The Incidence and Molecular Characterization of Banana Bunchy Top Virus (BBTV) Infecting Banana (Musa spp.) in Ismailia governorate, Egypt

H. Amin, Azza G. Farag, Abeer Abouelwafa
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Abstract

The banana is the most important fruit in Egypt. Banana bunchy top disease, caused by the banana bunchy top virus (BBTV), is one of the most damaging infections on banana plantations, resulting in significant crop losses. The current study was conducted to characterize the incidence of banana bunchy top disease (BBTD) and to study the molecular characteristics of BBTV from banana plantations in Ismailia. This study was conducted during the period 2019-2021 in three locations (Abu-Suwayr, El Tal-El kabir and Fayed) in Ismailia Governorate. The occurrence of BBTV was initially studied using visual inspection of external virus symptoms. TAS-ELISA, PCR and nucleic acid hybridization were used to detect the virus. Further molecular characterization of BBTV was performed based on Rolling Circle Amplification (RCA) and nucleotide sequencing. BBTD incidences ranging from 3.33 to 5.14% were observed in two localities Abu-Suwayr and El Tal-El kabir. While a relatively low incidence of 1.48% was observed in Fayed. Our results showed that the BBTD incidence is directly related to the percentage of winged aphids inside the aphid colonies regardless the percentage of the aphid's occurrence. The PCR product (973 bp) of BBTV-component 3 (DNA-S) was used for the creation of a non-radioactive Dig labeled-DNA probe for further use for BBTV detection using dot blot and Southern blot hybridization. The sequence comparison analysis of DNA-S component showed that the present Ismailia isolate (BBTV-3F) shared nucleotide identity 98.4%- 91.9 % with three isolates from Qalubya Egypt and shared 97.9% nt identity with east African isolates (Rwanda and Malawi). The phylogenetic analysis revealed that the DNA-component 3 (DNA-S) of the BBTV-3F isolate from Ismailia belongs to the South Pacific group (SPG) and is markedly separated from those of the Asian group (AG). This study is important for monitoring and early detection of BBTV threats and for rapid eradication of infected plants before the virus spreads and settles in a new area.
埃及伊斯梅利亚省香蕉束顶病毒(BBTV)的发病及分子特征
香蕉是埃及最重要的水果。香蕉束顶病是由香蕉束顶病毒(BBTV)引起的,是对香蕉种植园最具破坏性的感染之一,造成重大的作物损失。本研究旨在研究伊斯梅利亚地区香蕉束顶病(BBTD)的发病特征和BBTV的分子特征。这项研究于2019-2021年期间在伊斯梅利亚省的三个地点(Abu-Suwayr、El Tal-El kabir和Fayed)进行。BBTV的发生最初是通过外部病毒症状的目视检查来研究的。采用TAS-ELISA、PCR和核酸杂交检测病毒。基于滚动圈扩增(RCA)和核苷酸测序对BBTV进行了进一步的分子表征。Abu-Suwayr和El Tal-El kabir两个地区的BBTD发病率为3.33% ~ 5.14%。而法耶德的发病率相对较低,为1.48%。结果表明,无论蚜虫的发生比例如何,BBTD的发病率与蚜虫群体内有翅蚜虫的比例直接相关。利用BBTV-component 3 (DNA-S)的PCR产物(973 bp)构建无放射性Dig标记dna探针,进一步利用dot blot和Southern blot杂交技术进行BBTV检测。DNA-S序列分析表明,该Ismailia分离株(BBTV-3F)与埃及Qalubya分离株的核苷酸同源性为98.4% ~ 91.9%,与东非(卢旺达和马拉维)分离株的核苷酸同源性为97.9%。系统发育分析表明,Ismailia BBTV-3F分离株的dna组分3 (DNA-S)属于南太平洋群(SPG),与亚洲群(AG)明显分离。这项研究对于监测和早期发现BBTV威胁,以及在病毒传播和在新的地区定居之前迅速根除受感染的植物具有重要意义。
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