北方地区小西葫芦黄花叶病毒的酶联检测和西瓜品种抗病性鉴定

应用DAS-ELISA对分别于1998年在河南、陕西和1999年在河南、陕西、河北和山西采集的葫芦科病毒病样本84份、186份进行小西葫芦黄花叶病毒的检测,表明该病毒在这些地区广泛发生,ZYMV阳性检出率分别为79.8%和57.5%熏在北京和河南临颖县ZYMV发生率较低熏≤23.1%,在其余地区即河北石家庄,河南郑州、开封、孟津,山西运城,陕西西安其阳性检出率在40.0%～92.9%之间。除冬瓜样本没有ZYMV外,西瓜、甜瓜、南瓜、丝瓜、小西葫芦、苦瓜、黄瓜和瓠子均受到ZYMV的侵染。不同地区、年份和作物之间ZYMV阳性检出率存在差异。对18份西瓜品种的抗ZYMV温室人工接种鉴定的试验表明,目前推广的一些品种对ZYMV缺乏抗病性。     

Spread of potato virus Y in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity

The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus Y^NTN (PVY^NTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVY^NTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses.     

我国马铃薯主产区病毒病发生情况调查

为了解我国马铃薯病毒病发生情况,在马铃薯主产区黑龙江、内蒙古、甘肃、云南等马铃薯田采集了具有典型病毒病症状的样品、疑似样品和随机无症样品,试管苗和原原种为随机样品,共649份。应用DASELISA方法筛查6种马铃薯主要病毒:PVX、PVY、PVS、PLRV、PVM和PVA。结果表明:129份样品为阳性,PVY的检出率最高,为9.86%,PVS次之,为6.47%;试管苗PVS检出最多,为32份,原原种PVX检出最多,为10份,大田样品PVY病毒发生比例最高,为54份;有38份样品是由多种病毒复合侵染造成的,大田PVY+PVS复合侵染率最高,为2.93%,PVS+PVY+PLRV次之,为0.98%,试管苗PVS+PVM复合侵染率最高,为4.85%。通过历年的检测结果比较,试管苗中PVS病毒检出率最高,原原种中PVX和PLRV病毒成为危害最为严重的病害,大田样品中PVY检出率一直居高不下。     

贵州3种马铃薯病毒的DAS-ELISA检测与分析

应用DAS-ELISA法对从贵州省马铃薯种植区采集带症状的202份马铃薯叶片分别进行了PVX、PVY、PLRV 3种马铃薯病毒的检测。结果表明,3种马铃薯病毒在贵州马铃薯种植区表现出比较显著的田间症状,其病毒的发生与气候类型、种薯品种来源、种植方式存在相关性,并结合马铃薯实际生产对其病毒防治措施做了简要讨论。     

人工接种CMV后辣椒植株中病毒含量的时序变化及症状表型

以5份不同类型的辣(甜)椒为供试材料,人工接种CMV重花叶型株系,通过DAS-ELISA方法,分别检测接种5、10、15、20、25、30d后辣椒植株中CMV含量的时序变化,同时对供试材料进行人工接种抗性鉴定,并对2种方法得到的数据进行相关性分析。结果显示:不同类型供试材料接种5~15d后的病毒含量逐渐增加,并于15d时达到最高水平,之后略有回落;通过DAS-ELISA检测得到的植株病毒含量与人工接种抗性鉴定得到的病情指数呈正相关,说明2种鉴定方法获得的结果一致。     

In vitro characterization andin vivo detection ofRigidoporus lignosus,the causal agent of white root disease inHevea brasiliensis,by ELISA techniques

The aim of these studies is to develop a method for early detection ofRigidopoms lignosus (Basidiomycete, Polyporaceae), the causative agent of white root disease of rubber tree. Two polyclonal sera were produced against soluble mycelial proteins of twoR. lignosus isolates, one from Africa (FCI2), the other from Asia (FID2). The specificity of the antisera was tested using isoelectric focusing (IEF)/Western-blot and DAS-ELISA. The two sera recognized all 20R. lignosus isolates from various geographical origins. The banding patterns obtained by Western-blot enabled a distinction to be made between isolates from Africa and those from Asia. In DAS-ELISA and Western-blot analyses, strong cross reactions were observed withR. ulmarius. Only slight reactions were observed in Western-blot analysis toR. lineatus andP. noxius, both causative agents of root rot inHevea. These cross reactions were not observed under our DAS-ELISA analysis conditions. Finally, no cross reactions were obtained with 9 otherPolyporaceae orHevea root pathogen species. The sensitivity threshold of the DAS-ELISA method was 5 ng ml^–1 ofR. lignosus protein. An initial approach to using the DAS-ELISA test for the detection ofR. lignosus in infected plants was carried out on artificially inoculated root samples. The DAS-ELISA protocol enabled detection ofR. lignosus in the root systems of diseased plants. Moreover, no cross reaction was observed with healthy plant extracts.     

品种与季节和日龄对仔猪血清中IgG含量的影响

为了探索品种、季节和日龄对仔猪血清中IgG含量的影响,将280头试验猪按不同品种/类群(久仰香猪、剑白香猪和长白猪)、不同季节和不同日龄(0、4、7、14、21、28日龄)采样、然后用双抗体夹心ELISA法(DAS-ELISA)测定其血清中IgG含量。结果表明:剑白香猪IgG含量比久仰香猪和长白猪高,差异显著(P<0.05);冬季仔猪IgG含量比其他季节高,差异极显著(P<0.01),春季与夏、秋季差异显著(P<0.05);仔猪IgG含量随日龄而增加,但从21日龄开始下降。     

Improved detection of citrus psorosis virus using polyclonal and monoclonal antibodies

Citrus psorosis is a serious and widespread disease associated with citrus psorosis virus (CPsV), a novel filamentous negative-stranded virus in the genus Ophiovirus . Laborious and costly indexing on test plants has been the only routine diagnostic method available, but recently an antiserum usable in double antibody sandwich (DAS) ELISA has been prepared. Here, major improvements to the DAS-ELISA protocol, a new purification method, and production of two monoclonal antibodies (mabs) to CPsV, an IgG and an IgM are reported. A highly sensitive triple antibody sandwich (TAS) ELISA making use of the mabs is described. In glasshouse citrus the homologous virus was still detectable at a tissue dilution of 1/6250 in DAS and at 1/31250 in TAS-ELISA. Both the DAS and IgG mab-TAS formats detected all CPsV isolates so far tested (from Argentina, Italy, Lebanon, Spain and the USA). A few isolates were not detected by the IgM mab.     

建兰花叶病毒检测标准方法的建立

经过2004￣2005两年对上海口岸进境和进境后种植的260个兰花品种上建兰花叶病毒(CyMV)检测方法的研究,分别建立了检测和鉴定兰花上建兰花叶病毒的DAS-ELISA、电镜观察、鉴别寄主反应、RT-PCR和IC-RT-PCR的方法。     

3种检测方法在脱毒马铃薯种薯病毒检测中的应用及检测效果比较

为进一步明确适用于脱毒马铃薯种薯的病毒检测方法,为脱毒马铃薯生产提供更为有效和准确的病毒检测技术指导。应用反转录聚合酶联式反应(RT-PCR)、双抗体夹心-酶联免疫吸附法(DAS-ELISA)以及胶体金免疫层析技术(GICA)3种方法,对脱毒马铃薯生产中的马铃薯X病毒(PVX)、S病毒(PVS)进行检测,在不同品种及不同繁育等级的种薯中分别比较3种方法的检测效果。结果表明,RT-PCR对马铃薯原原种中的2种病毒检测效果显著优于其他2种方法;3种方法对一级种的2种病毒检测效果无显著差异。RTPCR非常适用于原原种病毒的检测;ELISA方法对原种及一级种大样本抽检更为适用,而GICA因成本高,更适宜一级种小样本的快速抽检。