广西岩溶区烟草黑胫病拮抗细菌的筛选鉴定及其抗病机理

从广西岩溶区靖西县优质烟叶生产区分离烟草黑胫病病原及土壤细菌,通过拮抗试验、离体及田间抗病能力测试等方法筛选拮抗细菌;利用形态观察、生理生化测试和16SrDNA序列分析三种方法相结合,对抗病效果良好的菌株进行分类鉴定;并对拮抗菌抗黑胫病机理进行初步研究。结果表明:从8份土壤样品中,共分离出土壤细菌340株,获得抗黑胫病效果良好的拮抗细菌3株,编号为8-23、6-70和13,它们分别属芽孢杆菌属、溶菌杆属和假单胞杆菌属细菌;三个拮抗细菌的抗黑胫病机理是通过胞外分泌一些可溶解病原菌细胞壁的酶或其它化学物质,破坏菌丝的细胞壁和细胞膜等生理结构,使细胞质渗漏、凝集,从而导致病原死亡;其中,菌株8-23和13的抗病活性物质主要为蛋白质类化合物;而菌株6-70除蛋白质外,还有其它一些非蛋白因子起作用。     

中国大陆地区曲霉病流行现状分析

目的了解中国大陆地区曲霉病流行状况及诊治现状。方法通过CNKI及Pubmed搜索1991年来中国大陆地区曲霉病及曲霉相关文献,进行数据统计和总结。结果 1991年1月～2009年10月报道曲霉病1457例,其中481例有病原学证据,258例进行菌种鉴定,其中烟曲霉153例。感染部位最常见于肺,为1047例。病原学诊断依据主要是镜检和培养;菌种鉴定主要依赖形态学特征,仅4例进行分子生物学鉴定;仅少数进行半乳甘露聚糖及葡聚糖检测;体外药敏试验少,仅报道2株黄曲霉对两性霉素B耐药,2株烟曲霉多药耐药。伊曲康唑为侵袭性曲霉感染经验治疗(抢先治疗)的首选药物。过敏性支气管肺曲霉病则多单用糖皮质激素或联合伊曲康唑,大部分慢性及腐生型曲霉病及鼻窦曲霉病患者行手术治疗。结论中国大陆地区曲霉病流行趋势与国际一致,非烟曲霉感染有所增加,感染部位仍以肺为主。应提高菌种形态学和分子生物学鉴定水平,积极开展非培养检测手段,保证正确和及时地诊断;加强药敏监测,为临床治疗提供有效信息。     

口腔黏膜真菌鉴定方法的比较

比较常见用于黏膜真菌菌种鉴别的多种方法,探寻最佳的鉴别方法。采集230例普通人群口腔黏膜样本,分别用玉米吐温-80培养观察厚膜孢子法、糖发酵生化反应法、CHROMagar假丝酵母菌显色培养基法、ITS基因的PCR-RFLP(聚合酶链反应-限制性片段长度多态性)法、ITS测序菌种鉴定法,鉴别真菌各菌株。结果显示:有56例菌株至少通过1种方法检出真菌;玉米吐温-80分离培养假丝酵母菌37株;50例菌株ITS基因测序共鉴定出8个菌种,白假丝酵母菌(C.albicans)29株,近平滑假丝酵母菌(C.parapsilosis)10株,热带假丝酵母菌(C.tropicalis)5株,Candida metapsilosis 1株,Lodderomyces elongisporus 1株,克柔假丝酵母菌(Candida krusei)1株,乙醇假丝酵母菌(C.ethanolica)1株,季也蒙毕赤酵母菌(Pichia guilliermondii)2株;CHROMagar假丝酵母菌显色培养基法鉴定出3种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌;PCR-RFLP法检出5种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌、季也蒙毕赤酵母菌、克柔假丝酵母菌,与基因的测序鉴定一致率为91%;糖发酵生化反应法阳性标本占被检出真菌例数的46.4%(26/56)。结果表明:ITS基因的测序法可以准确鉴定真菌各个菌种;PCR-RFLP法能鉴定常见的菌种,但操作繁琐;CHROMagar假丝酵母菌显色培养基法能快速准确鉴别3种常见假丝酵母菌菌种;玉米吐温-80可以准确培养鉴别白假丝酵母菌;糖发酵生化反应法,缺乏足够的敏感度和特异性,难以准确鉴别各个菌种。     

Pig identification and meat traceability by multiallelic amplification fragments with multiple single nucleotide polymorphisms

Compared with conventional identification methods, DNA-based genetic approaches such as single nucleotide polymorphisms (SNPs) and satellites are much more reliable for pig identification and meat traceability. In this study, multiallelic amplification fragments with multiple SNPs, incorporating the advantages of both SNPs and microsatellites, were explored for the first time for pig identification and meat traceability. Primer pairs for multiallelic fragments and their optimal SNPs were successfully selected and used for identification of individuals from Suzhong and Duroc populations. Meanwhile, the combined panel of the above mentioned primer pairs together with their optimal SNPs for Suzhong and/or Duroc pigs were validated for identification of the hybrids (Suzhong×Duroc). Therefore, we have successfully selected multiallelic amplification fragments with multiple SNPs to identify pigs and their meat samples from Suzhong, Duroc or their hybrids. Our study demonstrates that our method is more powerful for pig identification or meat traceability than SNPs or microsatellites.     

核糖体转录间隔子2应用于鱼类种属的鉴别

为了防止珍稀鱼类的非法捕捞和销售, 鱼类种属的鉴别就成为非常关键的问题, 特别是形态学方法无法区分的样品(如鱼苗、鱼鳞、鱼卵、鱼肉及其加工产品等)。为了帮助珍稀鱼类资源的管理和保护, 文章报道了一种利用核糖体基因的转录间隔子2鉴别鱼类种属的分子遗传学方法: (1) 利用同一目鱼类5.8S rRNA和28S rRNA基因的保守性, 设计出扩增鲤形目鱼类这两个基因间转录间隔子2 DNA片段, 测序获得它们的碱基排列顺序; (2) 再根据不同鱼类转录间隔子2序列的差异, 设计出每种鱼的种属特异引物、种属鉴别标准物, 构建鱼类分子分类图谱, 利用PCR复合扩增技术鉴别鱼类种属。通过对国内不同地方采集的5种鲤形目鱼类的210个单一品种样本和40个混合样本的鉴别检验, 该方法能够准确、灵敏和快速鉴别这5种鱼, 可用于鱼类资源保护和评估、管理和开发, 特别是在渔业管理人员渔业执法、海关打击珍稀鱼类走私、防止商业欺诈和外来有害生物入侵等方面非常有用     

基于赤水河流域生态补偿的政府和社会资本合作项目风险识别与分担

赤水河流域为生态脆弱区域,现行的流域生态补偿机制存在补偿资金来源单一、总量不足且持续性较差、补偿方式较为单一等问题。将政府和社会资本合作（Public-Private Partnership,PPP）模式应用于建立赤水河流域生态补偿机制,有助于拓宽补偿资金来源、增加资金总量、丰富补偿方式,推动各利益相关方收益共享、风险共担。与传统PPP项目相比,基于流域生态补偿的PPP项目具有更为复杂的风险结构,风险因素的正确识别和合理分担是成功运用PPP模式完善赤水河流域生态补偿机制的关键。识别基于赤水河流域生态补偿的PPP项目运作关键环节,甄别各环节面临的主要风险因素;基于云理论建立风险分担模型;将有关风险在政府和社会资本间进行分担。研究结果表明：（1）基于赤水河流域生态补偿的PPP项目运作过程共包括项目准备、项目实施和项目合同终结等三个阶段、共11个关键环节,各环节共面临26个主要风险因素。（2）分析了有关风险因素可能对赤水河流域生态补偿机制或PPP项目产生的不利影响,并指出了风险引致方。（3）在项目准备阶段,政府拥有绝对的资源优势,以政府为主承担主要风险;在项目实施阶段,项目风险总体上由政府承担为主向社会资本承担为主转移,80%的风险主要由社会资本承担;特许经营期满后,社会资本将项目的经营权（或所有权与经营权同时）向政府移交,在项目合同终结阶段,有关风险再次以政府承担为主。     

DNA probes to identify members of the Anopheles farauti complex

DNA probes have been constructed to distinguish between the members of the Anopheles farauti complex of mosquitoes known as species numbers 1, 2 and 3. Partial genomic libraries of the three known species were exposed to labelled total genomic DNA from each species. Colonies showing differential hybridization were selected for further testing. These probes were found which allow identification of the three known species: probe pAf1 (160 bp fragment) hybridizes to DNA from An. farauti nos. 1 and 2; probe pAf2 (95 bp fragment) hybridizes to DNA from An. farauti no. 2 only; and probe pAf3 (1.3 kb fragment) hybridizes strongly to DNA from An. farauti no. 3, less to no. 1 and faintly to no. 2. Increasing the stringency of hybridization reduced the cross-hybridization of probes pAf1 and pAf3. Only radioactively labelled probes were tested. Males and females and individuals from diverse habitats and localities showed the same species/probe hybridization characteristics. This technique allows faster identification of the sibling species than previous methods, and has the added advantage that it allows air-dried and alcohol stored specimens to be identified.     

Systematics of Mukdenia and Oresitrophe (Saxifragaceae): Insights from genome skimming data

Oresitrophe and Mukdenia (Saxifragaceae) are epilithic sister genera used in traditional Chinese medicine. The taxonomy of Mukdenia, especially of M. acanthifolia, has been controversial. To address this, we produced plastid and mitochondrial data using genome skimming for Mukdenia acanthifolia and Mukdenia rossii, including three individuals of each species. We assembled complete plastomes, mitochondrial CDS and nuclear ribosomal ETS/ITS sequences using these data. Comparative analysis shows that the plastomes of Mukdenia and Oresitrophe are relatively conservative in terms of genome size, structure, gene content, RNA editing sites and codon usage. Five plastid regions that represent hotspots of change (trnH-psbA, psbC-trnS, trnM-atpE, petA-psbJ and ccsA-ndhD) are identified within Mukdenia, and six regions (trnH-psbA, petN-psbM, trnM-atpE, rps16-trnQ, ycf1 and ndhF) contain a higher number of species-specific parsimony-informative sites that may serve as potential DNA barcodes for species identification. To infer phylogenetic relationships between Mukdenia and Oresitrophe, we combined our data with published data based on three different datasets. The monophyly of each species (Oresitrophe rupifraga, M. acanthifolia and M. rossii) and the inferred topology ((M. rossii, M. acanthifolia), O. rupifraga) are well supported in trees reconstructed using the complete plastome sequences, but M. acanthifolia and M. rossii did not form a separate clade in the trees based on ETS + ITS data, while the mitochondrial CDS trees are not well-resolved. We found low recovery of genes in the Angiosperms353 target enrichment panel from our unenriched genome skimming data. Hybridization or incomplete lineage sorting may be the cause of discordance between trees reconstructed from organellar and nuclear data. Considering its morphological distinctiveness and our molecular phylogenetic results, we strongly recommend that M. acanthifolia be treated as a distinct species.     

New species and records of Stigmaeidae (Acari: Prostigmata) from New Zealand - ID. Genus Stigmaeus Koch

Abstract

Stigmaeus arboricola, S. montanus, S. luxtoni, and S. novazealandicus are described as new species, and the larva of S. loadmani is described. Several new records are noted, and a key is given to the New Zealand species of Stigmaeus.