Molecular characterization and detection of recombinant isolates of potato virus Y from China
Molecular characterization and detection of recombinant isolates of potato virus Y from China
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DOI：
10.1007/s00705-009-0448-z
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发表时间：
2009-08-01
影响因子：
2.7
通讯作者：
Nie, Xianzhou
中科院分区：
文献类型：
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作者：
Hu, Xinxi;He, Changzheng;Nie, Xianzhou
马铃薯Y病毒（PVY）是我国马铃薯生产上最重要的病原菌之一，但国内对该病毒的研究却很少。利用逆转录-聚合酶链反应（RT-PCR）基因分型技术，在中国首次鉴定出两种重组PVY。一个类似于欧洲（Eu）型马铃薯块茎坏死菌株（Eu-PVYNTN），具有普通菌株（PVYO）和Eu型烟草脉坏死菌株（Eu-PVYN）的三个广泛认可的重组接头（RJs 1-3）。另一方面，另一个似乎只有RJ 1和RJ 2。随后对来自第二种类型的代表性分离物PVY-HN 2的全基因组进行测序。将“HN 2”与PVYO和Eu-PVYN的序列进行比较，不仅证实了“HN 2”的重组性质，而且还揭示了在分离物中存在与PVYNTN-Hun类似的三个重组事件。然而，两个分离物在RJ 1（PVYNTN-Hun对HN 2，nt 2419对nt 2521）和RJ 3（PVYNTN-Hun对HN 2，nt 9183对nt 8572）处显著不同，并且在RJ 2（PVYNTN-Hun对HN 2，nt 5844对nt 5867）处略有不同。开发了一对引物以便于检测替代RJ 3。使用新的和以前设计的RJ引物，所有的目标RJ检测。有趣的是，对可用PVY样品的测试表明，两个被两种类型的重组PVY双重感染，进一步证实了检测的有效性。利用酶联免疫吸附试验和生物测定对这些样品进行进一步分析，结果表明，HN 2具有PVYO血清型、烟草PVYN致病型和马铃薯PVYNTN致病型。
Although potato virus Y (PVY) is one of the most economically important pathogens of potatoes in China, few studies have been carried out to characterize the virus in that country. Using reverse transcription-polymerase chain reaction (RT-PCR)-based genotyping developed previously, two types of recombinant PVY were identified in China for the first time. One resembled the European (Eu) type of potato tuber necrosis strain (Eu-PVYNTN), possessing three widely recognized recombinant joints (RJs 1-3) of the common strain (PVYO) and the Eu-type tobacco veinal necrosis strain (Eu-PVYN). The other, on the other hand, appeared to have only RJ1 and RJ2. The complete genome of a representative isolate, PVY-HN2, from the second type was subsequently sequenced. Comparison of the sequence of 'HN2' with those of PVYO and Eu-PVYN not only confirmed the recombinant nature of 'HN2' but also revealed the existence of three recombinant events in the isolate, similar to that in PVYNTN-Hun. However, the two isolates differed significantly at RJ1 (PVYNTN-Hun vs. HN2, nt 2419 vs. nt 2521) and RJ3 (PVYNTN-Hun vs. HN2, nt 9183 vs. nt 8572) and slightly at RJ2 (PVYNTN-Hun vs. HN2, nt 5844 vs. nt 5867). A primer pair was developed to facilitate the detection of the alternative RJ3. Using newly and previously designed RJ primers, all targeted RJs were detected. Interestingly, tests of the available PVY samples indicated that two were doubly infected with both types of recombinant PVY, further confirming the effectiveness of the detection. Further analysis of these samples using enzyme-linked immunosorbent assay and bioassay revealed that 'HN2' possesses a PVYO serotype, a PVYN pathotype in tobacco and a PVYNTN pathotype in potato.