Legume Genomics and Genetics 2025, Vol.16, No.5, 215-224 http://cropscipublisher.com/index.php/lgg 215 Feature Review Open Access Gene Editing-Assisted Development of Herbicide-Resistant Lentils Hongpeng Wang, Shiying Yu Biotechnology Research Center, Cuixi Academy of Biotechnology, Zhuji, 311800, China Corresponding email: shiying.yu@cuixi.org Legume Genomics and Genetics, 2025 Vol.16, No.5 doi: 10.5376/lgg.2025.16.0022 Received: 16 Jul., 2025 Accepted: 02 Sep., 2025 Published: 22 Sep., 2025 Copyright © 2025 Wang and Yu, 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 H.P., and Yu S.Y., 2025, Gene editing-assisted development of herbicide-resistant lentils, Legume Genomics and Genetics, 16(5): 215-224 (doi: 10.5376/lgg.2025.16.0022) Abstract Lentil (Lens culinaris) is a globally important legume crop, but its yield is increasingly constrained by weed competition and limited access to selective herbicides. This study explores how gene editing technologies, particularly the CRISPR/Cas system, can revolutionize the development of herbicide-resistant lentil varieties. We first discuss the potential mechanisms of herbicide resistance in plants, including target site resistance (TSR) and non-target site resistance (NTSR), and compare transgenic approaches with endogenous gene editing strategies. We then review advances in gene editing platforms, such as base editing and primer editing, and examine delivery systems for lentil. We focus specifically on recent advances in editing key genes, such as ALS (acetolactate synthase) and EPSPS (5-enolpyruvylshikimate-3-phosphate synthase), followed by field evaluation of gene-edited lines. We also critically analyze regulatory frameworks, biosafety concerns, and public acceptance. A case study of imazethapyr-resistant lentil breeding in Canada showcases the practical applications and achievements of gene editing in lentil breeding. This study highlights the potential of combining gene editing with modern breeding tools to broaden the herbicide resistance spectrum, enhance sustainability, and ensure the sustainability of lentil production under changing agroecological conditions. Keywords Gene editing; Herbicide resistance; CRISPR/Cas; Lentil breeding; ALS gene 1 Introduction Among the many legumes, lentils (Lens culinaris Medik.) may not be the "star players", but they still firmly hold their place globally due to their rich protein content and unique position in sustainable agriculture. However, when it comes to actual cultivation, there are also quite a few problems. For instance, lentils grow slowly in the early stage, have shallow root systems and relatively weak competitiveness, which makes them almost powerless against weeds-once weeds invade, the yield loss can be very serious (Balech et al., 2023). In the past, farmers mainly relied on manual weeding or crop rotation to control weeds, but these methods were labor-intensive and time-consuming, and large-scale promotion was not realistic. So, people turned to chemical herbicides. The effect was obvious, but the plant toxicity problem that followed caused harm to the lentils themselves-in some extreme cases, the yield could even drop by half. No one is willing to lose their crops for weeding. Against this backdrop, cultivating lentil varieties that can withstand herbicides has become an unavoidable goal. This resistant variety can be used in combination with broad-spectrum herbicides, which can kill weeds without accidentally harming the lentils themselves. Naturally, it is expected to increase yields and reduce costs. However, traditional breeding methods have always been limited in this regard: there is too little natural variation, the traits are complex, and the progress is slow. Even mutant breeding or transgenic methods are often criticized-either for their low efficiency or being blocked by policies and public opinion (Singh et al., 2021). Nowadays, gene editing technology, especially CRISPR/Cas9, seems to have reached a turning point (Wang et al., 2024). It operates more precisely and efficiently, and has the potential to bypass the genetically modified label-a feature that has already shown initial success in many crops (Kuang et al., 2024). This also makes people look forward to its application on lentils. The focus of this study is precisely to sort out the latest progress of gene editing in the development of herbicide-resistant lentils. We will elaborate from several aspects: First, we will introduce the basic situation of lentil production and its vulnerability in the use of herbicides; Then explore the genetic basis of its resistance traits;
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