6株猪O型口蹄疫病毒VP1基因的克隆与序列分析

根据口蹄疫病毒（FMDV）VP1基因的序列，设计并合成了2对用于扩增VP1基因的引物。从组织中提取总RNA，首先用P1、P2引物对6株猪。型口蹄疫病毒进行RT—PCR扩增，获得1000bp的片段；再用P3、P4引物进行巢式PCR扩增，结果获得850bp的片段。将850bp的片段克隆到pMD18—-T载体中，通过PCR鉴定，将阳性重组质粒进行测序并分析。结果发现6株FMDV的核苷酸同源性为80．2％～99．4％，其推导的氨基酸序列同源性为86．9％～99．5％；构建遗传发生树，发现6株FMDV属于两个不同的基因型，其中的Shunde00、Sihui01、Shenzhen99、Fushan01株属一个基因型（与Hongkong93、广东86分离株属同一基因型）；Guangzhou99、Shenzhen00株属另一个基因型（与UKG-12—2001株、JPN2000株属同一基因型）。通过对口蹄疫病毒VP1基因的测序与分析，了解其变异情况，为科学地防控FMD提供分子水平的依据。     

山东地区三株鸽Ⅰ型副粘病毒的分离及鉴定

2005年在山东地区部分肉鸽养殖场出现鸽子大量死亡现象,从死亡鸽中分离到三株鸽Ⅰ型副粘病毒(SD05027,SD05028,SD05029),其鸡胚平均死亡时间(MDT)分别为69h,69.6h,81.6h;鸡脑内接种指数(ICPI)为1.31,1.43,1.48,证明此次分离的三株病毒株均属速发型新城疫病毒。应用RT-PCR技术对F基因重要功能区片段扩增后进行序列测定,分析表明,F蛋白裂解位点处的序列(112R/K-R-Q-K-R-117F)与新城疫强毒在这一区域的序列相符。与多株已报道的NDV参考株相应片段进行序列比对和基因进化树分析,将其鉴定为基因VIb型。     

资源型城市产业结构演变的环境效应研究——以鞍山市为例

产业结构是经济活动与生态环境相互作用的重要纽带,资源型城市产业结构演变的环境影响具有独特的研究价值。选取典型资源型城市-鞍山市为案例,从演变的视角,研究产业结构与环境质量的动态变化特征,并利用典型相关分析的方法探讨二者的相互作用。研究表明,第二产业为主体、黑色金属冶炼及压延工业为主导以及国有制为主要运行环境是鞍山市产业结构的主要特征;鞍山市大多数环境污染物排放量的波动下降趋势明显,但仍维持在较高水平上;产业结构演变的环境效应显著,不同类型的产业对不同污染物的环境影响不同。     

Virus impact at the interface of an ancient ecosystem and a recent agroecosystem: studies on three legume-infecting potyviruses in the southwest Australian floristic region

The southwest Australian floristic region (SWAFR) is an internationally recognized 'hot spot' of global biodiversity and has an endangered flora. It represents a unique interface between an ancient ecosystem and a recent agroecosystem, providing the opportunity to investigate encounters where the recipient of the virus is an introduced crop and the donor a native plant and vice versa. Phylogenetic analysis of the virus coat-protein genes was used to study isolates of three potyviruses representing different 'new encounter' scenarios at this interface. The incidence, symptomatology, host range, non-persistent aphid transmission and considerable genetic diversity of the first indigenous virus described from the SWAFR, where it infects the native legume Hardenbergia comptoniana , and its potential to damage lupin, a locally important, newly introduced cultivated grain legume, was studied. The name Hardenbergia mosaic virus is proposed for this virus. Two other examples of 'new encounter' scenarios involving other legume-infecting potyviruses studied were: Passion fruit woodiness virus , which has been found only in Australasia, where it damages recently introduced species of Passiflora and legumes; and Bean yellow mosaic virus, which is not indigenous to Australia and was introduced recently to the SWAFR, where it infects a number of introduced legumes, but also damages the local native legume Kennedia prostrata . Isolates of the former had considerable genetic diversity consistent with the virus being indigenous, while isolates of the latter virus from K. prostrata had a low genetic diversity consistent with recent arrival. This research illustrates how introduced viruses can damage indigenous plants and indigenous viruses can damage introduced cultivated plants within this unique ecosystem, and how human activities can facilitate damaging 'new encounters' between plants and viruses.     

变叶海棠变异类型的初步研究

应用形象化散点图研究变叶海棠的起源和进化。结果表明,其变异类型可能系变叶海棠和陇东海棠或花叶海棠之间的天然杂交种,该假设需要用现代新方法来证实。     

Use of Degenerate Primers in a RT-PCR Assay for the Identification and Analysis of Some Filamentous Viruses, with Special Reference to Clostero- and Vitiviruses of the Grapevine

RT-PCR with degenerate primers was used for the screening of the genome of some members of the Closterovirus, Vitivirus and Trichovirus genera. Two sets of primers, targeted to conserved sequences of the heat shock protein 70 homologue of closteroviruses or to the RNA dependent RNA polymerase genes of tricho- and vitiviruses, amplified the expected fragments from total RNA extracts or double-stranded RNAs of infected plants. Amplified cDNAs were cloned, sequenced and phylogenetically analyzed. Results support the allocation of grapevine viruses A, B, D and heracleum latent virus (HLV) in the genus Vitivirus, whereas, the detection of a HSP70 homologue in grapevine leafroll-associated viruses agrees with their assignment in the genus Closterovirus. The use of degenerate primers for the identification of grapevine viruses belonging to Vitivirus and Closterovirus genera is envisaged.     

岱海游乐场孔孢粉分析及其2500年来古气候演化

本文通过对岱海游乐场钻孔剖面沉积物孢粉的定性、半定量分析,并结合岩性、岩相变化,初步恢复了该地区２５００年来古植被、古气候演化过程。研究结果表明,岱海湖盆自２５００年来植被类型有典型草原—灌丛草原—森林草原—灌丛草原的演化过程,古气候有较暖湿—较暖干—较冷干—较冷湿—较冷干的变化过程。     

Succession and phylogenetic profile of eukaryotic communities in rice straw incorporated into a rice field: Estimation by PCR-DGGE and sequence analyses

Abstract

Succession and the phylogenetic profile of eukaryotic communities associated with rice straw decomposition in a rice field were studied using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by 18S rDNA sequencing. Nylon mesh bags containing leaf sheaths or blades were buried in the plow layer of a rice field under flooded conditions after transplanting (Experiment 1) and under drained conditions during the off-crop season (Experiment 2). In addition, rice straw samples in Experiment 2 were taken out before plowing in spring and re-placed in the rice field under flooded conditions at transplanting. Statistical analyses based on DGGE patterns showed that eukaryotic communities were divided into two groups, namely group A before the placement in soil, after the mid-season drainage in Experiment 1 and under the drained conditions in Experiment 2 and group B before the mid-season drainage in Experiment 1 and under the flooded conditions in Experiment 2. Based on the sequence analysis of DGGE bands, which characterized the eukaryotic communities, succession of the communities was revealed, that is, most of the bands in group A were closely related to fungi, whereas the bands in group B were closely related to protozoa. These results indicated that eukaryotic communities associated with rice straw decomposition in the rice field are mainly affected by soil conditions, such as oxic or reduced conditions, irrespective of rice straw parts (leaf sheaths and blades).     

云南省水稻稻瘟病菌无毒基因AVR-Pia变异及水稻抗性基因Pia的有效性

为进一步了解田间稻瘟病菌Magnaporthe oryzae群体中AVR-Pia基因的分布及变异,利用水稻单基因系IRBLa-C水稻品种对自云南省13个市(州)采集分离得到的471株稻瘟病菌菌株进行抗性基因Pia有效性测定;利用无毒基因AVR-Pia特异性标记对471株稻瘟病菌菌株进行PCR检测和测序,并分析稻瘟病菌群体中无毒基因AVR-Pia的分布及DNA结构变异;利用有效性结果和PCR检测结果对471株菌株进行反应型划分,筛选鉴定菌株;利用鉴定菌株对云南省112份地方稻种进行Pia基因鉴定。结果表明,在471株稻瘟病菌菌株中,对含有Pia基因的水稻单基因系IRBLa-C表现为抗病和感病的菌株数分别为139株和332株,所占比例分别为29.5%和70.5%;在471株稻瘟病菌菌株中,分别有244株和227株菌株含有无毒基因AVR-Pia和不含有无毒基因AVR-Pia,所占比例分别为51.8%和48.2%,无毒基因AVR-Pia主要为完全缺失变异;在471株稻瘟病菌菌株中,A-和V+反应型菌株数分别为56株和161株,共217株,占总菌株数的46.1%,在13个市(州)稻瘟病菌群体中,A-和V+反应型菌株所占比例差异较大,其中在普洱市、红河哈尼族彝族自治州、昭通市、玉溪市4个市(州)的比例较大,分别为77.8%、57.1%、52.1%和50.0%;在112份云南省地方稻种质资源中,有20份地方稻品种含有抗性基因Pia,主要分布在9个市(州)中。表明云南省13个市(州)绝大部分水稻产区水稻Pia基因已丧抗性,含Pia基因的水稻种质在云南省分布较广。