红裸须摇蚊幼虫肠道可培养细菌多样性及耐受重金属功能初步研究

红裸须摇蚊Propsilocerus akamusi为华北地区常见耐污种,广泛分布于各种水体中。本研究采用传统培养法对4龄红裸须摇蚊幼虫肠道内细菌种类进行分离,利用16S rRNA基因序列分析比对,结合菌落形态、系统进化分析等方法进一步鉴定菌株种类,同时探究可培养菌株对重金属（铜、镍）的耐受情况。结果表明,红裸须摇蚊幼虫在重金属铜暴露下的24 h-LC50、48 h-LC50、72 h-LC50、96 h-LC50分别为6304.86、403.59、29.33以及14.43 mmol/L,在重金属镍暴露下的24 h-LC50、48 h-LC50、72 h-LC50、96 h-LC50分别为339.52、182.18、102.59以及60.94 mmol/L。利用LB、NA、BHI、MC 4种培养基,从幼虫肠道分离纯化到16种不同遗传型细菌,分属2门 9科16属,其中优势菌门为变形菌门Proteobacteria。另外,丛毛单胞菌科Comamonadaceae丛毛单胞菌属Comamonas丛毛单胞菌Comamonas sp.对重金属铜和镍有良好耐受性。通过研究红裸须摇蚊幼虫肠道可培养细菌群落组成以及重金属胁迫下摇蚊幼虫肠道细菌群落生长情况,可为水生昆虫肠道菌群资源库的开发提供了一定的思路,也可为筛选出具备重金属代谢功能的细菌资源奠定基础。     

超积累植物伴矿景天镉耐受基因SpMT2的分离及功能鉴定

超积累植物由于其对重金属具有地上部超积累以及超耐受等特性,不仅是研究植物离子转运及毒性耐受的理想模式,而且在植物修复的发展和应用中具有不可替代的作用。伴矿景天是近年在我国境内发现的一种景天科镉(Cd)/锌(Zn)超积累植物。为鉴定其富集和耐受Cd的关键基因,笔者构建了其酵母表达cDNA文库,利用酵母的遗传互补系统筛选到一个极大提高了酵母抗Cd能力的基因SpMT2。SpMT2属于富含半胱氨酸(Cys)的金属硫蛋白(Metallothionein)家族。亚细胞定位表明SpMT2表达于酵母细胞质中,并特异地提高酵母对Cd的抗性。进一步研究发现SpMT2的表达显著降低了酵母液泡中Cd含量,但酵母吸收的总Cd含量无显著变化。推测抗性增加是由于SpMT2在酵母细胞质中通过螯合Cd从而降低Cd对酵母的毒害。qRT-PCR分析表明SpMT2在伴矿景天的根和地上部都高丰度表达,且不受Cd诱导变化。鉴于SpMT2也定位于植物细胞质中,结合上述结果,推测SpMT2可能在伴矿景天细胞质中螯合Cd,在降低Cd毒害的同时可能还保持Cd在细胞质中的流动性,从而在Cd长途转运过程中也发挥重要作用。     

Increased heat requirement for leaf flushing in temperate woody species over 1980–2012: effects of chilling,precipitation and insolation

Recent studies have revealed large unexplained variation in heat requirement‐based phenology models, resulting in large uncertainty when predicting ecosystem carbon and water balance responses to climate variability. Improving our understanding of the heat requirement for spring phenology is thus urgently needed. In this study, we estimated the species‐specific heat requirement for leaf flushing of 13 temperate woody species using long‐term phenological observations from Europe and North America. The species were defined as early and late flushing species according to the mean date of leaf flushing across all sites. Partial correlation analyses were applied to determine the temporal correlations between heat requirement and chilling accumulation, precipitation and insolation sum during dormancy. We found that the heat requirement for leaf flushing increased by almost 50% over the study period 1980–2012, with an average of 30 heat units per decade. This temporal increase in heat requirement was observed in all species, but was much larger for late than for early flushing species. Consistent with previous studies, we found that the heat requirement negatively correlates with chilling accumulation. Interestingly, after removing the variation induced by chilling accumulation, a predominantly positive partial correlation exists between heat requirement and precipitation sum, and a predominantly negative correlation between heat requirement and insolation sum. This suggests that besides the well‐known effect of chilling, the heat requirement for leaf flushing is also influenced by precipitation and insolation sum during dormancy. However, we hypothesize that the observed precipitation and insolation effects might be artefacts attributable to the inappropriate use of air temperature in the heat requirement quantification. Rather than air temperature, meristem temperature is probably the prominent driver of the leaf flushing process, but these data are not available. Further experimental research is thus needed to verify whether insolation and precipitation sums directly affect the heat requirement for leaf flushing.     

In Vitro,In Silico,ADME and Theoretical Analysis of Mn(II) and Co(II) Complexes Derived from Methyl-(Z)-N′-Carbamothioylcarbamohydrazonate Schiff Base Ligand

Mn(II) and Co(II) complexes of methyl-(Z)−N′-carbamothioylcarbamohydrazonate Schiff base ligand were synthesized. The ligand and metal salts were taken in 2 : 1 stoichiometric ratio. All the synthesized complexes were characterized using elemental analysis, molar conductance, magnetic moment and various spectroscopic techniques (FT-IR, UV/VIS, EPR) techniques. Elemental and spectroscopic results verified bidentate donor nature of the ligand and octahedral geometry of all the complexes. The non-electrolytic nature of Mn(II) and Co(II) complexes were suggested by conductivity data analysis. In vitro antibacterial (E. coli and S. aureus) and antifungal (C. albicans and C. tropicalis) screening were achieved by employing agar well diffusion method which revealed better antimicrobial activity of Co(II) complexes than Mn(II) complexes. In silico SwissADME study predicted the drug-likeness probability of ligand and complexes. The interaction of two bacterial proteins (E. coli and S. aureus) with compounds was also analyzed using molecular docking study, which corroborate the in vitro analysis.     

Association of headaches and the metals

Lack of specific markers constitutes a problem during diagnosis of headache syndromes. Recently, some metals have gained importance as biological parameters for the diagnosis and during treatment. Low-ionized Mg and high-ionized Ca/Mg in patients with daily migrainous headaches were noted. The blood Na level was shown to increase before and during headache. Headache is also a symptom of the common cold for which zinc may be an effective therapy. The existing relationship between genetic markers of the cluster headache and the efficacy of lithium salts therapy was noted. Headache was also found to be associated with toxic metals. When the health effects of mercury were investigated, the most frequently observed symptom was cephalalgia. Continuous exposure to lead was concomitant with the appearance of symptoms such as headache. In relation to some metabolic links, metals may be introduced as possible biological markers for the diagnosis and during the therapy of different headache syndromes in future clinical trials and laboratory measurements.     

Genes for all metals—a bacterial view of the Periodic Table

Bacterial chromosomes have genes for transport proteins for inorganic nutrient cations and oxyanions, such as NH₄ ⁺, K⁺, Mg²⁺, Co²⁺, Fe³⁺, Mn²⁺, Zn²⁺ and other trace cations, PO₄ ^3-, SO₄ ^2- and less abundant oxyanions. Together these account for perhaps a few hundred genes in many bacteria. Bacterial plasmids encode resistance systems for toxic metal and metalloid ions including Ag⁺ AsO₂ ^-, AsO₄ ^3-, Cd²⁺, Co²⁺, CrO₄ ²⁻, Cu²⁺, Hg²⁺, Ni²⁺, Pb²⁺, TeO₃ ²⁻, TI⁺ and Zn²⁺. Most resistance systems function by energy-dependent efflux of toxic ions. A few involve enzymatic (mostly redox) transformations. Some of the efflux resistance systems are ATPases and others are chemiosmotic ion/proton exchangers. The Cd²⁺-resistance cation pump of Gram-positive bacteria is membrane P-type ATPase, which has been labeled with ³²P from [γ-³²P]ATP and drives ATP-dependent Cd²⁺ (and Zn²⁺) transport by membrane vesicles. The genes defective in the human hereditary diseases of copper metabolism, Menkes syndrome and Wilson’s disease, encode P-type ATPases that are similar to bacterial cadmium ATPases. The arsenic resistance system transports arsenite [As(III)], alternatively with the ArsB polypeptide functioning as a chemiosmotic efflux transporter or with two polypeptides, ArsB and ArsA, functioning as an ATPase. The third protein of the arsenic resistance system is an enzyme that reduces intracellular arsenate [As(V)] to arsenite [As(III)], the substrate of the efflux system. In Gram-negative cells, a three polypeptide complex functions as a chemiosmotic cation/protein exchanger to efflux Cd²⁺, Zn²⁺ and Co²⁺. This pump consists of an inner membrane (CzcA), an outer membrane (CzcC) and a membrane-spanning (CzcB) protein that function together. Received 08 August 1997/ Accepted in revised form 01 November 1997     

A versatile and highly sensitive probe for Hg(II), Pb(II) and Cd(II) detection individually and totally in water samples

The detection of heavy metal ions using enzyme-linked immunosorbent assays (ELISA) has been reported by several research groups. However, highly sensitive and selective detection of total heavy metal ions using ELISA is a major technical limitation. Here we describe the development of a versatile and highly sensitive probe combining goat anti-mice IgG, colloidal gold nanoparticles (AuNPs) and horseradish peroxidase (HRP). We demonstrate the utility of this probe using three kinds of heavy metal complete antigens and three monoclonal antibodies (McAbs) in one ELISA system to establish a high-throughput screening protocol. The procedure was successfully applied to analysis of Hg(II), Pb(II) and Cd(II) individually and totally from different water samples. The sensitivities for the detection of Hg(II), Pb(II) and Cd(II) individually and totally are 27.4, 3.9, 15.8 and 18.2 nM, respectively. And all limit of detection (LODs) are lower than 1.2 nM. The recovery results obtained from the developed technique showed a good correlation (R² = 0.983) with those from ICP-MS. The major advantage of the probe is the versatility and high sensibility. The probe could be potentially used, upon demand, as a sensitive and versatile detector for a broad range of applications.     

一株虾池来源的螺旋拟柱孢藻藻株的分离鉴定及重金属离子Cu~(2+)、Cd~(2+)和Pb~(2+)对其生长的影响

【目的】探讨凡纳滨对虾养殖水体中入侵蓝藻拟柱孢藻的生长生理特性。【方法】从汕头澄海人工对虾养殖池分离纯化藻株,通过形态及其16SrRNA基因鉴定,之后在CT与BG11两种蓝藻通用培养基的基础上优化最佳培养条件,最后分析了不同浓度的3种重金属离子即Cu~(2+)(0–0.8 mg/L)、Cd~(2+)(0–4 mg/L)和Pb~(2+)(0–80 mg/L)对藻株生长的影响。【结果】澄海虾池来源的分离纯化藻株形态呈卷曲螺旋型,16S rRNA基因序列与多株其他来源的拟柱孢藻相似度均达98%以上。实验室培养,藻株最佳生长状态的培养条件是在BG11培养基的基础上调整氮浓度及氮磷比分别为N 62 mg/L,N︰P=9︰1,在此条件下,藻丝生物量可达(0.632±0.170)×107/L,藻丝比平均生长速率最高为(0.063±0.001)/d。本分离藻株活体对重金属Cu~(2+)、Cd~(2+)和Pb~(2+)具有一定的耐受性,其耐受浓度范围分别为0–0.2、0–0.5和1–40 mg/L,其中,Cu~(2+)和Cd~(2+)对藻的生长具有抑制作用,而且此抑制作用随着金属离子剂量的增加及作用时间的延长更加显著,Cu~(2+)和Cd~(2+)对藻体的半数抑制浓度(96 h EC50)分别为0.125和0.551 mg/L;而浓度范围为0–80 mg/L的Pb~(2+)对藻体的生长则表现为低剂量(≤40 mg/L)呈促进,高剂量(≥80 mg/L)则抑制。【结论】从凡纳滨对虾养殖池中分离鉴定出一株形态呈螺旋型的拟柱孢藻,命名为螺旋拟柱孢藻(Cylindrospermopsis raciborskii helix),本藻株活体能够在一定浓度的Cu~(2+)、Cd~(2+)和Pb~(2+)中生长,为螺旋拟柱孢藻活藻生物吸附重金属离子而改善虾池水体环境提供了可能性。     

黏土矿物中重金属离子的吸附规律及竞争吸附

采用等温吸附法,研究了重金属铜、铅、镉、镍在膨润土中的吸附特征,发现膨润土对铜、铅的吸附明显强于镉、镍,吸附强度大小顺序为Pb2 >Cu2 >Ni2 >Cd2 。Langmuir和Freundlich方程对这4种金属离子等温吸附的拟合均呈极显著关系。Pb2 、Cd2 、Ni2 分别与Cu2 的双组分竞争吸附表明,黏土矿物对4种离子具有"选择性吸附"。在Pb2 、Ni2 、Cd2 的存在条件下,黏土矿物对Cu2 的吸附产生不同程度的下降;100mg/LCu2 对Pb2 的影响不大,但可完全抑制Ni2 、Cd2 的吸附。建立了IAS和LCA模型来预测Pb2 与Cu2 的双组分竞争吸附,并对LCA模型进行修正,提出了更符合实际情况的竞争吸附模型。文章最后用LCA修正模型对Pb2 与Cu2 的双组分竞争吸附进行了模拟。     

镉胁迫对秋华柳根系活力及其Ca、Mg、Mn、Zn、Fe积累的影响

以秋华柳为试验材料,采用水培试验方式,设置CK(0 mg·L^-1 Cd²⁺)、T₁(2 mg·L^-1 Cd²⁺)、T₂(10 mg·L^-1Cd²⁺)、T₃(20 mg·L^-1Cd²⁺)、T₄(50 mg·L^-1Cd²⁺)5种镉处理浓度,通过对秋华柳根系活力,叶、韧皮部、木质部和根部的镉含量,以及Ca、Mg、Mn、Zn、Fe 5种常规金属元素含量的测定,研究了不同浓度镉胁迫下秋华柳根系活力及Ca、Mg、Mn、Zn、Fe 等金属元素含量的变化.结果表明:1)秋华柳根系活力随着镉处理浓度的增加而逐渐下降,当镉浓度≥10 mg·L^-1时,根系活力与对照相比显著下降.2)随着镉处理浓度的增加,秋华柳叶中Fe的积累受到显著抑制;韧皮部Mg、Mn、Fe的积累受到显著抑制;木质部Ca、Mg、Mn、Zn、Fe 等5种常规金属元素的积累无显著性差异;根部5种常规金属元素的吸收和积累受到显著抑制,表现出镉对其他金属元素积累的拮抗作用.其中,50 mg·L^-1的镉胁迫下,根部Zn的积累量降幅最大,受Cd的抑制最明显.3)各处理组Fe的转移系数与对照相比均无显著性差异;Ca、Mg、Mn、Zn的转移系数均高于对照,且在一定的镉处理水平上差异显著.4)镉胁迫下,秋华柳根部镉的积累量与Ca的积累量呈显著负相关,与其他4种常规金属元素的积累量呈极显著负相关,说明根部常规金属元素的变化可作为秋华柳受镉毒害程度的指示之一.