Identification and Characterization of Glycoproteins and Their Responsive Patterns upon Ethylene Stimulation in the Rubber Latex

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.

Molecular Mechanisms of Natural Rubber Biosynthesis

Natural rubber (NR), principally comprising cis-1,4-polyisoprene, is an industrially important natural hydrocarbon polymer because of its unique physical properties, which render it suitable for manufacturing items such as tires. Presently, industrial NR production depends solely on latex obtained from the Pará rubber tree, Hevea brasiliensis. In latex, NR is enclosed in rubber particles, which are specialized organelles comprising a hydrophobic NR core surrounded by a lipid monolayer and membrane-bound proteins. The similarity of the basic carbon skeleton structure between NR and dolichols and polyprenols, which are found in most organisms, suggests that the NR biosynthetic pathway is related to the polyisoprenoid biosynthetic pathway and that rubber transferase, which is the key enzyme in NR biosynthesis, belongs to the cis-prenyltransferase family. Here, we review recent progress in the elucidation of molecular mechanisms underlying NR biosynthesis through the identification of the enzymes that are responsible for the formation of the NR backbone structure.

Tobacco rattle virus -induced gene silencing in Hevea brasiliensis

BackgroundSince it is very difficult to obtain gene knockouts in rubber tree (Hevea Brasiliensis) due to low genetic transformation efficiency. Virus-induced gene silencing (VIGS) is a powerful gene silencing tool that has been intensively applied in plant. Up to now, the application of VIGS in rubber tree has not yet been reported.ResultsHevea brasiliensis phytoene desaturase (HbPDS) was identified in H. brasiliensis genome. The prediction of small interfering RNAs (siRNAs) from HbPDS and the silencing gene fragment (SGF) were predicted and a length of 409 bp SGF was chosen to be tested. We show that the tobacco rattle virus (TRV) -VIGS is able to induce effective HbPDS silencing in rubber tree. The TRV-VIGS system has the potential for functional gene studies in rubber tree.ConclusionsThis is the first time to report VIGS in rubber tree. The present TRV-VIGS method could be further applied to produce gene silenced rubber tree plants, to advance functional gene of rubber tree. The applied TRV-VIGS method will achieve deeper underground into the natural rubber biosynthesis and regulation in this important rubber-producing plant.

Biosynthesis of Natural Rubber: Current State and Perspectives

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.

NATURAL RUBBER BIOSYNTHESIS: STILL A MYSTERY

ABSTRACT Currently, Hevea brasiliensis (the Brazilian rubber tree) is the only commercially available source of natural rubber (NR) for use in many products, which vary from tires to medical products such as adhesive bandages. H. brasiliensis NR is used in these products because after vulcanization, superior properties, which include elasticity, abrasion resistance, and efficient heat dispersion, result. Issues, such as increasing demand and risk of a single source, make a synthetic (manmade) NR an attractive option. However, after years of research efforts, the exact structure of high-molecular-weight NR is still unproven, and a synthetic NR with similar properties to H. brasiliensis NR still has not been developed. To create a replacement synthetic rubber for H. brasiliensis NR, we believe an understanding of NR biosynthesis is necessary. We present a view of NR biosynthesis from a polymer chemistry viewpoint that is based on insight into the mechanism and kinetics of living carbocationic polymerization.