Search bioRxiv⌕ Search

Biology subjects

Redinbaugh, M. G.

Publications and source records attributed to Redinbaugh, M. G..

2 recordsLinked to original sources

Characterization of three resistance-breaking isolates of sugarcane mosaic virus from Rwanda and implications for maize lethal necrosis

Maize lethal necrosis (MLN) is a devastating disease of maize caused by synergy between two viruses: maize chlorotic mottle virus (MCMV) and a potyvirus, most often sugarcane mosaic virus (SCMV). Throughout the 2010s, severe MLN outbreaks occurred in sub-Saharan East Africa including Kenya, Rwanda, and Ethiopia. In this study, we assessed the virulence of SCMV isolates collected from Rwanda by screening a panel of maize near isogenic lines containing different combinations of major potyvirus resistance loci. We discovered that the three Rwandan SCMV isolates tested could overcome all three potyvirus resistance loci even when used in combination, including one isolate that could asymptomatically infect all resistant lines tested. To understand how SCMV virulence may contribute to MLN, each of the three isolates were co-inoculated with MCMV on a panel of SCMV and MCMV resistant maize lines. No significant differences in MLN severity were observed for the Rwandan isolates compared to the reference SCMV isolates, indicating that increased virulence in SCMV single infection does not necessarily correlate with increased MLN severity in co-infection with MCMV. For all SCMV isolates tested, at least two potyvirus resistance loci were needed to reduce MLN severity, and maize lines with a combination of SCMV and MCMV resistance were most effective. Surprisingly, in some cases co-infection with MCMV facilitated SCMV infection of potyvirus resistant lines that SCMV could not infect alone. These results underscore the challenges of developing durable MLN resistance and highlight the importance of incorporating strong, multigenic potyvirus resistance into MLN resistance breeding programs.

plant biology↗

Rescue of the first Alphanucleorhabdovirus entirely from cloned complementary DNA: an efficient vector for systemic expression of foreign genes in maize and insect vectors.

Recent reverse genetics technologies have enabled genetic manipulation of plant negative-strand RNA virus (NSR) genomes. Here, we report construction of an infectious clone for the maize-infecting Alphanucleorhabdovirus maydis, the first efficient NSR vector for maize. The full-length infectious clone was established using agrobacterium-mediated delivery of full-length maize mosaic virus (MMV) antigenomic RNA and the viral core proteins (nucleoprotein N, phosphoprotein P, and RNA-directed RNA polymerase L) required for viral transcription and replication into Nicotiana benthamiana. Insertion of intron 2 ST-LS1 into the viral L gene increased stability of the infectious clone in Escherichia coli and Agrobacterium tumefaciens. To monitor virus infection in vivo, a GFP gene was inserted in between the N and P gene junctions to generate recombinant MMV-GFP. cDNA clones of MMV-WT and MMV-GFP replicated in single cells of agroinfiltrated N. benthamiana. Uniform systemic infection and high GFP expression were observed in maize inoculated with extracts of the infiltrated N. benthamiana leaves. Insect vectors supported virus infection when inoculated via feeding on infected maize or microinjection. Both MMV-WT and MMV-GFP were efficiently transmitted to maize by planthopper vectors. The GFP reporter gene was stable in the virus genome and expression remained high over three cycles of transmission in plants and insects. The MMV infectious clone will be a versatile tool for expression of proteins of interest in maize and cross-kingdom studies of virus replication in plant and insect hosts.

plant biology↗