A new use of Agrobacterium plant growth regulator genes for plant bioengineering

Delivery of biomolecules into plant vascular tissues remains a barrier to managing diseases caused by insect vector-borne pathogens and to modifying phenotypes of established perennial crops. Inspired by the vascularized growth of crown galls induced by Agrobacterium tumefaciens , we repurposed the bacterium’s plant growth regulator (PGR) genes to engineer autonomously dividing, transgene-expressing plant cell structures termed symbionts. A plant transformation vector (pSYM) incorporating the IaaM, IaaH, Ipt and gene5 cassette from A. tumefaciens strain C58 together with a gene of interest on the same transfer DNA was delivered to stems of herbaceous and woody dicots using disarmed A. tumefaciens strain EHA105. Symbiont morphology, vascular differentiation, transgene expression, molecular mobility and protein secretion were evaluated using microscopy, fluorescent reporters, dye tracing, RNA silencing assays and mass spectrometry-based proteomics. pSym inoculation reproducibly generated symbionts across diverse host plant species that were vascularly integrated into their host plants and transgene expression ranging from heterogeneous niches to more uniform patterns. Small molecules moved between symbionts and host vascular tissues, whereas larger proteins exhibited more restricted mobility. Post-transcriptional gene silencing signals moved freely throughout the symbiont and slightly into adjacent stem tissue. Under tested field and greenhouse conditions in potato and tomato, respectively, gall or symbiont formation had no negative impacts on plant growth or tuber and fruit yield. In vitro , symbiont cultures abundantly secreted recombinant protein into surrounding media. Together, these results establish symbionts as a modular, plant bioengineering platform capable of producing and potentially delivering biomolecules without modifying the host plant genome, providing a foundation for vascular-targeted therapeutics and phenotype modulation in crops.

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Publication Details

Journal
Frontiers in Plant Science
Published
2026-03-16
DOI
https://doi.org/10.3389/fpls.2026.1754357
Primary Topic
Plant tissue culture and regeneration
Type
article
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article

A new use of Agrobacterium plant growth regulator genes for plant bioengineering

Randall P. Niedz, Stacy L. DeBlasio, A. Ian Scott, Richard Stange et al.
Frontiers in Plant Science
Plant tissue culture and regeneration
article

A new use of Agrobacterium plant growth regulator genes for plant bioengineering

Randall P. Niedz, Stacy L. DeBlasio, A. Ian Scott, Richard Stange, Liliana M. Cano, Brian H. Rhodes, Ali Nikoomanzar, Martin Stallone, Cindy L. McKenzie, Lauren Shrum, Magali Ferrari Grando, Redeat Tibebu, E.W. Stover, Michelle Heck, Brian K.W. Chang, Nicholas R. Larson, Lorenzo Rossi, Guilherme Locatelli, James E. Thomson, Douglas Stuehler, Wayne B. Hunter, Matthew G. Hentz, Rachel L. Cook, Robert G. Shatters, Ariana Makar, Kevin Howe, Luke Thompson, Robert A. Adair, Nursena Demirden, Chase Weeks-Purdy, Mark A. Ritenour, Samuel Coradetti, Samantha Sullivan, Lucy Bennett, Joanne Hodge, Meneka Ariyarante, W. Rodney Cooper, Ellen Cochrane, Nichole Gaza, Douglas Harper, Ketan Shende, Elijah Schechter, Marco Pitino, Li Wang
article en

Abstract

Delivery of biomolecules into plant vascular tissues remains a barrier to managing diseases caused by insect vector-borne pathogens and to modifying phenotypes of established perennial crops. Inspired by the vascularized growth of crown galls induced by Agrobacterium tumefaciens , we repurposed the bacterium’s plant growth regulator (PGR) genes to engineer autonomously dividing, transgene-expressing plant cell structures termed symbionts. A plant transformation vector (pSYM) incorporating the IaaM, IaaH, Ipt and gene5 cassette from A. tumefaciens strain C58 together with a gene of interest on the same transfer DNA was delivered to stems of herbaceous and woody dicots using disarmed A. tumefaciens strain EHA105. Symbiont morphology, vascular differentiation, transgene expression, molecular mobility and protein secretion were evaluated using microscopy, fluorescent reporters, dye tracing, RNA silencing assays and mass spectrometry-based proteomics. pSym inoculation reproducibly generated symbionts across diverse host plant species that were vascularly integrated into their host plants and transgene expression ranging from heterogeneous niches to more uniform patterns. Small molecules moved between symbionts and host vascular tissues, whereas larger proteins exhibited more restricted mobility. Post-transcriptional gene silencing signals moved freely throughout the symbiont and slightly into adjacent stem tissue. Under tested field and greenhouse conditions in potato and tomato, respectively, gall or symbiont formation had no negative impacts on plant growth or tuber and fruit yield. In vitro , symbiont cultures abundantly secreted recombinant protein into surrounding media. Together, these results establish symbionts as a modular, plant bioengineering platform capable of producing and potentially delivering biomolecules without modifying the host plant genome, providing a foundation for vascular-targeted therapeutics and phenotype modulation in crops.

Frontiers in Plant ScienceVol. 17
Oak Ridge Associated Universities (US), Agricultural Research Service (US), Indian Institute of Horticultural Research (IN), United States Department of Agriculture (US), Cornell University (US), Planta (RU), Carnegie Department of Plant Biology (US), Horticultural Research Institute (US), Tompkins-Seneca-Tioga Board of Cooperative Educational Services (US), Telesis Corporation (United States) (US), Institute for Sustainability (GB), Horticultural Crops Research Laboratory (US), U.S. Horticultural Research Laboratory (US), U.S. Vegetable Laboratory (US), Indian River State College (US)
Openalex Percentile: Top 21%
Plant tissue culture and regeneration
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