菜豆种子中菜豆普通花叶病毒的RT-PCR检测

为研究国家作物种质库中保存菜豆种质携带种传菜豆普通花叶病毒（BCMV）的状况，根据BCMV的印基因序列设计一对特异性引物，从BCMV侵染的菜豆叶片上扩增出与理论大小一致的714bp的片段。在此基础上，应用改进CTAB法从菜豆干种子中提取总RNA并特异性扩增出相同大小的片段。对目的片段进行克隆、测序和序列同源性比较，结果表明，扩增片段的序列与其它BcMV的印基因序列同源性达88％-98％，确证该方法可以从莱豆干种子申直接检测BCMV。利用建立的方法，分别从单粒菜豆种子样本和30粒菜豆种子样本中检测到BCMV，其检测灵敏度较ELISA方法更高。来自河北的紫芸豆单粒种子带毒率高达100％，而在不同地理来源的6份菜豆种质中，4份被检出携带BCMV，表明入库保存的菜豆种质和种子已普遍为BCMV所侵染。     

Identification of blackeye cowpea mosaic virus from germplasm of yard-long bean and from soybean,and the relationships between blackeye cowpea mosaic virus and cowpea aphid-borne mosaic virus

Two potyvirus isolates, one from germplasm of yard-long bean (Vigna unguiculata ssp.sesquipedalis) introduced into the Netherlands, and another one from soybean plants (Glycine max) in Indonesia, were compared with two virus isolates of blackeye cowpea mosaic virus (BICMV) from the USA and a Moroccan isolate of cowpea aphid-borne mosaic virus (CAMV). It is proposed that all five isolates be now considered BICMV on the basis of host ranges, symptoms and serology. From our results, and a reassessment of the literature it is suggested to drop the name CAMV in favour of BICMV.Samenvatting Twee potyvirussen, de een in Nederland ingevoerd met genenmateriaal vanVigna unguiculata ssp.sesquipedalis en de ander uit planten van sojaboon (Glycine max) in Indonesië, werden vergeleken met twee isolaten van blackeye cowpea mosiac virus (BICMV) en een Marokkaans isolaat van cowpea aphid-borne mosaic virus (CAMV). Op grond van waardplantenreeksen, symptomen en serologie stellen de auteurs voor om alle vijf isolaten te beschouwen als BICMV. Gebaseerd op de verkregen resultaten en een kritische beschouwing van de literatuur wordt de aanbeveling gedaan om de naam CAMV te laten vallen ten gunste van BICMV.     

The use of host resistance in disease management of rust in common bean

Bean rust, caused by Uramyces appendiculatus, is one of the major diseases in dry and snap bean production world-wide. Numerous advancements in disease management have been made to reduce rust losses. Host resistance is an important component of rust management. However, durability of disease resistance has often been short due to the use of single genes for resistance interacting with extremely high virulence diversity of the bean rust fungus. The challenge to increase durability of resistance has led to strategies such as gene pyramiding of race-specific resistance, selection and use of partial resistance, and investigation and discovery of leaf morphological features that may slow the rust epidemic. Germplasm with multiple sources of rust resistance has been developed in specific bean seed classes and released for public and commercial use in intensive production systems such as those in the United States. However, progress to develop rust resistant germplasm for the subsistence agriculture of Latin America and Africa where intercropping and mixed cultivars dominate the production system has been slow. Incorporation of high yielding, disease-resistant components as partial replacement in farmer's mixtures has the potential to reduce severity in the crop and increase yield in the presence of rust. This strategy would not erode the genetic diversity that is historically known to enhance resistance durability and for many years has given stability in production in the subsistent agriculture systems.     

超高效液相色谱-串联质谱法快速检测菜豆及土壤中的灭蝇胺残留

建立了采用超高效液相色谱-串联质谱仪(UPLC-MS/MS)快速检测分析菜豆及其土壤中灭蝇胺残留的方法。样品经5 mmol/L的乙酸铵-乙腈溶液[V(乙酸铵):V(乙腈)=1:4]提取、定容,N-丙基乙二胺(PSA)净化后用UPLC-MS/MS在正离子模式下以多反应监测扫描方式进行测定,外标法定量。结果表明:在0.05～1 mg/kg添加水平范围内,灭蝇胺在菜豆和土壤中的平均回 收率为80.1%～95.2%,相对标准偏差(RSD)为1.9%～9.1%,检出限(LOD)为3.8～5.9 μg/kg,定量限(LOQ)为50 μg/kg。该方法操作简单、灵敏度高、定量准确且测定浓度范围宽,适用于菜豆及土壤中灭蝇胺的残留分析。     

瓜蚜在西葫芦植株上传播获取西瓜花叶病毒的效率及西瓜花叶病毒病的发生规律

为明确瓜蚜Aphis gossypii Glover成蚜对西瓜花叶病毒(Watermelon mosaic virus,WMV)的传毒特征,应用RT-PCR技术检测了瓜蚜传播获取WMV的效率及其在成蚜体内的存留时间;同时调查了2015年山东省日光温室西葫芦瓜蚜种群动态与西瓜花叶病毒病的发生规律。结果表明,瓜蚜成蚜离开带毒西葫芦植株0 h后,带毒率为35.00%,3 h后成蚜带毒率随时间的延长开始呈下降趋势,24 h后仅为5.40%。26℃条件下,健康瓜蚜取食带毒植株5 min后即可获毒,0.5 h后获毒率达到62.50%;带毒瓜蚜取食健康植株0.5 h后植株即可带毒,24 h后植株带毒率达到94.44%。温度对瓜蚜成蚜的传毒率有一定影响,在21、26、31℃条件下,瓜蚜成蚜24 h传毒率分别为61.11%、94.44%和88.89%。调查发现,温室西瓜花叶病毒病的发生率与瓜蚜种群数量呈正相关,当温室瓜蚜成蚜种群数量为1.06～1.73头/叶时,西葫芦开始发病,当成蚜数量增至18.22头/叶时,全田发病率可达83.33%。表明瓜蚜成蚜具有较强的传播WMV的能力,有效控制瓜蚜发生对减少WMV的扩散具有重要意义。     

实时荧光RT-PCR一步法检测南方菜豆花叶病毒

南方菜豆花叶病毒（Southem bean mosaic virus，SBMV）是我国二类检疫性有害生物，以南方菜豆花叶病毒日本分离物（SBMV-J）总RNA为模板，采用RT-PCR方法扩增病毒外壳蛋白基因及其上游基因的cDNA片断并将其克隆到pMD18．T载体上。序列分析结果表明：SBMV-J cp基因由801个核苷酸组成，编码266个氨基酸，SBMV．J与其它分离物及株系印基因的核苷酸序列同源性为83％-97％，氨基酸序列同源性为86％-97％。由于SBMV各分离物及株系印基因的同源性较低，难于设计出较长的普通PCR引物。通过较短引物设计和TaqMan-MGB探针技术，建立了SBMV的实时荧光RT-PCR一步检测方法。该方法的检测低限是0．16pg，最佳检测总RNA的量是0．16ng。     

逆转录环介导等温扩增技术检测南方菜豆花叶病毒

 根据南方菜豆花叶病毒外壳蛋白基因序列设计并合成了4组RT-LAMP引物,通过引物筛选试验,确定SB1组引物为最佳引物,并进行了引物特异性与灵敏度检测试验,最终建立了南方菜豆花叶病毒的RT-LAMP检测方法。灵敏度检测试验显示,RT-LAMP方法比普通RT-PCR法灵敏度高10倍,而检测时间明显缩短,整个反应过程只需40 min。此外,体系中加入钙黄绿素,反应结束后,可裸眼观察颜色变化来判定结果。本研究所建立的SBMV RT-LAMP方法具有快速、稳定、灵敏、特异、操作简单的特点,适合于SBMV的现场快速检测。     

不同覆膜方式对山旱地菜豆土壤水分及利用效率的影响

以秋季和春季半膜平覆、全膜平覆、全膜双垄沟6种覆膜方式为处理,研究了不同覆膜方式对山旱地菜豆土壤水分及其利用率等的影响.结果表明,秋季全膜双垄沟和秋季全膜平覆较春季全膜双垄沟、秋季半膜平覆、春季全膜平覆和对照春季半膜平覆在播种时0～ 100 cm土层中土壤贮水量分别提高7.62％～14.20％、7.44％～14.02％...     

实时荧光RT-PCR-步法检测南方菜豆花叶病毒

南方菜豆花叶病毒(Southern bean mosaic virus,SBMV)是我国二类检疫性有害生物,以南方菜豆花叶病毒日本分离物(SBMV-J)总RNA为模板,采用RT-PCR方法扩增病毒外壳蛋白基因及其上游基因的cDNA片断并将其克隆到pMD18-T载体上。序列分析结果表明:SBMV-J cp基因由801个核苷酸组成,编码266个氨基酸,SBMV-J与其它分离物及株系cp基因的核苷酸序列同源性为83%～97%,氨基酸序列同源性为86%～97%。由于SBMV各分离物及株系cp基因的同源性较低,难于设计出较长的普通PCR引物。通过较短引物设计和TaqMan-MGB探针技术,建立了SBMV的实时荧光RT-PCR一步检测方法。该方法的检测低限是0．16 pg,最佳检测总RNA的量是0．16 ng。     

Broad bean wilt virus 2 in yams

We identified Broad bean wilt virus 2 (BBWV-2) in yams based on particle morphology, test plant symptoms, protein features, aphid transmission, and molecular classification using nucleotide sequences of coat protein genes.     

Incidence of viruses in <Emphasis Type="Italic">Alstroemeria</Emphasis> plants cultivated in Japan and characterization of <Emphasis Type="Italic">Broad bean wilt virus-2, Cucumber mosaic virus</Emphasis> and <Emphasis Type="Italic">Youcai mosaic virus</Emphasis>

Alstroemeria plants were surveyed for viruses in Japan from 2002 to 2004. Seventy-two Alstroemeria plants were collected from Aichi, Nagano, and Hokkaido prefectures and 54.2% were infected with some species of virus. The predominant virus was Alstroemeria mosaic virus, followed by Tomato spotted wilt virus, Youcai mosaic virus (YoMV), Cucumber mosaic virus (CMV), Alstroemeria virus X and Broad bean wilt virus-2 (BBWV-2). On the basis of nucleotide sequence of the coat protein genes, all four CMV isolates belong to subgroup IA. CMV isolates induced mosaic and/or necrosis on Alstroemeria. YoMV and BBWV-2 were newly identified by traits such as host range, particle morphology, and nucleotide sequence as viruses infecting Alstroemeria. A BBWV-2 isolate also induced mosaic symptoms on Alstroemeria seedlings.     

Identification and properties of a deviant isolate of the broad bean yellow band serotype of pea early-browning virus from faba bean (Vicia faba) in Algeria

A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.     

<Emphasis Type="Italic">Ulocladium atrum</Emphasis> as a biological control agent for white mold of bean caused by<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>

Field studies were conducted near Lethbridge, Alberta, Canada, in 2001, 2004 and 2005 to determine the efficacy of the antagonistic fungusUlocladium atrum for control of white mold of bean caused bySclerotinia sclerotiorum. Results of the 3 years of field trials showed that, compared with the untreated control, foliar application of a spore suspension ofU. atrum (300 ml m⁻² of 10⁶ spores ml⁻¹ suspension) significantly reduced incidence and severity of white mold, increased seed yield and reduced contamination of bean seed by sclerotia ofS. sclerotiorum. The level of control of white mold observed in the treatment ofU. atrum was similar to that of the mycoparasitic fungusConiothyrium minitans, but lower than the fungicide treatments of Ronilan (vinclozolin) at the rate of 1200 g ha⁻¹ per application in 2001, or Lance (boscalid) at the rate of 750 g ha⁻¹ per application in 2004 and 2005. The potential for use ofU. atrum as a biological control agent for sclerotinia diseases is discussed. http://www.phytoparasitica.org posting Nov. 12, 2006.     

Southern bean Mosaic Virus the Causal Agent of a New Disease of Phaseolus vulgaris beans in Spain

Southern bean mosaic virus (SBMV) has been identified as the cause of a new disease in greenhouse-cultivated common bean (Phaseolus vulgaris), in the south-east of Spain. The identification was based on host range comparisons, morphological and serological characteristics of the virus, the size of its dsRNA species and the nucleotide sequence of an 810-bp fragment from ORF2. The virus could be clearly discriminated from the related sobemovirus Southern cowpea mosaic virus. This is the first report of SBMV in Spain.     

Leaf-yellowing in combination with corm necrosis in freesia caused by bean yellow mosaic virus: Factors involved in syndrome development

In freesia cv. Aurora grown in the field for cutflower production, a disease occurred with symptoms of leaf-yellowing in combination with corm necrosis (LYCN). It is shown that this disease is caused by bean yellow mosaic virus (BYMV).No differences in symptoms of LYCN were observed between the freesia cultivars Aurora, Imperial and Rose Marie. Most BYMV isolates gave rise to LYCN; the isolates from crocus andIxia sp. did not. LYCN was stimulated by a high BYMV concentration in the inoculum, a temperature above 20°C, inoculation soon after emergence of the freesias, and by the absence of freesia mosaic virus. Freesias with mosaic symptoms and infected with a cross-protecting BYMV strain, did not show symptoms of leaf-yellowing and/or corm necrosis after inoculation with BYMV-Cm. The presence of the unknown agent causing leaf necrosis in freesias did not have an influence on symptom development after infection with BYMV.     

Stem and root rot of scarlet runner bean (Phaseolus coccineus) caused by Pythium myriotylum

A severe rot was found on the stems and roots of scarlet runner bean (Phaseolus coccineus) in Ibaraki Prefecture (Japan) in August 2004. The causal fungus was identified as Pythium myriotylum. We propose the name of stem and root rot of scarlet runner bean (“Kuki-negusare-byo” in Japanese) for this new disease.     

引起甘蔗花叶病的病原分子生物学进展

花叶病是最主要的甘蔗病毒病害之一,在全球种植甘蔗的国家或地区普遍发生,可导致甘蔗产量下降,糖分减少,给甘蔗生产带来严重的经济损失。引起甘蔗花叶病的病毒主要有甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)、高粱花叶病毒(Sorghum mosaic virus,Sr MV)和甘蔗条纹花叶病毒(Sugarcane streak mosaic virus,SCSMV)。本文综述了这3种病毒的生物学特性、鉴定与检测、基因组结构与基因功能、遗传变异与分子进化等方面的研究进展,并讨论了对甘蔗花叶病的生态防控措施。     

First report of <Emphasis Type="Italic">Bean common mosaic virus</Emphasis> in yam bean [<Emphasis Type="Italic">Pachyrhizus erosus</Emphasis> (L.) Urban] in Indonesia

Severe mosaic with leaf malformation and green vein banding was observed on yam bean in West and Central Java, Indonesia. Virions of the causal virus were flexuous filaments, about 700 nm in length, with a coat protein of 30 kDa. The virus was transmitted by mechanical inoculation and by aphids in a nonpersistent manner. The nucleotide sequence of the coat protein gene had the highest identity with that of Bean common mosaic virus (BCMV, genus Potyvirus) isolate VN/BB2-5. Based on demarcation criteria, including the genome sequence and host range, we tentatively designate this isolate as BCMV-IYbn (Indonesian yam bean). The nucleotide sequence reported is available in the DDBJ/EMBL/GenBank databases under accession number AB289438.