Journal of Energy Bioscience 2025, Vol.16, No.5, 238-247 http://bioscipublisher.com/index.php/jeb 238 Research Insight Open Access Enhancing Biofuel Production by Genetic Engineering of C4 Plant Photosynthesis Pathways Wenzhong Huang Biomass Research Center, Hainan Institute of Tropical Agricultural Resouces, Sanya, 572025, Hainan, China Corresponding email: wenzhong.huang@hitar.org Journal of Energy Bioscience, 2025, Vol.16, No.5 doi: 10.5376/jeb.2025.16.0023 Received: 04 Aug, 2025 Accepted: 10 Sep., 2025 Published: 24 Sep., 2025 Copyright © 2025 Huang, 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: Huang W.Z., 2025, Enhancing biofuel production by genetic engineering of C4 plant photosynthesis pathways, Journal of Energy Bioscience, 16(5): 238-247 (doi: 10.5376/jeb.2025.16.0023) Abstract This study mainly discusses how to use genetic engineering to improve the photosynthesis of C4 plants, thereby increasing the yield of biofuels. C4 crops, such as sugarcane, corn and sorghum, are regarded as good raw materials for biofuels because they can efficiently utilize carbon dioxide and accumulate more biomass. In recent years, genetic engineering methods have developed rapidly. Methods such as CRISPR/Cas editing, synthetic biology, and multi-omics analysis have all been employed to regulate enzymes, transcription factors, and metabolic pathways related to C4 photosynthesis. These methods make photosynthesis more efficient, nitrogen utilization better, and plants more resilient to adverse conditions. However, there are still many problems to be faced in truly applying these achievements to industries. For instance, the adaptive balance of plants in different environments, biosecurity and regulatory requirements, cost input and the difficulty of promotion, etc. In the future, C4 photosynthesis projects may be combined with the transformation of C3 crops. With the addition of systems biology modeling and collaboration among different disciplines, there is an opportunity to cultivate efficient and low-carbon fuel crops. This is also an important direction for promoting sustainable global energy development. The objective of this review is to summarize these advancements and provide references for subsequent research. Keywords C4 Photosynthesis; Genetic engineering; Biofuel; Synthetic biology; Multi-omics integration 1 Introduction The global energy crisis is deepening. Fossil fuels not only have a limited quantity but also bring about environmental problems such as greenhouse gas emissions and climate change. These contradictions make it urgent for people to find sustainable alternative energy sources. Biofuels are regarded as an important alternative to fossil fuels due to their high efficiency, economy and relative environmental friendliness. Especially in the context of the continuous increase in population and energy demand, the development of renewable energy is very important for energy security and environmental protection (Cui, 2021; Wang et al., 2021; Zafar et al., 2022). C4 plants, such as corn, sorghum, sugarcane and miscanthus, play an important role in biofuel production. They have a high photosynthetic efficiency, can make better use of resources, convert solar energy into chemical energy and produce a large amount of biomass. These plants can also grow normally under adverse conditions such as high temperature and drought, and are suitable for cultivation on marginal land, reducing competition with food crops. It is precisely because of its high yield and efficiency that C4 plants are regarded as ideal raw materials for the development of sustainable biofuels (Van Der Weijde et al., 2013; Zafar et al., 2022; Aggarwal et al., 2024). In recent years, with the unremitting efforts of researchers, molecular biology and genetic engineering have been vigorously developed, providing new opportunities for the improvement of C4 photosynthesis. During the research process, regulating genes related to cell cycle, hormone action, cell wall formation, etc., and using molecular tools such as transcription factors and mirnas on this basis can improve the photosynthetic efficiency of target plants (Von Caemmerer and Furbank, 2016). The use of new technologies such as gene editing and synthetic biology can also more precisely modify the photosynthetic pathway and optimize carbon fixation and energy utilization (Schuler et al., 2016; Ermakova et al., 2020; Nazari et al., 2024; Swift et al., 2024).
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