Bioscience Methods 2024, Vol.15, No.6, 289-301 http://bioscipublisher.com/index.php/bm 289 Research Insight Open Access Comparative Study of Rubber Biosynthesis Pathways in Eucommia ulmoides andHevea brasiliensis Degang Zhao1,2, , Shangmei Long1, Li Song1,YingZhu2, Ruoruo Wang2, DanZhao1, 1 National-local Joint Engineering Research Center of Karst Region Plant ResourcesUtilization & Breeding (Guizhou), College of Life Sciences/Institute of Agro- Bioengineering, Guizhou University, Guiyang, 550025, China 2 Plant Conservation & Breeding Technology Center, Guizhou Key Laboratory of Agricultural Biotechnology/Biotechnology Institute of Guizhou Province, Guizhou Academy of Agricultural Sciences, Guiyang, 550006, China Corresponding authors: dgzhao@gzu.edu.cn, dzhao@gzu.edu.cn Bioscience Methods, 2024, Vol.15, No.6 doi: 10.5376/bm.2024.15.0029 Received: 13 Sep., 2024 Accepted: 22 Oct., 2024 Published: 15 Nov., 2024 Copyright © 2024 Zhao 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: Zhao D.G., Long S.M., Song L., Zhu Y., Wang R.R., and Zhao D., 2024, Comparative study of rubber biosynthesis pathways in Eucommia ulmoides and Hevea brasiliensis, Bioscience Methods, 15(6): 289-301 (doi: 10.5376/bm.2024.15.0029) Abstract This study conducts a comparative analysis of the rubber biosynthesis pathways in two significant rubber-producing species, Eucommia ulmoides and Hevea brasiliensis. By examining the genetic and biochemical mechanisms underlying rubber production in these species, the study aims to uncover the evolutionary adaptations and potential biotechnological applications of their distinct biosynthetic pathways. The study reveals that Eucommia ulmoides primarily utilizes the methylerythritol-phosphate (MEP) pathway for isoprenyl diphosphate synthesis, which is a precursor for trans-polyisoprene rubber. In contrast, Hevea brasiliensis predominantly employs the mevalonate (MVA) pathway for cis-polyisoprene rubber production. Additionally, the farnesyl diphosphate synthase (FPS) gene families in both species show significant differences in expression patterns and gene expansions, which are crucial for their respective rubber biosynthesis processes. The study also identifies long non-coding RNAs (lncRNAs) and microRNAs that play regulatory roles in rubber biosynthesis, providing deeper insights into the molecular regulation of this process. The findings highlight the evolutionary divergence in rubber biosynthesis pathways between Eucommia ulmoides and Hevea brasiliensis. Understanding these differences not only enriches our knowledge of plant secondary metabolism but also opens up new avenues for genetic engineering to enhance rubber production in these and other species. The study underscores the potential for biotechnological advancements in the rubber industry by leveraging the unique biosynthetic pathways of these plants. Keywords Rubber biosynthesis; Eucommia ulmoides; Hevea brasiliensis; MEP pathway; MVA pathway; Farnesyl diphosphate synthase; Long non-coding RNAs; microRNAs; Genetic engineering 1 Introduction Natural rubber is a critical raw material with extensive applications in various industries, including automotive, medical, and defense. The primary source of natural rubber is the Para rubber tree, Hevea brasiliensis, which produces cis-1,4-polyisoprene. This biopolymer is valued for its high elasticity, flexibility, and resilience, making it indispensable for manufacturing over 50 000 rubber products, such as tires and medical gloves (Rahman et al., 2013; Cherian et al., 2019). The global demand for natural rubber continues to rise, driven by its unique properties that synthetic alternatives cannot fully replicate (Cherian et al., 2019). Consequently, understanding and improving rubber biosynthesis pathways is of paramount economic importance. Hevea brasiliensis is the most widely cultivated species for commercial natural rubber production. The biosynthesis of rubber in H. brasiliensis occurs through the mevalonate (MVA) pathway, which provides isopentenyl diphosphate (IPP) for cis-polyisoprene synthesis (Chow et al., 2007; Chow et al., 2011; Ambily et al., 2019). The genome of H. brasiliensis has been extensively studied, revealing a significant expansion of the rubber elongation factor (REF) and small rubber particle protein (SRPP) gene families, which are crucial for rubber biosynthesis (Lau et al., 2016; Tang et al., 2016). In contrast, Eucommia ulmoides, known for its medicinal applications, produces trans-1,4-polyisoprene, an isomer of natural rubber. The rubber biosynthesis in E. ulmoides primarily relies on the methylerythritol-phosphate (MEP) pathway rather than the MVA pathway. Recent genomic studies have provided high-quality assemblies of the E.
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