## 1 Introduction  
Since the approval of the first living modified (LM) tomato in 1996, the cultivation of LM crops has expanded significantly worldwide. The advantages of LM crops include their ability to overcome biological challenges such as pests, weeds, and diseases. This leads to increased crop yields and quality, a reduced need for chemical pesticides, and enhanced resistance to environmental stresses. By 2020, LM crops were planted across 185.6 million hectares (Mhas), with a significant increase in cultivation in developing countries [1; 2; 3]. Major LM crops, including canola, soybean, maize, and cotton, now account for 47.4% of the total global planting area for these four crops [1; 4]. Specifically, the global cultivation area of LM canola reached 10.1 Mhas in 2019. These crops typically carry genes conferring herbicide resistance (e.g., _cpu4-epsps_ genes from _Agrobacteriumium tunefaciens_ strain CP4, _patgenes from _Streptomyces viridochromogenes_, _bar_ genes from _S. hygroscopicus_, _dmo_ genes from _S. Stenotrophomonas maltoplilia_ strain DI-6, and _aad-12_ genes from _Delftia acidOVAs_) [5; 6; 7; 8; 9; 10] and insect resistance (e.g., _Cry_ toxin genes from _Bacillus thuringiensis_) to improve agricultural productivity and food security [11; 12; 13; 14; 15]. Most LM canola varieties have been developed for herbicide resistance to improve weed control in cropping systems. However, insect pests, such as flea beetles (_Phyllotrata cniciferase_ and _Phyllotrata striolata_), cutworms (family _Noctuidae_), and Eumylabris (_Mylabris sibrica_), continue to reduce crop yields by feeding on canola leaves and seeds in many countries [16]. Despite the benefits, the adoption of LM crops has raised concerns regarding potential environmental risks, particularly gene flow from LM crops to non-LM varieties or wild relatives [17; 18; 19]. In response to these risks, the Cartagena Protocol on Biosafety was adopted in 2000 to regulate LMOs resulting from modern biotechnology [20; 21; 22]. South Korea ratified this protocol in October 2007, with the Enforcement Decree of the Transboundary Movement of Living Modified Organisms Act (LMO Act) coming into force in January 2008 [23; 24]. Since the implementation of the LMO Act, South Korea has seen a steady increase in the importation of LM crops, rising from 8527 million tons in 2008 to 10,282 million tons in 2023 [25]. The unintentional release of LMOs into natural environments during transportation and utilization presents a significant regulatory challenge, particularly given the complexity of maintaining environmental biodiversity [26]. Rapeseed (_Brassica napus_ L.), commonly known as canola, is the second most important oilseed crop globally, following soybean [27; 28]. _B. napus_ (AACC, 2n = 38) is an amphidipoloid species in the Brassicaceae family, formed by the hybridization of _B. rapa_ (AA, 2n = 20) and _B. oleracea_ (CC, 2n = 18) [29]. Brassic species are important in agriculture, and these genetic relations are discovered by U's triangle [30; 31]. Many species within the Brassicaceae family are used as vegetable crops. Leaf mustard (_Brassica juncea_) is widely cultivated, while wild leaf mustard (_Brassica juncea_ [PERSON]) grows across South Korea, including in fields, along riversides, and by roadsides [32; 33]. _Brassica napus_ is primarily grown as a landscape crop for spring flower festivals [34]. Moreover, canola is a self-pollinating crop, but its pollen can be transported by wind and insects [35; 36]. Most canola pollen travels less than 10 m, but pollen is transferred 1.5 km by wind and 4 km by insects such as honeybees [37]. Additionally, canola is pollen-mediated, outcrossing ranges from 12% to 55% [38; 39]. The hybridization rate between canola and _B. juncea_ was reported to be 3% [40; 41], and it was reported that hybrids between _B. napus_ and _B. juncea_ have the potential to produce viable seeds [41; 42].