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绝对定量分析番茄曲叶病毒西班牙分离物(ToLCNDV-ES):以宿主特异性方式积累病毒。

Absolute Quantification of Tomato leaf curl New Delhi virus Spain strain, ToLCNDV-ES: Virus Accumulation in a Host-Specific Manner.

机构信息

IFAPA Centro La Mojonera, La Mojonera 04745, Almería, Spain.

IFAPA Centro Churriana, Málaga 29140, Spain.

出版信息

Plant Dis. 2018 Jan;102(1):165-171. doi: 10.1094/PDIS-06-17-0840-RE. Epub 2017 Oct 30.

DOI:10.1094/PDIS-06-17-0840-RE
PMID:30673467
Abstract

Tomato leaf curl New Delhi virus (ToLCNDV) (family Geminiviridae, genus Begomovirus) has recently been introduced in western Mediterranean countries. Isolates in Spain constitute a new strain, denominated ToLCNDV-ES, that is causing losses in commercial zucchini and melon crops; however, it is also, although less often, detected in commercial tomato crops. We developed a tissue-print hybridization test to detect the two genomic components of the virus and a TaqMan quantitative polymerase chain reaction (qPCR) test to estimate the number of genome copies in plants. qPCR was approximately 10 to 10 times more sensitive than tissue-print hybridization to detect viral genomic DNA-A and DNA-B, respectively. It also detected the virus in more experimentally and naturally ToLCNDV-ES-infected zucchini squash and tomato plants. ToLCNDV-ES DNA-A titers were significantly lower in tomato than in zucchini plants, often falling below the detection limits in the hybridization test. In addition, the DNA-B accumulation was impaired in tomato when compared with zucchini. According to the data obtained in this study, the differences in viral titers of both plant species contribute to explain the dissimilarities in symptom expression, capability of detection, and transmission of the virus.

摘要

番茄曲叶病毒(ToLCNDV)(双生病毒科,菜豆花叶病毒属)最近在西地中海国家出现。西班牙的分离物构成了一种新的毒株,称为 ToLCNDV-ES,它会导致商业西葫芦和瓜类作物的损失;然而,在商业番茄作物中也有较少的检测到。我们开发了一种组织印迹杂交检测法来检测病毒的两个基因组成分,以及一种 TaqMan 定量聚合酶链反应(qPCR)检测法来估计植物中的基因组拷贝数。qPCR 检测病毒基因组 DNA-A 和 DNA-B 的灵敏度分别比组织印迹杂交高约 10 倍和 10 倍。它还可以检测更多的实验和自然感染 ToLCNDV-ES 的西葫芦和番茄植物。与西葫芦相比,番茄中的 ToLCNDV-ES DNA-A 滴度明显较低,经常低于杂交检测的检测限。此外,与西葫芦相比,番茄中 DNA-B 的积累受到了损害。根据本研究获得的数据,两种植物的病毒滴度差异有助于解释症状表达、检测能力和病毒传播的差异。

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