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.
| Original language | English (US) |
|---|---|
| Journal | Journal of Advanced Research |
| DOIs | |
| State | Accepted/In press - 2023 |
Bibliographical note
Funding Information:Funding for the project was provided by the U.S. Department of Agriculture - Agricultural Research Service. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture. We thank Roger Casper, USDA-ARS Cereal Disease Lab, for excellent technical assistance and Dr. Daniel Voytas, University of Minnesota for providing the Cas9 expressing N. benthamiana lines. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. USDA is an equal opportunity provider and employer.
Funding Information:
Funding for the project was provided by the U.S. Department of Agriculture - Agricultural Research Service. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture. We thank Roger Casper, USDA-ARS Cereal Disease Lab, for excellent technical assistance and Dr. Daniel Voytas, University of Minnesota for providing the Cas9 expressing N. benthamiana lines. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. USDA is an equal opportunity provider and employer.
Publisher Copyright:
© 2023
Keywords
- AYLCB
- Gene silencing
- Genome editing
- Monocot-dicot barrier
- WDIV
PubMed: MeSH publication types
- Journal Article
