Pararamosis: Disease of the Rubber Plantations.

Pararamosis is a medical condition, described in the latex extracting areas of the Amazon (rubber tree regions), resulting from contact with the caterpillar of the Premolis semirufa moth. The disease can present itself in an acute form-similarly to other erucisms (injuries caused by moth larvae in humans)-or in a chronic form, typically characterized by the occurrence of changes in the joints of the hands. Because of its importance, in the context of tropical diseases, the objective of this article was to review the main facets of the disease, emphasizing the different pathogenic aspects of the interaction between the arthropod and man.

Adapting the Accelerated Solvent Extraction Method for Resin and Rubber Determination in Guayule Using the BÜCHI Speed Extractor.

Guayule (Parthenium argentatum Gray) is a promising alternative source to Hevea brasiliensis for the production of natural rubber, which can reach levels of 8-9% under industrialized farming conditions. The most common method for determining rubber concentration is by accelerated solvent extraction (ASE), a technique developed by the Dionex Corporation and almost exclusively performed with the Dionex ASE-200 or 350 systems. Herein, it is sought to apply and adapt the most common methods used in the literature for the Dionex system to another extraction platform, the BÜCHI Speed Extractor E-914. Results showed that using a sand sandwich method to confine the sample in the center and exploiting a larger cell volume (80 mL) for extraction prevents the occurrence of overpressure and problems with clogging. Under optimized conditions, the coefficient of variation was <15% for both resin quantification for samples containing 5.0-15.8% of resin and for rubber quantification for samples with 1.7-10.3% rubber content. The extraction time for resin (2 cycles of 5 min each) was smaller than for rubber (2 cycles of 20 min each). It would be interesting to carry out interlaboratory comparisons to standardize the method at an international level.

Effect of extraction methods on the molecular structure and thermal stability of kenaf (Hibiscus cannabinus core) biomass as an alternative bio-filler for rubber composites.

Lignin from kenaf (Hibiscus cannabinus) core was investigated as an alternative filler for rubber. Three types of extraction methods were used to isolate lignin from kenaf, namely kraft, soda and organosolv process. The particle size, surface area, functionalities changes, molecular weight and thermal properties of the lignin were characterized. The results showed that Kraft lignin (KL) has the smallest particle size (40.41 µm) compared to soda lignin (SL) (63.85 µm) and organosolv lignin (OL) (66.85 µm). This is in good agreement with the BET surface area of 9.52 m2/g, 1.25 m2/g and 2.40 m2/g respectively. However, the smaller surface area of SL compared to OL is due to the smaller pore size and pore volume of SL. KL also showed high hydroxyl content with corresponding high thermal stability as confirmed by NMR and TGA. The thermal stability of the lignin correlates well with the molecular weight (MW). From the overall characteristics, it can be concluded that KL, SL and OL can be used as an alternative filler in rubber compounds to substitute common fillers like silica and carbon.

Production of hydrophobic amino acids from biobased resources: wheat gluten and rubber seed proteins.

Protein hydrolysis enables production of peptides and free amino acids that are suitable for usage in food and feed or can be used as precursors for bulk chemicals. Several essential amino acids for food and feed have hydrophobic side chains; this property may also be exploited for subsequent separation. Here, we present methods for selective production of hydrophobic amino acids from proteins. Selectivity can be achieved by selection of starting material, selection of hydrolysis conditions, and separation of achieved hydrolysate. Several protease combinations were applied for hydrolysis of rubber seed protein concentrate, wheat gluten, and bovine serum albumin (BSA). High degree of hydrolysis (>50 %) could be achieved. Hydrophobic selectivity was influenced by the combination of proteases and by the extent of hydrolysis. Combination of Pronase and Peptidase R showed the highest selectivity towards hydrophobic amino acids, roughly doubling the content of hydrophobic amino acids in the products compared to the original substrates. Hydrophobic selectivity of 0.6 mol-hydrophobic/mol-total free amino acids was observed after 6 h hydrolysis of wheat gluten and 24 h hydrolysis of rubber seed proteins and BSA. The results of experiments with rubber seed proteins and wheat gluten suggest that this process can be applied to agro-industrial residues.

Effects of forests, roads and mistletoe on bird diversity in monoculture rubber plantations.

Rising global demand for natural rubber is expanding monoculture rubber (Hevea brasilensis) at the expense of natural forests in the Old World tropics. Conversion of forests into rubber plantations has a devastating impact on biodiversity and we have yet to identify management strategies that can mitigate this. We determined the life-history traits that best predict bird species occurrence in rubber plantations in SW China and investigated the effects of surrounding forest cover and distance to roads on bird diversity. Mistletoes provide nectar and fruit resources in rubber so we examined mistletoe densities and the relationship with forest cover and rubber tree diameter. In rubber plantations, we recorded less than half of all bird species extant in the surrounding area. Birds with wider habitat breadths and low conservation value had a higher probability of occurrence. Species richness and diversity increased logarithmically with surrounding forest cover, but roads had little effect. Mistletoe density increased exponentially with rubber tree diameters, but was unrelated to forest cover. To maximize bird diversity in rubber-dominated landscapes it is therefore necessary to preserve as much forest as possible, construct roads through plantations and not forest, and retain some large rubber trees with mistletoes during crop rotations.

Genetic and biochemical evaluation of natural rubber from Eastern Washington prickly lettuce (Lactuca serriola L.).

Alternative sources of natural rubber are of importance due to economic, biological, and political threats that could diminish supplies of this resource. Prickly lettuce (Lactuca serriola L.) synthesizes long-chain natural rubber and was studied to determine underlying genetic and phenotypic characteristics of rubber biosynthesis. Genotypic and phenotypic analysis of an F2 segregating population using EST-SSR markers led to the discovery of genetic regions linked to natural rubber production. Interval mapping (IM) and multiple QTL mapping (MQM) identified several QTL in the mapping population that had significance based on LOD score thresholds. The discovered QTL and the corresponding local markers are genetic resources for understanding rubber biosynthesis in prickly lettuce and could be used in marker-assisted selection (MAS) breeding. Prickly lettuce is an excellent candidate for elucidating the rubber synthesis mechanism and has potential as a crop plant for rubber production.

Natural rubber biosynthesis in plants: rubber transferase.

Rubber biosynthesis in plants is a fascinating biochemical system, which evolved at the dawn of the dicotyledoneae and is present in at least four of the dictolydonous superorders. Rubber biosynthesis is catalyzed by a membrane complex in a monolayer membrane envelope, requires two distinct substrates and a divalent cation cofactor, and produces a high-molecular-weight isoprenoid polymer. A solid understanding of this system underpins valuable papers in the literature. However, the published literature is rife with unreliable reports in which the investigators have fallen into traps created by the current incomplete understanding of the biochemistry of rubber synthesis. In this chapter, we attempt to guide both new and more established researchers around these pitfalls.

Proteome analysis of the large and the small rubber particles of Hevea brasiliensis using 2D-DIGE.

The rubber particle is a specialized organelle in which natural rubber is synthesised and stored in the laticifers of Hevea brasiliensis (para rubber tree). It has been demonstrated that the small rubber particles (SRPs) has higher rubber biosynthesis ratio than the large rubber particles (LRPs), but the underlying molecular mechanism still remains unknown. In this study, LRPs and SRPs were firstly separated from the fresh latex using differential centrifugation, and two-dimensional difference in-gel electrophoresis (2D-DIGE) combined with MALDI-TOF/TOF was then applied to investigate the proteomic alterations associated with the changed rubber biosynthesis capacity between LRPs and SRPs. A total of 53 spots corresponding to 22 gene products, were significantly altered with the |ratio|≥2.0 and T value ≤0.05, among which 15 proteins were up-regulated and 7 were down-regulated in the SRPs compared with the LRPs. The 15 up-regulated proteins in the SRPs included small rubber particle protein (SRPP), 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS), phospholipase D alpha (PLD α), ethylene response factor 2, eukaryotic translation initiation factor 5A isoform IV (eIF 5A-4), 70-kDa heat shock cognate protein (HSC 70), several unknown proteins, etc., whereas the 7 up-regulated proteins in the LRPs were rubber elongation factor (REF, 19.6kDa), ASR-like protein 1, REF-like stress-related protein 1, a putative phosphoglyceride transfer family protein, ß-1,3-glucanase, a putative retroelement, and a hypothetical protein. Since several proteins related to rubber biosynthesis were differentially expressed between LRPs and SRPs, the comparative proteome data may provide useful insights into understanding the mechanism involved in rubber biosynthesis and latex coagulation in H. brasiliensis.

Extraction and characterization of a natural rubber from Euphorbia characias latex.

A natural rubber was identified and characterized for the first time in the latex of the perennial Mediterranean shrub Euphorbia characias. Four different methods, i.e., acetone, acetic acid, trichloroacetic acid, and Triton® X-100, followed by successive treatments with cyclohexane/ethanol, were employed to extract the natural rubber. The rubber content was shown to be 14% (w/v) of the E. characias latex, a low content compared with that of Hevea brasiliensis (30-35%) but a similar content to other rubber producing plants. E. characias rubber showed a molecular weight of 93,000 with a M(w) /M(n) of 2.9. (1) H NMR, (13) C NMR, and FTIR analysis revealed the characteristic of the cis-1,4-polyisoprene typical of natural rubber. These results provided novel insight into latex components and will ultimately benefit the broader understanding of E. characias latex composition.

Extraction and characterization of latex and natural rubber from rubber-bearing plants.

Consecutive extraction of latex and natural rubber from the roots of rubber-bearing plants such as Taraxacum kok-saghyz (TKS), Scorzonera tau-saghyz (STS), and Scorzonera Uzbekistanica (SU) were carried out. Latex extraction was carried via two methods: Blender method and Flow method. The results of latex extraction were compared. Cultivated rubber-bearing plants contained slightly higher latex contents compared to those from wild fields. Several creaming agents for latex extraction were compared. About 50% of total natural rubber was extracted as latex. The results of the comparative studies indicated that optimum latex extraction can be achieved with Flow method. The purity of latex extracted by Blender method ( approximately 75%) was significantly lower than that extracted by Flow method (99.5%). When the latex particles were stabilized with casein, the latex was concentrated significantly. Through concentrating latex by flotation, the latex concentration of 35% was obtained. Bagasse contained mostly solid natural rubber. The remaining natural rubber in the bagasse (left after the latex extraction) was extracted using sequential solvent extraction first with acetone and then with several nonpolar solvents. Solid natural rubber was analyzed for gel content and characterized by size exclusion chromatography (SEC) for molecular weight determinations. SEC of solid natural rubber has shown that the molecular weight is about 1.8E6 and they contain less gel compared to TSR20 (Grade 20 Technically Specified Rubber), a commercial natural rubber from Hevea brasiliensis.

Characterization of natural rubber using size-exclusion chromatography with online multi-angle light scattering. Study of the phenomenon behind the abnormal elution profile.

Natural and synthetic poly(cis-1,4-isoprene) were characterized by size-exclusion chromatography coupled with an online multi-angle light scattering detector (SEC-MALS). Unlike synthetic poly(cis-1,4-isoprene) (SR), natural rubber (NR) samples showed anomalous elution profiles. The beginning of elution was very similar to SR but, after a certain elution volume, the molar masses of the eluting macromolecules increased with elution volume instead of continuing to decrease, which resulted in an upturn curve profile. Adding tetrabutylammonium bromide (TBABr) to THF (solvent and mobile phase) removed this phenomenon. In addition, using different concentrations of TBABr showed that TBABr had two simultaneous actions. TBABr reduced the abnormal elution profiles and the quantity of aggregates (insoluble part or gel). These results mean that the main phenomenon involved in abnormal elution was delayed entities adsorbing on the column packing. Their delayed elution was responsible for the artificial increase in molar masses, especially at high elution volumes. The results obtained suggest that these entities are very compact and have a sphere-like structure.

Establishment of new crops for the production of natural rubber.

Natural rubber is a unique biopolymer of strategic importance that, in many of its most significant applications, cannot be replaced by synthetic alternatives. The rubber tree Hevea brasiliensis is the almost exclusive commercial source of natural rubber currently and alternative crops should be developed for several reasons, including: a disease risk to the rubber tree that could potentially decimate current production, a predicted shortage of natural rubber supply, increasing allergic reactions to rubber obtained from the Brazilian rubber tree and a general shift towards renewables. This review summarizes our knowledge of plants that can serve as alternative sources of natural rubber, of rubber biosynthesis and the scientific gaps that must be filled to bring the alternative crops into production.

Periploca sepium Bunge as a model plant for rubber biosynthesis study.

Periploca sepium Bunge (Chinese silk vine) is a woody climbing vine belonging to the family Asclepiadaceae. It originally comes from Northwest China. Periploca resembles the Para-rubber tree, Hevea brasiliensis, regarding a similar body plan to produce a milky exudate containing rubber latex. The Periploca plant was assessed as a rubber-producing plant by rubber structure elucidation and its molecular weight distribution. A rubber fraction purified from the milky exudate was subjected to 1H NMR analysis, and a characteristic signal derived from cis-polyisoprene was observed. In addition, when the molecular weight distribution of rubber components in the exudate was measured (using size-exclusion chromatography), the number-average molecular weight (Mn), weight-average molecular weight (Mw), and polydispersity (Mw/Mn) were estimated to be Mn = 1.3 x 10(5), Mw = 4.1 x 10(5), and Mw/Mn = 3.1, respectively. Furthermore, the presence of polyisoprene, with Mn = 4.0 x 10(4), Mw = 7.6 x 10(4), and Mw/Mn = 2.5, was also confirmed in plantlets obtained from shoots as a result of tissue culture.

Relationship between latex yield of Hevea brasiliensis and antecedent environmental parameters.

A study on the relationship between latex yield and antecedent environmental data was undertaken for five clones (RRII203, RRII118, RRIM600, RRII105 and GT1) of Hevea brasiliensis (rubber) in Agartala, northeast India, a region in which rubber is not traditionally cultivated. The explained variance for the regression equations based on parameters determined on the day of tapping and up to 3 days prior to it, varied from 72% to 37% during the NWT period and 94-83% during the WT period. Soil moisture storage, 1 and 3 days prior to tapping, was found to be the primary parameter affecting yield for the NWT and WT periods, respectively. It was observed that the clone RRII105, with a comparatively lower yield to that of RRIM600, was more susceptible to daily WD conditions during the non-winter season. RRIM600 and RRII105 being high-yielding clones were also found to be fairly dependent on the AT of the day prior to tapping. The mean lag period correlation of this parameter with yield was also found to be higher during the WT period than during the NWT period. As a whole, the mean lag period based on prior measurements of environmental variables showed optimum correlation with yield at 15-20 days prior to the day of tapping. The study also confirms that varied responses of yield with environmental factors in this non-traditional region of rubber cultivation depend on clonal character.

Purification and characterization of NAD(P)H quinone reductase from the latex of Hevea brasiliensis Müll.-Arg. (Euphorbiaceae).

NAD(P)H quinone reductase [NAD(P)H-QR] present in the latex of Hevea brasiliensis Müll.-Arg. (Euphorbiaceae) was purified to homogeniety from the B-serum fraction obtained by freeze-thawing of the bottom fraction of ultracentrifuged fresh latex. The purification protocol involved acetone fractionation, heat treatment, ion exchange chromatography and affinity chromatography. The M(r) determined by SDS-PAGE for the protein subunit was 21 kDa, and the molecular mass of the native enzyme estimated by gel filtration was 83 kDa, indicating that the native enzyme is a homotetramer. The enzyme showed pH stability over a range of 6 to at least 10 (with an optimum at pH 8) and thermal stability up to 80 degrees C. High NAD(P)H-QR activity (70%) was still retained after 10 h of preincubation at 80 degrees C. A comparable substrate specificity for this enzyme was observed among menadione, p-benzoquinone, juglone, and plumbagin, with only duroquinone generating a lower activity. Positive correlations between latex NAD(P)H-QR activity and rubber yield per tapping [fresh latex (r=0.89, P<0.01), dry rubber (r=0.81, P<0.01)] together with flow time (r=0.85, P<0.01) indicated that enzyme activity could possibly be used as a marker to predict the yield potential of selected clones.

Identification of natural rubber and characterization of rubber biosynthetic activity in fig tree.

Natural rubber was extracted from the fig tree (Ficus carica) cultivated in Korea as part of a survey of rubber producing plants. Fourier transform infrared and (13)C nuclear magnetic resonance analysis of samples prepared by successive extraction with acetone and benzene confirmed that the benzene-soluble residues are natural rubber, cis-1,4-polyisoprene. The rubber content in the latex of fig tree was about 4%, whereas the rubber content in the bark, leaf, and fruit was 0.3%, 0.1%, and 0.1%, respectively. Gel-permeation chromatography revealed that the molecular size of the natural rubber from fig tree is about 190 kD. Similar to rubber tree (Hevea brasiliensis) and guayule (Parthenium argentatum Gray), rubber biosynthesis in fig tree is tightly associated with rubber particles. The rubber transferase in rubber particles exhibited a higher affinity for farnesyl pyrophosphate than for isopentenyl pyrophosphate, with apparent K(m) values of 2.8 and 228 microM, respectively. Examination of latex serum from fig tree by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed major proteins of 25 and 48 kD in size, and several proteins with molecular mass below 20 and above 100 kD. Partial N-terminal amino acid sequencing and immunochemical analyses revealed that the 25- and 48-kD proteins were novel and not related to any other suggested rubber transferases. The effect of EDTA and Mg(2+) ion on in vitro rubber biosynthesis in fig tree and rubber tree suggested that divalent metal ion present in the latex serum is an important factor in determining the different rubber biosynthetic activities in fig tree and rubber tree.

Structure of cis-polyisoprene from Lactarius mushrooms.

Sporophores from five species of Lactarius mushrooms had a liquid rubber content of 0.1% to 7% based on the dry weight. Rubber from L. volemus, L. chrysorrheus and L. hygrophoroides was found to be a homologue of polyprenol being composed of dimethylallyl group, two trans isoprene units, 160-300 cis isoprene units, and terminal hydroxyl or ester group aligned in that order by 13C-NMR analysis. The ratio of fatty acid ester group to hydroxyl group was about 9/1 to 5/5. The number of both terminal groups and trans units decreased during aging of sporophores. Rubber from L. piperatus, L. vellereus and L. subpiperatus was found to be cis polyisoprene having very small quantities of both terminal groups and trans units. The biosynthesis of cis polyisoprene in Lactarius mushrooms was found to start from trans, trans-farnesyl pyrophosphate. The termination was assumed to occur by esterification of polyisoprenyl pyrophosphate. Occurrence of some chemical modifications on both terminal groups was presumed during aging of sporophores.