Utilization of infectious clones to visualize Cassava brown streak virus replication in planta and gain insights into symptom development

Virus Genes
Katie R TomlinsonGary D Foster

Abstract

Cassava brown streak disease (CBSD) is a leading cause of cassava yield losses across eastern and central Africa and is having a severe impact on food security across the region. Despite its importance, relatively little is known about the mechanisms behind CBSD viral infections. We have recently reported the construction of Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) infectious clones (IC), which can be used to gain insights into the functions of viral proteins and sequences associated with symptom development. In this study, we perform the first reporter gene tagging of a CBSV IC, with the insertion of green fluorescent protein (GFP) sequence at two different genome positions. Nicotiana benthamiana infections with the CBSV_GFP ICs revealed active CBSV replication in inoculated leaves at 2-5 days post inoculation (dpi) and systemic leaves at 10-14 dpi. We also constructed the chimera CBSV_UCP IC, consisting of the CBSV genome with a UCBSV coat protein (CP) sequence replacement. N. benthamiana infections with CBSV_UCP revealed that the CBSV CP may be associated with high levels of viral accumulation and necrosis development during early infection. These initial manipulations pave the way for...Continue Reading

References

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Citations

Aug 24, 2020·Pesticide Biochemistry and Physiology·Lin CaiXianchao Sun
Jul 17, 2021·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Herieth Rhodes MeroErik Bongcam-Rudloff

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Datasets Mentioned

BETA
MK409379
MK409380
MK409381

Methods Mentioned

BETA
PCR
reverse transcription PCR
Enzyme Linked Immunosorbent Assay
Fluorescence
amplicon sequencing
Confocal microscopy

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