Abstract
Geminiviruses and their diseases are a considerable economic threat to a vast number of crops worldwide. Investigating how and where these viruses replicate and accumulate in their hosts may lead to novel molecular resistance strategies. In this study, we used the Rep-inducible In Plant Activation (INPACT) expression platform, based on the genome of tobacco yellow dwarf virus (TYDV), to determine where this model mastrevirus replicates in its host tobacco. By developing an infectious clone of TYDV and optimizing its delivery by agroinfiltration, we first established an efficient artificial infection process. When delivered into transgenic tobacco plants containing a TYDV-based INPACT cassette encoding the β-glucuronidase (GUS) reporter, we showed the virus activates GUS expression. Histology revealed that reporter gene expression was limited to phloem-associated cell types suggesting TYDV replication has a restricted tissue tropism.
Highlights
Understanding virus–host interactions and the processes that occur during pathogenesis is fundamental to the development of virus control measures and resistance
We developed a deconstructed virus vector based on the genome of Tobacco yellow dwarf virus (TYDV) termed In Plant Activation as a means of biopharming a commercially important non-therapeutic protein, human vitronectin, and to express an industrial enzyme and a ribonuclease in Nicotiana species
Agrobacterium strains (AGL1, C58, and GV3101) harboring pART-TYDV-2mer were compared for their ability to initiate virus infection via agroinfiltration
Summary
Understanding virus–host interactions and the processes that occur during pathogenesis is fundamental to the development of virus control measures and resistance. While many studies have examined the tissue and cellular localization of different geminiviruses [3,4,5,6,7,8,9,10,11,12,13], relatively few have sought to determine whether replication of these viruses is limited to specific cell types. These latter studies have predominantly used three different approaches, in situ hybridization to detect DNA forms indicative of rolling circle replication (RCR) [14], immuno-localization based on the incorporation of the thymidine analogue, 5-bromo-2-deoxyuridine, into newly synthesized viral.
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