Transmission studies with Cassava brown streak Uganda virus (Potyviridae: Ipomovirus) and its interaction with abiotic and biotic factors in Nicotiana benthamiana
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Date
2010
Journal Title
Journal ISSN
Volume Title
Publisher
Journal of Virological Methods
Abstract
Cassava brown streak disease (CBSD), caused by two distinct species, Cassava brown streak Uganda virus
(CBSUV) and Cassava brown streak virus (CBSV), is a major constraint to cassava (Manihot esculenta Crantz)
production in Africa. Absence of infectious clones of CBSUV or CBSV and the lack of efficient means of
mechanical transmission of CBSD has hampered laboratory studies of this disease. Mechanical transmission,
achieved mainly by plant sap inoculation, is a widely used technique for characterizing plant
viruses. Efficient sap transmission of CBSUV/CBSV to the common laboratory host Nicotiana benthamiana
is essential for both basic and applied studies of the virus. We report here the development of an efficient
protocol for sap transmission of CBSUV to N. benthamiana and N. debneyi. Several factors affecting
transmission efficiency were identified such as the effects of buffer composition, antioxidants, inoculum
concentration, plant age and temperature. Higher temperatures (30 ◦C) favored rapid symptom initiation
compared to lower temperatures (21 ◦C) when sap prepared in phosphate buffer of pH 7.0 was applied
on the leaves of N. benthamiana dusted with the abrasive (carborundum). We demonstrated the usefulness
of the transmission method in transient evaluation of CBSUV[UG:Nam:04]-derived RNA interference
constructs for CBSD resistance and also in studying the interaction of CBSUV[UG:Nam:04] with cassava
mosaic geminiviruses, another important group of viruses infecting cassava.
Description
Keywords
Cassava brown streak virus, Potyviridae, Ipomovirus, Sap inoculation, Synergism, Transient studies
Citation
Ogwok, E., Patil, B. L., Alicai, T., & Fauquet, C. M. (2010). Transmission studies with Cassava brown streak Uganda virus (Potyviridae: Ipomovirus) and its interaction with abiotic and biotic factors in Nicotiana benthamiana. Journal of Virological Methods, 169(2), 296-304. doi:10.1016/j.jviromet.2010.07.030