植物对水中菲和芘的吸收

以菲和芘为多环芳烃（PAHs）代表物,采用水培体系研究了黑麦草（Lolium multiflorum Lam）对水中PAHs的吸收作用,重点研究了植物吸收菲和芘的时间动态.水中菲和芘起始浓度分别为1.00mg/L和0.12mg/L.0～288h内,黑麦草根和茎叶中菲和芘含量均先快速增加而后降低,积累量不断增大,植物根系和茎叶富集系数则先快速升高而后趋于稳定.茎叶中菲和芘含量、茎叶对菲和芘的富集系数比根低1～3数量级,积累量也明显小于根系.黑麦草根系对水中芘有更强的富集能力,其根系富集系数比菲大85%～179%;而其茎叶对菲的富集作用则略强.菲和芘在植物体内有明显的传导作用.0～288h,传导系数（TF）先显著升高而后趋于恒定;但实验条件下,菲和芘的TF值均很小,分别不高于0.031和0.009,且芘的TF值明显小于菲,表明供试植物对芘的传导能力更弱.     

植物根对土壤中PAHs的吸收及预测

研究了多种植物根对土壤中多环芳烃(PAHs)的吸收作用,阐述了根系吸收与土壤污染强度、污染物性质、植物组成等的关系,并用实验数据检验了限制分配模型对植物吸收土壤中PAHs的预测性能。供试土壤中菲和芘的起始浓度分别为0~457和0~489mg/kg;45d后,随土壤中菲和芘浓度提高,根中菲和芘含量明显增大,根系富集系数则减小。不同植物根中菲和芘含量和根系富集系数与根的脂肪含量呈显著正相关。由于芘的Kow较大,同种植物根中芘含量、芘的根系富集系数则远大于菲。经45d处理,尽管土壤中菲浓度变化很大(从不足1mg/kg到约45mg/kg),限制分配模型能较好地预测供试植物根中菲的含量,黑麦草和菜心根中菲含量的预测误差低于81%。作为限制分配模型预测植物吸收的关键参数,不同植物根吸收菲的αpt值与根脂肪含量显著正相关。     

AMF和PGPR联合修复菲和芘污染土壤的效应

丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）与根围促生细菌（plant growth-promoting rhizobacteria,PGPR）联合降解有毒有机物、修复污染土壤和促进植物生长的作用倍受关注。本试验旨在探究AMF与PGPR联合降解土壤中菲和芘的效应,以菲和芘1:1混合处理浓度各0、50mg/kg、100mg/kg和150mg/kg下对高羊茅Festuca elata接种AMF根内根孢囊霉Rhizophagus intraradices（Ri）、变形球囊霉Glomus versiforme（Gv）、PGPR荧光假单胞菌Pseudomonas fluorescens Ps2-6、芽孢杆菌Bacillus velezensis Ps3-2、Ri+Ps2-6、Ri+Ps3-2、Gv+Ps2-6、Gv+Ps3-2和不接种对照共36个处理。结果表明,供试AMF增加了PGPR的定殖数量;接种PGPR则显著提高AMF的侵染率。AMF、PGPR或AMF+PGPR处理均显著降低土壤中菲和芘含量,促进植物对土壤中菲和芘的吸收,显著提高高羊茅根系和叶片内的菲和芘含量。在土壤中菲和芘100mg/kg和150mg/kg水平下,Gv与Ps2-6及Ri与Ps2-6能相互促进对土壤中菲和芘的去除效应,其中接种Gv+Ps2-6组合处理的去除率最高,达到95%-98%,土壤中多酚氧化酶、脱氢酶和过氧化氢酶活性显著高于单接种处理和不接种对照,而酸性磷酸酶活性变化则表现为相反趋势。其中以Gv+Ps2-6组合处理的多酚氧化酶活性最高,为0.17mg/g,是不接种对照的1.9倍;脱氢酶和过氧化氢酶活性分别达到1.32µg/(g·h)和1.81mL/g;酸性磷酸酶活性则比不接种对照土壤降低27%-45%;易提取球囊霉素相关土壤蛋白含量和总球囊霉素相关土壤蛋白含量分别是不接种对照的1.6倍和1.5倍。     

固定化微生物对多环芳烃污染土壤的降解

利用微生物固定化技术,研究了微生物固定化菌剂对土壤中菲、蒽、芘、(艹屈)和苯并(a)芘的降解动态,并且采用Michaelis-Menton和Monod动力学模型对结果进行拟合.结果显示,4种处理(TB02、TB07、TBB03、TBB08)均有降解菲、蒽、芘、(艹屈)和苯并(a)芘的能力.其中,处理TB02的降解能力强、降解速率快、半衰期短且处理成本低,而处理TB07则需要较长时间作用于PAHs污染土壤,其降解能力才能充分发挥出来.当菲、蒽、芘、(艹屈)和苯并(a)芘的初始浓度均为20 mg·kg-1时,42 d后,TB02对菲、蒽、芘、(艹屈)和苯并(a)芘的降解率分别为84.32%、85.24%、82.59%、43.75%和62.25%; 133 d后,TB07对5种污染物的降解率分别为95.00%、95.24%、90.93%、74.82%和72.20%.通过比较5种污染物半衰期,其可降解性由大到小依次为菲、蒽、芘、苯并(a)芘、(艹屈).     

蚯蚓在植物修复芘污染土壤中的作用

采用盆栽试验法,研究了蚯蚓(Pheretima hupeiensis)在植物修复芘污染土壤中的作用。结果显示,试验浓度(20.24-321.42 mg/kg) 范围内,蚯蚓活动促进了芘污染土壤中修复植物黑麦草(Lolium multiforum)黑麦草的生长,其根冠比明显增大。添加蚯蚓72 d后,种植黑麦草的土壤中芘的去除率高达60.01%-86.26%,其平均去除率(74.66%)比无蚯蚓活动的土壤-植物系统(64.55%)提高10.11%,比无植物对照组(18.24%)提高56.42%。各种生物、非生物修复因子中,植物-微生物交互作用对芘去除的平均贡献率(51.75%)最为突出,比无蚯蚓活动时(44.94%)提高6.81%。说明蚯蚓活动可强化土壤-植物系统对土壤芘污染的修复作用。     

多环芳烃降解菌的筛选与降解能力测定

从本溪多环芳烃(PAHs)污染土壤中经富集培养筛选出8株PAHs降解菌,研究了8株菌及其等比例混合培养对菲、芘和苯并[a]芘的降解能力。结果表明,在28℃,培养基中菲、芘和苯并[a]芘的浓度分别为50、50和5mg·L-1的复合底物条件下,培养28d后,菌株B3的降解效果最好,对菲、芘和苯并[a]芘的降解率分别为88.4%、54.0%和68.4%,8株菌的混合培养对菲、芘和苯并[a]芘的降解率分别为87.7%、35.3%和42.0%;经生理生化实验和16SrRNA序列比对,初步鉴定B3菌为假单胞菌属(Pseudomonas sp.)。     

玉米幼苗根际土壤微生物活性对芘污染的响应

用根际袋法土培试验研究了玉米幼苗根际与非根际土壤微生物量碳、微生物熵、代谢熵和土壤酶活性对不同芘污染水平(50、200、800mg·kg-1,记为T1、T2、T3)的响应差异。结果表明,较低浓度芘可适当的刺激玉米幼苗的生长,而较高浓度芘则抑制幼苗生长,其抑制作用随芘处理浓度的提高而增强;芘对玉米根系的影响要大于对茎叶的影响。玉米幼苗能够明显促进土壤中芘的去除。根际和非根际土壤中芘的去除率分别为56.67%-76.18%和32.64%-70.44%,根际土壤中芘的平均去除率比非根际土壤高16.06%。同处理中根际土壤芘含量显著低于非根际土壤,随着芘处理浓度的提高其差异更加显著。根际土壤微生物量碳、微生物熵、多酚氧化酶活性、脱氢酶活性和磷酸酶活性均高于非根际土壤,代谢熵低于非根际土壤,且其差异随芘处理浓度的提高而增大。在不同芘污染水平下,微生物量碳、微生物熵和脱氢酶活性根际和非根际土壤为T1T2T3,代谢熵为T3T2T1;多酚氧化酶活性根际土壤为T2T1T3,非根际土壤为T1T2T3;磷酸酶活性根际土壤为T3T1T2,非根际土壤为T1T2T3。土壤中残余芘含量与土壤微生物量碳、微生物熵、多酚氧化酶、脱氢酶和磷酸酶活性呈显著负相关,与代谢熵呈显著正相关。     

蚯蚓对细菌降解土壤中菲的作用

通过30d室内培养试验,分别研究了接种蚯蚓(E)、细菌(B)以及同时接种细菌和蚯蚓(BE)对土壤中菲降解的影响.结果表明: 在土壤中菲的初始污染浓度为50 mg*kg-1的条件下,各处理间菲的降解率差异显著,其降解率的大小顺序依次为:BE》B》E》CK(对照); 在150 mg*kg-1菲的初始污染浓度下,BE处理中菲的降解率高达98.86%,显著高于CK和E处理.B处理中细菌的双加氧酶活性在3种菲初始污染浓度下没有显著差异,而BE处理中双加氧酶的活性随着土壤中菲的初始污染浓度的升高而增加.在相同菲污染浓度下BE处理中蚯蚓体内的菲含量明显高于E处理.表明蚯蚓能够通过生物富集作用降低土壤中菲的浓度,而蚯蚓与细菌的相互作用能够进一步促进土壤中菲的降解.     

植物CytP450和抗氧化酶对土壤低浓度菲、芘胁迫的响应

以小麦(Triticum acstivnm)为供试植物,草甸棕壤为供试土壤,以微粒体细胞色素P450及抗氧化酶SOD、POD和CAT酶活性为指标,进行了土壤中菲、芘单一及复合胁迫响应研究.结果初步表明,菲、芘胁迫引起植物体内解毒代谢和抗氧化防御酶反应.菲、芘单一胁迫浓度为1mg kg-1时对细胞色素P450产生显著诱导;4 mg kg-1时P450酶含量明显被抑制,表明低浓度菲、芘单一胁迫对植物代谢解毒系统产生损伤;而菲、芘复合1mg kg-1时P450酶含量明显被抑制,说明菲、芘复合胁迫对植物的代谢解毒具有协同毒性效应.土壤中菲、芘单一胁迫未引起SOD酶活性的明显改变,复合胁迫下SOD酶活性出现微弱下降;菲、芘单一胁迫对CAT和POD酶活性具有显著抑制作用;复合胁迫对CAT产生抑制作用,而POD酶活性并未对菲、芘复合产生增强毒性响应.研究从代谢解毒和抗氧化防御酶系统两方面,为土壤低浓度PAHs污染诊断提供了实验依据.     

多环芳烃降解菌X20的鉴定及降解特性

从多环芳烃降解高效的混合菌群中分离筛选到1株多环芳烃降解菌X20,经形态观察和16SrRNA序列分析,属于假单胞菌(Pseudomonas sp.)。采用室内摇瓶培养的方法,研究了该菌在不同环境条件下对菲和芘的降解。结果表明:弱碱环境有利于菌株X20对菲和芘的降解,最适pH为8.0;葡萄糖对菲芘降解率的影响呈抛物线变化,当葡萄糖浓度为0.2%时,X20对菲和芘的降解达到最高;X20对菲和芘的降解率随其初始浓度的上升而降低,菲和芘在初始浓度为10、20和40mg.L-1时的7d降解率分别为56.3%、39.25%、29.75%和41.8%、29.55%、23.50%,芘对X20降解的抑制强度高于菲。本研究结果将为构建高效的多环芳烃降解菌群,提高多环芳烃原位污染土壤的生物修复效果奠定基础。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

Synthesis of hapten and conjugates of coumestrol and development of immunoassay

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Effect of soil salinity and water stresses on growth and content of nitrogen,chloride and phosphate of wheat and triticale

Summary Three wheats and one triticale were grown, up to flowering stage, in pots on calcareous soil adjusted to a range of salinities (S₁=3.5, S₂=6, S₃=8.5, and S₄=11 mmhos/cm, 20°C, soilpaste extract) by adding solution consisting of 3∶2∶1 of Na-, Ca- and Mg chlorides in chemical equivalent amounts. Moisture in the pots was kept at 100% (W₁), 40% (W₂) and 20% (W₃) of the available water. The vegetative growth, nitrogen and phosphate were affected by S and W treatments, chloride was affected only by S. The interaction S×W affected only dry weight. Varietal effect was observed between wheat as a group and triticale. Multiple quadratic regression equations of these properties on salinity and water revealed that the higher the available water the wider the range of tolerable salinity. Triticale was relatively more tolerant to water stress. Salinity increases Cl and decreases N, whereas water stress enhances N accumulation to a certain extent. However, in triticale at S₃ and S₄ the effect of water stress on N was overshadowed by the excessive salinity. This did not occur for the wheat (Florence). P trends were described. R² for P was low (0.7435–0.3603) which made interpretations rather difficult.     

放牧对牧草光合作用、呼吸作用和氮、碳吸收与转运的影响

研究放牧对草地植物生理活动的影响,对于揭示草地放牧演替的生理机制有重要意义.大量研究表明,家畜放牧对牧草光合作用、呼吸作用以及C和N吸收与转运的影响,可以分为生理伤害和生理恢复2个阶段.放牧通过改变草地冠层结构影响牧草光合作用,净光合作用速率短期内迅速下降,随着叶面积指数增加又逐渐上升,呼吸作用有相似的变化趋势.牧草放牧后再生长所需的C和N最初主要来自根系和留茬中的贮藏物质,此后随着牧草生长恢复逐渐由同化作用供给,C代谢与土壤N水平负相关.放牧后牧草生理活动变化与牧草遗传特性、种间竞争、家畜放牧特征、非生物环境等因素密切相关.