Complete Nucleotide Sequence of Australian Tomato spotted wilt virus Isolate TSWV-QLD2

Abstract

The complete nucleotide (nt) sequence of an Australian isolate of Tomato spotted wilt virus was determined by deep RNA sequencing and deep small RNA sequencing. The tripartite genome consists of an 8,914-nt L segment, a 4,851-nt M segment, and a 2,987-nt S segment.

Investigation of virus-induced defence modulations during plant-virus interactions

“…That is why I dedicate this thesis to you, because you have made me who I am today and I would never have achieved as much as I have if it wasn't for you! Moyle, Carvalhais et al (2017). The pGrDL_SPb plasmid is used as a quantitative reporter system for validating possible viRNAs target transcripts via transient expression.…”

“…benthamiana leaves were infiltrated as previously described in Moyle et al (2017). Plants were left in normal growing condition for three days.…”

“…Selected predicted transcript target sequences were cloned into the dual luciferase plasmid pGrDL_SPb (Moyle et al 2017 Table 1a and b). Approximately 7 nM of each primer pair were combined into a 1.5 mL microfuge tube in a volume of ~ 50-60 µL and annealed by heating to 95°C in a waterbath, followed by slowly cooling down to room temperature (at a rate of ~ 1 degree per minute).…”

“…Dual luciferase assays were performed using a modified transient Agroinfiltration system based on the protocol described by Moyle et al (2017). Plasmids were transformed into Agrobacterium strain GV3101 and preparation of cultures to OD 0.5 for Agroinfiltration was performed as previously described (Moyle et al 2017).…”

“…Through deep small-RNA sequencing I characterise and profile the viRNAs generated from TuMV. A recent report described an optimised transient dual luciferase assay used to assess predicted miRNA-target sequence interactions (Moyle et al 2017). I investigated the utility of a similar transient dual luciferase assay system to experimentally validate predicted target sequences of viRNAs.…”