Temporal distribution and pathogenicity of the predominant tomato‐infecting begomoviruses in Taiwan

Between 1998 and 2009, the four tomato‐infecting begomovirus species detected in Taiwan were Ageratum yellow vein Hualien virus (AYVHuV), Tomato leaf curl Taiwan virus (ToLCTWV), Tomato yellow leaf curl Thailand virus (TYLCTHV) and a newly defined species Tomato leaf curl Hsinchu virus (ToLCHsV). AYVHuV was detected occasionally in 2003 and ToLCHsV only in 2000–2001, whilst ToLCTWV was detected throughout the period. TYLCTHV was first detected in 2005. Between 1998 and 2005, >99% of the begomovirus‐positive samples were infected with ToLCTWV. In 2007 in western Taiwan, 16% of the positive samples were infected with ToLCTWV, 35% with TYLCTHV and 49% with mixed infection (ToLCTWV/TYLCTHV). In contrast, in eastern Taiwan the proportions were 84% ToLCTWV, 2% TYLCTHV and 14% mixed infection. However, throughout Taiwan in 2008–2009, most positive samples were either identified as TYLCTHV (51%) or mixed infection (ToLCTWV/TYLCTHV; 41%), and only 8% were ToLCTWV. This shows a clear trend of shifting from ToLCTWV to TYLCTHV and mixed infection over a short time period in Taiwan. Sequence analyses indicated that tomato‐infecting AYVHuV, an apparent recombinant between ToLCTWV and AYVHuV from Ageratum, represents a new strain Hsinchu. TYLCTHV Taiwan isolates were highly similar to each other, whereas ToLCTWV isolates had greater diversity and were classified into three strains which had one country‐wide and two local distributions. ToLCTWV and TYLCTHV were confirmed as monopartite and bipartite begomoviruses, respectively, by agroinfection followed by transmission with Bemisia tabaci biotype B. In addition, TYLCTHV was found to be mechanically transmissible together with viral DNA‐B.     

湖北省棉花主产区烟粉虱生物型分布及系统发育分析

［目的］ 明确武汉、荆州、孝感、随州、武穴5个棉花主产区烟粉虱生物型组成。［方法］ 通过以线粒体细胞色素氧化酶I（mitochondrial cytochrome oxidase I, mtCOI）为基础的PCR RFLP和基因测序的方法,鉴定湖北5个棉花主产区烟粉虱地理种群的生物型组成,并分析5个地理种群与其他省种群的系统发育关系。［结果］ Q型烟粉虱为湖北省棉花主产区的优势种群,而武汉温室种群则均为B型烟粉虱,土著生物型ZHJ1型在大田中略有分布;系统发育分析表明,湖北省棉花主产区B型和Q型烟粉虱种群均与中国其他多个省的烟粉虱种群亲缘关系较近。［结论］ 本文研究结果可以为湖北省5个棉花主产区烟粉虱的有效防治提供参考。     

河南隐孢子虫分离株的系统发生

以线粒体功能性蛋白替代氧化酶(alternative oxidase,AOX)为研究对象,对隐孢子虫(Cryptosporidium)分离株进行扩增和测序,并对测得的序列进行种系发育关系研究,以确定不同隐孢子虫分离株之间的进化关系,并以18SrRNA和HSP70基因构建的进化树作参照,评价AOX基因是否适合作隐孢子虫基因分型和进化关系分析的基因座。结果表明,在AOX序列构建的进化树上Cryptosporidum分为两大类:Cryptosporidium baileyi和C.meleagridis处于一个进化枝上,C.hominis、C.suis、C.parvum牛基因型和C.parvum鼠基因型处于另一个进化枝上。在C.parvum内,鼠基因型分离株和猪基因I型(即C.suis)分离株与牛基因型和人基因型(即C.hominis)在AOX基因序列上都存在着差异。鹌鹑源C.meleagridis分离株与禽源C.baileyi3个分离株在AOX基因序列的同源性显著高于前者与C.parvum的同源性,在进化树上C.meleagridis与C.baileyi的亲缘关系也较其与C.parvum密切。AOX基因不仅可作为隐孢子虫分离株种系发育的遗传标记,又为发现隐孢子虫新的遗传特性提供了一种途径。     

7株解有机磷细菌的分离和鉴定

从土壤中分离筛选出7株解有机磷的微生物。对这7株解磷细菌进行了形态、生理生化性状测定及16SrDNA序列分析(GenBankaccessionNo:S2,AY651922;S3,AY661923;X1,AY651925;Y1,AY651924;H1,AY663435;H2,AY663436andHe,AY663436)。其中S2、S3、X1和He属于假单胞菌属(Pseudomonas),Y1属于芽孢杆菌属(Bacillus),H1属于不动杆菌属(Acinetobacter),H2属于寡养单胞菌属(Stenotrophomonas)。进一步通过G C含量和DNA-DNA杂交研究,结果表明,S2、S3和X1为产碱假单胞菌(Pseudomonasalcaligenes),Y1为蜡状芽孢杆菌(Bacilluscereus)。     

Molecular Monitoring of SRB Community Structure and Dynamics in Batch Experiments to Examine the Applicability of in situ Precipitation of Heavy Metals for Groundwater Remediation (15 pp)

Background, Aims and Scope   Sulfate-reducing bacteria (SRB) are known for their capacity to reduce and precipitate heavy metals (HM) as metal sulfides, offering the opportunity to create an in situ reactive zone for the treatment of heavy metal-contaminated groundwater, a process called in situ metal precipitation (ISMP). The applicability of the ISMP technology first has to be investigated at a laboratory scale before going into an on site application. The evaluation and optimization of the ISMP process is facilitated when physical/chemical analysis techniques are combined with molecular tools that specifically monitor the abundance, diversity and dynamics of the indigenous sulfate reducing microbial community. In this study, batch experiments were conducted in order to investigate the feasibility of ISMP as a groundwater remediation strategy for an industrial site contaminated with elevated levels of Zn, Cd, Co and Ni. Methods   The potential of different types of carbon source/ electron donor (lactate, acetate, methanol, ethanol, Hydrogen Release Compound?, molasses) to stimulate the sulfate reduction and metal precipitation activity of the naturally present (or indigenous) SRB community was explored. In addition, the effect of amending vitamin B12 and yeast extract was evaluated. The ISMP process was monitored by combining analytical analyzes of process parameters (SO42-concentration, heavy metal concentrations, pH, Eh) with molecular tools such as SRB subgroup and genus specific PCR, denaturing gradient gel electrophoresis (DGGE), and phylogenetic analysis of clone sequences, based on either the 16S rRNA or the dsr (dissimilatory sulfite reductase) gene. Results and Discussion   The efficiency of different carbon-sources to stimulate the ISMP process followed the order HRC 〉 molasses 〉 methanol 〉 lactate 〉 ethanol 〉 acetate. Within 10 weeks, the highest sulfate and metal removal efficiencies ranged from 85% to 99%. Addition of yeast extract boosted the ISMP process, whereas vitamin B12 negligibly affected SRB activity. Analysis of the sulfate reducing population by SRB subgroup and genus specific PCR demonstrated that members of the genus Desulfosporosinus dominated in all batch tests, while 16S rDNA DGGE profiles additionally revealed the presence in the microbial communities of non-sulfate reducing bacteria within the family Clostridium and the -proteobacteria. The dsrB-based DGGE profiles allowed us to assess the diversity and dynamics of the sulfate reducing community and added to a better understanding of the effects of different batch conditions on the ISMP process. Remarkably, all dsrB sequences affiliated with the dsrB gene sequence cluster found in Desulfotomaculum, which received their xenologous dsrB gene from the -proteobacteria. Conclusions   The batch experiments, which aimed at stimulating the activities of the indigenous SRB communities, demonstrated that these communities were present and that their activities could be used to obtain efficient in situ precipitation of the contaminating heavy metals. This opens the possibility to test this concept in the future as an on site demonstration as part of the groundwater strategy for the heavy metal contaminated site. Although batch setups are suitable for preliminary feasibility studies for ISMP, they do not reflect the in situ situation where sulfate and heavy metal and metalloid polluted groundwater are supplied continuously. A sulfate reducing strain JG32A was isolated from whose 16S rRNA gene affiliated with the genus Desulfosporosinus, while its dsrB gene sequence clustered with Desulfotomaculum dsrB gene sequences, which received their xenologous dsr genes from -proteobacteria. Therefore we hypothesize that the batch experiments enrich members of the Desulfosporosinus genus that possess a non-orthologous dsrB gene. Recommendation and Perspective   The next step towards an on site pilot test for ISMP will be the setup of a series of column experiments, with process conditions that are selected based on the above mentioned results. This will allow to define optimal ISMP process conditions and to test its long-term efficacy and sustainability before going into an on site bioremediation application. By applying the described molecular tools together with physical-chemical analyzes, it can be investigated whether the same SRB community is enriched and which type of C-source is most effective in promoting and sustaining its growth and sulfate-reduction activity.     

瓜类褪绿黄化病毒山东分离物全基因组序列扩增及分析

为明确分离自山东省寿光市甜瓜上的瓜类褪绿黄化病毒(cucurbit chlorotic yellows virus,CCYV)分离物的全基因组序列信息和遗传变异情况,利用毛形病毒属Crinivirus简并引物进行RTPCR检测,利用RACE技术结合RT-PCR方法克隆CCYV山东分离物2条RNA链的全基因组序列,通过与GenBank中其它地区CCYV分离物的全长序列进行比对分析其同源性,并基于CP基因序列构建系统进化树分析其遗传变异情况。结果表明,山东分离物经RT-PCR检测和测序后确定为CCYV。CCYV山东分离物与其它CCYV分离物的RNA1链和RNA2链的全基因组序列一致性的平均值分别为99.82%和99.88%,且2条链的5′末端均比较保守,没有碱基突变的情况发生;RNA1链3′末端存在2个碱基变异,RNA2链3′末端存在1个碱基变异。CCYV不同地区分离物主要分为3个簇群,其中山东分离物和中国其它地区分离物、日本分离物、苏丹分离物、黎巴嫩分离物和塞浦路斯分离物聚类在一起。研究表明CCYV基因组序列比较保守,该病毒的分化可能与地理来源存在一定的相关性。     

茶树叶斑病病原菌可可毛色二孢菌的鉴定

The tea leaf disease, which led to a large loss of production and decrease of quality, was found in the tea region in Huishui county, Guizhou province. Some strains were isolated from the diseased samples, and the representative strains fulfilled Koch’s postulate. Then the isolates were further identified as Lasiodiplodia theobromae based on morphological characters and phylogenetic analysis with internal transcribed spacer (ITS) and elongation factor 1-α (EF-1α) regions. The results showed that the pathogen of the leaf disease occurred in Huishui county, Guizhou province was L. theobromae.     

烧成温度对C—B4C—SiC复合材料结构与性能的影响

本文就烧成温度对Ｃ－Ｂ＿４Ｃ－ＳｉＣ碳／陶复合材料结构与性能的影响进行了初步研究，发现复合材料密度、强度随烧结温度的升高而明显提高，电阻率随烧成温度的升高而下降，这和烧成体显微结构（气孔率、晶粒大小与分布、晶界状况等）密切相关。     

Phoma–Didymella complex on hybrid arctic bramble with wilting symptoms

Pycnidia containing conidia characteristic of Phoma spp. and pseudothecia containing ascospores characteristic of Didymella applanata were isolated from edges of expanding stem lesions and dead stems of wilted cultivated hybrid arctic bramble plants ( Rubus arcticus nothossp. stellarcticus ) in Sweden in 1998 and 1999. The fungi were morphologically similar when grown on culture media, but some differences in growth rate were observed. They were also similar to the reference isolates of D. applanata (anamorph Phoma argillacea ). However, they were different from an isolate of Phoma sp. (HPP 38) isolated from cultivated arctic bramble ( R. arcticus ssp. arcticus ) in Finland in 1980, and from reference isolates of Phoma glomerata isolated from other hosts. Multivariate analysis of growth rate data and conidial dimensions measured in vitro indicated that the fungi isolated from hybrid arctic bramble in Sweden were not distinguishable from D. applanata , but were clearly distinct from P. glomerata and P. exigua. Furthermore, they had identical ITS1 and ITS2 sequences, and were placed in a phylogenetic clade very closely related to the clade that contained isolates of D. applanata isolated from raspberry ( Rubus idaeus ). In contrast, isolate HPP 38 from Finland was placed in a clade with P. exigua. These data indicate that the Swedish isolates infecting arctic bramble belong to a strain of D. applanata that differs from the isolate infecting raspberry only by two common nucleotide substitutions in ITS2. Fungi of the Phoma–Didymella complex on wild and cultivated arctic bramble (a total of 291 plants showing symptoms sampled from 37 sites) were detected by a PCR-based assay and found to be common in northern Sweden, but rare, albeit widely distributed, in Finland.     

部分栽培葱属植物叶绿体基因组的RAPD分析

  提取我国栽培的几种主要葱属植物的叶绿体DNA , 利用RAPD 技术进行遗传关系分析。从56 个10 bp 随机引物中筛选出18 个多态性引物进行扩增分析, 14 个材料共扩增出270 条带, 其中246 条具多态性。UPGMA 聚类将供试葱属品种分成4 个组。葱属在进化上存在两个方向, 分别形成了管状叶和扁平叶两种类型。