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ARS Home » Midwest Area » Wooster, Ohio » Corn, Soybean and Wheat Quality Research » Research » Research Project #440982

Research Project: In Planta Movement Dynamics of Maize-Infecting Potyviruses from the Subcellular to Organismal Level

Location: Corn, Soybean and Wheat Quality Research

Project Number: 5082-22000-002-004-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Sep 1, 2021
End Date: Dec 31, 2022

Objective:
Our overall objective is to understand potyvirus and potyviral protein subcellular and systemic movement. Objective 1 will examine the subcellular movement dynamics of two potyviral proteins of maize-infecting potyviruses. Objective 2 will examine cell-to-cell and long-distance movement of maize-infecting potyviruses in the context of recently identified resistance genes.

Approach:
Objective 1. Subcellular movement dynamics of two proteins of maize- infecting potyviruses, such as sugarcane mosaic virus (SCMV, Family Potyviridae: Genus Potyvirus), maize dwarf mosaic virus (MDMV, Potyviridae: Potyvirus), or wheat streak mosaic virus (WSMV, Potyviridae: Tritimovirus), will be observed in a heterologous system using the model plant Nicotiana benthamiana. Recombinant fluorescently tagged forms of these proteins will be transiently expressed singly or doubly in N. benthamiana via agroinfiltration and subcellular localization observed via laser-scanning confocal microscopy. Further investigation may include observing these proteins in the context of natural infection of maize using fluorescently tagged infectious clones of the viruses or immunostaining of wild- type forms of the viruses in singly infected maize. Different combinations of these 2 proteins from SCMV, MDMV, and WSMV may be considered. Objective 2. Cell-to-cell and long-distance movement of SCMV, MDMV, and WSMV will be observed in maize containing a wheat streak mosaic resistance gene which confers resistance to WSMV but not the other two potyviruses. Previously developed fluorescently tagged infectious clones of each of these viruses will be used to mechanically inoculate resistant (OH28wsm3) and susceptible (OH28) near-isogenic lines of maize. Virus movement will be visualized using epifluorescent and/or laser scanning confocal microscopy at various time points during infection with each virus. Double or single virus infections may be considered as well as germplasm containing other known potyvirus resistance genes/loci in maize.