全基因组重测序研究长期铬胁迫对希瓦氏菌MR-1的影响机制

在实验室纯培养条件下,设计了希瓦氏菌MR-1在恒定浓度Cr（VI）和Cr（Ⅲ）环境中长期胁迫120d实验,探讨了胁迫后的菌株对Cr（VI）的还原能力和生长情况,采用电化学分析仪表征其电化学性质的变化,并且利用全基因组重测序技术分析了不同条件胁迫后不同阶段的菌株基因变异信息.结果表明,Cr（VI）长期胁迫下明显提高了菌株对Cr（VI）的耐受性和还原能力,在38h左右能完全还原初始Cr（VI）浓度55mg/L;在电位-0.2V还原峰附近,细胞色素c与核黄素的结合作用有明显差异;通过重测序结果发现,Cr（VI）长期胁迫下的菌株发生突变的基因点位明显高于Cr（Ⅲ）环境.Cr（Ⅲ）环境胁迫下的非同义突变基因主要集中在葡萄糖转化及合成相关功能的基因、编码呼吸链酶和合成ATP酶的相关基因、编码信号转导和跨膜转运蛋白相关基因,Cr（VI）环境胁迫120d后,主要涉及细胞膜组分基因,转运蛋白、信号转导相关基因,DNA合成、修复相关基因和氧化还原活性相关.     

榨菜线粒体DNA雄性不育相关片段克隆及序列分析

研究了榨菜(Brassica juncea var．tumida Tsen and Lee)细胞质雄性不育性(CMS)与线粒体基因组的关系,以榨菜细胞质雄性不育系及其保持系线粒体DNA(mtDNA)为模板,设计合成引物进行PCR扩增,从榨菜胞质雄性不育系mtDNA上扩增得到与雄性不育相关的特异片段T1170,点杂交分析表明,T1170仅与榨菜胞质雄性不育系mtDNA有杂交信号,而保持系的mtDNA则无,序列测定该片段全长1173bp,编码389个氨基酸,其中疏水氨基酸占35％。图4参9。     

Genome size of termites (Insecta, Dictyoptera, Isoptera) and wood roaches (Insecta, Dictyoptera, Cryptocercidae)

The evolution of genome size has been discussed in relation to the evolution of various biological traits. In the present study, the genome sizes of 22 dictyopteran species were estimated by Feulgen image analysis densitometry and 6-diamidino-2-phenylindole (DAPI)-based flow cytometry. The haploid genome sizes (C-values) of termites (Isoptera) ranged from 0.58 to 1.90 pg, and those of Cryptocercus wood roaches (Cryptocercidae) were 1.16 to 1.32 pg. Compared to known values of other cockroaches (Blattaria) and mantids (Mantodea), these values are low. A relatively small genome size appears to be a (syn)apomorphy of Isoptera + Cryptocercus, together with their sociality. In some phylogenetic groups, genome size evolution is thought to be influenced by selective pressure on a particular trait, such as cell size or rate of development. The present results raise the possibility that genome size is influenced by selective pressures on traits associated with the evolution of sociality.     

宏基因组学分析深度处理阶段污水中细菌的赋存特征及其功能

为探究深度处理阶段污水中细菌的赋存特征及其功能,采集了污水深度处理阶段沿程各单元的进出水样品,并基于宏基因组学对污水中细菌的群落结构及功能进行了解析.结果表明,不同深度处理单元出水中细菌的多样性存在差异,臭氧接触池出水中细菌的多样性最低;相比夏季,冬季深度处理阶段污水中细菌的丰富度和多样性较低.不同季节深度处理阶段污水中的细菌群落结构变化较大,反硝化滤池出水中的细菌群落结构与其它样品存在较大差异;变形菌门（41.5%～71.0%）是深度处理阶段污水中的主要优势菌门,其次是拟杆菌门（3.8%～16.2%）;反硝化滤池出水中主要菌属有脱氯单胞菌（4.1%～7.4%）、弓形杆菌（3.0%～8.3%）和不动杆菌（2.3%～3.0%）.在深度处理阶段各工艺出水中共发现了29种与氮代谢有关的功能基因,并且在各工艺出水中均检测到了与反硝化有关的功能基因,如nosZ、napA、nirK和norB等,表明深度处理阶段污水中的细菌具有持续脱氮的潜力.糖苷转移酶和糖苷水解酶是深度处理阶段主要的碳水化合物活性酶,深度处理阶段污水中的细菌表现出了对多种有机物的降解潜力.     

Modeling genetic benefits and financial costs of integrating biobanking into the conservation breeding of managed marsupials

Managed breeding programs are an important tool in marsupial conservation efforts but may be costly and have adverse genetic effects in unavoidably small captive colonies. Biobanking and assisted reproductive technologies (ARTs) could help overcome these challenges, but further demonstration of their potential is required to improve uptake. We used genetic and economic models to examine whether supplementing hypothetical captive populations of dibblers (Parantechinus apicalis) and numbats (Myrmecobius fasciatus) with biobanked founder sperm through ARTs could reduce inbreeding, lower required colony sizes, and reduce program costs. We also asked practitioners of the black-footed ferret (Mustela nigripes) captive recovery program to complete a questionnaire to examine the resources and model species research pathways required to develop an optimized biobanking protocol in the black-footed ferret. We used data from this questionnaire to devise similar costed research pathways for Australian marsupials. With biobanking and assisted reproduction, inbreeding was reduced on average by between 80% and 98%, colony sizes were on average 99% smaller, and program costs were 69- to 83-fold lower. Integrating biobanking made long-standing captive genetic retention targets possible in marsupials (90% source population heterozygosity for a minimum of 100 years) within realistic cost frameworks. Lessons from the use of biobanking technology that contributed to the recovery of the black-footed ferret include the importance of adequate research funding (US$4.2 million), extensive partnerships that provide access to facilities and equipment, colony animals, appropriate research model species, and professional and technical staff required to address knowledge gaps to deliver an optimized biobanking protocol. Applied research investment of A$133 million across marsupial research pathways could deliver biobanking protocols for 15 of Australia's most at-risk marsupial species and 7 model species. The technical expertise and ex situ facilities exist to emulate the success of the black-footed ferret recovery program in threatened marsupials using these research pathways. All that is needed now for significant and cost-effective conservation gains is greater investment by policy makers in marsupial ARTs.     

Vitamins E and C reduce oxidative stress and mitochondrial permeability transition caused by camptothecin – an in vitro study

Camptothecin (CPT), a broad spectrum antineoplastic agent, is known to induce oxidative stress and mitochondria are among the main sources of intracellular reactive oxygen species (ROS). We investigated the merit of vitamins E and C supplementation on CPT-induced mitochondrial alterations in vitro. Following treatment of isolated liver mitochondria with CPT, we assessed the mitochondrial membrane permeability transition (MPT), concentration of malondialdehyde, antioxidants and activities of the enzymes of the respiratory chain and Krebs cycle. Our results provide evidence that CPT caused mitochondrial swelling, increased lipid peroxidation and transition of mitochondrial permeability. The CPT lowered the levels of reduced mitochondrial thiols suggesting that thiol oxidation is the mechanism underlying CPT-induced MPT. Identical experiments were also performed after preincubating the mitochondria with vitamins E and C. It was found that vitamins E and C pretreatment inhibited the deleterious effects of CPT and loss of enzyme activity was restored by antioxidant supplementation. Our results suggest that the toxicity of CPT was mediated by an increase in ROS production by mitochondria. However, the addition of vitamins E or C ameliorated the oxidative stress. We propose that an attempt to counteract the deleterious consequences of chemotherapy with nutritional therapies may be a rational approach in superior patient care especially in a disease like cancer.     

Toxicity of cigarette smoke on isolated lung,heart, and brain mitochondria: induction of oxidative stress and cytochrome c release

Cigarette smoking is one of the main risk factors for premature human death which is associated with a variety of respiratory and vascular diseases, and cancer due to exposure to hundreds of toxicants. Rat mitochondria were obtained by differential ultracentrifugation and incubated with different concentrations (1%, 10%, or 100%) of standardized cigarette smoke extract (CSE). Our results showed that CSE induced a rise in mitochondrial reactive oxygen species (ROS) formation, lipid peroxidation, and mitochondrial membrane potential (MMP) collapse before mitochondrial swelling ensued in isolated pulmonary mitochondria. Disturbance in oxidative phosphorylation was also confirmed by decrease in ATP concentration in the CSE-treated mitochondria. In addition, collapse of MMP and mitochondrial swelling produced release of cytochrome c via outer membrane rupture or mitochondrial permeability transition (MPT) pore opening. Our results suggested that CSE-induced toxicity in lung tissue is the result of disruptive effect on mitochondrial respiratory chain that leads to ROS formation, lipid peroxidation, MMP decline, and cytochrome c expulsion which results in apoptosis signaling and cell loss.     

玉米多胞质系线粒体数量和结构特征的比较分析

以玉米多胞质系为材料,采用透射电镜的观察方法,比较观察了１１种材料中黄化幼芽细胞的线粒体结构特征的变化．观察表明,１１种材料黄化幼芽线粒体形态及外膜、内膜、泡状嵴和基质颗粒等没有什么显著的差异,而线粒体大小和每个细胞内的线粒体数量却是明显不同的．Ｆ检验说明,线粒体大小和每个细胞线粒体数量上有极显著的差异,也即是说,１１种不同细胞质类型的材料细胞质作用和核质（基因）互作是明显的．试验还观察到线粒体、分裂现象和线粒体、淀粉粒聚集在胞间连丝附近的有趣现象．     

林丹通过MAPK和UPR通路与干扰线粒体功能诱导黑腹果蝇(Drosophila melanogaster)肿瘤细胞迁移

环境污染物增加癌症死亡率的潜在机理研究尚需开展。选择与癌症致死显著相关的林丹作为目标物质,将黑腹果蝇作为受试生物进行暴露,对1 000μg·L~(-1)和对照组的果蝇样本进行实时聚合酶链式反应(RT-PCR),检测发现参与调控癌症细胞迁移的丝裂原活化蛋白激酶(MAPK)信号通路中的MEKK和p38α、未折叠蛋白反应(UPR)中的ATF4、Ire1、PERK和XBP1等基因的表达量呈现显著下调(P0.05)。通过肿瘤迁移品系果蝇统计林丹暴露下肿瘤细胞迁移率,结果显示肿瘤细胞迁移率随林丹浓度升高而增加,1 000μg·L~(-1)林丹暴露组与对照组的果蝇肿瘤细胞迁移率分别为69.2%和45.9%,具有显著性差异(P0.05)。进一步的RNA测序结果表明,高浓度暴露组与对照组之间存在380个基因具有显著性差异(P0.05),其中含169个上调基因和211个下调基因。差异基因主要涉及水解酶催化活性(42个基因)、神经系统(20)、对化学刺激的感知(6)和线粒体(5)。差异基因的富集分析表明林丹的暴露主要改变蛋白水解过程(83个基因)、胞外区域(84)和催化活性(311),其对应的平均富集因子分别为1.59、1.52和1.24(P0.001)。差异基因的表达解释了林丹的暴露不仅致使线粒体功能障碍,而且影响蛋白水解酶活性,加速胞外基质降解,为肿瘤细胞迁移供能并提供微环境。     

离子液体氯化1-辛基-3-甲基咪唑对EMT6细胞的毒性及其机理研究

为评估咪唑类离子液体的生物毒性,研究了氯化1-辛基-3-甲基咪唑([C8mim] [C1])对EMT6细胞的毒性作用和可能的机制.不同浓度(0.06、0.25、1 mmol·L-)的[C8 mim][Cl]对EMT6细胞染毒12h后,采用MTT方法检测细胞活力,二乙酸荧光素(FDA)方法检测细胞膜通透性的变化,Rhodamine 123染色方法检测线粒体膜电位的变化,ELISA方法检测了Caspase-3的活性,并测定了细胞内活性氧(ROS)的含量.结果表明,经[Csmim] [Cl]染毒12h后,EMT6细胞活力下降,并呈剂量依赖关系.当[Csmim] [Cl]浓度高于0.25 mmol·L-1时,细胞活力与对照相比,差异显著.研究还发现,[Csmim][Cl]染毒增加了EMT6细胞膜通透性,降低了线粒体膜电位并诱导产生过量的活性氧,增强了Caspase-3活性.实验结果表明,[C8mim] [Cl]染毒造成了EMT6细胞膜通透性的改变、活性氧的过量产生和凋亡分子表达的增强,这可能是[Csmim] [Cl]导致细胞凋亡和活力下降的主要原因.