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Natural rubber is a kind of indispensable biopolymers with great use and strategic importance in human society. However, its production relies almost exclusively on rubber-producing plants Hevea brasiliensis, which have high requirements for growth conditions, and the mechanism of natural rubber biosynthesis remains largely unknown. In the past two decades, details of the rubber chain polymerization and proteins involved in natural rubber biosynthesis have been investigated intensively. Meanwhile, omics and other advanced biotechnologies bring new insight into rubber production and development of new rubber-producing plants. This review summarizes the achievements of the past two decades in understanding the biosynthesis of natural rubber, especially the massive information obtained from the omics analyses. Possibilities of natural rubber biosynthesis in vitro or in genetically engineered microorganisms are also discussed.

Enhanced natural rubber biosynthesis is associated with the activation of the COI1-JAZ3-MYC2 jasmonate signalling module in laticifer cells of rubber trees. Abstract Rubber trees are the world's major source of natural rubber. Rubber-containing latex is obtained from the laticifer cells of the rubber tree (Hevea brasiliensis) via regular tapping. Rubber biosynthesis is a typical isoprenoid metabolic process in the laticifer cells; however, little is known about the positive feedback regulation caused by the loss of latex that occurs through tapping. In this study, we demonstrate the crucial role of jasmonate signalling in this feedback regulation. The endogenous levels of jasmonate, the expression levels of rubber biosynthesis-related genes, and the efficiency of in vitro rubber biosynthesis were found to be significantly higher in laticifer cells of regularly tapped trees than those of virgin (i.e. untapped) trees. Application of methyl jasmonate had similar effects to latex harvesting in up-regulating the rubber biosynthesis-related genes and enhancing rubber biosynthesis. The specific jasmonate signalling module in laticifer cells was identified as COI1-JAZ3-MYC2. Its activation was associated with enhanced rubber biosynthesis via up-regulation of the expression of a farnesyl pyrophosphate synthase gene and a small rubber particle protein gene. The increase in the corresponding proteins, especially that of farnesyl pyrophosphate synthase, probably contributes to the increased efficiency of rubber biosynthesis. To our knowledge, this is the first study to reveal a jasmonate signalling pathway in the regulation of rubber biosynthesis in laticifer cells. The identification of the specific jasmonate signalling module in the laticifer cells of the rubber tree may provide a basis for genetic improvement of rubber yield potential.

[This corrects the article DOI: 10.3389/fpls.2025.1655155.].

Background Tapping panel dryness (TPD) in rubber tree has become the most severe restricting factor of natural rubber production. To date, there is no effective measures to prevent and control TPD. Previous studies primarily focused on analyzing the molecular mechanism underlying TPD occurrence. However, there is no research on the molecular mechanism of TPD recovery. Results In this study, the TPD trees were recovered by treatment with TPD rehabilitation nutrient agents that could promote the recovery of latex flow on the tapping panel of TPD trees. The genes and proteins involved in TPD recovery were first identified by employing integrated transcriptomics and proteomics analyses. In total, 2029 differentially expressed genes (DEGs) and 951 differentially expressed proteins (DEPs) were detected in the bark of recovery trees compared to that of TPD trees. Among them, 19 DEPs and 11 DEGs were found to be involved in the starch and sucrose metabolism pathway, suggesting their important roles in regulating the syntheses of sucrose and D-glucose, which were the key precursors of natural rubber biosynthesis. Furthermore, 16 DEPs and 15 DEGs were identified in the rubber biosynthesis pathway. Interestingly, almost all the DEPs and DEGs related to rubber biosynthesis exhibited significantly up-regulated expressions in the recovery trees, indicating that latex biosynthesis were probably markedly enhanced during TPD recovery. Conclusions These results provide new insights into the molecular mechanisms underlying TPD recovery, as well as excellent supplements to the mechanisms of TPD occurrence, which will contribute to the development of more effective agents for the prevention and treatment of TPD in the future.

The increase in demand for natural rubber has led to the search for alternative sources. Lactuca serriola is emerging as a promising candidate, as the quality of the natural rubber it produces is comparable to that of the Pará Rubber Plant, Hevea brasiliensis . This study examines the effect of methyl jasmonate (MeJA), a known elicitor, on the expression of key rubber biosynthesis pathway genes ( HMGR1 , HMGS1 , CPT2 , and SRPP1 ) in the latex of L. serriola plants. The expression levels of these genes increased significantly after the foliar application of 200 and 400 µM MeJA. The highest relative expression level for HMGR1 , HMGS1 , CPT2 and SRPP1 was 3.74, 18.56, 11.91and 16.59 fold respectively. Furthermore, the rubber content in L. serriola showed a significant rise post-treatment compared to the control with increasing the level of MeJA (6.19%, 7.24% and 7.85% which correspond to 0, 200 and 400 µM). Gel permeation chromatography revealed an augmentation in the molecular weight of extracted natural rubber from treated plants. Samples treated with 400 µM of MeJA had the highest molecular weight (1570 kg mol −1 ) compared to control (1186 kg mol −1 ). This study has demonstrated that MeJA, through the regulation of rubber biosynthesis genes, is capable of enhancing the quality and quantity of natural rubber extracted from alternative sources, such as L. serriola .

Main conclusionNineTkOSCgenes have been identified by genome-wide screening. Among them,TkOSC4-6might be more crucial for natural rubber biosynthesis inTaraxacum kok-saghyzroots.Taraxacum kok-saghyz Rodin (TKS) roots contain large amounts of natural rubber, inulin, and valuable metabolites. Oxidosqualene cyclase (OSC) is a key member for regulating natural rubber biosynthesis (NRB) via the triterpenoid biosynthesis pathway. To explore the functions of OSC on natural rubber producing in TKS, its gene family members were identified in TKS genome via genome-wide screening. Nine TkOSCs were identified, which were mainly distributed in the cytoplasm. Their family genes experienced a neutral selection during the evolution process. Overall sequence homology analysis OSC proteins revealed 80.23% similarity, indicating a highly degree of conservation. Pairwise comparisons showed a multiple sequence similarity ranging from 57% to 100%. Protein interaction prediction revealed that TkOSCs may interact with baruol synthase, sterol 1,4-demethylase, lupeol synthase and squalene epoxidase. Phylogenetic analysis showed that OSC family proteins belong to two branches. TkOSC promoter regions contain cis-acting elements related to plant growth, stress response, hormones response and light response. Protein accumulation analysis demonstrated that TkOSC4, TkOSC5 and TkOSC6 proteins had strong expression levels in the root, latex and plumular axis. Comparison of gene expression patterns showed TkOSC1, TkOSC4, TkOSC5, TkOSC6, TkOSC7, TkOSC8 and TkOSC9 might be important in regulating NRB. Combination of gene and protein results revealed TkOSC4-6 might be more crucial, and the data might contribute to a more profound understanding of the roles of OSCs for NRB in TKS roots.

Natural rubber is an important industrial material, which is obtained from the only commercially cultivated rubber tree, Hevea brasiliensis. In rubber latex production, ethylene has been extensively used as a stimulant. Recent research showed that post-translational modifications (PTMs) of latex proteins, such as phosphorylation, glycosylation and ubiquitination, are crucial in natural rubber biosynthesis. In this study, comparative proteomics was performed to identify the glycosylated proteins in rubber latex treated with ethylene for different days. Combined with Pro-Q Glycoprotein gel staining and mass spectrometry techniques, we provided the first visual profiling of glycoproteomics of rubber latex and finally identified 144 glycosylated protein species, including 65 differentially accumulated proteins (DAPs) after treating with ethylene for three and/or five days. Gene Ontology (GO) functional annotation showed that these ethylene-responsive glycoproteins are mainly involved in cell parts, membrane components and metabolism. Pathway analysis demonstrated that these glycosylated rubber latex proteins are mainly involved in carbohydrate metabolism, energy metabolism, degradation function and cellular processes in rubber latex metabolism. Protein–protein interaction analysis revealed that these DAPs are mainly centered on acetyl-CoA acetyltransferase and hydroxymethylglutaryl-CoA synthase (HMGS) in the mevalonate pathway for natural rubber biosynthesis. In our glycoproteomics, three protein isoforms of HMGS2 were identified from rubber latex, and only one HMGS2 isoform was sharply increased in rubber latex by ethylene treatment for five days. Furthermore, the HbHMGS2 gene was over-expressed in a model rubber-producing grass Taraxacum Kok-saghyz and rubber content in the roots of transgenic rubber grass was significantly increased over that in the wild type plant, indicating HMGS2 is the key component for natural rubber production.

Rubber particles are a specific organelle for natural rubber biosynthesis (NRB) and storage. Ethylene can significantly improve rubber latex production by increasing the generation of small rubber particles (SRPs), regulating protein accumulation, and activating many enzyme activities. We conducted a quantitative proteomics study of different SRPs upon ethylene stimulation by differential in-gel electrophoresis (DIGE) and using isobaric tags for relative and absolute quantification (iTRAQ) methods. In DIGE, 79 differentially accumulated proteins (DAPs) were determined as ethylene responsive proteins. Our results show that the abundance of many NRB-related proteins has been sharply induced upon ethylene stimulation. Among them, 23 proteins were identified as rubber elongation factor (REF) and small rubber particle protein (SRPP) family members, including 16 REF and 7 SRPP isoforms. Then, 138 unique phosphorylated peptides, containing 129 phosphorylated amino acids from the 64 REF/SRPP family members, were identified, and most serine and threonine were phosphorylated. Furthermore, we identified 226 DAPs from more than 2000 SRP proteins by iTRAQ. Integrative analysis revealed that almost all NRB-related proteins can be detected in SRPs, and many proteins are positively responsive to ethylene stimulation. These results indicate that ethylene may stimulate latex production by regulating the accumulation of some key proteins. The phosphorylation modification of REF and SRPP isoforms might be crucial for NRB, and SRP may act as a complex natural rubber biosynthetic machine.

The rubber grass Taraxacum kok-saghyz (TKS) contains large amounts of natural rubber (cis-1,4-polyisoprene) in its enlarged roots and it is an alternative crop source of natural rubber. Natural rubber biosynthesis (NRB) and storage in the mature roots of TKS is a cascade process involving many genes, proteins and their cofactors. The TKS genome has just been annotated and many NRB-related genes have been determined. However, there is limited knowledge about the protein regulation mechanism for NRB in TKS roots. We identified 371 protein species from the mature roots of TKS by combining two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Meanwhile, a large-scale shotgun analysis of proteins in TKS roots at the enlargement stage was performed, and 3545 individual proteins were determined. Subsequently, all identified proteins from 2-DE gel and shotgun MS in TKS roots were subject to gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses and most proteins were involved in carbon metabolic process with catalytic activity in membrane-bounded organelles, followed by proteins with binding ability, transportation and phenylpropanoid biosynthesis activities. Fifty-eight NRB-related proteins, including eight small rubber particle protein (SRPP) and two rubber elongation factor(REF) members, were identified from the TKS roots, and these proteins were involved in both mevalonate acid (MVA) and methylerythritol phosphate (MEP) pathways. To our best knowledge, it is the first high-resolution draft proteome map of the mature TKS roots. Our proteomics of TKS roots revealed both MVA and MEP pathways are important for NRB, and SRPP might be more important than REF for NRB in TKS roots. These findings would not only deepen our understanding of the TKS root proteome, but also provide new evidence on the roles of these NRB-related proteins in the mature TKS roots.

Eucommia ulmoides Oliv., a Tertiary period relict tree species endemic to China, is a rubber-producing plant valued for both medicinal and edible applications. E.ulmoides rubber is a high-quality natural rubber prized for its excellent elasticity, abrasion resistance, and insulation properties, leading to broad industrial applications. Previous research identified the EuUSP16 gene, encoding a protein containing E.ulmoides rubber particle protein peptides. While overexpression of EuUSP16 in tobacco enhanced drought tolerance, its role in E.ulmoides rubber biosynthesis remained undefined. In this study, we identified 29 EuUSP genes at the whole-genome level in E.ulmoides . Following low-temperature and drought treatments, the expression level of the EuUSP16 gene was found to be positively correlated with changes in rubber content ( p<0.05 ), suggesting its potential regulatory role in rubber synthesis. In E.ulmoides subjected to Agrobacterium-mediated EuUSP16 gene overexpression or silencing, the expression levels of key E.ulmoides rubber biosynthesis enzyme genes, such as EuFPS1 , exhibited corresponding increases and decreases, respectively. Furthermore, rubber content in EuUSP16 -overexpressing callus increased by 254.51% compared to wild-type callus. These findings indicate that EuUSP16 regulates E.ulmoides rubber biosynthesis by modulating the expression of these genes. The 1,967 bp promoter region upstream of the EuUSP16 ATG start codon contains several responsive elements, including MBS (MYB-binding site; CAACTG), LTR (low-temperature responsive element; CCGAAA), ABRE (ABA-responsive element; ACGTG), and a Dof transcription factor binding motif (AAAG). Promoter activity assays showed that EuUSP16 promoter activity was induced by low temperature and drought but repressed by abscisic acid (ABA) treatment. Furthermore, using yeast one-hybrid screening, we identified a Cys2-Cys2 zinc finger domain-containing transcription factor, designated EuDof. Interaction analysis revealed that the EuDof transcription factor enhances the activity of the EuUSP16 promoter. The binding of EuDof to the EuUSP16 promoter was enhanced under low temperature and drought stress but inhibited by ABA. Collectively, this study provides crucial insights into the regulatory mechanism of EuUSP16 in E.ulmoides rubber biosynthesis.