微生物降解磺胺甲恶唑的研究进展

抗生素是一类难降解、低浓度就有高生态毒性效应的化合物,近年来被归为新型环境污染物,其环境残留与去除备受关注。作为广泛使用的抗生素之一,磺胺甲恶唑在水土环境中的残留量不断增加,检出率也越来越高。研究表明,磺胺甲恶唑是少数几种可被微生物降解的抗生素之一,微生物降解法是最具潜力的残留磺胺甲恶唑去除手段。本文总结了磺胺甲恶唑在土壤、沉积物、活性污泥、混合菌群、酶等条件下的降解及已分离的具有降解能力的单菌株对磺胺甲恶唑的降解情况,包括其降解效率、降解条件等,归纳了目前磺胺甲恶唑微生物降解的主要分类,并讨论了影响磺胺甲恶唑降解的两个特有因素。指出从分子生物学及生物信息学角度研究其降解途径,降解菌、降解菌群的人工构建及其在含磺胺甲恶唑污水处理中的应用与效果评价等应为今后磺胺甲恶唑生物降解与应用研究的重点。     

磺胺甲恶唑高效降解菌群的多样性分析和降解微生物的分离表征

为了解决养殖水体中抗菌药物磺胺甲恶唑残留污染的问题,以抗菌药物磺胺甲恶唑为唯一碳源,通过连续4次传代富集,获得了磺胺甲恶唑降解菌群。通过16S V3-V4区高通量测序分析了每个代次的群落组成和丰度变化,推测了与磺胺甲恶唑降解相关的菌株信息,进一步连续分离纯化到了磺胺甲恶唑降解菌株LLE3。经16S rDNA及生理生化鉴定,初步将LLE3鉴定为鞘氨醇杆菌属（Sphingobacterium）菌株。对该菌进行了降解条件优化,在最优降解条件下（30 ℃,pH 7,接种浓度为5×107 CFU·mL-1）,该菌7 d内对50 mg·L-1的磺胺甲恶唑降解效率可达93.7%。运用高效液相色谱与质谱联用鉴定了菌株LLE3降解后的代谢产物为N-\[4-(1,2-恶唑-3-基氨磺酰基)苯基\]乙酰胺。研究结果为磺胺甲恶唑污染的水体和土壤环境修复提供了优良的微生物资源。     

抗磺胺对甲氧嘧啶单克隆抗体的研制及其特性分析

目的制备针对磺胺对甲氧嘧啶的单克隆抗体,建立对该物质的免疫学检测方法。方法以BSA-磺胺对甲氧嘧啶为免疫原,免疫BALB／c小鼠,取脾细胞与小鼠Sp-2／0骨髓瘤细胞融合后,经筛选和亚克隆,建立杂交瘤细胞株。结果获得2株能稳定分泌抗磺胺对甲氧嘧啶抗体的细胞株。对抗体进行了特性分析,抗体的效价分别为1：400000和1：1630000,抗体类型及亚类都为IgGl。其中,单克隆抗体1H10的亲和力为1．4×109L／mol,利用该抗体采用竞争间接ELISA法检测磺胺对甲氧嘧啶的范围是1025—16μg/mL,最低检测浓度是8μg/mL。单抗1H10与其他6种磺胺药（SMM、SMZ、SM2、SD、SulfaquinoxalineSodium、Sulfametetyrazine）无交叉反应。结论单克隆抗体1H10可用于研制免疫学方法检测磺胺对甲氧嘧啶残留的产品。     

海水养殖水体周丛纤毛虫的群落结构及其对环境因子的响应

为了解周丛纤毛虫在特定海水养殖环境巾的功能与地位,作者以青岛近郊一封闭式海水对虾养殖池为采集地,采用人工基质（载玻片）法对周丛类纤毛虫的群落结构进行了为期140d（以全养殖周期为时限）的调查,同时开展了环境理化因子的采集和分析。在此基础上,探讨了周丛纤毛虫群落结构参数和环境因子的相关性关系。本工作共涉及41种周丛纤毛虫,其中以缘毛类Zoothamnium和Pseudovorticella等属种类为优势类群。采集期间纤毛虫丰度和物种数量呈上升趋势,丰度分别在第100天和第120天达到高峰,物种数量则在第80天达最大值,缘毛类纤毛虫是微型群落中丰度／物种数量峰值的主要组成成分。物种丰富度指数、均匀度指数和多样性指数变化趋势基本一致,在第100天出现峰值。多元统计分析结果表明,周丛纤毛虫群落结构中的大部分参数与养殖水体的环境因子具有显著的相关性：物种数量和均匀度指数均与氨态氮浓度呈极显著的正相关（P〈0．01）;丰度和多样性指数均与氨态氮浓度呈显著正相关（P〈0．05）;丰度和物种数均与可溶性磷酸盐浓度呈极显著正相关（P〈0．01）;多样性指数与细菌密度呈显著的负相关（P〈0．05）。本研究结果初步显示,周丛纤毛虫群落具有良好的自我维持和修复能力,并能较好的响应集约化海水养殖水体环境的变化。     

黄连木对干旱胁迫的生理响应

研究了自然干旱条件下黄连木（Pistacia chinensis Bunge）的生理变化。结果表明,随土壤含水量的减少,叶绿素b含量、光合速率、叶片相对含水量与叶水势均下降;叶绿素a和可溶性糖含量、叶绿素a和b的比值及总叶绿素含量呈现上升的趋势;超氧化物歧化酶活性先升后降;丙二醛含量干旱胁迫前期升高,后期变化不明显;净光合速率、气孔导度和蒸腾速率随土壤含水量的减少逐步降低。气孔和可溶性糖含量都是影响黄连木光合速率的关键因子,干旱胁迫前12d光合速率主要受气孔限制,之后为非气孔限制。干旱胁迫前期渗透调节物质以可溶性糖为主,干旱胁迫较重时脯氨酸含量急剧升高,与可溶性糖同时起渗透调节作用。     

一株霍氏肠杆菌对四环素的降解作用及其降解产物的毒性评估

【目的】抗生素作为新兴污染物,已经引起社会的极大关注。针对四环素有效降解菌株缺乏这一现状,本研究旨在筛选和鉴定具有降解四环素功能的菌株,分析其降解特性和降解作用类型、初步探讨其降解活性物质定位并评估其降解产物的生理毒性。【方法】以四环素为唯一碳源,从受四环素污染的猪场污泥中筛选四环素降解菌株;结合菌落形态学特征、生理生化特征、扫描电镜观察和16S rRNA基因测序鉴定菌株,通过不同外源碳、pH及去除动力学阐明菌株对四环素的降解特性,提取菌株不同成分探讨其去除四环素的作用类型,并进一步从细胞内液和细胞外液开展生物降解的活性物质定位,最后评估降解产物的生理毒性。【结果】筛选鉴定得到一株霍氏肠杆菌(Enterobacter hormaechei) MEH2305,pH为7.0和添加10 g/L外源碳胰蛋白胨是其发挥降解作用的最适条件。MEH2305通过非生物降解和生物降解的共同作用,在培养第7天对四环素总去除率达到68% (对土霉素和盐酸强力霉素去除率分别为53%和56%),其分泌的细胞内液和细胞外液对四环素的去除率分别为40.77%和31.18%。同时,与未经MEH2305处理的四环素对照组比较,MEH2305降解四环素的产物对革兰氏阴性大肠杆菌(Escherichia coli) K88和革兰氏阳性枯草芽孢杆菌(Bacillus subtilis) 168的生理毒性作用显著降低。【结论】MEH2305可以作为一株潜在的有效且安全的四环素降解菌株,应用于抗生素的环境治理领域。     

捕食者对空心莲子草叶甲种群的生物胁迫

广食性捕食者广泛捕食植食性昆虫,常被用于有害生物的生物防治,也因此影响植食性昆虫对杂草的生物效果。空心莲子草叶甲(Agasicles hygrophila)(鞘翅目:叶甲科Chrysomelidae)作为入侵恶性杂草空心莲子草(Alternanthera philoxeroides)(苋科:莲子草属Alternanthera)的专性天敌,从美国的弗罗里达州引入中国,在释放地防治空心莲子草取得了较好的防治效果。虽然空心莲子草叶甲在引入地均已建立田间种群并有一定程度的自然扩散,但丰富的食物资源,并未使空心莲子草叶甲的自然种群数量变得繁荣,因此其未能有效抑制空心莲子草的扩散蔓延。在野外调查时发现空心莲子草生境中存在大量广食性捕食者。这些广食性捕食者是抑制空心莲子草叶甲种群数量扩张的生物胁迫因子吗?为此,选择捕食性昆虫龟纹瓢虫(Propylaea japonica)(鞘翅目:瓢虫科Coccinellidae)、蜘蛛类捕食者拟水狼蛛(Pirata subpiraticus)(蜘蛛目:狼蛛科Lycosidae)与斜纹猫蛛(Oxyopes sertatus)(蜘蛛目:猫蛛科Oxyopidae)为捕食者,分别以空心莲子草叶甲各虫态为猎物,构建简单的捕食者-猎物系统,在室内检测了上述3种捕食者对空心莲子草叶甲各虫态在不同密度下的日捕食量,以期了解捕食者对空心莲子草叶甲的捕食作用,客观评估空心莲子草叶甲的生物防治效能。研究结果表明:捕食者龟纹瓢虫、斜纹猫蛛与拟水狼蛛均捕食空心莲子草叶甲的卵粒及1龄、2龄幼虫,斜纹猫蛛与拟水狼蛛捕食3龄幼虫,捕食者的捕食量均随着猎物密度的升高而增加,寻找效应降低。三者均不捕食成虫。除拟水狼蛛对3龄幼虫的捕食用Holling II模型拟合不呈显著相关关系外,其余捕食反应均拟合Holling II模型并显著相关。通过拟合方程得出捕食者对空心莲子草叶甲卵粒的理论日最大捕食量为:斜纹猫蛛10.9粒,拟水狼蛛为6.2粒,龟纹瓢虫为5.6粒;对1龄幼虫的理论日最大捕食量为:斜纹猫蛛为17.1头;拟水狼蛛为35.8头,龟纹瓢虫为10.4头;对2龄幼虫的理论日最大捕食量为:斜纹猫蛛为6.6头,拟水狼蛛为11.2头,龟纹瓢虫为2.9头;对3龄幼虫的理论日最大捕食量为:斜纹猫蛛捕食12.3头,拟水狼蛛为1.1头。研究结果证实了捕食者可通过捕食作用降低空心莲子草叶甲种群密度,削弱空心莲子草叶甲对空心莲子草的控害效能,是空心莲子草叶甲种群存活的生物胁迫因子。建议在提高空心莲子草叶甲田间种群数量,达到对空心莲子有效的持续控制效果方面开展进一步研究。     

辣椒对镉胁迫的生理响应

以福湘5号、兴蔬215和川腾1号这3个辣椒品种为材料,用含有0mg/L、100mg/L、200mg/L、300mg/L重金属镉的1/2霍格兰培养液对具10片真叶的辣椒苗进行处理,7d后测定各项生理指标。结果表明,镉对辣椒的生长有很强的毒害作用,随着镉胁迫浓度的升高,3个辣椒品种的生物量、叶片叶绿素含量、电导率、丙二醛含量和活性氧清除系统酶活性都受到了不同程度的影响。在镉胁迫下,辣椒植株的株高、根长、鲜重受到明显抑制,根系发育畸形,叶片黄化。镉胁迫的毒害作用在3个辣椒品种间有差异。其中川腾1号耐镉胁迫能力较强,福湘5号次之,兴蔬215最差。     

水稻花药对高温胁迫的生理响应

为了探明水稻花药在高温胁迫下的生理反应及适应机理,以耐热水稻品系996和热敏感水稻品系4628为材料,在人工气候室进行高温（9：00-17：00,37℃;17：00-9：00,30℃）和适温处理（9：00-17：00,30℃;17：00～9：00,25℃）,研究高温胁迫对水稻花药可溶性糖、可溶性蛋白质、热稳定蛋白、维生素C含量、膜透性及内源激素含量等生理特性的影响。结果表明,高温使水稻花药中可溶性糖含量、可溶性蛋白质含量和维生素C含量下降,热稳定蛋白含量上升;热敏感品系4628花药中丙二醛含量和相对电导率在高温胁迫下增幅大于耐热品系996;高温胁迫下,水稻花药中GA3和IAA含量降低,ABA含量上升,耐热品系996花药中GA3、IAA含量降低幅度小于热敏感品系4628,ABA含量增加幅度小于热敏感品系4628。     

发状念珠藻对盐胁迫的响应

探讨了发状念珠藻(NostocflagelliformeBornetFlah)对盐胁迫的耐受适应机制,采用含不同浓度NaCl(0、01、02、04、06、08、10mol/L)的BG110培养液处理具有正常生理活性的丝状体,25±05℃,40μmol/m2/s下照光培养12h,测定藻体光合作用、呼吸作用等生理活性以及体内一些物质的含量,结果表明:随培养液中NaCl浓度的升高藻体光合作用、呼吸作用以及PSⅡ活性(Fv/Fm)降低;质膜透性不断增大,丙二醛含量升高,自由水含量、自由水/束缚水比值下降,类胡萝卜素、可溶性糖含量增加,脯氨酸含量变化不大。由此可知,盐胁迫下发状念珠藻正常生理活性受到抑制而表现出一定的抗逆能力;该藻对盐胁迫具有一定的耐受能力,类胡萝卜素的增加有助于清除藻体内的氧自由基,可溶性糖可能是其主要渗透调节物质之一,脯氨酸在盐胁迫中的渗透调节作用不大。       

群体和单细胞微囊藻对短期高光胁迫的生理响应

为了探讨不同形态的微囊藻（Microcystis）对光的耐受能力及其应对机制,研究比较了短期高光强条件下群体微囊藻和单细胞微囊藻的生理响应,结果表明,在高光强胁迫下,群体和单细胞微囊藻的叶绿素含量、最大电子传递速率（ETR_max）均降低,但与单细胞微囊藻相比,群体微囊藻的下降幅度较小;在高光强胁迫下,群体微囊藻的过氧化氢酶（CAT）与超氧化物歧化酶（SOD）的活性均显著增加,而单细胞微囊藻只有CAT活性增加;在短期高光胁迫下,群体微囊藻的死亡率没有显著变化。这些结果表明群体微囊藻比单细胞微囊藻能耐受更高的光强,也暗示了群体微囊藻在野外高光强条件下更具竞争优势。     

南方4种草本植物对铝胁迫生理响应的研究

 不同的植物对铝胁迫的生理响应不同, 因而对铝毒的耐性也不相同。设置5种铝浓度,进行砂培法处理,研究了4种我国南方红壤广泛分布的草本植物——牵牛(Pharbitis nil)、望江南(Cassia occidentlis)、光头稗(Echinochloa colonum)和合萌(Aeschynomene indica)的种子萌发、光合色素、脯氨酸含量、丙二醛(MDA)含量、可溶性糖(SS)含量、质膜透性(MP)、过氧化氢酶 (CAT) 活性以及过氧化物酶 (POD)活性的变化。结果表明铝对4种植物的生理特性都有明显的影响。4种植物的种子在10 000 mg·L－1 Al3+处理条件下都不能萌发。2 000 mg·L－1 Al3+处理都不利于4种植物的生长,与对照相比,2 000 mg·L－1 Al3+处理时4种草本植物叶绿素和叶绿素总含量显著降低(p<0.05);MDA含量和MP显著增加(p<0.05);脯氨酸含量极显著增加(p<0.01);POD和CAT活性极显著降低(p<0.01)。中低铝(80和400 mg·L－1)处理时,牵牛和合萌与对照相比,MP和MDA含量降低,POD和CAT活性升高;望江南的反应与牵牛和合萌的反应相反;光头稗在80 mg·L－1 Al3+处理时,与牵牛和合萌的变化一致,在400 mg·L－1 Al3+处理时,则相反。植物在中低铝处理条件下,通过维持较高的POD和CAT活性和脯氨酸、叶绿素含量,较低的MP和MDA含量来增加其对铝的耐性。     

共生与非共生爪哇伪枝藻对高温胁迫的响应

研究以自由生长的爪哇伪枝藻(Free-living S. javanicum, fs)和分离自地衣的爪哇伪枝藻(Symbiotic S. javanicum, ss)为研究对象, 探究了不同生长状态爪哇伪枝藻对高温(45℃)胁迫的响应。结果发现在高温胁迫下, 爪哇伪枝藻光合活性、叶绿素a及类胡萝卜素含量下降; 丙二醛(MDA)、胞外多糖及可溶性蛋白含量上升。在高温处理下, 与fs相比, ss光合活性下降较慢, 且高温处理后ss的叶绿素a及类胡萝卜素含量也明显高于fs。高温处理下, 与fs相比, ss的MDA含量和增长速度均较低; 并且在面临高温胁迫时, ss能够更快的分泌胞外多糖和可溶性蛋白质, 从而在一定程度上达到自我保护的目的。研究结果表明, 在暴露于高温胁迫时, 相较于自由生长状态, 来自地衣的爪哇伪枝藻具有更高的自我保护效率。     

SUBSIDY AND STRESS RESPONSES OF STREAM PERIPHYTON TO GRADIENTS IN WATER VELOCITY AS A FUNCTION OF COMMUNITY GROWTH FORM

Previous studies have shown major differences in the way biomass of stream periphyton is controlled by spatial variations in velocity. We hypothesize that these differences may be the result of different growth forms within the community. Some dense and coherent growth forms (e.g. mucilaginous diatom/cyanobacterial mats) may be resistant to diffusion and also resistant to dislodgment by shear stress. Higher velocities applied to such communities could therefore be expected to enhance biomass accrual by increasing rates of mass transfer, but without greatly increasing losses through sloughing. Conversely, other growth forms (e.g. long filamentous green algae) have an open matrix, and high rates of diffusion into the mats can potentially occur even at low velocity. However, as velocities increase, high skin friction and form drag should lead to higher rates of sloughing. The overall result of these processes should be that maximum biomass occurs at low velocities. This “subsidy-stress” hypothesis was tested twice with each of three different periphytal growth forms: a coherent, mucilaginous, diatom community; a moderately coherent, stalked/ short, filamentous diatom community; and an open-weave, long, filamentous green algal community. A monotonic increase in chl a biomass occurred as a function of near-bed velocities for the first of the two mucilaginous diatom communities investigated. No biomass-velocity relationship was found, however, with the second mucilaginous community, probably because the waters were highly enriched and mass transfer driven by molecular diffusion was probably high throughout the velocity gradient. Biomass was moderate at low velocities, peaked at near-bed velocities from 0.18 to 0.2 m·s^?1 (～0.40–0.45 m·s^?1 mean column velocity), and then decreased at higher velocities in both of the stalked/ short filament communities of diatoms analyzed. With the long filamentous green algal communities, a monotonic reduction in biomass occurred as a function of increases in velocity. Proliferations greater than 100 mg·m^?2 chl a occurred at low near-bed velocities (i.e. <0.2 m·s^?1), after which biomass declined nearly exponentially as a function of increasing velocity to less than 10 mg·m^?2 chl a at velocities greater than 0.4 m·s^?1. These biomass-velocity trends support our hypothesis that community growth form determines periphytal responses to spatial variations in velocity within stream reaches.     

植物对水分胁迫响应的分子机制与抗逆基因工程的研究进展（综述）

对近年来植物对水分胁迫（包括干旱、盐害和低温）响应的分子机制的研究进展以及水分胁迫中诱导的植物基因产物的种类、功能及其相关的基因工程作一简要的综述,同时对今后的研究方向提出了一些看法。     

FLEXURAL RIGIDITY OF CHIVE AND ITS RESPONSE TO WATER POTENTIAL

The biomechanical relationship between the ability of a plant organ to resist bending and the extent to which tissues are hydrated is illustrated for the cylindrical leaves of chive (Allium schoenoprasnum var. schoenoprasnum L.). The flexural rigidity (EI), which measures the ability to resist bending, is maximum when leaves are fully turgid and decreases monotonically as a function of water potential (r² = 0.99). Dehydration results in a reduction in the elastic modulus (E) of leaves. Reductions in E are correlated with geometric distortion in the transverse geometry of leaves which influences their second moment of inertia (I). The traditional theory of elastic stability (developed on the basis of the mechanical behavior of nonbiological systems) is shown to be inadequate to distinguish the behavior of E as plant organs geometrically distort during dehydration. This inadequacy results from the violation of a principal assumption made by the theory (= uniform cross-sectional geometry). A derivation is presented that accommodates the localized geometric distortions in cylindrical plant organs and permits a valid estimate of reductions in E as tissues dehydrate. Based on this derivation, the Young's modulus of chive leaves just before mechanical failure due to buckling is shown to be less than 50% of that calculated for fully turgid leaves.     

水分胁迫对羊草光合产物分配及其气体交换特征的影响

 通过对典型草原优势植物种羊草(Leymus chinensis)的盆栽实验,模拟5个土壤水分梯度（分别为土壤持水量的75%～80%(对照)、60%～65%、50%～55%、35%～40%和25%～30% ）对羊草叶片相对含水量、光合速率、光合产物分配和种群CO2交换速率的影响。结果表明：随着土壤水分胁迫的增加,羊草叶片相对含水量呈先增加而后下降的单峰型变化,且在50%～55%处理下达到最大;叶片光合速率随着水分胁迫的增加而减小,且75%～80%、60%～65%、50%～55%的水分处理与35%～40%、25%～30%的水分处理的叶片光合速度日动态规律不同。羊草总生物量及根、鞘、叶生物量均随着水分胁迫的增加呈下降趋势。干旱促进早期羊草根的分配和根冠比增加, 但到后期却使它们降低, 表明羊草在受到较长期的持续干旱后通过增加根部的比重来提高抗旱性的能力逐渐降低。羊草根茎的生物量和分配随着土壤水分含量降低均呈现出先增加而后下降的趋势,羊草根茎的生物量在50%～55%处理下达最大（1.28 g·株-1）,而羊草根茎的分配在35%～40%处理下达最大（48.5%）。羊草种群CO2的净交换速率随着水分胁迫的增加而减小,其日交换量随着水分胁迫的增加而增加,且在60%～65%处理下达到最高,而后呈下降趋势,并在25%～30% 处理下为负值。研究结果表明,土壤持水量的40%可能是羊草对于水分变化响应的阈值。     

RESPONSE OF PERIPHYTON COMMUNITIES TO CLAY AND PHOSPHATE LOADING IN A SHALLOW RESERVOIR1

Enclosures in a small piedmont reservoir in the southeastern United States were used to determine the effects of loading by phosphate and two clay sediments on periphyton community structure and production during the summer growing season. The experimental design included replicated controls and the following treatments added at 2-to 3-day intervals: phosphate (PHOS), kaolinite (KAOL), montmorillonite (MONT), kaolinite with phosphate (K+P), and montmorillonite with phosphate (M+P). Periphyton were sampled from polyethylene strips of the same material as the enclosure walls, suspended at depths of 0.5 m and 2.0 m to assess treatment effects on shallow (epilimnetic) vs. deep (hypolimnetic) communities. Colonization after 40 days was sparse, dominated in biovolume by blue-green algae in all but the epilimnion of controls, in which small gelatinous green algae and chrysophyte flagellates were abundant. The biovolume of shallow periphyton after 79 days was lowest in KAOL and MONT, intermediate in controls and PHOS, and greatest in clay + P. Hence, P enrichment alleviated the deleterious effects of clay on periphyton production, and clay + P appeared to act synergistically in stimulating benthic algal growth. Periphyton biovolume was comparable among shallow and deep controls, PHOS, and treatments with the rapidly settling kaolinite. Lowest biovolume occurred at depth in treatments with montmorillonite under extreme light attenuation imposed by the suspended, finely particulate clay. The relative contribution of blue-green algae to total periphyton production was highest in clay + P treatments and lowest in controls. Heterotrophic dinoflagellates increased in the shallow periphyton under clay loading, and in the deep communities under PHOS and M + P. Periphyton contributed only 20–32% of total mesocosm productivity in controls, PHOS, and KAOL, increasing to about 40% in MONT, and to 72–91% of the total in M + P and K + P. The data indicate that benthic microalgae can represent a major proportion of the primary production in shallow reservoirs under high phosphate and sediment loading.     

烤烟伸根期的光合特性及其对土壤水分的响应

烤烟NC89、LJ851、LJ911伸根期的生长中心在地下部分。合理地控制土壤水分含量,协调地上部分和地下部分的生长是此期生产管理的重要环节。土壤相对含水量在75%~85%的光合速率最高,但水分利用效率最高的土壤相对含水量在65%~75%之间,且此土壤水分含量有利于烤烟烟苗的蹲苗、壮苗,是烤烟生长的最佳土壤水分条件。