Three sensitive and reliable serological assays for detection of potato virus A in potato plants

Vegetative propagation of seed potato often allows passaging of viruses to seed tubers,resulting in significant yield losses and reduction of potato tuber quality.Thus,virus detection approach is crucial for effective virus management programs and the production of virus-free seed potatoes.Among the reported potato-infecting viruses,potato virus A(PVA)is considered as one of the most important viruses in potato-growing regions worldwide.This study prepared four hybridoma lines secreting PVA-specific monoclonal antibodies(MAbs)(2D4,8E11,14A6 and 16H10)using purified PVA virions as an immunogen.Western blotting results indicated that all the four MAbs reacted strongly and specifically with the putative capsid protein of PVA.Using these four MAbs,this study developed antigen-coated plate enzyme-linked immunosorbent assay(ACP-ELISA),Dot-ELISA and Tissue print-ELISA for detection of PVA infection in potato plants.The results indicated that PVA can be detected in crude tissue extracts from infected potato plants diluted up to 1:327680(w/v,g m L^(-1))by ACP-ELISA or up to1:10240 by Dot-ELISA.The Tissue print-ELISA is the quickest and easiest approach among the three serological assays,and is more suitable for onsite large-scale potato screening programs.Further analyses of field-collected potato samples showed that the sensitivities and specificities of the three serological approaches were similar to those of RT-PCR in PVA detection and confirmed that PVA is currently widespread in Yunnan and Zhejiang provinces of China.Hence,the results strongly suggest that these highly sensitive serological approaches based on PVA-specific MAbs are useful and powerful for PVA-free seed potato production programs and PVA field surveys.

Prevalence and Spatial Distribution of Badnavirus in the Banana (Musa spp) Major Growing Areas in Burkina Faso

Banana streak virus (BSV) and Sugarcane bacilliform virus (SCBV) are two badnaviruses commonly found in all banana growing areas of the world. It is a threat to the production and improvement of Musa germplasm. In Burkina Faso, the presence of badnaviruses was reported in banana producing regions. The objective of this study was to determine the prevalence of BSV and SCBV in banana production areas of Burkina Faso. A survey followed by a symptomatologic study was conducted in banana plantations in 27 localities of the nine main banana producing regions from July to October 2018 and September to December 2020. In all, 251 leaf samples were collected and analysed for BSV and SCBV infection by Indirect Antigen Coated Plate Assay-ELISA followed by amplification of the RT/RNase H region using Polymerase chain reaction with Badna FP/RP and SCBV F/R primers, respectively. A variety of symptoms were observed on almost all plant organs which were revealed due to BSV by symptomatologic study. The results of serological and molecular diagnosis revealed a high overall prevalence of BSV in 80.48% of the samples tested. BSV was distributed in seven survey regions out of nine with prevalence ranging from 10% to 100% in North, Centre, Centre West, Hauts Bassins, Cascades, Centre East and Boucle of Mouhoun regions. Very low prevalence was recorded for SCBV in Cascades and East Centre region with 4.35 and 12.5%, respectively. Species detection using specific primers to each species revealed three main species: Banana streak Obino l’ewaï virus (BSOLV), Goldfinger virus (BSGFV) and Imové virus (BSIMV) in the samples tested, respectively in the proportions of 23%, 8% and 0.8%. Co-infection between BSV species was also detected.