绿色荧光蛋白分子标记在环境微生物学研究中的应用

绿色荧光蛋白(green fluorescent protein,GFP)分子标记是现有遗传标记中最简单方便的一种方法，GFP及GFP突变体在微生物降解污染物、生物膜菌群构架、环境生态学和环境检测生物传感器等研究领域取得了很好的应用效果。     

生长调节剂对春玉米氮素积累、转运及籽粒蛋白质含量的影响

为探讨生长调节剂对玉米的影响,文章通过不同生长调节剂(主要成分为羧甲基壳聚糖)处理玉米,研究了生长调节剂对春玉米功能叶片、茎杆、籽粒中氮素含量的变化。结果表明,不同配方的生长调节剂能够促进春玉米功能叶中全氮、蛋白氮的转运;提高春玉米茎秆中氮素的转运量以及玉米的产量;加速籽粒中蛋白质的积累,从而提高玉米的品质。并以施生长调节剂配方2 吐温80(PGR2)的配方效果最好。实验中应用的生长调节剂是一种有效的促进玉米氮白质积累的生长调节剂。     

根际微生物耦合降解系统的构建及其对蒽污染土壤的修复

摘要将增强型绿色荧光蛋白(EGFP)标记的黄麻土生根际优势菌Tu-1B分别与葸高效降解菌An-2和生物表面活性剂产生菌P7-50进行原生质体电融合，得到两株具有荧光标记的融合子Tu-An和Tu-P．将二融合子接种于植物根际土壤，构建根际微生物耦合降解系统，：在40d时该系统的最大降解率为96％．对根际土壤中的Tu-An融合子进行了检测，结果表明融合子在根际能稳定旺盛生长．图3表4参18     

大气细颗粒物对CHO细胞的损伤作用及天然物质的拮抗机制

PM2.5引起的健康危害日益受到广泛关注,但其确切的损伤机理仍不完全清楚,目前也缺乏有效拮抗PM2.5损伤的天然活性物质。因此本文对北京市PM2.5造成CHO细胞的损伤作用及天然物质的拮抗作用进行研究,结果可为PM2.5污染治理和疾病的预防提供科学依据。通过CCK-8法测定细胞存活率;通过流式细胞仪测定细胞凋亡;用荧光探针DCFH-DA法测定ROS;提取细胞内总蛋白并测定其中SOD含量;以及通过Western Blot法分别测定了p-akt/akt、p65及Bad的相对表达量。结果显示:(1)10~40μg·m L-1的PM2.5可明显降低CHO细胞存活率,并呈现极显著的剂量-效应关系(r=-0.964,P0.01);15μg·m L-1PM2.5可以显著增加细胞凋亡、引起细胞内ROS增加、SOD含量下降;p-akt/akt、p65表达量增加说明Akt通路与NF-κB通路均被激活。(2)20μmol·L-1阿魏酸、50μmol·L-1绿原酸和10μmol·L-1荭草素预处理可拮抗PM2.5引起的细胞存活率下降以及细胞凋亡率增加,同时降低细胞内ROS并增加SOD含量,下调p-akt/akt、p65及Bad的相对表达量。其中20μmol·L-1阿魏酸的保护作用最佳。结果说明,PM2.5引起的CHO细胞损伤与细胞凋亡与Akt和NF-κB通路的激活有关;三种天然活性物质具有拮抗PM2.5造成的细胞损伤的能力,其保护机理是通过其降低PM2.5引起的细胞内氧化应激反应,进而降低被PM2.5激活的Akt通路与NF-κB通路和下调Bad蛋白表达量来实现的。     

铜绿微囊藻生物钟蛋白KaiC的自激活活性和自身相互作用研究

通过PCR扩增了铜绿微囊藻的生物钟主控基因KaiC,并将其分别克隆到酵母双杂交系统的诱饵质粒pGBKT7和猎物质粒pGADT7中,然后将重组质粒pGBKT7-kaiC/pGADT7- kaiC和pGBKT7-kaiC/pGADT7分别共转化酵母菌AH109,经营养缺陷生长和β-半乳糖苷酶印迹检测表明,KaiC蛋白不具有毒性不会影响酵母细胞的生长,也不具有自激活活性,不会激活报告基因的表达,并且KaiC蛋白自身存在较强的相互作用.诱饵质粒pGBKT7-kaiC可用于从基因组文库中筛选KaiC的相互作用蛋白.     

Imperfect Replacement of Native Species by Non‐Native Species as Pollinators of Endemic Hawaiian Plants

Native plant species that have lost their mutualist partners may require non‐native pollinators or seed dispersers to maintain reproduction. When natives are highly specialized, however, it appears doubtful that introduced generalists will partner effectively with them. We used visitation observations and pollination treatments (experimental manipulations of pollen transfer) to examine relationships between the introduced, generalist Japanese White‐eye (Zosterops japonicus) and 3 endemic Hawaiian plant species (Clermontia parviflora, C. montis‐loa, and C. hawaiiensis). These plants are characterized by curved, tubular flowers, apparently adapted for pollination by curve‐billed Hawaiian honeycreepers. Z. japonicus were responsible for over 80% of visits to flowers of the small‐flowered C. parviflora and the midsize‐flowered C. montis‐loa. Z. japonicus‐visited flowers set significantly more seed than did bagged flowers. Z. japonicus also demonstrated the potential to act as an occasional Clermontia seed disperser, although ground‐based frugivory by non‐native mammals likely dominates seed dispersal. The large‐flowered C. hawaiiensis received no visitation by any birds during observations. Unmanipulated and bagged C. hawaiiensis flowers set similar numbers of seeds. Direct examination of Z. japonicus and Clermontia morphologies suggests a mismatch between Z. japonicus bill morphology and C. hawaiiensis flower morphology. In combination, our results suggest that Z. japonicus has established an effective pollination relationship with C. parviflora and C. montis‐loa and that the large flowers of C. hawaiiensis preclude effective visitation by Z. japonicus. Remplazo Imperfecto de Especies Nativas por Especies No‐Nativas como Polinizadores de Plantas Endémicas de Hawaii     

The malaria parasite Plasmodium relictum in the endemic avifauna of eastern Cuba

Island populations are vulnerable to introduced pathogens, as evidenced by extinction or population decline of several endemic Hawaiian birds caused by the malaria parasite, Plasmodium relictum (order Haemosporida). We analyzed blood samples from 363 birds caught near Guantánamo Bay, Cuba, for the presence of haemosporidian infections. We characterized parasite lineages by determining nucleotide variation of the parasite's mitochondrial cyt b gene. Fifty‐nine individuals were infected, and we identified 7 lineages of haemosporidian parasites. Fifty individuals were infected by 6 Haemoproteus sp. lineages, including a newly characterized lineage of Haem. (Parahaemoproteus) sp. CUH01. Nine individuals carried the P. relictum lineage GRW4, including 5 endemic Cuban Grassquits (Tiaris canorus) and 1 migratory Cape May Warbler (Setophaga tigrina). A sequence of the merozoite surface protein gene from one Cuban Grassquit infected with GRW4 matched that of the Hawaiian haplotype Pr9. Our results indicate that resident and migratory Cuban birds are infected with a malaria lineage that has severely affected populations of several endemic Hawaiian birds. We suggest GRW4 may be associated with the lack of several bird species on Cuba that are ubiquitous elsewhere in the West Indies. From the standpoint of avian conservation in the Caribbean Basin, it will be important to determine the distribution of haemosporidian parasites, especially P. relictum GRW4, in Cuba as well as the pathogenicity of this lineage in species that occur and are absent from Cuba.     

蛋白质及无机离子对纳滤膜去除PFOS的影响

以全氟辛烷磺酸（PFOS）为目标去除物,选取牛血清蛋白（BSA）为典型蛋白质类有机物,考察了BSA及其浓度,以及BSA与无机离子共存时,离子强度、离子种类对聚酰胺纳滤膜去除水中PFOS的影响.研究发现,原液中存在BSA时,PFOS的去除率有显著提高,而且BSA浓度越高,PFOS的去除率越高;当BSA与无机离子共存时,离子强度越大,PFOS去除率越高.这可能是因为BSA不但会吸附一部分PFOS,还会造成膜污染,BSA浓度越大,膜污染越严重,膜的筛分能力越强,且膜面与PFOS之间的静电排斥力越大,从而提高了PFOS的截留率.而无机离子的存在减小了BSA分子之间及与膜面之间的静电排斥力,使BSA污染层更加厚实,进一步增强了膜的筛分能力.此外,Ca²⁺提高PFOS去除率的能力优于Na⁺.     

营养失衡下的分层分质EPS对异型膨胀污泥及其沉降性差异影响

为探索EPS对膨胀污泥沉降性的影响,通过减少进水P源、N源的方式培养异型膨胀活性污泥,分析不同层、不同组分EPS对异型膨胀污泥沉降性影响.结果表明：异型膨胀（非丝状菌粘性膨胀、丝状菌膨胀）活性污泥EPS多糖组分Total-PS、中层组分（L-PS）含量均高于正常污泥.非丝状菌粘性膨胀污泥EPS各层多糖含量高于丝状菌膨胀污泥,而各层蛋白质（PN）含量均低于丝状菌膨胀污泥.非丝状菌粘性膨胀污泥Total-EPS（（229±94）mg/g MLSS）、PS/PN值不仅显著高于正常活性污泥（（86±16）mg/g MLSS）,也高于丝状菌膨胀污泥（（108±30）mg/g MLSS）.非丝状菌粘性膨胀污泥EPS多糖总量Total-PS、蛋白质总量Total-PN越高,SVI值越大,污泥沉降性越差,越易膨胀,且EPS蛋白质组分对污泥膨胀作用大于多糖;从不同层EPS看,松散外层（S层）EPS对非丝状菌粘性膨胀作用最大（S-PN与SVI值相关性最大,r为0.881,P<0.05）,是主要影响因子.丝状菌膨胀污泥,除了内层紧密型T-PN对SVI值影响较大外,其他各层、各组分EPS含量与SVI均呈微弱负相关.     

四溴双酚A的鲫鱼血清蛋白生物标志物研究

以鲫鱼为供试生物,对四溴双酚A(TBBPA)的急性毒性、血清蛋白潜在生物标志物以及相应的毒性靶器官组织病理进行了研究,结果表明TBBPA对鲫鱼96 h的半致死浓度LC50是3.13 mg/L,TBBPA诱导下鲫鱼血清蛋白可区分为11个组分,各组分随时间和浓度的变化规律存在差异。组分11条带清晰,表达量变化幅度最大,在低浓度TBBPA暴露下表达量先降低后升高,第16天达到最高;高浓度时表达量在第2天达到最高。组分11的分子量为24 kDa,可能为鲫鱼斯坦尼小体(CS)中降血钙因子的解聚物。组织病理切片结果显示,鲫鱼肝脏和肾脏均表现出时间-剂量依赖性,且与组分11的诱导差异性表达呈现相应的病理损伤。因此,鲫鱼血清蛋白组分11对TBBPA的诱导具有浓度和时间响应敏感性,且易于识别和鉴定,可作为TBBPA暴露的鲫鱼血清蛋白潜在生物标志物。