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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。
Uneven leaf curl leaf disease is a new disease found in Haikou City, Hainan Province in 2012. The diseased plants showed curly leaf upward, swollen veins and dark green veins. PCR test results showed that the disease-like samples are present in golden bean mosaic virus. The results of gene cloning and sequence analysis showed that the HN08 genome contained only part A (DNA-A), which had a total length of 2 738 nt, which was closely related to the isolates of cotton leaf curl virus (CLCuMuV) Similarities were greater than 89.0%, of which the similarity with all isolates in China were greater than 99.0%. The virus isolate was also accompanied by a β-satellite molecule with a total length of 1 346 nt. The similarity to the CLCuMuB sequence of the CLCuMuV isolates was more than 83.0%. The sequence similarity with the Fz1 isolates was the highest, Is 99.7%. HN08 DNA-A and its β satellite molecularly invasive clones were constructed and inoculated into kenaf by Agrobacterium tumefaciens injection. After 30 days of inoculation, new leaves of kenaf plants inoculated with HN08 DNA-A and β satellite molecules began to produce koji Leaf symptom. At 60 days after inoculation, most of the leaves of the mixed plants showed severe leaf curl symptoms, which were the same as those of the natural plants in the field. However, the kenaf plants separately inoculated with them had no obvious symptoms. PCR and Southern blot analysis further confirmed that these symptoms were caused by co-infection of HN08 DNA-A and its beta satellite. Therefore, Hainan kenaf leaf curl disease is caused by co-infection of CLCuMuV and the accompanying satellite β (CLCuMuB). This paper reports for the first time that kenaf is a natural new host of CLCuMuV.