速效钾、速效磷和有机碳均存在显著的相关关系,其中厚壁菌门(Firmicutes)、Myxococcota和Desulfobacterota与土壤pH、速效磷、速效钾和有机碳存在显著的正相关关系(P0.05),Bacterdidota与Patescibacteria与土壤pH、速效钾、速效磷和有机碳存在显著负相关关系;生物炭施用通过改善根系周围矿质营养和微生物群落进而提高烟叶质量。生物炭有利于成熟期土壤养分固持及根际促生细菌群落增加。     

一组木质纤维素分解菌复合系的筛选及培养条件对分解活性的影响

以麦秸垛下的土壤和麦秸堆肥为材料 ,利用限制性培养技术 ,经过多代淘汰及其不同系之间的组配 ,最终筛选构建了一组木质纤维素分解菌复合系。用羧甲基纤维素 (CMC)糖化力法和纤维素减重法研究了不同培养条件对复合系分解纤维素能力的影响。结果表明 ,以滤纸、棉花、稻草、CMC Na和葡萄糖为碳源时 ,复合系在天然纤维素含量高的碳源 (如滤纸、棉花 )存在时表现出高的纤维素分解活性 ;利用蛋白胨和酵母粉作氮源时的纤维素分解活性远高于硝酸铵、尿素等无机氮源 ;以滤纸和酵母粉作为惟一碳、氮源时 ,最适发酵质量浓度分别为 2 5和10 0 g·L-1,其分解活性最高峰均出现在发酵第 5天。最适纤维素分解的 pH为 8 0。最适纤维素分解温度为 6 0℃ ,大于 6 0℃的高温抑制复合系对纤维素分解 ;复合系的适宜溶氧量 (DO)为 0 0 7～ 0 16mg·L-1,过高过低的溶氧量均抑制纤维素分解。复合系的微好氧特点对堆肥工程降低生产成本具有重要意义。     

乳酸菌Lact.1和Lact.2降解胆固醇的研究

从发酵菠萝、桔子中分离选育到2菌株,经鉴定为乳酸菌,将菌株接种到含有胆固醇MRS-CHOL液体培养基中,在38℃条件下,恒温培养3d后,测定胆固醇的降解率、pH值和菌数的变化．结果表明：2菌株都具有降解胆固醇能力,随着培养时间的延长和接种量的增加,降解率都呈现增加趋势;在基质中胆固醇浓度较低时,胆固醇降解率都逐渐增高,而当浓度不断增加时,胆固醇降解率都出现下降趋势．但2菌株之间又存在不同变化幅度．在降解过程中,pH值和菌数也相应发牛了变化。