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香石竹斑驳病毒,一种感染伊朗马哈拉特香石竹切花作物的重要病毒病原体。

Carnation mottle virus, an important viral agent infecting carnation cut-flower crops in Mahallat of Iran.

作者信息

Safari M, Koohi Habibi M, Mosahebi G, Dizadji A

机构信息

Department of Plant Protection, College of Horticultural Science and Plant Protection, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.

出版信息

Commun Agric Appl Biol Sci. 2009;74(3):861-5.

Abstract

One of the most important cut-flower crops grown worldwide on commercial scale is Carnation (Dianthus caryophyllus L.). It's the main production of Mahallat where is one of the most important ornamental plants production centers of Iran. Infection of carnation with pathogens Like viral agents causes economic losses in carnation cut-flower crop. One of the viral agents of this flower is Carnation mottle virus (CarMV) which is the type member of genus Carmovirus and belongs to the Tombusviridae family. It is naturally transmitted by grafting and contacting between plants. Although its infection lead to mild symptims, it weakens the plant to infection by other pathogens. The carnation greenhouses of Mahallat were visited during 2008 January to April and 100 samples with mild mosaic symptom were collected and tested by DAS-ELISA using CarMV specific polyclonal antibody. The results showed that 75% of samples wrere infected with this virus. Mechanical inocubation of Chenopodium quinoa, C. amaranticolor and Spinacea oleracea with extracted crude sap of CarMV infected carnation Leaves in phosphate buffer (pH, 7) resulted in appearance of chlorotic and necrotic local lesions on inoculated leaves 4-7 days after incubation. The virus was partially purified using C. amaranticolor locally symptomatic leaves. Total soluble proteins were extracted from healthy and CarMV infected C. amaranticolor plants and beside partially purified preparation electrophoresed through 15% poly acrylamide get according to SDS-PAGE standard procedure. Protein bands were electroblotted onto nitrocelluse membrane and incubated with CarMV polyclonal during western immunoblot analysis according to standard method. The result revealed a distinc protein band with Mr of 35.5 kDa in total protein preparation of infected plant and viral partial pure preparation, without any reaction in those of healthy plant. RT-PCR carried out using total RNA extracted from infected plant by Rneasy Plant Mini Kit (Qiagen)and a pair of primers, CPu, CPd, corresponding to the flanking region of the virus CP resulted in amplification of a DNA fragment in expected size around 1 kbp.

摘要

全球范围内商业种植的最重要的切花作物之一是康乃馨(香石竹,学名Dianthus caryophyllus L.)。它是马哈拉特的主要农产品,马哈拉特是伊朗最重要的观赏植物生产中心之一。康乃馨受到病毒等病原体感染会给康乃馨切花作物带来经济损失。这种花卉的病毒之一是康乃馨斑驳病毒(CarMV),它是香石竹病毒属的模式成员,属于番茄病毒科。它通过嫁接和植株间接触自然传播。虽然其感染会导致轻微症状,但会使植株易受其他病原体感染。2008年1月至4月期间,对马哈拉特的康乃馨温室进行了走访,采集了100份有轻微花叶症状的样本,并使用CarMV特异性多克隆抗体通过双抗夹心酶联免疫吸附测定(DAS-ELISA)进行检测。结果显示,75%的样本感染了这种病毒。用磷酸盐缓冲液(pH 7)中感染CarMV的康乃馨叶片提取的粗汁液对接种藜(Chenopodium quinoa)、千日红(C. amaranticolor)和菠菜(Spinacea oleracea)进行机械接种,接种后4至7天,接种叶片上出现褪绿和坏死局部病斑。利用千日红的局部症状叶片对病毒进行了部分纯化。从健康和感染CarMV的千日红植株中提取总可溶性蛋白,并按照十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)标准程序,将部分纯化制剂在15%聚丙烯酰胺凝胶上进行电泳。蛋白质条带被电转移到硝酸纤维素膜上,并按照标准方法在蛋白质免疫印迹分析过程中与CarMV多克隆抗体一起孵育。结果显示,在感染植株的总蛋白制剂和病毒部分纯制剂中出现了一条分子量为35.5 kDa的明显蛋白条带,而健康植株的制剂中没有任何反应。使用Rneasy植物微型试剂盒(Qiagen)从感染植株中提取总RNA,并使用一对对应于病毒外壳蛋白(CP)侧翼区域的引物CPu、CPd进行逆转录聚合酶链反应(RT-PCR),扩增出了预期大小约1 kbp的DNA片段。

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