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- 조회223
Plant Cell Environ., Feb 13 2025, | https://doi.org/10.1111/pce.15431
Combining the CowPEAsy Web Application With in Planta Agroinfiltration for Native Promoter Validation in Vigna unguiculata
Jooyeon Jeong 1 †, Jake Harris 1 2 †, Larissa Larocca de Souza 1, Lauriebeth Leonelli 1 2 *
1Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana Champaign, Champaign, Illinois, USA.
2Department of Agricultural and Biological Engineering, University of Illinois at Urbana Champaign, Champaign, Illinois, USA.
2Department of Agricultural and Biological Engineering, University of Illinois at Urbana Champaign, Champaign, Illinois, USA.
†Jooyeon Jeong and Jake Harris contributed equally to this work.
*Corresponding author: correspondence to Lauriebeth Leonelli
*Corresponding author: correspondence to Lauriebeth Leonelli
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 data set. 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.
논문정보
- Research article
- 2025년 02월 (BRIC 등록일 2025-02-18)
- 국외 연구진
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