Samarfard Samira, McTaggart Alistair R, Sharman Murray, Bejerman Nicolás E, Dietzgen Ralf G
Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, Queensland 4072, Australia.
Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, Ecosciences Precinct, Dutton Park, Queensland 4102, Australia.
Pathogens. 2020 Mar 13;9(3):214. doi: 10.3390/pathogens9030214.
Alfalfa plants in the field can display a range of virus-like symptoms, especially when grown over many years for seed production. Most known alfalfa viruses have RNA genomes, some of which can be detected using diagnostic assays, but many viruses of alfalfa are not well characterized. This study aims to identify the RNA and DNA virus complexes associated with alfalfa plants in Australia. To maximize the detection of RNA viruses, we purified double-stranded RNA (dsRNA) for high throughput sequencing and characterized the viromes of ten alfalfa samples that showed diverse virus-like symptoms. Using Illumina sequencing of tagged cDNA libraries from immune-captured dsRNA, we identified sequences of the single-stranded RNA viruses, alfalfa mosaic virus (AMV), bean leafroll virus, a new emaravirus tentatively named alfalfa ringspot-associated virus, and persistent dsRNA viruses belonging to the families and . Furthermore, rolling circle amplification and restriction enzyme digestion revealed the complete genome of chickpea chlorosis Australia virus, a mastrevirus (family ) previously reported only from chickpea and French bean that was 97% identical to the chickpea isolate. The sequence data also enabled the assembly of the first complete genome (RNAs 1-3) of an Australian AMV isolate from alfalfa.
田间的苜蓿植株可能会表现出一系列类似病毒的症状，尤其是在多年种植用于种子生产时。大多数已知的苜蓿病毒具有RNA基因组，其中一些可以通过诊断检测方法检测到，但许多苜蓿病毒的特征尚不明确。本研究旨在鉴定与澳大利亚苜蓿植株相关的RNA和DNA病毒复合体。为了最大限度地检测RNA病毒，我们纯化了双链RNA（dsRNA）用于高通量测序，并对十个表现出不同类似病毒症状的苜蓿样本的病毒组进行了特征分析。通过对免疫捕获的dsRNA的标记cDNA文库进行Illumina测序，我们鉴定出了单链RNA病毒苜蓿花叶病毒（AMV）、菜豆卷叶病毒、一种暂命名为苜蓿环斑相关病毒的新型埃玛病毒以及属于 和 科的持久性dsRNA病毒的序列。此外，滚环扩增和限制性内切酶消化揭示了澳大利亚鹰嘴豆褪绿病毒的完整基因组，该病毒属于微病毒科（ 科），此前仅在鹰嘴豆和法国豆中报道过，与鹰嘴豆分离株的同源性为97%。序列数据还使得我们能够组装出一株来自澳大利亚苜蓿的AMV分离株的首个完整基因组（RNA 1-3）。
