基于苋菜转录组的ARF基因家族鉴定及表达

ARFs(auxin response factors)是重要的生长素响应因子,在植物的生长发育过程中起着关键的作用.基于'大红'苋菜转录组数据,通过在线分析软件SMART和NCBI-Blastp的注释筛选苋菜ARF基因家族成员(AtrARF);利用生物信息学分析软件,对AtrARFs蛋白的理化性质、二级结构、亚细胞定...     

Optimization of the T3-induced Xenopus metamorphosis assay for detecting thyroid hormone signaling disruption of chemicals

T3-induced Xenopus metamorphosis is an ideal model for detecting thyroid hormone(TH)signaling disruption of chemicals. To optimize the T3-induced Xenopus assay and improve its sensitivity and reproducibility, we intend to develop quantitatively morphological endpoints and choose appropriate concentrations and exposure durations for T3 induction.Xenopus laevis at stage 52 were exposed to series of concentrations of T3(0.31–2.5 nmol/L)for 6 days. By comparing morphological changes induced by T3, we propose head area,mouth width, unilateral brain width/brain length, and hindlimb length/snout-vent length as quantitative parameters for characterizing T3-induced morphological changes, with body weight as a parameter for indicating integrated changes. By analyzing time-response curves, we found that following 4-day exposure, T3-induced grossly morphological changes displayed linear concentration–response curves, with moderate morphological changes resulting from 1.25 nmol/L T3 exposure. When using grossly morphological endpoints to detect TH signaling disruption, we propose 4 days as exposure duration of T3, with concentrations close to 1.25 nmol/L as induction concentrations. However, it is appropriate to examine morphological and molecular changes of the intestine on day 2 due to their early response to T3. The quantitative endpoints and T3 induction concentrations and durations we determined would improve the sensitivity and the reproducibility of the T3-induced Xenopus metamorphosis assay.     

大肠杆菌植酸酶在毕赤酵母中的表达与纯化

为探讨植酸酶的结构与功能,研究了来源于大肠杆菌的植酸酶基因在酵母中的表达和纯化条件.将含有大肠杆菌植酸酶基因的毕赤酵母工程菌在不同甲醇浓度和不同诱导时间下培养,检测植酸酶的表达情况.结果表明:诱导培养基中一次性添加2.5%的甲醇,诱导96 h后,蛋白浓度达到1.36 mg mL-1,酶活力达到6 530 U mL-1;将在毕赤酵母中表达的植酸酶粗酶液经过硫酸铵盐析、Resource S柱和Superdex-75三步纯化,得到单峰纯的蛋白,比活力为137 280 Umg-1.用圆二色谱分析纯化后的蛋白在pH 5.0和8.0环境中的结构变化,结果显示pH 8.0环境使大肠杆菌植酸酶发生了变性.图3参16     

五氯苯酚对污泥性能和功能基因表达水平的影响

氯酚废水是一类广泛存在于环境的典型工业废水,具有显著的生态毒性,为探讨废水中氯酚类化合物对污泥性能和功能基因表达水平的影响,采用HRT（水力停留时间）为8 h、SBR（间歇曝气的序批式生物反应器）处理进水浓度为2 mg/L的PCP（五氯苯酚）废水,并与处理不含PCP的废水作对比分析,探讨PCP对污染物去除和微生物菌群的影响,并分析PCP诱导下污泥中Pseudomonas功能基因的表达水平.结果表明:①1~90 d,进水浓度为2 mg/L的PCP严重抑制污泥活性,出水ρ（COD_Cr）超过100 mg/L,PCP降解去除缓慢,而对照组出水ρ（COD_Cr）在31.6 mg/L左右波动.90 d后,降解PCP的优势菌群富集,水相、泥相PCP被降解去除,但PCP的毒性作用使其出水ρ（COD_Cr）仍高于对照组.②经PCP长期驯化污泥富集的优势菌属为Chondromyces、Ignavibacterium、Thauera、Pseudoxanthomonas、Bosea、Achromobacter、Comamonas和Salimesophilobacter,均归属于变形菌门和厚壁菌门,在降解氯酚类污染物方面起着重要作用.③当处理PCP的SBR处于稳定运行阶段时,SBR运行周期末污泥中降解PCP的菌属Pseudomonas调控ATP水解酶（ATPase）、过氧化氢酶（CAT）和细胞色素p450（CYP450）的功能基因表达被激活,而调控超氧化物歧化酶（SODb）、氨单加氧酶（AM）、脱氯水解酶（HDLH）和甘油三磷酸脱氢酶（GAPDH）的功能基因表达被抑制,即活性污泥中微生物调控不同基因的表达受进水PCP的抑制或激活.研究显示:进水PCP诱导SBR富集的优势菌属可实现废水中PCP的去除,起到削减废水毒性的作用;通过分析不同优势菌属的相关功能基因表达,可加强对不同微生物降解污染物机理的认识.      

岩溶坡地土壤空间异质性的表述与调查方法——以贵州清镇市王家寨坡地为例

讨论了岩溶坡地土壤空间异质性的表述与调查方法,选取了地表石质土地面积比例及空间分布和土壤厚度及空间分布两个方面作为土壤空间异质性的表述内容,引入了地块破碎度、离散度、变异函数等指标;以王家寨为例,研究发现岩溶坡地土石比、破碎度、离散度、土壤厚度等指标从坡顶到坡下逐渐增大;石质地块破碎度在微观地块上表现为值越大地块越完整,是否与研究尺度有关还需要进一步论证;土壤厚度变异系数基本在1~1.5之间,与土壤厚度变程和土壤平均厚度有关;最后用土壤侵蚀的原理解释了岩溶坡地土壤空间异质性。     

抗柑桔溃疡病菌可溶性单链抗体的表达及鉴定

前期采用核糖体展示技术筛选了抗柑桔溃疡病菌(Xanthomonas axonopodis pv.citri,Xac)高亲和力单链抗体(Single chain variable fragment,scFv)GX13、GX44和GX95,为高效表达具有生物活性的单链抗体,本研究将前期筛选的高亲和力单链抗体基因转入大肠杆菌HB2151中进行可溶性表达,点印迹和Western印迹检测可溶性单链抗体的表达水平,亲和层析纯化表达的单链抗体,酶联免疫吸附法(ELISA)检测纯化单链抗体的抗原结合活性.结果显示,3株抗柑桔溃疡病菌可溶性单链抗体在大肠杆菌HB2151中均获得成功表达,表达产物分子量(Mr)约为32.0×103,主要集中于细菌周质腔中.ELISA检测纯化的单链抗体都具备抗原结合活性,可进一步应用于柑桔溃疡病菌的免疫诊断和病害防治.图4参14     

Mechanisms of photosynthetic inactivation on growth suppression of Microcystis aeruginosa under UV-C stress

This study aims to investigate the effects of UV-C irradiation on photosynthetic processes of Microcystis aeruginosa to unravel the mechanism(s) involved in how and in what ways UV-C mediates growth suppression and cellular recovery. Changes in the concentration of photosynthetic pigments, photochemical efficiency, PS II core protein (D1) content, and the coding genes expressions were measured. The results indicate that UV-C doses at 20–200 mJ cm⁻² lead to rapid reduction in gene expression of both psbA (for D1) and cpc (for phycocyanin), but the suppression was short term and recoverable within 3 d of post-UV incubation. Conversely, UV-C doses at ?50 mJ cm⁻² could induce marked decline in photochemical efficiency (represented by the optimal PS II quantum yield, F_V/F_M, and the effective PS II quantum yield, Y) as well as decreases in D1 content and water soluble pigments (phycoerythrins, phycocyanins, allophycocyanins) in M. aeruginosa during the post UV-C incubation period. The results suggest that interruption of both the light energy harvesting apparatus (especially the water soluble pigments) and the photochemical process mainly accounted for the growth suppression effect in UV-C irradiated M. aeruginosa.     

Estrogenic Compounds Downstream From Three Small Cities in Eastern Nebraska: Occurrence and Biological Effect1

Abstract: Recent studies have detected estrogenic compounds in surface waters in North America and Europe. Furthermore, the presence of estrogenic compounds in surface waters has been attributed, in some cases, to the discharge of wastewater treatment plant (WWTP) effluent. The primary objective of the current study was to determine if WWTP effluent contributes estrogens to the surface waters of Nebraska. A second objective of this study was to determine if estrogens were found in concentrations sufficient enough to manifest feminizing effects on fish. These objectives were satisfied by deploying polar organic chemical integrative samplers (POCIS) and caged fathead minnows at eight field sites. Deployment sites included: three reference sites (Pawnee Creek, the Little Blue River, and the Middle Loup River), two sites upstream of the WWTPs at Grand Island and Columbus, and three sites downstream of the WWTPs at Grand Island, Columbus, and Hastings. Following the seven day deployments, POCIS extracts were analyzed for estrone, 17β‐estradiol, estriol and 17α‐ethinylestradiol using liquid chromatography tandem mass spectrometry (LC/MS/MS). 17β‐estradiol was detected in POCIS from six of the eight field sites with the greatest quantities recovered in POCIS deployed downstream from the Grand Island and Hastings WWTPs. Estrone was detected only in the POCIS deployed downstream from the Grand Island and Hastings WWTPs. Estrogenic effects were detected in caged minnows analyzed for the hepatic mRNA expression of two estrogen‐responsive genes, vitellogenin (vg1) and estrogen receptor α (ERα). Fish deployed at the site where the greatest quantities of estrogens were recovered (Hastings) had significantly higher expression of both vg1 and ERα than fish deployed at any of the other sites. These results confirm that WWTP effluent contributes biologically significant levels of estrogens to Nebraska surface waters.     

植物滞尘分析及其数学表达模式

在植被的树冠结构分析、植物滞尘机理和粉尘沉降速度半经验公式的基础上,建立了植物滞留大气颗粒物的数学表达式。利用Beta功能函数φθ(X)、叶面的投影面积、叶面积密度a(z)、叶投影面积分布参数Kx、Kz等描述不同植被冠层的结构特征,并分析了冠层内的空气动力变化和树冠内气溶胶的平衡公式。借鉴Slinn的颗粒物沉降半经验公式、Petroffa等的阔叶滞尘计算模式,植物滞尘沉降通量数学表达式以树冠中树叶的位置为变量、由颗粒物在单片树叶上的各物理机制的沉降速度积分而成。结合国内外植物上沉降速度的试验结果和植物滞尘沉降通量公式的分析结果对影响植物滞尘的主要因素进行了比较分析。     

Protective effect of Spirulina platensis against aluminium-induced nephrotoxicity and DNA damage in rats

The protective effect of Spirulina platensis (SP) powder against aluminium-induced nephrotoxicity and DNA damage in rats was studied. Male rats receiving daily 40 mg/kg b.wt. aluminium chloride (AlCl₃) orally had increased serum levels of urea and creatinine, up regulated kidney injury molecule-1 and tissue inhibitor of metalloproteinase-1 genes, down regulated catalase and glutathione peroxidase genes, and increased all parameters of kidney DNA damage using comet assay. Treatment with SP alleviated all AlCl₃-induced effects of toxicity, especially when the animals were pre-treated.