Molecular Identification for Sweet Potato Leaf Curl Virus Isolated in Fujian
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摘要: 为了鉴定福建省甘薯曲叶病毒Sweet potato leaf curl virus，SPLCV，并分析其病原的分子特征，根据已报道的SPLCV基因组序列设计特异性引物，对获得的特异性片段进行回收、克隆、测序。根据测序结果设计了扩增SPLCV基因组全长引物（SPLCVQ），同时利用已报道的DNA-B的通用引物PCRc1/PBLlv进行扩增，克隆获得SPLCV分离物基因组全长为2 827 bp，该序列含有6个开放阅读框，只含有DNA-A组分，不含有DNA-B组分（GenBank登录号：KT202303）。核苷酸同源性分析结果表明，该分离物与河南、江苏、浙江、韩国、美国亲缘关系最近，处于同一亚组的不同分支中。Abstract: Molecular characteristics of the sweet potato leaf curl virus (SPLCV) in Fujian province were studied. Specific segments of the genome were recovered, cloned and sequenced using selected primers. A sequence analysis positively confirmed the segments to be a part of SPLCV. Then, the full-length SPLCV was constructed with a pair of primers Q, and sequenced. The previously reported general purpose primers, PCRc1/PBLlv, for DNA-B were used for the subsequent amplification. The genome of SPLCV was found to be 2 827 bp containing 6 ORFs with DNA-A but not DNA-B (GenBank accession number:KT202303). The nucleotide homology between the isolated genome and the reported 6 other geminiviridae genomes was less than 85%;and, between it and SPLCV was the highest at 95.94%. In addition, the isolate shared a close genetic relationship with other isolates from Henan, Jiangsu, Zhejiang, South Korea, and the United States belonging to a same subgroup but in different branches.
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Key words:
- sweet potato /
- SPLCV /
- sequence analysis /
- genome
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图 1 感染甘薯曲叶病毒甘薯植株症状
注：A为曲叶，B为对照。
Figure 1. Symptoms on sweet potatoes infected by SPLCV
图 3 甘薯曲叶病毒福建分离物全长序列PCR产物电泳结果
Figure 3. Agarose gel electrophoresis on PCR products of full-length SPLCV isolate from Fujian
图 4 基于SPLCV核苷酸序列NJ树
Figure 4. Neighbor-joining tree based on nucleotide sequence of SPLCV
表 1 用于病毒鉴定引物
Table 1. Primers used for begomovirus characterization
引物 序列 引物用途 产物大小/bp SPLCVF CCGGTAAGACGGAGGCTGAACTTCG 扩增CP基因引物 0.6 SPLCVR AAATAAGCCCTGCAACGCAG SPLCVQF GCTGCCCGAAGCTATGTCCC 扩增全长引物 2.8 SPLCVQR AATTGGGACAGCATTCCCAG Beta0lF GGTACCACTACGCTACGCAGCAGCC 检测卫星DNA分子DNAβ 1.3 Beta02R GGTACCTACCCTCCCAGGGGTACAC PCRc1F CATATTTACRARWATGCCA 检测双生病毒DNA-B组分的存在 2.8 PBLlvR CARTGRTCKATCTTCATACA 
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表 2 甘薯曲叶病毒福建分离物和已报道的双生病毒基因组同源性分析
Table 2. Homology between isolates from Fujian and reported geminiviridae
病毒名称 DNA-A 病毒链编码 互补链编码 AV1 AV2 AC1 AC2 AC3 AC4 SPLCCaV 84.56 93.60 90.20 82.21 89.04 88.28 75.68 SPLCCNV 81.04 87.60 75.37 81.83 78.20 78.9 89.53 SPLCGoV 78.27 89.28 76.94 78.19 78.28 74.89 72.09 SPLCSPV 78.52 85.12 71.29 81.92 76.67 73.29 77.27 SPLCSCV 80.48 86.55 75.94 84.84 80.00 74.43 83.72 SPLCV 95.94 95.69 86.47 97.54 90.60 88.97 98.84 SPLCUV 83.17 86.16 73.97 84.22 90.83 87.13 81.82 SPMoV 79.34 86.03 71.78 82.01 - 74.20 - SPLCV-FJ - - - - - - -
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