Soil salinity is one of the most pressing challenges facing agriculture worldwide. Excess salt disrupts water and nutrient uptake, severely restricting crop productivity. Bok choy, a widely cultivated leafy vegetable, is particularly vulnerable due to its shallow root system, making it sensitive to drought and saline soils. Previous studies have shown that WRKY transcription factors play pivotal roles in regulating stress responses and root architecture. However, the precise pathways linking these regulators to root growth and salt adaptation remained elusive. Due to these challenges, it is necessary to conduct in-depth research on the genetic mechanisms driving root development and stress tolerance.
A research team from Nanjing Agricultural University, Beijing University of Agriculture, and North Carolina State University has uncovered a key genetic pathway that strengthens Bok choy roots under salt stress. The study, published (DOI: 10.1093/hr/uhae280) on September 28, 2024, in Horticulture Research, reveals that the transcription factors BcWRKY25 and BcWRKY33A directly regulate downstream genes BcLRP1 and BcCOW1 to promote root elongation and hair formation. This newly identified module offers a molecular blueprint for breeding crops with stronger roots and enhanced resilience to environmental stress.
The researchers began by examining the expression of BcWRKY33A in Bok choy roots under different stress treatments. Salt exposure triggered a 53-fold increase in BcWRKY33A expression, while overexpression experiments in both Arabidopsis and Bok choy confirmed its role in extending primary roots and stimulating root hair formation. Conversely, silencing BcWRKY33A suppressed root growth and caused abnormal hair morphology.
To uncover the molecular mechanism, DNA affinity purification sequencing (DAP-seq) and yeast one-hybrid assays demonstrated that BcWRKY33A directly binds to the promoters of BcLRP1 and BcCOW1. Functional tests showed that BcLRP1 overexpression promoted primary root elongation, while BcCOW1 enhanced root hair development, confirming their complementary roles. Moreover, BcWRKY25 was identified as an upstream regulator, activated by salt stress and directly inducing BcWRKY33A. Together, this cascade—BcWRKY25–BcWRKY33A–BcLRP1/BcCOW1—was shown to coordinate multiple aspects of root growth, creating a stronger root system under saline conditions.
This integrative model not only explains how WRKY transcription factors fine-tune root development but also highlights the modular nature of plant genetic networks, where stress-responsive genes activate downstream effectors to shape adaptive growth.
“Our study demonstrates how a single transcriptional module can reprogram root development in response to salt stress,” said Professor Tongkun Liu, senior author of the study. “By linking BcWRKY25 and BcWRKY33A with the downstream regulators BcLRP1 and BcCOW1, we were able to map a complete pathway that enhances root elongation and stabilizes root hairs. This provides a powerful target for crop improvement. The findings not only deepen our understanding of root biology but also open new doors for breeding salt-tolerant vegetables and other crops.”
The discovery of the BcWRKY25–BcWRKY33A–BcLRP1/BcCOW1 module provides valuable molecular tools for developing crops resilient to salinity. In practical terms, manipulating these genes could lead to varieties with deeper, stronger root systems that efficiently absorb water and nutrients even in degraded soils. For Bok choy and other leafy vegetables, this translates into stable yields under climate-driven stress. Beyond immediate applications, the insights offer a framework for cross-species improvement, enabling breeders to design salt-tolerant traits in cereals, legumes, and horticultural crops. As soil salinization intensifies globally, such genetic strategies will be critical for securing sustainable food production.
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References
DOI
10.1093/hr/uhae280
Original Source URL
https://doi.org/10.1093/hr/uhae280
Funding information
This work was supported by National Natural Science Foundation of China (32372698, 32072575), Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX22_0752), and National Vegetable Industry Technology System (CARS-23-A16) to T.L., and the USDA National Institute of Food and Agriculture (NIFA) Hatch project 02913 to W.L.
About Horticulture Research
Horticulture Research is an open access journal of Nanjing Agricultural University and ranked number one in the Horticulture category of the Journal Citation Reports ™ from Clarivate, 2023. The journal is committed to publishing original research articles, reviews, perspectives, comments, correspondence articles and letters to the editor related to all major horticultural plants and disciplines, including biotechnology, breeding, cellular and molecular biology, evolution, genetics, inter-species interactions, physiology, and the origination and domestication of crops.