Cloning and characterization of the promoters of the key genes CPT, SRPP and REF involved in Periploca sepium rubber biosynthesis / 生物工程学报

Hevea brasiliensis is the main source of natural rubber. Restricted by its tropical climate conditions, the planting area in China is limited, resulted in a low self-sufficiency. Periploca sepium which can produce natural rubber is a potential substitute plant. cis-prenyltransferase (CPT), small rubber particle protein (SRPP) and rubber elongation factor (REF) are key enzymes involved in the biosynthesis of cis-1, 4-polyisoprene, the main component of natural rubber. In this study, we cloned the promoter sequences of CPT, SRPP and REF through chromosome walking strategy. The spatial expression patterns of the three promoters were analyzed using GUS (β-glucuronidase) as a reporter gene driven by the promoters through Agrobacterium-mediated genetic transformation. The results showed that GUS driven by CPT, SRPP or REF promoter was expressed in leaves and stems, especially in the leaf vein and vascular bundle. The GUS activity in stems was higher than that in leaf. This study provided a basis for analyzing the biosynthesis mechanism of natural rubber and breeding new varieties of high yield natural rubber.

Atmospheric evaporative demand and water deficit on the ecophysiology of Rubber seedlings

The search for genetic materials resistant to adverse weather conditions has been a major focus in studies on species of economic interest. The objective of the present study was to assess the growth and photosynthesis of rubber seedlings clones under two conditions of atmospheric evaporative demand, characterized by fluctuations in temperature (TEMP) and vapor pressure deficit (VPD), associated to two water regimens. Hevea brasiliensis Muell. Arg (RRIM 600 and FX 3864) clones were assessed in two microclimates, at low (TEMP 21.2 ºC and VPD 0.29 Kpa) and high (TEMP 26.9 ºC and VPD 1.49 Kpa) atmospheric evaporative demand, under two water regimens: water deficit and well-watered. Water deficit 50% water availability was sufficient to reduce the net CO2 assimilation rate, leaf area and total chlorophyll of the clones studied that impacted growth in both microclimates. The effects of water deficit on growth and net carbon assimilation rate were intensified under high atmospheric evaporative demand. However, when comparing the two clones studied, RRIM 600 showed greater growth and photosynthesis without water restriction. The FX 3864 clone, despite the high CO2 assimilation values under high atmospheric demand and without water restriction, showed a reduced growth. The results of this study form an important basis for the selection of genotypes with the potential to develop in adverse climatic conditions. In this sense, the RRIM 600 genotype is recommended as a promising material that would best adapt under adverse climatic conditions.

Rubber gloves biodegradation by a consortium, mixed culture and pure culture isolated from soil samples

Abstract An increasing production of natural rubber (NR) products has led to major challenges in waste management. In this study, the degradation of rubber latex gloves in a mineral salt medium (MSM) using a bacterial consortium, a mixed culture of the selected bacteria and a pure culture were studied. The highest 18% weight loss of the rubber gloves were detected after incubated with the mixed culture. The increased viable cell counts over incubation time indicated that cells used rubber gloves as sole carbon source leading to the degradation of the polymer. The growth behavior of NR-degrading bacteria on the latex gloves surface was investigated using the scanning electron microscope (SEM). The occurrence of the aldehyde groups in the degradation products was observed by Fourier Transform Infrared Spectroscopy analysis. Rhodococcus pyridinivorans strain F5 gave the highest weight loss of rubber gloves among the isolated strain and posses latex clearing protein encoded by lcp gene. The mixed culture of the selected strains showed the potential in degrading rubber within 30 days and is considered to be used efficiently for rubber product degradation. This is the first report to demonstrate a strong ability to degrade rubber by Rhodococcus pyridinivorans.

Membrana de látex natural de Hevea brasiliensis auxilia no processo de reparação tecidual em bovinos / Natural latex rubber from Hevea brasiliensis aids tissue repair of bovine

Feridas cutâneas em bovinos são um constante desafio clínico cirúrgico por desencadearem perdas econômicas bastante significativas. O látex proveniente da seiva da seringueira (Hevea brasiliensis) apresenta potencial terapêutico para incrementar o processo de reparação tecidual. Portanto, pretendeu-se com esse estudo avaliar o tipo de reação tecidual e os possíveis mecanismos de angiogênese desencadeados pelo implante de uma membrana de látex natural em bovinos. Para tal, foram utilizados seis bovinos da raça Nelore, submetidos ao implante subcutâneo experimental de três fragmentos de membranas de látex natural. Foram coletadas amostras de tecido e da membrana aos 15, 30 e 45 dias após a implantação, para avaliações histológicas, ultraestruturais por microscopia eletrônica de varredura e imunoistoquímicas com anticorpos antimarcador de macrófagos (MAC), CYR 61 e VEGF. O implante de látex proporcionou aumento da angiogênese e reparação tecidual em bovinos, não mediada pela expressão do VEGF e CYR 61.(AU)

Cattle wounds are a constant surgical and clinical challenge, leading to important economical losses. The latex from the sap of the rubber tree (Hevea brasiliensis) has therapeutic potential to enhance tissue repair process. Therefore, we evaluated the type of tissue reaction and possible mechanisms of angiogenesis triggered by implanting natural latex rubber in bovine species. Six Nelore bovines were subjected to subcutaneous experimental implant of three fragments of natural rubber latex membranes. Tissue and rubber membrane samples were harvested at 15, 30 and 45 days implantation for histology, scanning electron microscopy and immunohistochemical evaluation with anti macrophage marker (MAC), anti CYR 61, anti VEGF antibodies. The latex membrane estimulates tissue reaction and repair and significant angiogenesis stimuli without activating CYR 61 and VEGF pathways.(AU)

Evaluation of Rubber Tree Clones Under Different Tapping Systems in Northwestern Paraná State, Brazil

ABSTRACT This study evaluates the potential of rubber tree clones (Hevea spp.) under four tapping systems for the Northwestern region of the State of Paraná with the objective of enhance genetic diversification and increase natural rubber production in this new and promising region for rubber tree cultivation by selecting the better combination of clone and tapping system. The experiment was installed at Guanabara Farm, municipality of Paranapoema, Paraná State, in a split-plot randomized complete blocks experimental design with three replications. In the plots 12 clones were tested, namely RRIM 600 (control), IRCA 18, IRCA 111, IRCA 130, PB 235, PB 252, PB 260, PB 330, IAC 15, IAC 35, IAC 41 and RO 38. The subplot treatments consisted of four tapping systems: 1) ½ S d/4 ET 2.5%, 2) ½ S d/4, 3) ½ S d/7 ET 2.5% and 4) ½ S d/7. Dry rubber production was evaluated over a period of two years. Due to the higher productivity the clones IRCA 130, IRCA 111, IRCA 18, IAC 35, IAC 41, IAC 15, PB 260 and PB 235 are suggested to be planted at a proportion of 5% to 10% of the total area in commercial plantings for more detailed evaluation. Because of their significantly lower productivity, the clones PB 330, PB 252 and RO 38 should not be recommended, as well as the tapping system S d/7 ½ without stimulation.

Novel sustained-release of Stryphnodendron obovatum leaves extract using natural rubber latex as carrier

Natural rubber latex biomembranes (NRL), obtained from rubber tree Hevea brasiliensis (Willd. ex A. Juss.) Mull. Arg., have been used as sustained drug release of drugs and plant extracts with medicinal properties. The Stryphnodendron obovatum Bench (Fabaceae), popularly known as "barbatimão" has anti-inflammatory and healing properties already described in literature. Thus, the aim of this work were to study the release behavior of the hydroethanolic extract from the leaves of S. obovatum loaded in the NRL by ultraviolet­visible spectroscopy (UV-VIS). The release followed a biexponential pattern and the mechanism of release was Super Case II (n > 1). FTIR analyses did not show reaction between NRL and extract, only intermolecular interaction. From SEM was possible to observe the extract at the surface, responsible for the initial fast release, which the concentrations at 5.0 mg/mL released 2.4% and at 0.1 mg/mL released 96.8%; both reached the plateau in 7 days.

Casearia sylvestris Swartz Extract Release Using Natural Rubber Latex Biomembranes as Carrier.

The Natural Rubber Latex (NRL) from Hevea brasiliensis has shown promise in biomedical applications due to its low cost, easy handling, mechanical properties and biocompatibility, being used for bone regeneration and wound healing due to its natural stimulus to angiogenesis. The aim of this work was to incorporate Casearia sylvestris Sw. extract in NRL biomembranes and study its release behavior. The complex membraneextract has as object of study a new approach of using C. sylvestris extract in the treatment of wounds, for possessing antiseptic activity, anti-inflammatory and analgesic properties. The C. sylvestris species (Salicaceae), popularly known as "guaçatonga", presents great distribution and is used in folk medicine as antiulcer, wound healing, anti- snake venom, properties which have been proven and related to clerodane diterpenes (casearins A-X). The release rate of C. sylvestris compounds from extract-membrane complex was monitored and analyzed using the method of optical spectroscopy (UV-VIS). The release varied with temperature ranging from 14 to 33 days, releasing more than 90%, with an interesting and promising biomedical application, such as wound healing and burns.

Liberação de componentes do extrato de Casearia sylvestris Swartz empregando membranas de látex natural como suporte / Analysis of Casearia sylvestris Swartz extract release from natural rubber latex membranes

A membrana de látex natural (NRL - Natural Rubber Latex), manipulada a partir do látex extraído da seringueira Hevea brasiliensis, tem apresentado grande potencial de aplicação no campo da biomedicina e dos biomateriais. Graças a sua biocompatibilidade e baixa taxa de rejeição, ela tem sido utilizada para próteses e enxertos, atuando como estimulante da angiogênese, adesão celular e barreira física a agentes infecciosos. Além dessas aplicações, as membranas são utilizadas como matriz de sistemas de liberação para avaliar o comportamento da liberação de fármacos e extratos de origem vegetal que apresentam propriedades medicinais. O sistema extrato-membrana tem como objeto de estudo uma nova abordagem dessas substâncias no tratamento de feridas visando à cicatrização e regeneração do tecido envolvido. Casearia sylvestris, conhecida popularmente como guaçatonga, pertence à família Salicaceae, conhecida na medicina popular pelos seus efeitos antiulcerogênicos, cicatrizantes, antiofídicos, anti-inflamatórios e antissépticos, propriedades já comprovadas por estudos. Trabalhos recentes demonstraram que a liberação controlada de fármacos e extratos utilizando membranas de látex natural é uma alternativa interessante e promissora para aplicações biomédicas. Assim, o objetivo deste projeto foi estudar a liberação do extrato de Casearia sylvestris incorporado à membrana de látex em soluções com diferentes valores de pH, com o propósito de estudar seu comportamento e sua liberação de forma controlada. A taxa de liberação do extrato do sistema extrato-membrana foi monitorada e analisada utilizando-se o método de espectroscopia ótica (UV). O pH básico age desestabilizando a membrana indicando que o polímero possui a habilidade de proteger o extrato de ser liberado em valores de pH ácidos, direcionando sua aplicação para o sítio de melhor absorção. Além disso, a liberação segue uma função bi-exponencial...

Natural Rubber Latex (NRL) membrane, made from latex extracted from the rubber tree Hevea brasiliensis, has shown great potential for use in the biomedicine and biomaterials area. Thanks to its biocompatibility and low rejection rate, it has been used for implants and grafts, acting as a stimulant of angiogenesis and cell adhesion and as a barrier against infectious agents. Besides these applications, the membranes are used as model release systems, to assess the release behavior of drugs and plant extracts that exhibit medicinal properties. The extract-membrane system represents a new approach to studying these substances, as aids to wound healing and tissue regeneration. Casearia sylvestris, popularly known as guaçatonga, belongs to the family Salicaceae, known in popular medicine for its anti-ulcerogenic, wound healing, anti-ophidian, antiinflammatory and antiseptic properties, all of which are proven by scientific studies. Recent studies have also shown that the controlled release of drugs and extracts from natural latex membranes is an interesting and promising process for biomedical applications. The objective of this project was thus to study the release of Casearia sylvestris extract incorporated into natural rubber membranes. The main concern was to study and optimize the controlled release of the extract at various pHs. The rate of release was monitored and analyzed by the method of optical spectroscopy (UV). Basic pHacts to destabilize the membrane, indicating that the polymer has the ability to protect the extract from being released at acidic pH values. The controlled release follows a bi-exponential function...

Biodegradation of a blended starch/natural rubber foam biopolymer and rubber gloves by Streptomyces coelicolor CH13

Background: The growing problem of environmental pollution caused by synthetic plastics has led to the search for alternative materials such as biodegradable plastics. Of the biopolymers presently under development, starch/natural rubber is one promising alternative. Several species of bacteria and fungi are capable of degrading natural rubber and many can degrade starch. Results: Streptomyces coelicolor CH13 was isolated from soil according to its ability to produce translucent halos on a mineral salts medium, MSM, supplemented with natural rubber and to degrade starch. Scanning electron microscope studies showed that it colonized the surfaces of strips of a new starch/natural rubber biopolymer and rubber gloves and caused degradation by forming holes, and surface degradation. Starch was completely removed and polyisoprene chains were broken down to produce aldehyde and/or carbonyl groups. After 6 weeks of cultivation with strips of the polymers in MSM, S. coelicolor CH13 reduced the weight of the starch/NR biopolymer by 92 percent and that of the rubber gloves by 14.3 percent. Conclusions: This study indicated that this bacterium causes the biodegradation of the new biopolymer and natural rubber and confirms that this new biopolymer can be degraded in the environment and would be suitable as a ‘green plastic’ derived from natural sources.

Efecto de la incculación con hongos formadores e micorrizas arbusculares sobree plántulas de caucho / Effects Of Inoculation With Arbuscular Mycorrhizal Fungi On Rubber Seedlings

Hongos formadores de micorrizas arbusculares (HFMA) obtenidos a partir de suelos caucheros se multiplicaron en plantas de Lolium sp., con 73% de colonización radical luego de cuatro meses. Se obtuvieron siete morfotipos de HFMA, con los cuales se inocularon dos grupos de plántulas de Hevea brasiliensis: 1) producidas in vivo a partir de semilla; 2) producidas in vitro por rescate de embrión, para determinar efectos sobre mortalidad, crecimiento, micorrización y contenido de fósforo foliar. Los niveles de colonización por HFMA para las plántulas obtenidas in vitro e in vivo fueron de 12,6% y de 44,7%, respectivamente. La biomasa media total acumulada por las plántulas producidas in vitro fue de 0,41 y de 1,40 g por las procedentes del material in vivo, en comparación con los controles no inoculados, los cuales acumularon 0,37 y 0,40 g , respectivamente. El tratamiento con HFMA disminuyó la mortalidad en las plántulas obtenidas in vitro, aunque no tuvo un efecto significativo sobre el contenido de fósforo foliar. La respuesta del crecimiento de las plántulas inoculadas fue diferente dependiendo del origen del material vegetal y del tipo de inóculo (nativo o no nativo). La simbiosis entre HFMA y H. brasiliensis se desarrolló en condiciones controladas de crecimiento, aunque su avance dependió del estado de desarrollo de la plántula. No obstante, influyó en el crecimiento y en la disminución de la mortalidad de las plántulas, lo que abre la posibilidad de utilizarla como alternativa de inoculación en las fases tempranas de obtención del material vegetal.

Hevea brasiliensis rubber plants were inoculated with native arbuscular mycorrhizal fungi (AMF) during their greenhouse acclimatization. The AMF were multiplied for 4 months associated with Lolium sp. plants having 73% root colonization. Seven morphotypes were obtained. Two different groups of H. brasiliensis plants were inoculated with these morphotypes to determine AMF effect on mortality rate, growth, mycorrhization and leaf P content: those produced in vivo by seed germination and those produced in vitro by micropropagation. Plantlets produced in vitro returned 12.6% AMF colonization and 0.41 g accumulated dry biomass, whereas seedlings produced in vivo had 44.7% and 1.40 g , respectively, compared to control plants with no AMF inoculation which accumulated 0.37 g (in vitro) and 0.40 g (in vivo). AMF inoculation reduced the plants’ mortality rate regarding in vitro produced material; however, it did not influence foliar P content. Inoculated plants’ growth response was different, depending on plant material and the kind of inoculum used (native or foreign). Symbiosis developed between AMF and H. brasiliensis in controlled growth conditions although its advance was limited by the plants’ growth stage. Nevertheless, such association influenced inoculated plants’ biomass accumulation and decreased their mortality rate and could be used as an alternative treatment during the earliest stages of H. brasiliensis plantlet production.

Study on the Necessity of Clearing Butyl Rubber Stoppers for Infusion Bottles / 中国药房

OBJECTIVE:To discuss the necessity of clearing the pharmaceutical butyl rubber stoppers for infusion bottles before use.METHODS:Pharmaceutical butyl rubber stoppers for infusion bottles produced by manufacturer A and B were se-lected and washed at least once before a preliminary test was performed.Then product A was chosen for further test.The stoppers of two different batch numbers by manufacturer A were selected,one washed with injecting water and another not washed,all of which were used in the production of all kinds of transfusions in the hospital where the authors worked,then infusible motes in these products were measured.RESULTS:The infusible motes in the products with unwashed stoppers was not in accordance with the requirements in China Pharmacopoeia,while those with washed ones was in accordance.CONCLU_ SION:The pharmaceutical butyl rubber stoppers for infusion bottles must be washed by injecting water at least once before use to ensure the quality of the products.