Pepper mild mottle virus: A plant pathogen with a greater purpose in (waste)water treatment development and public health management.

During 2014 and 2015, commercial crops and germplasm collections of chilli ( Capsicum spp.) in Italy were surveyed for the presence of viral diseases. In 2015, 2% of plants from two fields located in different provinces of the Lazio region…

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First report of Pepper vein yellows virus infecting chilli pepper ( Capsicum spp.) in Italy

Pepper mild mottle virus (PMMoV), a tobamovirus of family Virgaviridae affects the quality and quantity of Capsicum. PMMoV is highly contagious, capable of transmitting through infected seeds and soil. Symptoms are more severe when crop is infected at young stage but remain unnoticed when infection takes place at maturity. Therefore, cost effective diagnostic techniques are required for timely and accurate detection of virus. In present study, coat protein encoding region of PMMoV-HP1 isolate was cloned into expression vector system, pET28a and expressed in BL21, a protease deficient strain of Escherichia coli. The PMMoV-HP1 pathotype was identified as PMMoV-P12 on the basis of coat protein amino acid sequence analysis in our previous study. The overexpression of recombinant coat protein of 26 kDa, corresponding to the expected 6X Histidine tag fused recombinant protein was purified using Ni–NTA columns from insoluble fraction. For antisera production, the purified recombinant protein was dialyzed ~ 24 h to remove urea and then used for raising polyclonal antisera. The specificity and sensitivity of antiserum obtained was demonstrated using different dilutions of antiserum for western blot assay and direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA). In Western blot assay, the test antiserum reacted strongly both with PMMoV-CP in purified protein and native CP in crude sap from PMMoV infected pepper plants, whereas no reaction was observed with healthy plant sap. In DAC-ELISA antiserum dilution up to 1:1000 was capable of detecting the virus in infected sample. The absence of any cross reactivity of test antiserum was confirmed against tobacco mosaic virus, cucumber mosaic virus, tomato spotted wilt virus, pepper veinal mottle virus, potato virus Y and tomato yellow leaf curl virus antigen, known to infect capsicum.

Heterologous expression of pepper mild mottle virus coat protein encoding region and its application in immuno-diagnostics.

Revealing new information from existing genomic data for pepper mild mottle virus pathotype determination

This datasheet on Pepper vein yellows virus covers Identity, Distribution, Hosts/Species Affected, Further Information.

Pepper vein yellows virus

Chilli pepper ( Capsicum annuum ) is an important cash crop in Pakistan which is cultivated on an area of 62,500 ha with an annual yield of 145,100 tonnes (Farooq, 2014). Pepper mild mottle virus (PMMoV) is a single-stranded RNA virus…

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First report of Pepper mild mottle virus infecting chilli pepper in Pakistan

During June 2014, approximately 65% of hot pepper (Capsicum annum) plants in a commercial field in Multan district of Pakistan showed symptoms of curling, deformation, and interveinal leaf yellowing. Such symptoms are usually produced by either nutritional disorders or polerovirus infection (Tomassoli et al. 2016). Twenty randomly collected symptomatic and two asymptomatic (control) leaf samples were tested by double antibody sandwich (DAS)-ELISA using antiserum of Cucurbit aphid-borne yellows virus (Sediag, Longvic, France), which is known to cross-react with Pepper vein yellows virus (PeVYV) (Afouda et al. 2017). Seven of the 20 symptomatic samples were positive in DAS-ELISA. Total RNA from all ELISA-positive samples isolated by TRIzol reagent (Life Technologies, Carlsbad, CA) was used in one-step RT-PCR using the generic primers targeting the RdRp coding conserved sequence of Polerovirus (Lotos et al. 2014). The resulting amplicons were purified using Amicon Ultra kit (EMD Millipore, Billerica, MA) and directly sequenced in both orientations (BioFab, Rome, Italy). The sequences were identified as 594 bp of the P2 protein gene of the PeVYV genome using BLASTn analysis. The sequences of four isolates were found to be identical, so the sequences of three isolates were submitted to GenBank under accession numbers KX121043–45. Pakistani isolates shared 98 to 99% nucleotide sequence identities with each other and 93 to 96% with previously reported PeVYV isolates. All isolates shared 96% identity with Japanese (LC126041–45) and Israeli (HM439608) isolates and 94% identity with isolates from Australia (KU999109) and Greece (LT559484). To our knowledge, this is the first report of PeVYV from Pakistan. As in other countries around the world, the presence of PeVYV in Pakistan is threatening hot pepper cultivation. The fact that poleroviruses are aphid transmitted (Dombrovsky et al. 2010) supports the hypothesis that this virus may be present in other crops in various parts of the country. Therefore, further studies regarding incidence and distribution of this virus are needed.

L. Tomassoli

Papers

First report of Pepper mild mottle virus infecting chilli pepper in Pakistan

First Report of Pepper vein yellows virus Infecting Hot Pepper in Pakistan