海南红麻曲叶病的病原检测与鉴定  

Detection and identification of the pathogen causing kenaf(Hibiscus cannabinus)leaf curl disease in Hainan Province of China

作　　者：汤亚飞[1,2] 何自福[1,2] 杜振国[1,2] 佘小漫[1,2] 蓝国兵[1,2] 

机构地区：[1]广东省农业科学院植物保护研究所,广州510640 [2]广东省植物保护新技术重点实验室,广州510640

出　　处：《植物病理学报》2015年第6期561-568,共8页Acta Phytopathologica Sinica

基　　金：广东省科技计划项目(2012A020200015);广州市科技计划项目(2013J4500032);广东省农业科学院院长基金(201424)

摘　　要：红麻曲叶病是2012年在海南省海口市发现的一种新病害,病株表现为叶片向上卷曲、叶脉肿大、叶脉变深绿色等症状。PCR检测结果显示,该病样中均存在菜豆金色花叶病毒属病毒。基因克隆及序列分析结果表明,该病毒分离物(HN08)基因组仅含A组分(DNA-A),其全长为2 738 nt,与木尔坦棉花曲叶病毒(CLCuMuV)各分离物的相似性均大于89.0%,其中与中国各分离物的相似性均大于99.0%。该病毒分离物也伴随有β卫星分子,其全长为1 346 nt,与CLCuMuV各分离物伴随的β卫星分子(CLCuMuB)序列相似性大于83.0%,其中与分离物Fz1的序列相似性最高,为99.7%。构建了HN08 DNA-A及其β卫星分子侵染性克隆,通过农杆菌注射接种红麻,接种后30 d,HN08 DNA-A及其β卫星分子混合接种的红麻植株新出叶片开始产生曲叶症状;接种后60 d,二者混合接种的植株大部分叶片表现为严重的曲叶症状,且与田间自然病株症状相同,而二者各自单独接种的红麻植株没有产生明显的症状。PCR及Southern blot检测进一步证实这些症状是由HN08 DNA-A及其β卫星分子的共同侵染引起的。因此,海南红麻曲叶病是由CLCuMuV及其伴随的β卫星分子(CLCuMuB)共同侵染引起的。本文首次报道了红麻是CLCuMuV自然新寄主。Kenaf leaf curl disease is a new disease occurred in Haikou, Hainan Province of China. The symptoms of the infected plants exhibits leaf curling upwards, vein swelling and vein dark green. PCR analysis indicates that the symptomatic plants were infected by begomoviruses and sequence analysis indicates that the isolate HN08 only contains the DNA-A component.The full length of DNA-A was 2 738 nucleotides. Multiple sequence alignment showed that the DNA-A of HN08 had more than 89.0% sequence identity with that of all other isolates of Cotton leaf curl Multan virus （CLCuMuV） deposited in GenBank, and more than 99.0% sequence identity with that of all isolates from China. The virus was also accompanied by betasatellite molecular,which was determined to be 1 346 nucleotides. Pairwise comparison indicated that the betasatellite molecular of HN08 had more than 83.0% sequence identity with that of Cotton leaf curl Multan betasatellite （CLCuMuB）, and shared the highest sequence identity （99.7%） with that of isolate Fz1 from Fujian. The infectious clones of HN08 DNA-A and its betasatellite molecular, pGreenⅡ049-1.6A and pGreenⅡ049-2.0β were constructed, and were agro-inoculated into Hibiscus cannabinus plants. At 30 dpi, the new leaves of the H. cannabinus plants inoculated with the recombination of pGreenⅡ049-1.6A and pGreenⅡ049-2.0β displayed leaf curl symptoms. At 60 dpi, most of the leaves of the inoculated H. cannabinus plants showed severe leaf curl symptoms, similar to that observed in the fields. However, neither the plants inoculated with pGreenⅡ049-1.6A or pGreenⅡ049-2.0β showed visible symptoms. The results of PCR and Southern blot detection confirmed these symptoms caused by co-infection of HN08 DNA-A and its betasatellite molecular. Taken together, Kenaf leaf curl disease in Hainan province was caused by co-infection of CLCuMuV and its betasatellite （CLCuMuB）. This is the first report of CLCuMuV on H. cannabinus .

关 键 词：红麻曲叶病 木尔坦棉花曲叶病毒 卫星分子 病原鉴定 

分 类 号：S432.41[农业科学—植物病理学]