Journal of Energy Bioscience 2025, Vol.16, No.2, 75-84 http://bioscipublisher.com/index.php/jeb 76 mainly studied several different agricultural practices (such as fertilization methods and variety selection) to observe their effects on rapeseed yield and oil content, and at the same time figure out the principles of emerging processing technologies to improve oil extraction and oil production efficiency. At the end of the study, we combined the environmental and economic effects of these methods to comprehensively evaluate the potential of these improvement methods. This study covers all aspects from field planting to factory processing, hoping to find a set of optimal solutions that can both increase yield and ensure quality, and enhance the sustainability of biodiesel production. 2 Genetic Improvements for Higher Oil Content 2.1 Advances in breeding techniques In the past, people usually used some more traditional methods (such as hybridization and mutation breeding) to improve rapeseed. These methods have been used for many years, mainly to increase the oil content of rapeseed. Through breeding from generation to generation, researchers screened and retained good traits and cultivated many excellent improved varieties. There is a chemical mutagen called EMS. After treating rapeseed with EMS, new rapeseed varieties with high oleic acid content and low linolenic acid content can be obtained. At first, the agricultural performance of these varieties was not good. In order to improve the agricultural performance of this variety, scientists used "marker-assisted selection" technology to improve these varieties, which not only increased the oil yield of the new varieties, but also reduced the cost of planting (Spasibionek et al., 2020). 2.2 Identification of key yield-related genes Recent studies have found that some key genes directly affect rapeseed oil content (SOC) and yield-related traits. For example, there is a gene called SFAR, which is related to the synthesis of fatty acids. The researchers used CRISPR-Cas9 technology to modify this gene, which increased the oil content without affecting seed germination and vitality (Karunarathna et al., 2020). Zhang et al. (2023) used the GWAS (genome-wide association analysis) method in their study and found a number of genes related to yield. These genes affect the number of siliques, the number of seeds in each silique, and the weight of seeds. They also combined transcriptome analysis to further find candidate genes such as RNA helicase and lipase, which also have a certain effect on yield. These findings provide very useful gene targets for subsequent variety improvement (Zhang et al., 2023). 2.3 Application of genomic selection and CRISPR-Cas9 The emergence of CRISPR-Cas9 technology has made the improvement of rapeseed much faster. It can modify certain genes very accurately, helping us to breed excellent varieties faster. In the study by Liu et al. (2022), researchers used this technology to modify the BnFAD2 gene, resulting in a significant increase in the oleic acid content in rapeseed. Zhang et al. (2019) knocked out multiple copies of the BnLPAT2 and BnLPAT5 gene families, which also resulted in higher oil production. This technology was also used to modify the EPSPS gene to breed rapeseed varieties that are resistant to herbicides (glyphosate). This shows that CRISPR can not only increase yield, but also improve quality (Wang et al., 2021). In polyploid crops such as Brassica napus, there are still some problems, such as the easy occurrence of non-target modifications (off-target effects) or multiple genes with duplicate functions. However, with the advancement of technology, these problems are gradually being solved, and the efficiency and scope of application are constantly improving (Figure 1) (Sandgrind, 2022; Tian et al., 2022; Ali and Zhang, 2023). 3 Agronomic Practices Enhancing Oil Yield 3.1 Soil and nutrient management To increase the yield of rapeseed, soil and nutrient management are particularly important. Studies have found that adding biochar to the soil can bring many benefits. For example, it can increase the pH of the soil, increase the available phosphorus and organic carbon in the soil, and help the soil better retain water. These improvements can make rapeseed grow better and have higher yields. These effects will not last forever. Over time, the effect of biochar will weaken, so the soil needs to continue to be managed regularly (Jin et al., 2019). In addition to adding biochar, the rational application of fertilizers such as nitrogen, phosphorus, potassium, sulfur and boron can also
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