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Deep sequencing of banana bract mosaic virus from flowering ginger (Alpinia purpurata) and development of an immunocapture RT-LAMP detection assay

机译:从开花姜中提取香蕉br片花叶病毒的深度测序和免疫捕获RT-LAMP检测方法的开发

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Banana bract mosaic virus (BBrMV) has never been reported in banana plants in Hawaii. In 2010, however, it was detected in a new host, flowering ginger (Alpinia purpurata). In this study, we characterize the A. purpurata isolate and study its spread in flowering ginger in Hawaii. A laboratory study demonstrated that BBrMV could be transmitted from flowering ginger to its natural host, banana, therefore raising a serious concern about the potential risk to the rapidly growing banana industry of Hawaii. To quickly monitor this virus in the field, we developed a robust immunocapture reverse transcription loop-mediated isothermal amplification (IC-RT-LAMP) assay. Deep sequencing of the BBrMV isolate from A. purpurata revealed a single-stranded RNA virus with a genome of 9,713 nt potentially encoding a polyprotein of 3,124 aa, and another predicted protein, PIPO, in the +2 reading-frame shift. Most of the functional motifs in the Hawaiian isolate were conserved among the genomes of isolates from one found in the Philippines and India. However, the A. purpurata isolate had an amino acid deletion in the Pl protein that was most similar to the Philippine isolate. Phylogenetic analysis of an eastern Pacific subpopulation that included A. purpurata was closest in genetic distance to a Southeast Asian subpopulation, suggesting frequent gene flow and supporting the hypothesis that the A. purpurata isolate arrived in Hawaii from Southeast Asia.
机译:夏威夷的香蕉植物中从未报道过香蕉片花叶病毒(BBrMV)。然而,在2010年,在新寄主开花姜(Alpinia purpurata)中发现了它。在这项研究中,我们表征了紫癜的分离物,并研究了其在夏威夷开花姜中的传播。一项实验室研究表明,BBrMV可以从开花的姜中传播到其天然寄主香蕉中,因此引起了人们对夏威夷迅速发展的香蕉产业的潜在风险的严重关注。为了在现场快速监测该病毒,我们开发了一种强大的免疫捕获逆转录环介导的等温扩增(IC-RT-LAMP)分析。对紫花苜蓿的BBrMV分离株进行深度测序后,发现单链RNA病毒的基因组为9,713 nt,可能编码3,124 aa的多聚蛋白,而另一个预测的蛋白PIPO则处于+2阅读框移位。夏威夷分离物中的大多数功能性基序在来自菲律宾和印度的一种分离株的基因组中是保守的。然而,紫癜曲霉分离物在P1蛋白中具有与菲律宾分离物最相似的氨基酸缺失。对东太平洋亚群(包括紫癜)的系统发育分析在遗传距离上最接近东南亚亚群,这表明基因流频繁,并支持了紫癜菌从东南亚到达夏威夷的假说。

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