中文摘要：柑桔碎叶病毒（CTLV） 是一种嫁接传播的毛状病毒组病毒，可引起柑橘碎叶病，导致严重的经济损失。本研究开发了一种基于TaqMan（R）化学的实时逆转录聚合酶链反应（RT-qPCR）分析检测柑橘上的CTLV。从基因库购买的8个CTLV分离株的全基因组序列的核苷酸序列分析显示，不同CTLV分离株病毒基因组3'区域比其他区域更保守。在CTLV基因组3'末端设计一组新的引物和TaqMan（R）探针，利用RT-qPCR 进行CTLV检测。研究结果表明，该检测方法稳定可靠。
外文摘要：Citrus tatter leaf virus (CTLV) is a graft-transmissible capillovirus that causes the tatter leaf disease of citrus and bud union incompatibility between scion and trifoliate hybrid rootstocks which can result in serious economic losses. Although several CTLV detection protocols have been described, there is a need for a highly sensitive protocol that can be used in routine diagnostic tests for virus-free budwood certification programs. To address this need, a TaqMan (R) chemistry based real time reverse transcription-polymerase chain reaction (RT-qPCR) assay was developed to detect CTLV from citrus plants. The nucleotide sequence analysis based on full genomic sequences of eight CTLV isolates available from GenBank showed that 3' region of the viral genome is more conserved among different CTLV isolates than the rest of the viral genome. A new set of primers and a TaqMan (R) probe were designed from a conserved 500 nt-long fragment at the 3' end of the CTLV genome for the detection of CTLV by RT-qPCR. The newly designed primers for CTLV RT-qPCR assay had ca. 98.6% amplification efficiency and showed 100% CTLV detection rate when they were tested for previously known CTLV-positive trees infected with different CTLV isolates. The sequencing of 132 bp-long RT-qPCR amplicon followed by BLASTn search showed that it had 94% to 98% sequence identity to those of various CTLV isolates available in GenBank, confirming the specificity of the assay to reliably detect CTLV.
外文关键词：Citrus; Virus; Capillovirus; Quantitative polymerase chain reaction
作者：Park, Jong-Won; Kunta, Madhurababu; McCollum, Greg; 等
作者单位：德克萨斯A&M大学
期刊名称：JOURNAL OF PLANT PATHOLOGY
期刊影响因子：0.944
出版年份：2018
出版刊次：4
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