Isolation of Olive latent virus 1 from Tulip in Toyama Prefecture

A virus whose coat protein gene had a high sequence homology with the coat protein gene of Olive latent virus 1 was isolated from diseased tulip in Toyama Prefecture. Received 1 June 2001/ Accepted in revised form 1 August 2001     

The spore coat of the bean anthracnose fungus Colletotrichum lindemuthianum is required for adhesion, appressorium development and pathogenicity

The spores (conidia) of the bean anthracnose fungal pathogen, Colletotrichum lindemuthianum, adhere to the aerial parts of plants to initiate the infection process. In previous studies we have shown that the Colletotrichum spores are surrounded by a fibrillar spore coat, comprising several major glycoproteins. Previous evidence showed that a monoclonal antibody (UB20) that recognised these glycoproteins was able to inhibit adhesion of spores to a hydrophobic surface. In this paper we have further studied the role of the spore coat in adhesion, germination and fungal development by studying the effects of UB20 and protease treatment of spores. The latter treatment has previously been shown to remove the spore coat. Spores germinate on glass, polystyrene and water agar, however, appressoria only develop on glass or polystyrene, showing a requirement for a hard surface. Removal of the spore coat with protease inhibits adhesion at 30 min, before the secretion of ECM glycoproteins. Protease treatment also inhibits the development of appressoria and reduces pathogenicity on leaves. Incubation of spores with the MAb UB20 inhibits adhesion at 30 min, but does not affect appressorium formation or pathogenicity. The results suggest that an intact spore coat has two functions; it is required for adhesion to a hydrophobic surface and for the detection of a hard surface necessary for appressorium formation. We suggest that contact with a hard surface, rather than adhesion, is the key event leading to appressorium formation.     

<Emphasis Type="Italic">Amaranthus leaf mottle virus</Emphasis>: 3′-end RNA sequence proves classification as distinct virus and reveals affinities within the genus <Emphasis Type="Italic">Potyvirus</Emphasis>

Amaranthus leaf mottle virus (AmLMV) was classified as a member of the genus Potyvirus on the basis of its particle morphology, serology, and biological properties (Casetta et al., 1986). Based on these properties, an Amaranthus viridis-infecting virus isolated in Spain, causing mottle and leaf blistering as well as reduced growth has been identified as AmLMV. The 3′ terminal genomic region of this and a reference isolate from Italy has been sequenced and reveals a 95% nucleotide identity between the two isolates. The sequenced part comprises the coat protein with 281 amino acids and 315 nucleotides of the 3′ untranslated region (UTR) preceding a polyadenylated tail. Pairwise comparisons and phylogenetic analysis of the nucleotide and deduced amino acid sequences of the CP and 3′ UTR of the cloned cDNAs with those of other potyviruses shows that AmLMV is a distinct potyvirus closely related to Potato virus Y.     

蚕豆染色病毒CPS基因的序列测定及RT-PCR检测方法的建立

 The nucleotide sequence of small coat protein (CPS) gene of Broad bean stain virus (BBSV) was determined and compared with other comoviruses. The CPS gene of BBSV consisted of 687 nucleotides and encodes a putative protein of 228 amino acid residues. The CPS sequence of BBSV and those of other comoviruses shared identities of 36.5%-58.9% and 35.2%-70.3% at the nucleotide and amino acid levels, respectively. The RT-PCR method specific for BBSV detection was developed based on the determined CPS sequence. The RT-PCR assay presented here allows, for the first time, rapid and specific detection of BBSV.     

Isolation of two strains of a new Tombusvirus (Havel river virus, HaRV) from surface waters in Germany

Two virus isolates from water samples — one from a small stream in South Western Germany and another one from the Havel river in North Eastern Germany c. 500 km away, proved to be strains, named S and H, respectively, of a new Tombusvirus for which the name Havel river virus (HaRV) had been suggested previously in a brief account. Immunoelectron microscopical decoration tests and sequence comparisons of the coat proteins indicated that the two HaRV strains are only distantly related to known Tombusviruses. The closest relationships were found to Cucumber necrosis virus. Nothing is known about their natural hosts. Because the S strain of HaRV was isolated in a woody area from a small stream close to its origin, they may be pathogens of trees or wild plants in such habitats.     

芜菁花叶病毒不同分离物3'-cDNA片段的克隆及序列分析

 从河北、北京、山东泰安和潍坊等地区的萝卜、大白菜、甘蓝、油菜上得到8个芜菁花叶病毒(Turnip mosaic virus,TuMV)分离物,测定了它们3'-末端cDNA片段的序列。根据衣壳蛋白基因(cp)核苷酸序列可以将这8个分离物与另外2个TuMV分离物分为3组:泰安旧镇大白菜(JZBC)、甘蓝(JZGL)和萝卜(JZLB)分离物为一组;北京(BJILB)、潍坊萝卜分离物(WFLB)与引起萝卜红心病的TuMV分离物(WFLB99)为一组;泰安范镇、河北、潍坊(WFLB04)萝卜和泰安旧镇油菜(JZYC)上的TuMV分离物为一组。农壳蛋白氨基酸序列比较结果与此基本一致,所不同的是分离物WFLB99与所有分离物的差异均较大,形成单独一个分支。有必要对TuMV的变异情况和致病机理进行深入研究。     

侵染葫芦的黄瓜绿斑驳花叶病毒广西分离物分子鉴定

从广西南宁市郊温室大棚中的葫芦[Lagenaria siceraria(Molina)Stand.]上采集到一个表现脉绿、花叶症状的病毒样品,ELISA检测表明,该样品与黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)有密切的血清学关系,利用RT-PCR方法从样品中扩增获得约500bp的DNA片段,序列分析表明,该片段是CGMMV的外壳蛋白基因,暂将该病毒分离物定名为GX-BG。外壳蛋白基因核苷酸序列系统进化树分析表明,已报道的CGMMV主要分为3大群体,GX-BG与中国辽宁分离物(CGMMV-LN)分别属于不同的群体。     

甘薯褪绿斑病毒外壳蛋白的分子变异及其特异抗血清制备

 甘薯褪绿斑病毒(Sweet potato chlorotic fleck virus,SPCFV)是侵染甘薯的主要病毒之一。本研究利用RT-PCR方法克隆了SPCFV中国4个分离物的外壳蛋白(CP)基因。序列分析表明,cp基因全长900 bp,编码299个氨基酸残基。4个分离物cp基因的核苷酸序列一致性为78.3%～89.9%,推导的氨基酸序列一致性为91.3%～95.7%,存在较大的分子变异。不同分离物CP氨基酸序列N末端的第3-32位氨基酸为多变区。将四川分离物的cp基因克隆到原核表达载体pET-28a(+)上,SDS-PAGE分析表明,经IPTG诱导,cp基因在大肠杆菌BL21(DE3)中得到了高效表达。以表达的蛋白为抗原免疫家兔,制备了SPCFV CP的特异性抗血清。ACP-ELISA检测结果表明,制备的抗血清效价达1∶128 000,可用于田间甘薯样品的检测。     

Pancytopenia in a cat with visceral leishmaniasis

Abstract: A 4‐year‐old, domestic shorthair, female spayed cat was presented for decreased appetite and depression. Severe pancytopenia with erythrocyte autoagglutination was found. The cat was seronegative for feline immunodeficiency and leukemia viruses. Immune‐mediated hemolytic anemia was suspected but no response to treatment with a blood transfusion, enrofloxacin, and prednisone was observed. Blood and bone marrow smears obtained 11 days later contained Leishmania amastigotes in the cytoplasm of neutrophils and macrophages, respectively. Serologic and PCR testing of peripheral blood confirmed infection with Leishmania infantum. Despite treatment, the cat worsened clinically and was euthanized. At necropsy, visceral dissemination of the parasite was confirmed. The findings in this case indicate that visceral leishmaniasis should be considered as a differential diagnoses in cats with pancytopenia in areas endemic for Leishmania. In addition, amastigotes may be observed in peripheral blood neutrophils.     

脱落酸与种皮对杜梨种子休眠与萌发的影响

以当年新采收杜梨种子和自然贮藏1 a种子为试验材料,研究了种皮和不同浓度的脱落酸(ABA)对诱导杜梨种子休眠与萌发的影响.结果表明:杜梨种皮和脱落酸均能够抑制杜梨种子的萌发,种皮的抑制萌发效应显著大于ABA的作用,杜梨种子去除种皮后,休眠性可被部分解除.脱落酸对杜梨种子抑制萌发的效应在去皮和不去皮种子中表现明显差异,5 mg/L ABA浓度对未去皮种子产生明显抑制效应,20 mg/L ABA浓度对去皮种子产生明显抑制效应,说明未去皮种子对ABA的敏感性大于去皮种子.种皮与ABA的双重抑制效应,使种子对ABA浓度水平差异的敏感性增强.不同激素组合试验说明了GA3、6-BA和ABA在杜梨种子休眠与萌发中的作用.