A geminivirus crosses the monocot-dicot boundary and acts as a viral vector for gene silencing and genome editing

Jitendra Kumar; Anshu Alok; Brian J Steffenson; Shahryar Kianian

doi:10.1016/j.jare.2023.09.013

A geminivirus crosses the monocot-dicot boundary and acts as a viral vector for gene silencing and genome editing

J Adv Res. 2023 Sep 18:S2090-1232(23)00262-X. doi: 10.1016/j.jare.2023.09.013. Online ahead of print.

Authors

Jitendra Kumar¹, Anshu Alok², Brian J Steffenson², Shahryar Kianian³

Affiliations

¹ Department of Agronomy and Plant Genetics, University of Minnesota, Saint Paul, MN 55108, United States.
² Department of Plant Pathology, University of Minnesota, Saint Paul, MN 55108, United States.
³ USDA-ARS Cereal Disease Laboratory, Saint Paul, MN 55108, United States. Electronic address: shahryar.kianian@usda.gov.

PMID: 37730118
DOI: 10.1016/j.jare.2023.09.013

Abstract

Introduction: Members of the family Geminiviridae have been reported to infect either a monocot plant or a dicot plant, but not both. This study reports a geminivirus, Wheat Dwarf India Virus (WDIV), first identified in wheat, that is capable of infecting both monocot and dicot plants and acting as a viral vector.

Objectives: This study was aimed at developing a broad host range viral vector system for reverse genetics and genome editing.

Methods: Here we used a wheat isolate of WDIV and Ageratum yellow leaf curl betasatellite (AYLCB) for infectivity assays and vector development. We performed Agrobacterium-mediated inoculation of WDIV and AYLCB in wheat, oat, barley, corn, soybean, and tobacco. To examine the potential of WDIV to act as a viral vector, we modified the WDIV genome and cloned DNA fragments of the phytoene desaturase (PDS) genes from wheat and tobacco, separately. For gene editing experiments, tobacco lines expressing Cas9 were infiltrated with a WDIV-based vector carrying gRNA targeting the PDS gene.

Results: About 80 to 90% of plants inoculated with infectious clones of WDIV alone or WDIV together with AYLCB showed mild symptoms, whereas some plants showed more prominent symptoms. WDIV and AYLCB were detected in the systemically infected leaves of all the plant species. Furthermore, the inoculation of the WDIV vector carrying PDS fragments induced silencing of the PDS gene in both wheat and tobacco plants. We also observed high-efficiency genome editing in the Cas9-expressing tobacco plants that were inoculated with WDIV vector-carrying gRNA.

Conclusion: Detection of WDIV in naturally infected wheat, barley, and sugarcane in the field and its ability to systemically infect wheat, oat, barley, corn, soybean, and tobacco under laboratory conditions, provides compelling evidence that WDIV is the first geminivirus identified with the capability of infecting both monocot and dicot plant species. The wide host range of WDIV can be exploited for developing a single vector system for high-throughput genome editing in many plant species.

Keywords: AYLCB; Gene silencing; Genome editing; Monocot-dicot barrier; WDIV.