Infectivity of Tomato yellow leaf curl virus isolated from imported tomato fruit in Estonia

Abstract

There is a risk that Tomato yellow leaf curl virus (TYLCV) and its vector, whiteflies of the Bemisia tabaci species complex, will become established in greenhouses in temperate regions of the world, including northern Europe. In this study, TYLCV isolated from imported tomato fruit in Estonia (originating from Spain) was shown to be able to infect plants of tomato and Nicotiana benthamiana using Agrobacterium-mediated inoculation with an infectious clone as well as using biolistic delivery of products from rolling circle amplification (RCA). A 1.8-mer genomic construct of TYLCV was engineered and efficiently agroinfiltrated into plants of tomato and N. benthamiana, and induced symptoms characteristic of natural infection. With Agrobacterium-mediated inoculation, the infection efficiency was 100% for both tomato and N. benthamiana, whereas biolistic inoculation using RCA products resulted in efficiencies of 57% and 36%, respectively. Particle bombardment with monomeric linear genome failed to produce any infection in tomato or N. benthamiana. The genome of TYLCV amplified from tomato fruit was infectious confirming that tomato fruit may serve as a source of virus inoculum. This is the first report of agroinfiltration and particle bombardment assay using TYLCV DNA derived from infected tomato fruit tissue.

The Peptide AmPep1 Derived from Amaranth Recognizes the Replication Hairpin of TYLCV Disturbing Its Replication Process in Host Plants

“…HF548826) was prepared for agroinoculation as previously described. 22 2.2. Oligonucleotide and Peptides for Biochemical Studies.…”

“…The antiviral activity of the AmPep1 peptide was evaluated in the natural host tomato, by inoculating tomato seedlings (containing 2 true leaves) with TYLCV-IL. 22 At 10 dpi, new apical leaves started to show symptoms, such as curling. Tissue from apical leaves was then collected and DNA was extracted to verify systemic infection using PCR.…”

“…Agrobacterium tumefaciens C58C1 containing a 1.8-mer construct of the TYLCV genome (Accession no. HF548826) was prepared for agroinoculation as previously described …”

In Planta Interaction and Transreplication of Distinct Begomoviruses and their Associated Components

“…Infectious constructs of ToLCNDV DNA-A ([hereafter referred to as TV]; accession# U15015), ToLCNDV DNA-B ([hereafter referred to as TB]; accession# U15016), PeLCV ([hereafter referred to as PV]; accession# AM712436) and TbLCuB ([hereafter referred to as TbB]; accession# AM955608) (Padidam et al 1995;Ilyas et al 2010). All the constructs were transformed into GV3101 Agrobacterium strain using electroporation (Just et al 2017).…”

“…After electrophoresis, the DNA were transferred onto Hybond nylon membranes (Amersham, the Netherlands), probed with radioactively [P-32P] dCTP labelled PCR-amplified CP gene fragment for TV and PV, MP fragment for TB, and βC1 gene fragment for TbB, respectively. The protocol from Just et al (2017) was followed for Southern blot hybridization and phosphorimager (Bio-Rad) was used to detect hybridization signals.…”