Triticeae Genomics and Genetics, 2025, Vol.16, No.3, 120-129 http://cropscipublisher.com/index.php/tgg 120 Feature Review Open Access Stacking of Multiple Resistance Genes in Wheat via Transgenic Approaches Delong Wang, Pingping Yang, Shujuan Wang Hainan Provincial Key Laboratory of Crop Molecular Breeding, Sanya, 572025, Hainan, China Corresponding email: shujuan.wang@hitar.org Triticeae Genomics and Genetics, 2025, Vol.16, No.3 doi: 10.5376/tgg.2025.16.0013 Received: 06 Apr., 2025 Accepted: 17 May, 2025 Published: 03 Jun., 2025 Copyright © 2025 Wang et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Wang D.L., Yang P.P., and Wang S.J., 2025, Stacking of multiple resistance genes in wheat via transgenic approaches, Triticeae Genomics and Genetics, 16(3): 120-129 (doi: 10.5376/tgg.2025.16.0013) Abstract Wheat diseases continue to threaten global food security, leading to yield losses and reduced grain quality. Consequently, effective and sustainable disease resistance strategies are urgently needed. This study explores the stacking of multiple resistance genes in wheat through transgenic approaches as a promising solution to these challenges. We first outline the principles of gene stacking, including the necessity to overcome pathogen evolution, ensure broad and durable resistance, and meet environmental and agricultural needs. We then discuss various transgenic strategies, such as direct genetic transformation, synthetic multigene constructs, and CRISPR/Cas-mediated genome editing, highlighting their potential for assembling and integrating multiple resistance genes. We also detail specific resistance genes commonly used in transgenic wheat, including those targeting rusts (e.g., Lr34, Sr22, Yr36), fungal and viral pathogens, and genes involved in broad-spectrum defense (e.g., pathogenesis-related proteins). Using the case study of transgenic stacking for rust resistance, specifically against Ug99 rust, we illustrate the practical applicability and global impact of this approach. We also explore the technical challenges, biosafety regulations, and genetic complexity that hinder its implementation. Looking ahead, we explore innovations in synthetic biology, precision gene editing, and breeding for climate resilience. Finally, we summarize recent advances in gene stacking, identify key gaps, and highlight the future potential of transgenic technology for enhancing durable disease resistance in wheat. Keywords Transgenic wheat; Gene stacking; Disease resistance; CRISPR/Cas genome editing; Rust resistance genes 1 Introduction In recent years, the problem of frequent wheat diseases has shown no sign of abating. The "familiar faces" such as stem rust and powdery mildew still cause considerable production losses worldwide. Although people had high hopes for the single resistance gene (R gene) in the past, reality soon poured cold water on it-pathogens are not herbivores either; they evolve, and resistance thus becomes ineffective. Thus, "multi-gene combination" has become a more reliable option. However, it's easier said than done. Traditional gene stacking breeding is not only cumbersome but also often makes the breeding process complex and time-consuming because the resistance loci are not closely adjacent (Athiyannan et al., 2022). The development of technology has indeed brought about a turning point. Nowadays, plant genomics and transformation technologies enable us to clone multiple resistance genes at a single site and then transfer them into the wheat genome. This is known as transgenic resistance stacking (RTGS). Compared with the traditional way, this method not only saves time but is also more efficient. In addition to expanding the range and duration of resistance, it is also possible to discover high-quality disease-resistant genes in wild or distant species that cannot be obtained through traditional means. For instance, there have been studies that have successfully cultivated wheat strains carrying up to five resistance genes through this approach, which still perform strongly against various pathogenic bacteria (Koller et al., 2023). So the focus of this study lies here-we want to see how far the current research on achieving the superposition of disease-resistant genes through genetic modification in wheat has come. In addition to summarizing the technical principles and application effects, the problems encountered will also be discussed. Of course, recent cases and related genomic resource integration work will not be absent. Finally, some ideas and prospects on how to achieve more durable resistance in breeding in the future will also be put forward.
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