loading page

Combining the CowPEAsy web application with in planta agroinfiltration for native promoter validation in Vigna unguiculata
  • +1
  • Jooyeon Jeong,
  • Jake Harris,
  • Larissa Larocca de Souza,
  • Laurie Leonelli
Jooyeon Jeong
University of Illinois Urbana-Champaign Carl R Woese Institute for Genomic Biology
Author Profile
Jake Harris
University of Illinois Urbana-Champaign Carl R Woese Institute for Genomic Biology
Author Profile
Larissa Larocca de Souza
University of Illinois Urbana-Champaign Carl R Woese Institute for Genomic Biology
Author Profile
Laurie Leonelli
University of Illinois Urbana-Champaign Carl R Woese Institute for Genomic Biology

Corresponding Author:lbl@illinois.edu

Author Profile

Abstract

Cowpea ( Vigna unguiculata) is an important protein source in Sub-Saharan Africa. Optimizing resilience and productivity through genetic engineering in cowpea has been slow due in part to a lack of defined species-specific regulatory elements and difficulty testing gene function within the native system. In many plant species, Agrobacterium-mediated transient gene expression is widely used to validate constructs before investing in transgenic lines, but its implementation in legumes has been challenging. In this study, we optimized an in planta agroinfiltration assay in trifoliate cowpea leaves using a betalain reporter. To demonstrate the “intact plant” aspect of this system, we used this assay to characterize drought-inducible promoters by challenging cowpea plants with drought stress. Subsequently, to identify and broaden the pool of native promoters known in cowpea, we developed a user-friendly web application, CowPEAsy, allowing users to interrogate gene expression from our canopy-level, developmental-series RNA-Seq dataset. Finally, using CowPEAsy, we identified six promoters that showed constitutive expression across all conditions and verified these promoters with our transient system. This work provides an in vivo platform for preliminary validation of regulatory elements in cowpea and other legumes and enhances current genetic resources by identifying a suite of physiologically relevant promoters of varying strengths.
27 Nov 2024Submitted to Plant, Cell & Environment
28 Nov 2024Submission Checks Completed
28 Nov 2024Assigned to Editor
28 Nov 2024Review(s) Completed, Editorial Evaluation Pending
09 Dec 2024Reviewer(s) Assigned