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首页> 外文期刊>Journal of Plant Pathology >CITRUS TRISTEZA VIRUS ON THE ISLAND OF CRETE: A SURVEY AND DETECTIONTI CITRUS TRISTEZA VIRUS ON THE ISLAND OF CRETE: A SURVEY AND DETECTION PROTOCOL APPLICATIONS
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CITRUS TRISTEZA VIRUS ON THE ISLAND OF CRETE: A SURVEY AND DETECTIONTI CITRUS TRISTEZA VIRUS ON THE ISLAND OF CRETE: A SURVEY AND DETECTION PROTOCOL APPLICATIONS

机译：CRETE TRISTEZA病毒在克里特岛上的研究与检测方法CITRUS TRISTEZA病毒在克里特岛上的研究和检测方法应用

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摘要

Over a period of two years, more than 5,000 citrus trees were tested for the presence of Citrus tristeza virus (CTV) on the island of Crete, resulting in thirty eight positives. Comparisons of the relative transcript levels of CTV p23, coat protein (CP), polymerase (POL) and an intergenic (POL/p33) region using quantitative RT-PCR, revealed consistent differences in abundance for each of these RNAs among flowers, stems, young fruits and leaves of infected orange trees. CTV p23 RNAs accumulated at highest levels, reaching a maximum in the flowers and the lowest levels in the leaves, while POL RNAs consistently accumulated at low levels in all tissues tested. A PCR-amplified dig-labelled CTV p23 DNA probe was applied to stem and leaf prints, and to crude and total RNA leaf extracts, using non isotopic hybridization. This technique, when applied to stem or leaf prints, and particularly to total RNA, unequivocally provided strong signals with minimal backgrounds. Moreover, an antiserum with high sensitivity and specificity of CTV detection as shown by DAS and immuno-print ELISA was produced against bacterially-expressed CTV CP. In the former method, stems and flowers contained higher levels of CTV CP when compared to leaf extracts. Taking into account Cretan geography and the importance of citrus to the island, systematic surveys for CTV eradication, sustainable control measurements and epidemiological studies need to be undertaken. The observations, materials and methods presented here may assist all three tasks at local and national level.
机译：在两年的时间里，测试了超过5,000棵柑橘树在克里特岛上是否存在柑桔柑橘（CTV），产生38例阳性。使用定量RT-PCR比较CTV p23，外壳蛋白（CP），聚合酶（POL）和基因间（POL / p33）区域的相对转录水平，发现花朵，茎，年轻的果实和感染的橙树的叶子。 CTV p23 RNA的积累水平最高，在花中达到最大值，在叶子中达到最低水平，而POL RNA在所有测试的组织中始终以低水平积累。使用非同位素杂交，将PCR扩增的带有dig标记的CTV p23 DNA探针应用于茎和叶印迹以及粗制和总RNA叶片提取物。当将这种技术应用于茎或叶的印刷品，特别是总RNA时，无疑会提供具有最小背景的强信号。而且，针对细菌表达的CTV CP产生了如DAS和免疫印迹ELISA所示的具有高灵敏度和CTV检测特异性的抗血清。在前一种方法中，与叶提取物相比，茎和花中CTV CP含量更高。考虑到克里特岛的地理环境和柑橘在岛上的重要性，需要进行系统的CTV根除调查，可持续控制措施和流行病学研究。此处介绍的观察结果，材料和方法可能会在地方和国家级协助完成所有这三个任务。

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