Pb胁迫对马蔺种子萌发和幼苗根尖细胞有丝分裂的影响

采用水培法研究了不同浓度Pb胁迫对马蔺[Iris lactea Pall. var. chinensis (Fisch. ) Koidz. ]种子萌发及根尖细胞有丝分裂的影响.结果显示,在500～1 500 mg·L-1浓度范围内,随Pb浓度的提高,马蔺种子的发芽势、发芽率、发芽指数、活力指数及幼苗根尖细胞有丝分裂指数均呈现出逐渐降低的趋势.经500、1 000和1 500 mg·L-1Pb胁迫处理后,种子发芽势分别仅为对照的52.8%、51.3%和47.2%,种子发芽率分别仅为对照的55.8%、54.3%和46.5%, 种子发芽指数分别比对照降低了55.1%、 56.7%和57.8%, 种子活力指数分别比对照降低了65.4%、 72.0%和79.2%,幼苗根尖细胞有丝分裂指数分别降至对照的28.1%、21.9%和13.5%,且各处理组各指标与对照的差异均达到显著水平(P<0.05),但各处理组间的种子发芽势、发芽率、发芽指数及幼苗根尖细胞有丝分裂指数差异均不显著.研究结果表明,Pb胁迫对马蔺种子萌发及幼苗根尖有丝分裂有显著抑制作用,但马蔺种子萌发和幼苗根尖有丝分裂对浓度高于500 mg·L-1的Pb胁迫反应不敏感.     

镉、锌、铜胁迫对向日葵早期幼苗生长的影响

分析了3种重金属离子(Cd2+、Cu2+、Zn2+)对向日葵种子胚根伸长和早期幼苗生长的影响.结果表明,3种重金属离子对向日葵胚根伸长的抑制作用依次为:Cd2+>Cu2+>Zn2+.3种重金属胁迫明显降低了幼苗生长和叶绿素含量,并显著提高了H2O2水平.其中Cd2+胁迫引起幼苗H2O2爆发高于Cu2+和Zn2+胁迫.进一步分析植株抗氧化系统的变化发现,随着重金属离子浓度的增加,向日葵幼苗酶类抗氧化物质SOD和CAT的活性表现为先增加后降低的趋势;重金属胁迫提高了非酶类的抗氧化物质脯氨酸和GSH的含量.其中Cd2+和Zn2+胁迫对脯氨酸含量变化的影响大于Cu2+胁迫;而Cu2+胁迫对GSH含量变化的影响大于Cd2+和Zn2+胁迫.     

早熟禾对4种重金属胁迫生长响应特征

采用砂培法研究了早熟禾对不同浓度Cu2 、Zn2 、Cd2 和Pb2 胁迫的生长响应特征.结果表明,当Cu2 、Zn2 、Cd2 的浓度达100 m g/L时,早熟禾的种子萌发率、幼苗株高均有所下降,并随着胁迫浓度的增加,下降幅度增大;Pb2 对两指标的影响不太明显.Cu2 对根系及地上生物量具有抑制作用,尤其对根系生长抑制效应极为明显,当浓度增加到600 m g/L时,与对照相比,根长最高下降了96.67%.Zn2 、Cd2 和Pb2 在浓度超过200 m g/L时对早熟禾根系及地上生物量表现出抑制效应,并随着浓度的增加抑制效应增加.4种重金属对叶绿素的影响表现出一致的规律,即当浓度小于200 m g/L时对叶绿素的合成起促进作用,当浓度超过200 m g/L时,叶绿素的含量又随着处理浓度的增加而下降.     

盐胁迫下黄芩种子萌发及幼苗对外源抗坏血酸的生理响应

以黄芩(Scutellaria baicalensis)种子为材料,用氯化钠(Na Cl)模拟盐胁迫条件,采用纸上发芽,研究外源抗坏血酸(As A)对盐胁迫下黄芩种子萌发及幼苗的影响。结果表明,在80 mmol·L-1 Na Cl胁迫下,黄芩种子萌发及幼苗生理指标受到显著抑制;0.50 mmol·L-1 As A处理可显著提高盐胁迫下黄芩种子的发芽势(GE)、发芽率(GR)、发芽指数(GI)、简化活力指数(SVI)以及根系总黄酮、可溶性糖和脯氨酸(Pro)含量,同时也可显著提高幼苗根系活力和超氧化物歧化酶(SOD)活性,显著降低丙二醛(MDA)含量。这说明适宜浓度的抗坏血酸能提高黄芩种子的萌发能力和幼苗对盐胁迫的适应能力,从而起到缓解盐胁迫对种子萌发及幼苗生长的抑制作用。     

甘露醇对海水胁迫下小麦种子萌发及幼苗生长的影响

本文研究了不同浓度甘露醇(MA)对海水胁迫下冬小麦种子萌发及幼苗生长的影响.我们采用不同浓度的甘露醇溶液处理海水胁迫的小麦种子及幼苗,记录小麦种子发芽率、发芽势及发芽指数并在幼苗生长期测定幼苗相关生理指标.结果表明低浓度甘露醇(≤0.5 mmol/L)可提高种子的发芽率、发芽势及发芽指数,高浓度的甘露醇溶液(＞1 mmol/L)对种子的萌发有抑制作用,一定浓度的甘露醇溶液(≤1 mmol/L)可以提高海水胁迫下幼苗的叶绿素含量,降低游离脯氨酸(Pro)和游离丙二醛(MDA)含量,并可通过分析结果观察施加海水胁迫与甘露醇后叶绿素、类胡萝卜素、脯氨酸(Pro)和丙二醛(MDA)的含量随时间在冬小麦幼苗中的积累情况.低浓度的甘露醇对海水胁迫下的冬小麦萌发及幼苗生长具有一定的保护作用,为缓解农业生产中的盐害问题提供了新的思路.本研究为小麦的盐渍化土壤种植提供了新思路,并为甘露醇进一步开发利用于农业提供了一定的理论依据.     

镉胁迫下玉米幼苗生理生态的变化

用不同浓度Cd2 + 处理玉米种子 ,在室内常规培养 ,研究了种子萌发和幼苗生理生态的变化。结果表明 ,镉影响玉米种子的萌发和幼苗的生长。当Cd2 + 浓度高于 5 0mg·L-1时 ,显著抑制种子的发芽率 ;随Cd2 + 浓度的增加 ,根系长度和侧根数减小 ;在Cd2 + 浓度小于 5mg·L-1时 ,镉刺激苗高和根系及地上部干物质量增加 ,当Cd2 + 浓度超过相应浓度时 ,苗高和根系及地上部的生长量随Cd2 + 浓度提高而降低。镉胁迫下幼苗根系活力和叶绿素含量明显降低 ,根内丙二醛含量增加。镉影响玉米幼苗对矿质元素的吸收。根系和茎叶中Ca、Mg、Fe、Cu的吸收量随Cd2 + 浓度提高而增加 ,K、Zn的吸收量随Cd2 + 浓度提高而减少。     

不同处理对沙生针茅种子萌发的影响

沙生针茅是重要的荒漠植物,研究其种子萌发特性及其对主要影响因子的响应对于认识其生存适应性以及荒漠植被恢复具有重要意义。鉴于沙生针茅种子常处休眠状态、不易萌发,而现有研究尚未充分揭示各种因素和处理对沙生针茅种子萌发的影响,本论文报道了不同因素、不同处理对沙生针茅种子萌发的影响。结果表明:(1)60℃蒸馏水浸泡种子24 h,种子吸水率、发芽率、发芽势分别达到26%、38%和32%;(2)-20/20℃变温处理96 h,发芽率、发芽势分别达到48%和46%;(3)切除绝大部分胚乳(3/4)后,种子发芽率、发芽势分别可达到58%和54%;(4)用80 mg·L~(-1)赤霉素(GA3)水溶液浸泡种子24 h,发芽率、发芽势分别达到54%和48%;(5)用聚乙二醇(PEG-6000)处理,随浓度增加,种子发芽率、发芽势、发芽指数均呈下降趋势,种子萌发水分胁迫的临界值和极限值分别为15.9%和26.3%;(6)盐胁迫处理下,种子发芽率、发芽势、发芽指数均随盐碱浓度增加呈下降趋势,其中Na2CO3处理下的种子萌发率下降幅度明显大于Na Cl,表明碱性盐胁迫对沙生针茅种子萌发的抑制作用大于中性盐胁迫;(7)采用综合处理方法,种子发芽率、发芽势分别达到78%和76%。     

Cd胁迫对紫花苜蓿种子发芽及幼苗生理生化特性的影响

以‘甘农3号’紫花苜蓿(Medicago sativa L.cv.‘Gannong 3’)品种为试验材料,研究不同浓度Cd胁迫对紫花苜蓿种子萌发及幼苗生理生化特性的影响。结果显示:(1)Cd胁迫显著抑制紫花苜蓿种子萌发及幼苗生长,种子发芽势、发芽率及幼苗的胚芽长、胚根长和干重均随着Cd处理浓度提高显著降低,且对发芽势的抑制作用大于发芽率,对胚根生长的抑制作用大于胚芽。(2)Cd胁迫第4d时,萌发种子中蛋白水解酶的活性受到显著抑制,可溶性蛋白和可溶性糖含量显著降低。(3)Cd胁迫第10d时,随着Cd胁迫浓度的增加,苜蓿幼苗的超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)、愈创木酚过氧化物酶(GPX)活性均先升高后降低,过氧化氢酶(CAT)活性持续下降,超氧阴离子自由基产生速率、H2O2含量和丙二醛(MDA)含量均显著增加。研究表明,低浓度Cd胁迫下,苜蓿幼苗抗氧化系统清除活性氧的能力上升,高浓度Cd胁迫下,苜蓿幼苗抗氧化系统活性下降,膜脂过氧化程度加剧,活性氧的过量积累是Cd伤害苜蓿幼苗和其生物量下降的主要原因。     

水杨酸对锌胁迫下小麦幼苗生长抑制的缓解效应

以小麦品种‘新麦18’为材料,采用室内水培实验研究了不同浓度水杨酸(SA)处理对300 mg.L-1锌胁迫下小麦种子萌发和幼苗生长的影响。结果表明:在Zn2+胁迫下,小麦种子的发芽势和发芽率、幼苗根长、芽长以及幼苗叶片的可溶性蛋白含量、根系活力显著降低,而脯氨酸和丙二醛(MDA)含量显著增加(P<0.05);外施SA显著提高了Zn2+胁迫下小麦种子的发芽势和发芽率,同时也使Zn2+胁迫7 d后的小麦幼苗的根长、芽长,幼苗叶片的脯氨酸和可溶性蛋白含量以及根系活力显著升高,膜脂过氧化产物MDA含量却显著降低(P<0.05)。由此可见,外施SA可通过提高小麦幼苗根长和芽长来增加幼苗根系活力,通过提高小麦幼苗可溶性蛋白含量、脯氨酸含量来维持细胞膜的稳定性,降低膜脂过氧化伤害程度,从而缓解了Zn2+胁迫对幼苗生长的抑制,并以14 mg.L-1外源水杨酸缓解效果最好。     

匍茎翦股颖对Cu2+、Zn2+、Cd2+与Pb2+ 胁迫的生长响应与阈限浓度

采用砂培法,研究了匍茎翦股颖对Cu2+、Zn2+、Cd2+与Pb2+胁迫的生长响应及阈限浓度,结果表明:种子萌发率随着4种重金属浓度的增加而下降。对株高的影响是当重金属浓度小于100mg/L时会促进株高生长,高于100mg/L则产生抑制作用。Cu2+显著抑制根系生长,并随浓度的增加抑制效应愈加显著;在Cu2+浓度为600mg/L时匍茎翦股颖的根长比对照下降了93.75%。Cu2+、Zn2+、Pb2+浓度小于200mg/L时会促进地上生物量的增加,但高于200mg/L时,地上生物量会随着3种重金属的增加而减少。Cu2+、Zn2+浓度小于100mg/L或Cd2+、Pb2+浓度小于200mg/L会增加叶绿素的含量,高浓度会降低叶绿素的含量;Cd2+在浓度为600mg/L时显著降低叶绿素含量,与对照相比,下降了43.55%。匍茎翦股颖生长的综合效应分析表明,匍茎翦股颖对Cu2+胁迫最敏感,具有较低的阈限浓度,而Zn2+胁迫对匍茎翦股颖的生长影响最小,阈限浓度相对较高。     

重金属铅与两种淡水藻的相互作用

为了研究重金属铅与淡水藻类之间的相互作用,采用不同Pb2+浓度处理铜绿微囊藻(Microcysis aeruginosa Kutz.)和斜生栅藻[Scenedesmus obliquus(Turp.)Kutz.],分别对两种藻的生物量、藻液电导率、O-·2含量、过氧化物酶(POD)、过氧化氢酶(CAT)活性以及藻对Pb2+的吸收作用等进行了测定,并通过扫描电镜观察了不同Pb2+浓度处理下两种藻细胞的表面结构。结果显示:(1)Pb2+浓度低于3 mg/L促进铜绿微囊藻生长,高于9 mg/L抑制其生长;但在3—12 mg/L范围内,Pb2+均明显抑制了斜生栅藻的生长,说明斜生栅藻对Pb2+毒性的敏感程度要高于铜绿微囊藻。(2)受到铅离子的胁迫,两种藻细胞膜通透性均有一定改变,扫描电子显微镜的照片观察,两种藻细胞表面的絮状物随着Pb2+的升高而增多,尤其是斜生栅藻细胞结构改变明显,多数细胞变形破裂;同时,O-·2含量升高,POD、CAT活性早期均可随Pb2+的增加而上升,表明氧自由基的产生增多以及由其引起的细胞生理生化改变可能是铅离子作用于藻细胞的主要机制。(3)两种淡水藻对Pb2+均有吸收作用,单位量藻细胞内,斜生栅藻对Pb2+的吸收能力好于铜绿微囊藻。所有结果提示:斜生栅藻不仅可以作为对重金属敏感的指示生物来监测水体Pb2+污染程度;同时由于斜生栅藻比铜绿微囊藻具有更好的Pb2+吸收能力,因此还可以利用斜生栅藻作为处理水体Pb2+的生物材料。     

Risedronate metal complexes potentially active against Chagas disease

In the search for new metal-based drugs for the treatment of Chagas disease, the most widespread Latin American parasitic disease, novel complexes of the bioactive ligand risedronate (Ris, (1-hydroxy-1-phosphono-2-pyridin-3-yl-ethyl)phosphonate), [M^II(Ris)₂]·4H₂O, where M═Cu, Co, Mn and Ni, and [Ni^II(Ris)₂(H₂O)₂]·H₂O were synthesized and characterized by using analytical measurements, thermogravimetric analyses, cyclic voltammetry and infrared and Raman spectroscopies. Crystal structures of [Cu^II(Ris)₂]·4H₂O and [Ni^II(Ris)₂(H₂O)₂]·H₂O were solved by single crystal X-ray diffraction methods. The complexes, as well as the free ligand, were evaluated in vitro against epimastigotes and intracellular amastigotes of the parasite Trypanosoma cruzi, causative agent of Chagas disease. Results demonstrated that the coordination of risedronate to different metal ions improved the antiproliferative effect against T. cruzi, exhibiting growth inhibition values against the intracellular amastigotes ranging the low micromolar levels. In addition, this strong activity could be related to high inhibition of farnesyl diphosphate synthase enzyme. On the other hand, protein interaction studies showed that all the complexes strongly interact with albumin thus providing a suitable means of transporting them to tissues in vivo.     

Structural and thermodynamic characterization of metal ion binding in Streptococcus suis Dpr

The use of protein cages for the creation of novel inorganic nanomaterials has attracted considerable attention in recent years. Ferritins are among the most commonly used protein cages in nanoscience. Accordingly, the binding of various metals to ferritins has been studied extensively. Dps (DNA-binding protein from starved cells)-like proteins belong to the ferritin superfamily. In contrast to ferritins, Dps-like proteins form 12-mers instead of 24-mers, have a different ferroxidase center, and are able to store a smaller amount of iron atoms in a hollow cavity (up to ∼ 500, instead of the ∼ 4500 iron atoms found in ferritins). With the exception of iron, the binding of other metal cations to Dps proteins has not been studied in detail. Here, the binding of six divalent metal ions (Zn²⁺, Mn²⁺, Ni²⁺, Co²⁺, Cu²⁺, and Mg²⁺) to Streptococcus suisDps-like peroxide resistance protein (SsDpr) was characterized by X-ray crystallography and isothermal titration calorimetry (ITC). All metal cations, except for Mg²⁺, were found to bind to the ferroxidase center similarly to Fe²⁺, with moderate affinity (binding constants between 0.1 × 10⁵ M^− 1 and 5 × 10⁵ M^− 1). The stoichiometry of binding, as deduced by ITC data, suggested the presence of a dication ferroxidase site. No other metal binding sites were identified in the protein. The results presented here demonstrate the ability of SsDpr to bind various metals as substitutes for iron and will help in better understanding protein-metal interactions in the Dps family of proteins as potential metal nanocontainers.     

Activation and inhibition of rubber transferases by metal cofactors and pyrophosphate substrates

Metal cofactors are necessary for the activity of alkylation by prenyl transfer in enzyme-catalyzed reactions. Rubber transferase (RuT, a cis-prenyl transferase) associated with purified rubber particles from Hevea brasiliensis, Parthenium argentatum and Ficus elastica can use magnesium and manganese interchangably to achieve maximum velocity. We define the concentration of activator required for maximum velocity as [A](max). The [A](max)(Mg2+) in F. elastica (100 mM) is 10 times the [A](max)(Mg2+) for either H. brasiliensis (10 mM) or P. argentatum (8 mM). The [A](max)(Mn2+) in F. elastica (11 mM), H. brasiliensis (3.8 mM) and P. argentatum (6.8 mM) and the [A](max)(Mg2+) in H. brasiliensis (10 mM) and P. argentatum (8 mM) are similar. The differences in [A](max)(Mg2+) correlate with the actual endogenous Mg(2+) concentrations in the latex of living plants. Extremely low Mn(2+) levels in vivo indicate that Mg(2+) is the RuT cofactor in living H. brasiliensis and F. elastica trees. Kinetic analyses demonstrate that FPP-Mg(2+) and FPP-Mn(2+) are active substrates for rubber molecule initiation, although free FPP and metal cations, Mg(2+) and Mn(2+), can interact independently at the active site with the following relative dissociation constants K(d)(FPP) 相似文献   

Functional assessment of Nramp-like metal transporters and manganese in Caenorhabditis elegans

Nramp1 (natural resistance-associated macrophage protein-1) is a functionally conserved iron-manganese transporter in macrophages. Manganese (Mn), a superoxide scavenger, is required in trace amounts and functions as a cofactor for most antioxidants. Three Nramp homologs, smf-1, smf-2, and smf-3, have been identified thus far in the nematode Caenorhabditis elegans. A GFP promoter assay revealed largely intestinal expression of the smf genes from early embryonic through adult stages. In addition, smf deletion mutants showed increased sensitivity to excess Mn and mild sensitivity to EDTA. Interestingly, these smf deletion mutants demonstrated hypersensitivity to the pathogen Staphylococcus aureus, an effect that was rescued by Mn feeding or knockdown of the Golgi calcium/manganese ATPase, pmr-1, indicating that Mn uptake is essential for the innate immune system. This reversal of pathogen sensitivity by Mn feeding suggests a protective and therapeutic role of Mn in pathogen evasion systems. We propose that the C. elegans intestinal lumen may mimic the mammalian macrophage phagosome and thus could be a simple model for studying Mn-mediated innate immunity.     

Social evolution of toxic metal bioremediation in Pseudomonas aeruginosa

Bacteria are often iron-limited, and hence produce extracellular iron-scavenging siderophores. A crucial feature of siderophore production is that it can be an altruistic behaviour (individually costly but benefitting neighbouring cells), thus siderophore producers can be invaded by non-producing social ‘cheats’. Recent studies have shown that siderophores can also bind other heavy metals (such as Cu and Zn), but in this case siderophore chelation actually reduces metal uptake by bacteria. These complexes reduce heavy metal toxicity, hence siderophore production may contribute to toxic metal bioremediation. Here, we show that siderophore production in the context of bioremediation is also an altruistic trait and can be exploited by cheating phenotypes in the opportunistic pathogen Pseudomonas aeruginosa. Specifically, we show that in toxic copper concentrations (i) siderophore non-producers evolve de novo and reach high frequencies, and (ii) producing strains are fitter than isogenic non-producing strains in monoculture, and vice versa in co-culture. Moreover, we show that the evolutionary effect copper has on reducing siderophore production is greater than the reduction observed under iron-limited conditions. We discuss the relevance of these results to the evolution of siderophore production in natural communities and heavy metal bioremediation.     

Tracking the evolution of porphobilinogen synthase metal dependence in vitro

Metal ions are indispensable cofactors for chemical catalysis by a plethora of enzymes. Porphobilinogen synthases (PBGSs), which catalyse the second step of tetrapyrrole biosynthesis, are grouped according to their dependence on Zn(2+). Using site-directed mutagenesis, we embarked on transforming Zn(2+)-independent Pseudomonas aeruginosa PBGS into a Zn(2+)-dependent enzyme. Nine PBGS variants were generated by permutationally introducing three cysteine residues and a further two residues into the active site of the enzyme to match the homologous Zn(2+)-containing PBGS from Escherichia coli. Crystal structures of seven enzyme variants were solved to elucidate the nature of Zn(2+) coordination at high resolution. The three single-cysteine variants were invariably found to be enzymatically inactive and only one (D139C) was found to bind detectable amounts of Zn(2+). The double mutant A129C/D139C is enzymatically active and binds Zn(2+) in a tetrahedral coordination. Structurally and functionally it mimics mycobacterial PBGS, which bears an equivalent Zn(2+)-coordination site. The remaining two double mutants, without known natural equivalents, reveal strongly distorted tetrahedral Zn(2+)-binding sites. Variant A129C/D131C possesses weak PBGS activity while D131C/D139C is inactive. The triple mutant A129C/D131C/D139C, finally, displays an almost ideal tetrahedral Zn(2+)-binding geometry and a significant Zn(2+)-dependent enzymatic activity. Two additional amino acid exchanges further optimize the active site architecture towards the E.coli enzyme with an additional increase in activity. Our study delineates the potential evolutionary path between Zn(2+)-free and Zn(2+)-dependent PBGS enyzmes showing that the rigid backbone of PBGS enzymes is an ideal framework to create or eliminate metal dependence through a limited number of amino acid exchanges.     

Crystal structure of the stationary phase survival protein SurE with metal ion and AMP

The stationary phase survival protein SurE is a metal ion-dependent phosphatase distributed among eubacteria, archaea, and eukaryotes. In Escherichia coli, SurE has activities as nucleotidase and exopolyphosphatase, and is thought to be involved in stress response. However, its physiological role and reaction mechanism are unclear. We report here the crystal structures of the tetramer of SurE from Thermus thermophilus HB8 (TtSurE) both alone and crystallized with Mn(2+) and substrate AMP. In the presence of Mn(2+) and AMP, differences between the protomers were observed in the active site and in the loop located near the active site; AMP-bound active sites with the loops in a novel open conformation were found in the two protomers, and AMP-free active sites with the loops in a conventional closed conformation were found in the other two protomers. The two loops in the open conformation are entwined with each other, and this entwining is suggested to be required for enzymatic activity by site-directed mutagenesis. TtSurE exists as an equilibrium mixture of dimer and tetramer in solution. The loop-entwined structure indicates that SurE acts as a tetramer. The structural features and the absence of negative cooperativity imply the half-of-the-sites reactivity mechanism resulting from a pre-existing tendency toward structural asymmetry.     

Seasonal variation in trace metal concentrations in three talitrid amphipods from the Gulf of Gdansk, Poland

Talitrid amphipod crustaceans are increasingly being used as biomonitors of trace metal bioavailabilities in coastal waters, and it is important to understand how other factors, in addition to metal pollution, might affect their accumulated body concentrations. Seasonal variation (April-October 2000) in body concentrations of copper, zinc, cadmium, iron, manganese, lead and nickel has been investigated in three species, Orchestia cavimana, Talitrus saltator and Talorchestia deshayesii, from the region of the Gulf of Gdansk (including the Vistula Lagoon), Poland, an area with considerable temperature variation between summer and winter. Seasonal variation in the body metal concentrations of talitrid amphipod crustaceans was confirmed in each population, but only copper showed a consistent pattern. Copper concentrations fell from early to late summer, perhaps either as a result of the replacement of the older overwintering generation by the younger spring one, or as a result of the loss of haemocyanin in autumn amphipods entering a period of low metabolic activity. Metal concentration data for T. saltator from Gdynia-Osada Rybacka compared with equivalent data for 1996 and 1998 show a significant drop in metals associated with the influx of the Vistula river water, which occurred in 1998, following a major flood event in 1997.