作物秸秆材料处理养殖废水中氮的周年去除效果及其对氮循环微生物丰度的影响

基于前期研究,利用秸秆材料处理养殖废水,能够有效降低养殖废水中氮的浓度,但周年去除效果及其对氮循环微生物丰度的影响尚不清楚.设置麦秸、玉米秆、稻草和空白对照这4个处理,试验周期为1 a,研究秸秆材料处理养殖废水中氮的周年去除效果及其氮循环微生物功能基因丰度动态变化.结果表明：① 3种作物秸秆对养殖废水TN和NH₄⁺-N等主要污染物的去除效率均以前6个月为最佳,后6个月去除效率明显下降;稻草及麦秸对TN和NH₄⁺-N的周年去除效果较好,其中稻草和麦秸对TN去除率分别为（32.81±11.34）%和（32.99±9.60）%,对NH₄⁺-N去除率分别为（35.3±13.23）%和（34.97±12.00）%;②添加秸秆材料生物基质消纳系统中氮循环微生物功能基因nirK、nirS和hzsB基因丰度为6.45×10⁹、6.18×10⁹和2.31×10⁹copies·L^-1,AOA和AOB基因丰度分别为6.12×10¹⁰copies·L^-1和4.93×10⁹copies·L^-1;16S rRNA高达8.90×10¹⁰copies·L^-1,均显著高于空白处理（P<0.05）.其中添加稻草和麦秸处理中hzsB基因和nirS基因丰度相对较高,表明添加稻草和麦秸显著增强了生物基质消纳系统厌氧氨氧化和反硝化微生物作用.同时,相较于其他处理,添加麦秸增加了生物基质池AOA和AOB基因丰度,表明麦秸能够促进生物基质消纳系统的硝化作用.以上研究结果为秸秆材料处理养殖废水中氮去除分子机制提供了数据支撑.     

氨态氮对菲律宾蛤仔毒理机制研究的内参基因筛选

为了准确和系统地分析NH₃-N对菲律宾蛤仔(Ruditapes philippinarum)毒性的分子机制,并对相关基因的转录组学数据进行验证,利用geNorm和Normfinder这2种软件,以暴露30 d中不同时间点的鳃组织cDNA为模板,从18S rRNA (18S)、beta-actin (Actin)、beta-tubulin (Tubu)、elongation factor 1-alpha (EF1α)、cyclophilin A (CyPA)、Ubiquitin (Ub)等6个备选管家基因中,筛选可以稳定表达的内参基因,作为转录组学分析的内参基因。以稀释50倍的鳃组织cDNA模板作为检测模板,geNorm软件分析获得6对备选内参基因的表达稳定性依次为：EF1α>CyPA>Actin>Tubu>Ub>18S,较优内参为EF1α和CyPA;NormFinder软件分析获得6对备选内参基因的表达稳定性为：EF1α>Actin>Tubu>CyPA>Ub>18S,最优内参基因为EF1α。为了验证上述筛选结果,分别以EF1α和CyPA作为内参基因研究unigene1的表达趋势,发现以EF1α为内参基因时,unigene1的qRT-PCR的表达趋势与其转录组表达倍数的变化趋势一致。因此确定NH3-N暴露菲律宾蛤仔后,鳃组织中表达最为稳定的内参基因为EF1α。     

The metabolism of neonicotinoid insecticide thiamethoxam by soil enrichment cultures,and the bacterial diversity and plant growth‐promoting properties of the cultured isolates

A soil enrichment culture (SEC) rapidly degraded 96% of 200 mg L^?1 neonicotinoid insecticide thiamethoxam (TMX) in MSM broth within 30 d; therefore, its metabolic pathway of TMX, bacterial diversity and plant growth‐promoting rhizobacteria (PGPR) activities of the cultured isolates were studied. The SEC transformed TMX via the nitro reduction pathway to form nitrso, urea metabolites and via cleavage of the oxadiazine cycle to form a new metabolite, hydroxyl CLO‐tri. In addition, 16S rRNA gene‐denaturing gradient gel electrophoresis analysis revealed that uncultured rhizobacteria are predominant in the SEC broth and that 77.8% of the identified bacteria belonged to uncultured bacteria. A total of 31 cultured bacterial strains including six genera (Achromobacter, Agromyces, Ensifer, Mesorhizobium, Microbacterium and Pseudoxanthomonas) were isolated from the SEC broth. The 12 strains of Ensifer adhaerens have the ability to degrade TMX. All six selected bacteria showed PGPR activities. E. adhaerens TMX‐23 and Agromyces mediolanus TMX‐25 produced indole‐3‐acetic acid, whereas E. adhaerens TMX‐23 and Mesorhizobium alhagi TMX‐36 are N₂‐fixing bacteria. The six‐isolated microbes were tolerant to 200 mg L^?1 TMX, and the growth of E. adhaerens was significantly enhanced by TMX, whereas that of Achromobacter sp. TMX‐5 and Microbacterium sp.TMX‐6 were enhanced slightly. The present study will help to explain the fate of TMX in the environment and its microbial degradation mechanism, as well as to facilitate future investigations of the mechanism through which TMX enhances plant vigor.