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。     

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.     

角蛋白酶产生菌的分离鉴定及其kerC的克隆表达

以四川农业大学养鸡场堆砌废弃羽毛处土壤为样品,筛选出一株具有较强降解羽毛能力的细菌B-3菌株.经形态学、生理生化特性和16S rRNA基因序列分析,鉴定其为枯草芽孢杆菌(Bacillus subtilis),命名为枯草芽孢杆菌B-3.并成功克隆到该菌株的角蛋白酶基因kerC(GenBank No.:JN021789),在大肠杆菌BL21(Escherichia coli BL21)中获得了高效表达.该基因全长1146bp,GC含量46.5%,编码381个氨基酸,与已报道的枯草芽孢杆菌YYW-1的kerC基因(GenBank No.: EU362730)同源性达到100%.重组菌株经IPTG诱导后角蛋白酶酶活力达14.8U/mL,经His-Tag纯化和SDS-PAGE分析表明,重组角蛋白酶分子量约为60kDa(融合了硫氧还蛋白,Trx).重组角蛋白酶最适反应温度和pH值分别为65℃与7.0.     

蚯蚓体内过氧化物还原蛋白PRDX基因对土壤PAHs污染胁迫的转录响应

在前期建立的苯并[a]芘(benzo[a]pyrene,BaP)诱导上调表达的蚯蚓cDNA文库中,发现一种过氧化物还原蛋白peroxiredoxin6(PRDX6)基因片段,并提交到GenBank.Blastx比对与基因进化分析结果表明,该序列与已知PRDX6的最高相似率为86%,期望值为4E-30.序列分析表明,该序列具有PRDX6基因含Cys编码的一个特征motif,表明该序列属于PRDX6基因.为进一步验证蚯蚓PRDX6基因对多环芳烃污染的响应情况,使用OECD推荐方法,进行了土壤菲、芘、荧蒽和苯并芘污染对蚯蚓的毒性响应的试验(周期28 d),并采用定量PCR方法,检测了各实验组蚯蚓PRDX6基因的表达差异.结果发现,1.0 mg·kg-1的芘和苯并[a]芘均可显著上调蚯蚓PRDX6基因的表达,表明PRDX6基因可作为土壤污染引起蚯蚓抗氧化应激检测的分子标记.     

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.      

Gene expression responses of paper birch (Betula papyrifera) to elevated CO2 and O3 during leaf maturation and senescence

Gene expression responses of paper birch (Betula papyrifera) leaves to elevated concentrations of CO₂ and O₃ were studied with microarray analyses from three time points during the summer of 2004 at Aspen FACE. Microarray data were analyzed with clustering techniques, self-organizing maps, K-means clustering and Sammon's mappings, to detect similar gene expression patterns within sampling times and treatments. Most of the alterations in gene expression were caused by O₃, alone or in combination with CO₂. O₃ induced defensive reactions to oxidative stress and earlier leaf senescence, seen as decreased expression of photosynthesis- and carbon fixation-related genes, and increased expression of senescence-associated genes. The effects of elevated CO₂ reflected surplus of carbon that was directed to synthesis of secondary compounds. The combined CO₂ + O₃ treatment resulted in differential gene expression than with individual gas treatments or in changes similar to O₃ treatment, indicating that CO₂ cannot totally alleviate the harmful effects of O₃.     

牛蛙生长激素基因在COS7细胞中的转染表达

应用RT-PCR技术克隆牛蛙生长激素(BfGH)的cDNA,T-A克隆法构建反向插入的pMD18-T/BfGH重组质粒,回收BamHⅠ酶切的BfGH cDNA片段,并与去磷酸化处理的真核表达载体VR1020/BamHⅠ,酶切鉴定方法筛选BfGH正向插入的重组真核表达质粒VBfGH.脂质体法介导质粒VBfGH转染哺乳动物细胞COS7,对转染后的COS7细胞进行RT-PCR、ELISA和免疫荧光检测,分别在转录和翻译水平证实BfGH基因在COS7细胞中得到了正确的转染表达.图4表1参17     

豆豉纤溶酶在枯草杆菌WB800中的高水平表达

一个来源于豆豉产生菌DC12的新的纤溶酶基因序列被克隆测序.为了实现在枯草杆菌中高水平表达豆豉纤溶酶,将来源于枯草杆菌168的重叠启动子P43序列以及转录终止信号序列通过重叠延伸融合,将融合序列克隆到大肠杆菌-枯草杆菌穿梭载体pBE3中,构建了pBEP43载体.将豆豉纤溶酶基因插入到载体pBEP43后转化枯草杆菌WB800.结果表明,在P43启动子驱动下,豆豉纤溶酶基因在重组菌中的对数生长期和平衡期均获得了表达且分泌到培养基中.重组菌经培养后上清液中的纤溶酶活性最高达1270U/mL,是豆豉纤溶酶基因在自身启动子驱动下表达量的1.87倍和野生菌株枯草杆菌DC12表达量的4.1倍.图5参16