Siriluck Liengprayoon

Glycolipid composition of Hevea brasiliensis latex.

Glycolipids of fresh latex from three clones of Hevea brasiliensis were characterized and quantified by HPLC/ESI-MS. Their fatty acyl and sterol components were further confirmed by GC/MS after saponification. The four detected glycolipid classes were steryl glucosides (SG), esterified steryl glucosides (ESG), monogalactosyl diacylglycerols (MGDG) and digalactosyl diacylglycerols (DGDG). Sterols in SG, ESG and total latex unsaponifiable were stigmasterol, β-sitosterol and Δ⁵-avenasterol. The latter was found instead of fucosterol formerly described. Galactolipids were mainly DGDG and had a fatty acid composition different from that of plant leaves as they contained less than 5% C18:3. Glycolipids, which represented 27-37% of total lipids, displayed important clonal variations in the proportions of the different fatty acids. ESG, MGDG and DGDG from clone PB235 differed notably by their higher content in furan fatty acid, which accounted for more than 40% of total fatty acids. Clonal variation was also observed in the relative proportions of glycolipid classes except MGDG (8%), with 43-51% DGDG, 30-34% SG and 7-19% ESG. When compared with other plant cell content, the unusual glycolipid composition of H. brasiliensis latex may be linked to the peculiar nature of this specialized cytoplasm expelled from laticiferous system, especially in terms of functional and structural properties.

Rubber particle proteins REF1 and SRPP1 interact differently with native lipids extracted from Hevea brasiliensis latex

Rubber particle membranes from the Hevea latex contain predominantly two proteins, REF1 and SRPP1 involved in poly(cis-1,4-isoprene) synthesis or rubber quality. The repartition of both proteins on the small or large rubber particles seems to differ, but their role in the irreversible coagulation of the rubber particle is still unknown. In this study we highlighted the different modes of interactions of both recombinant proteins with different classes of lipids extracted from Hevea brasiliensis latex, and defined as phospholipids (PL), glycolipids (GL) and neutral lipids (NL). We combined two biophysical methods, polarization modulated-infrared reflection adsorption spectroscopy (PM-IRRAS) and ellipsometry to elucidate their interactions with monolayers of each class of lipids. REF1 and SRPP1 interactions with native lipids are clearly different; SRPP1 interacts mostly in surface with PL, GL or NL, without modification of its structure. In contrast REF1 inserts deeply in the lipid monolayers with all lipid classes. With NL, REF1 is even able to switch from α-helice conformation to β-sheet structure, as in its aggregated form (amyloid form). Interaction between REF1 and NL may therefore have a specific role in the irreversible coagulation of rubber particles.

Effect of Rubber Clone on Fatty Acid Composition and Properties of Air Dried Sheet

Properties of air dried sheet (ADS) made from four different clones of Hevea brasiliensis (BPM24, RRIM600, PR255 and PB235) were investigated. BPM24, RRIM600 and PR255 rubbers show a bimodal molar mass distribution (MMD) while PB235 rubber has quasi-unimodal molar mass distribution. It was found that ADS from rubber clone with bimodal MMD (BPM24, RRIM600 and PR255) displayed lower original plasticity (P0) but higher plasticity retention index (PRI). Those rubber samples with bimodal MMD show higher accelerated storage hardening test (ASHT) values than ADS from PB235. Within bimodal rubbers, ADS from PR255 displayed lower PRI and P than the 2 other clones, i.e. RRIM600 and BPM24. ADS made from BPM24 and RRIM600 presented similar fatty acid composition with 3 major fatty acids namely linoleic acid (C18:2), stearic acid (C18:0) and oleic acid (C18:1). In contrast, major fatty acids of ADS made from PR255 were linoleic acid (C18:2) and furan fatty acid. Moreover, ADS from PB235 showed the highest amount of furan fatty acid which confirmed the results of Liengprayoon et al. [1]. Relationship between mesostructure, fatty acids composition and ADS properties are discussed.

A rapid quantitative analysis of native antioxidants in natural rubber (#Hevea brasiliensis#) during maturation

Natural rubber (NR) obtained from H. brasiliensis is known to be susceptible to oxidative degradation according to the amount of double bonds in the structure of the polymer, i.e. poly (cis-1,4-isoprene) [1]. However NR has been reported to contain native antioxidants such as phytosterols, phospholipids and tocotrienols [2]. Among those, γ-tocotrienol, present in hevea latex was reported to exhibit “in-vitro” antioxidant activity [3]. However the direct involvement of γ-tocotrienol or other antioxidants naturally present in NR in the protection of NR against oxidation is difficult to assess as the works were carried out with different of rubber types and in different conditions [4,5]. In the framework of a work on the dynamics of γ-tocotrienols in NR samples obtained from coagula maturated during several durations (0 to 15 days) in parallel with measurement of standard properties such as P0 and PRI, methodological development of the analysis of tocotrienols and derivatives is presented in this study.HPLC-MS has been shown to be an accurate technique for lipid analysis in NR [6]. However, this is a time-consuming technique especially with large number of samples due to a necessary step of saponification. Moreover, the structures of γ-tocotrienol and its dimers were found to be affected by the strong alkali condition of saponification (data not shown). Therefore a rapid quantitative method for γ-tocotrienol from NR using high performance thin layer chromatography (HP-TLC) has been developed. Lipid extracts from NR samples could be simply analyzed by HP-TLC without any derivatization and the detectable quantity could be in nanogram range. Statistical analysis of the data showed that the method is precise, accurate, reproducible and sensitive. Thus the proposed HP-TLC method can be successfully used for the quantification of γ-tocotrienol from NR samples. This technique will be useful to conduct further experiments on antioxidant activity of NR lipids and to relate the results with physical properties of NR, especially those linked to resistance to oxidation.

Starch Synthesis and Mobilization in Wood and Bark of Rubber Tree, in Relation with Latex production, (1) Methodological approach

In rubber tree, starch reserves are necessary for growth and latex regeneration when the demand exceeds supply from photosynthesis. It tends to accumulate in the wood near the tapping cut [1,2] whereas sucrose remains rather stable in the wood and decreases in the latex vessels where it is used to regenerate the exported latex [3].Thus higher starch ability could sustain higher latex yield. However the enzymatic processes driving the dynamics of starch synthesis and hydrolysis as related to tapping are not known. The objective of the study is to analyze the effects of tapping on the enzymes involved in starch and sucrose metabolism in the wood of rubber trees. The first approach of this study was to set up the most adapted methodology on measurement of total nonstructural carbohydrates (NSC) and related enzymes activities. The experiment was conducted in Heveabrasiliensis (rubber tree), clone RRIM600. Treatments include untapped trees (Control) and yielding trees tapped with Ethephon stimulation (ET). Each treatment includes 6 trees. Samples have been collected along the trunk and separated into 2 parts, wood and bark, from both side of the tree tapped and untapped panel. The activity of acid invertase (AI), amylase (AMY), sucrose phosphate synthase (SPS) and sucrose synthase (SuSy) were assessed in the part of soft bark with amethod of sample preparation recoveringmore protein and concentrated enzyme with acetone precipitation. The method has increased enzyme activities of SuSy and SPS. On another hand, starch, sucrose, fructose and glucose concentrations have been enzymatically measured.The results showed that starch was the major component in wood and sucrose was mostly found in bark. There was no difference between the former drainage area and resting area after 2 untapped years before restarting tapping.