De Novo Transcriptome Analysis in Hevea brasiliensis to Unveil Genes Involved in Low Temperature Stress Response

Low temperature is one of the adversities threatening the growth and development and reduces the yield of rubber trees.However,molecular mechanisms toward rubber trees in response to low temperature are largely unclear.In this study,7,159 and 7,600 differentially expressed genes(DEGs)were identified in‘Reyan 73397’rubber trees.Through GO analysis,the catalytic activity was the representative of the GO term in the only DEGs at the two studied temperatures(room temperature and 4°C,respectively),while KEGG analysis showed that carbon metabolism was the most important grouping under the comparison of these two temperatures.In addition,expression of 9 members of transcription factor MYB family genes were further verified by qRT-PCR,and MYB family genes may play important roles in the regulation of rubber trees under low temperature stress.This study provided a theoretical foundation for(1)revealing the molecular mechanisms of rubber trees in response to low temperature and(2)breeding of tolerant varieties of rubber trees.

Evaluation of unconstrained and constrained mathematical functions to model girth growth of rubber trees (Hevea brasiliensis) using young agemeasurements

No attempt has been made to date to model growth in girth of rubber tree （Hevea brasiliansis）. We evaluated the few widely used growth functions to identify the most parsimonious and biologically reasonable model for describing the girth growth of young rubber trees based on an incomplete set of young age measurements. Monthly data for girth of immature trees （age 2 to 12 yearsi from two locations were sub- jected to modelling. Re-parameterized, unconstrained and constrained growth functions,of Richards （RM）, Gompertz （GM） and the monomo- lecular ＇model ^（MM） were fitted to data. Duration of growth was the firsf constraint introduced. In the stagel We attempted a population aver- age （PA） model to capture the trend in growth. The best PA model was fitted as a subject specific （SS） model. We used appropriate error vari- ance-covariance structure to account for correlation due to repeated measurements over time. Unconstrainecl functions underestimated the asymptotic maximum that did not reflective carrying capacity of the locations. Underestimafions were attributed to the partial set＇ of meas- urements made during the early growth phase of the trees. MM proved superior to RM and GM. In the randomcoefficient models, both Gf and Go appeared to be influenced by tree level effects. Inclusion of diagonal definite positive matrix removed the correlation between random effects. The results were similar at both locations. In the overall assessment MM appeared as the candidate model for studying the girth-age relationships in Hevea trees. Based on the fitted model we conclude that, in Hevea trees, growth rate is maintained at maximum value at to, then decreases until the final state at dG/dt 〉 0, resulting in yield curve with no period of accelerating growth. One physiological explanation is that photosynthetic activity in Hevea trees decreases as girth increases and constructive metabolism is larger than destructive metabolism.

Preliminary Study on Seedling Growth Rhythm and Grey Correlation Analysis of Rubber (Hevea brasiliensis) Seedlings in Danzhou District, Hainan

The rubber tree physiological and ecological process quantitatively described by using mathematical method is an important means to the analysis of rubber tree growth process and mechanism. The study on growth simulation model of rubber tree will lay the foundation for the application of rubber tree cultivation intelligent decision system. A Richards equation was formulated to describe the height and stem diameter growth dynamics of the annual rubber seedlings. An area correlation analysis was done according to the closeness of the observed parameters to the dynamic curve on the gray system composed of the seedling growth increment and the meteorological factors including aerial temperature, precipitation and solar radiation hours that influence upon the seedling growth. The results showed that rubber seedling response fitted the Richards equation quite well. The growth increment displayed a distinct alternation of 'slow—fast—slow— fast—slow' rhythm. The growth course of the seedlings might be partitioned into three periods of time by the sequential clustering analysis, namely pre-growing, fast-growing, late-growing stage. The tray correlation analysis revealed that air temperature had the most significant influence while precipitation had the least impact on height growth of the rubber seedlings. In conclusion, the air temperature had the most significant influence while solar radiation hours had the least impact on stem diameter growth of the rubber seedlings.

Effect of natural and synthetic antioxidants on oxidative stability of FAMEs obtained from hevea brasiliensis

Biodiesel(fatty acid alkyl esters),an alternate to fossil fuel,has the tendency of autoxidation and hence requires antioxidants for long term storage.The influence of synthetic and natural antioxidants on the oxidative stability was analysed for fresh FAMEs(fatty acid methyl esters)obtained from hevea brasiliensis at 140 C.Higher activity was observed for synthetic antioxidants following the order of GA】BHT】DTBP】Q】GT-M】PH-M】GT-C,whereas the oxidative stability of stored FAMEs samples measured at 110 C reveals a nearly inverse trend.Storage stability was tested for the FAMEs obtained from hevea brasiliensis stored at 30 C,after addition of synthetic and natural antioxidants—butylated hydroxytoluene(BHT),2,6-di-tert-butylphenol(DTBP),quercetin(Q),gallic acid(GA),methanol extracts from green tea(GTM),pomegranate hull(PH-M),and chloroform extract of green tea(GT-C).Antioxidant activities above 1500 ppm was in the order of DTBP】BHT】GA】GT-C】GT-M】Q】PH-M.Synthetic antioxidants have been found more efficient to improve the storage stability of FAMEs obtained from hevea brasiliensis.DTBP in particular has the highest protection factor.

Proximate Composition and Fatty Acid Profile of Raw and Roasted Salt-Dried Sardines (<i>Sardinella Brasiliensis</i>)

The proximate composition and fatty acid profile of five lots of samples of raw and roasted salt-dried sardines (Sardi-nella brasiliensis) bought locally in Maringá, Paraná State, Brazil were determined. Significant differences (P < 0.05) were observed between samples of raw and roasted sardines, both in relation to moisture, total lipids, proteins, and ashes. The major fatty acids in raw and roasted sardine samples were docosahexaenoic acid (DHA, 22: 6n-3, 35.98%, 12.46%);palmitic acid (16: 0, 37.59%;24.18%), and eicosapentaenoic acid (EPA, 20: 5n-3, 6.62%;2.95%), respec-tively. The ratios of polyunsaturated to saturated fatty acid (PUFA/SFA) were 1.32 and 0.33, and the n-6/n-3 ratios were 0.07 and 0.13 in raw and roasted sardines, respectively. The results showed that roasting increased the SFA and reduced the PUFA in sardines, which still were rich in PUFA and remained a low-cost and nutritionally healthy food.

Molecular Cloning and Analysis of LEA14L2 Gene from Rubber Tree( Hevea brasiliensis Muell. Arg. )

Using an isotig encoding a putative polypeptide with high similarity to Arabidopsis LEA14 as a query, a 613 bp long cDNA was in silico cloned from the transeriptome data of rubber tree. The sequence nominated as HbLEA14L2 contains an ORF of 456 bp with 3 bp 5＇ UTR and 154 bp 3＇ UTR. Subsequently, a 464 bp eDNA and an 834 bp genome sequence containing this ORF was amplified and sequenced. Sequence analysis suggested that HbLEA14L2 has one intron and encodes 151 amino acids with a theoretical molecular weight of 16.55 kDa, isoleetric point of 4.93 and GRAVY value of -0.022, indicating a cytoplasmle localization pattern; HbLEA14L2 protein contains a conserved LEA_2 domain and belongs to LEA_2 subfamily, sharing 91%, 76%, 75%, 72% and 63% similarity with the homologous proteins in castor bean, leafy spurge, poplar, cotton, and Arabidopsis, respectively. Swiss-Model indicated that the tertiary structure of HbLEA14L2 is consisted of one α-helix and seven β-sheets, which was proposed to serve as a regulatory protein to prevent cellular desiccation.

Corneal ulcer caused by nocardia brasiliensis in a patient with diabetes

Dear Sir,Iam Dr.Sheng-Li Mi,from the Biomanufacturing engineering laboratory,Graduate School at Shenzhen,Tsinghua University,Shenzhen,China.I write to present a case report of nocardia brasiliensis in a patient with diabetes.Nocardia is part of a group of aerobic actinomycetes,widely distributed in soil.Nocardia asteroids and nocardia brasiliensis are the most common human pathogens.Humans infected by nocardia asteroids through the respiratory tract mainly suffer from primary suppurative pulmonary conditions.Infection of nocardia brasiliensis often occurs in the advanced stages of a progressive disease or immune disorder,especially Cushing syndrome,diabetes,or in patients using corticosteroids,immunosuppressive agents and broad-spectrum antibiotics for long time[1].To our knowledge,the reports of nocardia keratitis was very rare

Intra-and inter-clonal tree growth variations of Hevea brasiliensis

We evaluated the effects of planting densities （500, 1,000, 1,500 and 2,000 trees.ha-1） on tree growth performance （diameter at base, diameter at breast height, and clear bole height） of two clones （RRIM 2020 and PRIM 2025） of nine years old plantations of rubber tree （Hevea brasiliensis Muell. Arg） in Malaysia. For the four planting densities of the two clones, basal area and diameter at breast height declined with increasing planting density. Clear bole heights were greatest at 1,500 trees.haland lowest at 500 trees.ha-1 for the clone RRIM 2020, and at 2,000 trees.ha-1and 500 trees.halfor clone RRIM 2025. We conclude that the ideal planting density is 2,000 trees.ha＂l for obtaining high volume of wood production and 500 trees-ha-1 for high wood quality.

Habitat Preference and Effects of Coastal Fragmentation in the Sand-Dwelling Spider <i>Allocosa brasiliensis</i>(Lycosidae, Allocosinae)

Allocosa brasiliensis is a sand-dwelling wolf spider considered a good bioindicator to evaluate the quality of coastal dune ecosystems from Uruguay. Habitat fragmentation and human activities have impacted and reduced the Southern Uruguayan coast during the last decades. The aim of the present study was to evaluate the density, surface activity and habitat preference of A. brasiliensis in beaches with different human impact. For that purpose, we sampled during the night with head lamps and applied the capture-mark-recapture method. Females were more abundant than males and were the most recaptured sex. The most fragmented beaches showed lower number of burrows and individuals, especially in immatures stages. We found that the most suitable sandy habitat for A. brasiliensis should present a vegetation cover optimum of 25% - 50% of the surface. This study could provide tools for the implementation of future management conservation plans of the Southern Uruguayan coastline.

Immunophenotyping of peripheral blood mononuclear cells and intracellular detection of IL-2, IFNγ and IL-4 by flow cytometry in patients with actinomycetoma by Nocardia brasiliensis and Actinomadura madurae. Findings in six patients

Mycetoma is a chronic, granulomatous, progressive inflammatory disease;it usually involves the subcutaneous tissue after a traumatic inoculation of the causative organism. Mycetoma may be caused by true fungi or bacteria (actinomycetes) and hence it is classified as eumycetoma and actinomycetoma respectively. Mycetoma immunological studies have fre- quently addressed humoral aspects. Few reports have addressed the role of cellular immunity in humans, for this reason, we were interested in finding differences in the circulating population of mononuclear cells or in the ability of this pathology to produce cytokines after mitogen stimulation. In this study, immunophenotyping of peripheral blood mononuclear cells (PBMN) and intracellular detection by flow cytometry of IL-2, IFNγ and IL-4 in patients with actino- mycetoma by Nocardia brasiliensis and Actino- madura madurae were evaluated. We investi- gated the expression of T-cells (CD3, CD4 and CD8), B-cells (CD19), monocytes (CD14), and natural killer cells (CD16 and CD56) markers. CD69 and CD25 were used to monitor individual activated cell subsets. The percentage values of the cells were calculated. Our results indicated that PBMC from patients with mycetoma show similar percentages of circulating cells when compared with healthy donors. The expression of IL-2 receptor (IL-2R) by mitogen activation was similar in these two groups. These findings suggest that circulating lymphocytes are not affected by this pathology. Intracellular IL-4 was increased only in patients with mycetoma by N. brasiliensis, suggesting a TH2 profile;this ob- servation has been reported by other authors.

An Analysis of the Patents concerning Hevea brasiliensis

This paper mainly analyzes the application data of patents concerning Hevea brasiliensis in the world and China from the perspectives of time,research field,and the geographical distribution of applicants. It also interprets the patents for invention in China from time,research field,and types of applicants. Based on the study,it shows that although the patent applications fluctuate,the general trend is rising. And the patents are mostly concentrated in plant protection,and agricultural biotechnology. The Chinese applicants of patents for invention largely work in the research institutes,and most of these patents focus on the field of plant protection,agricultural biotechnology,and agro-processing.However,the foreign applicants mainly work in companies,and most of these patents only focus on plant protection. These findings will provide a reference for formulating development policies about natural rubber industry in China.

Studies on Rubber (<i>Hevea brasiliensis</i>) Trees Exist Plant Type after Planting and Available Tapping Tree of Rubber Plantation in China

Existing plant types of rubber tree after planting and available tapping tree were investigated, and there were about 28 rubber plantations with different tapping years of 8 varieties “CATAS7-33-97”, “CATAS8-79”, “CATAS7-20-59”, “PR107”, “RRIM600”, “GT1”, “INA873”, “93-114”in South China. The results showed that there were six kinds of existing plant types of rubber tree after planting of rubber plantations, which were available tapping trees, wind damaged trees, cold damaged trees, tapping panel dryness trees, absent trees and weak trees, respectively. These data investigated also showed rubber trees under available tapping, stoppage due to tapping panel dryness, absence, wind damage, cold damage and weakness were counted and calculated and made up for 72.21%, 14.75%, 5.61%, 3.86%, 2.68% and 1.89%. Tapping panel dryness trees, wind damage and absent trees are major factors for the loss of tapping rubber trees in the rubber plantations. Of these investigated varieties, available tapping trees per 100 trees of rubber plantation of “PR107”at the 1st, 12th, 14th, 16th, 20th, 24th tapping year were 96, 67, 70, 75, 66, 46 trees in Hainan planting zone, respectively. Available tapping trees per 100 trees of rubber plantation of “RRIM600”at the 9th, 15th, 20th, 22nd tapping year were 88, 62, 55, 36 trees in Yunnan planting zone, respectively. Available tapping trees per 100 trees of rubber plantation of “93-114” at the 10th, 19th, tapping year were 94, 62 trees in Guangdong planting zone. These results showed that available tapping trees of rubber plantation decreased with increasing tapping age under different planting zones in China.

Molecular Cloning and Sequence Analysis of an Aquaporin Gene Related to Tapping Panel Dryness in Rubber Tree( Hevea brasiliensis Muell. Arg. )

As a syndrome with tapping incision blocked partly or entirely during latex exploiting in rubber tree （Hevea brasiliensis ）, ＂Tapping panel dryness （TPD）＂ causes great yield losses, thereby becoming the most important factor limiting rubber production. On the basis of an EST down-regulated in TPD-affected rubber trees, a 903 bp cDNA denoted HbPIP2;2 was isolated from the bark tissue with a combination of in silico cloning and RT-PCR. The cDNA contains an 867 hp ORF, 13 bp 5＇UTR and 23 bp 3＇ UTR. Sequence analysis indicated that HbPIP2;2 encodes 288 amino acids with a theoretical molecular weight （Mw） of 30. 71 kDa and isolectric point （Pi） of 8.20. Bioinformatics analysis suggested that the deduced protein is predicted to have six transmembrane helices located to the plasma membrane and harbor one conserved MIP domain that can be grouped into plasma membrane intrinsic proteins （PIPs） of aquaporin （AQP） family. Homolo- gy search revealed that the protein shares a similarity of more than 90% with the homologues in Ricinus communis, Popttlus trichocarpa, Juglans regia and Theobro- ma cacao, suooorting a hie.hly conserved evolution. This study provided basis for further uncovering the regulatory role of AOPs in TPD occurrence.

Advances in Tissue Culture of Rubber( Hevea brasiliensis L. )

Natural rubber, which is produced as a secondary plant metabolite, has been well recognized as an important raw industrial material due to its unique properties that can not be substituted by any alternative synthetic materials. Although more than 2000 plant species have been shown to possess the ability to synthe- size natural rubber, Hevea brasiliensis L. is yet the only one qualified as a commercial source for natural rubber production, which not only has important economic value, but ＂also leads to significant ecological and social benefits to maintain ecological forest microclimate and ease current ecological issues caused by dwindling natural forests. With the growing demand for natural rubber, providing a large number of high-quality rubber planting materials has become a major issue to be solved. In this paper, advances on tissue culture of H. brasiliensis were summarized, which laid the foundation for breeding high-quality rubber planting materials.

Molecular Cloning and Sequence Analysis of a Tonoplast Aquaporin Gene TIP1 Associated with Tapping Panel Dryness in Hevea brasiliensis

[Objective]To address the role of aquaporins（AQPs）in the occurrence of Tapping Panel Dryness（TPD）,a phenomenon with tapping incision blocked partly or entirely during latex exploiting in the rubber tree（Hevea brasiliensis）that causes great losses on rubber production,a tonoplast aquaporin gene associated with TPD occurence was cloned and analyzed.[Method]Based on an EST down-regulated in TPD-affected rubber trees,a 774 bp cDNA designated HbTIP1 was isolated from the bark tissue of Hevea brasiliensis with a combination of in silico cloning and RT-PCR.And the gene structure and sequence characteristics were analyzed using bioinformatics tools.[Result]The cDNA includes a 759 bp ORF,8 bp 5’UTR and 7 bp 3’UTR.Sequence analysis indicated that HbTIP1putatively encodes 252 amino acids with a theoretical molecular weight（Mw）of 25.88 kDa and isolectric point（pI）of 4.96.Bioinformatics analysis suggested that the deduced protein is predicted to have six transmembrane helices located to the vacuolar membrane and to harbor one conserved MIP domain that can be grouped into the tonoplast intrinsic proteins（TIPs）of aquaporin family.Homology search revealed that the protein shares a similarity of more than 90%with the homologues in Theobroma cacao,Prunus persica,Citrus sinensis and Ricinus communis,supporting a highly conserved evolution.[Conclusion]This study provided basis for further revealing the regulatory role of AQPs in the TPD occurrence.

Seasonal Changes Impact on Growth of Rubber (<i>Hevea brasiliensis</i>) Seedlings under Different Cultivation

The vegetative growth of rubber tree is an important basis for rubber tree cultivation and economic management. Stem diameter and height growth patterns of rubber seedlings and the seasonal variability are still poorly understood. Studying the annual growth patterns and seasonal variation of rubber seedling will help understanding biological characteristics of rubber tree and provide a reference for field cultivation and management. Height growth and stem diameter of field-sown rubber seedlings cultivation and polybag-raised cultivation all displayed a distinct annual pattern of “slow-fast-slow”. Height growth and stem diameter growth response fitted the logistic equation preferably, supporting the observed pattern. Fast-growth of plant height for polybag-raised rubber seedlings cultivation and field-sown rubber seedlings cultivation occurred from June to November and May to November, respectively. This period saw the greatest accumulations of plant height growth for polybag-raised rubber seedlings cultivation and field-sown rubber seedlings cultivation accounting for 83.99% and 86.65% of total growth, respectively. Fast-growth of stem diameter for polybag-raised rubber seedlings cultivation and field-sown rubber seedlings cultivation occurred from June to November and May to November, respectively. This period saw the greatest accumulations of stem growth for polybag-raised rubber seedlings cultivation and field-sown rubber seedlings cultivation accounting for 86.71% and 84.60% of total growth, respectively. Polybag-raised rubber seedlings cultivation and field-sown rubber seedlings cultivation for the whole year had nine extension units and each month had one extension unit from May to November. Extension unit of for the whole year did not stop period and plant height growth rate has the seasonal difference. However, leaf phenophase of field-sown rubber seedlings cultivation was earlier than that of polybag-raised rubber seedlings cultivation and each extension unit of field-sown rubber seedlings cultivation was greater than that of poly-bag-raised rubber seedlings cultivation. The precipitation had the greatest impact on seedlings growth under different cultivation. There was no period annually in which height growth and stem diameter growth did not occur and the fastest growth occurred during the rainy season. Plant height and stem diameter of rubber seedlings under different cultivation showed isogony phenomenon. Extension unit and leaf phenophase of polybag-raised rubber seedlings and field-sown rubber seedlings showed consistency and synchronization characteristic. Different cultivation of rubber seedling should take corresponding measures tending management and operation.

Methanolysis of Mixed Crop Oils (<i>Hevea brasiliensis</i>and <i>Jatropha curcas</i>L.) into Biodiesel: Kinetics Study

In this study, the methanolysis of mixed crop oils (Hevea brasiliensis and Jatropha curcas L.) was investigated. Due to the oils contain high impurities, a particular pre-treatment of which was carried out. For alkali-catalyzed methanolysis, few parameters (such as: molar ratio, catalyst percentage, and reaction time) were studied. The results showed that the methanolysis optimum conditions were 5.5 of molar ratio, 0.9% w/w of catalyst, and 60 minutes of reaction time with the highest yield of 98.9%. The kinetics study indicated that first order reaction mechanism with reaction rate constant and activation energy were 1.5 × 10-2 minute-1 and 23.93 KJ?mol-1 respectively. As fuel, the physicochemical properties of biodiesel met the standard values required by ASTM D6751.

Oral Use of an Infusion of Leaves of <i>Solanum paniculatum</i>L., </i>Jacaranda brasiliensis</i>and </i>Sonchus oleraceus</i>for Treatment of Vitiligo

Background: A healthy normal skin is essential for a person’s physical and mental well being. It is an important aspect of their sexual attractiveness, a sense of well being and a sense of self confidence. Vitiligo is an acquired depigmentation disorder of skin affecting 1% - 4% of the world population. Neither life threatening nor symptomatic (except that depigmented patches burn easily when exposed to the sun) the effects of vitiligo can be cosmetically and psychologically devastating. Because the disease is still not understood, there is a plethora of different treatments approaches, but they are largely unsatisfactory from patient’s perspective. Objective: To report the outcomes from oral use of an infusion of leaves of Solanum paniculatum L., Jacaranda brasiliensis and Sonchus oleraceus for treatment of vitiligo. Ethical issues: Study approved by the Research Ethics Committee of Hospital Federal dos Servidores do Estado do Rio de Janeiro (CAAE: 32143314.9.0000.5252). All participants confirmed the condition of volunteer by signing the Informed Consent Form. Methods: From October 2014 to October 2015 twelve subjects consumed daily four glasses (800 ml) of proposed phytotherapeutic preparation. Results: All subjects were highly satisfied about their outcomes. Total depigmentation was considered as a positive outcome because it is cosmetically acceptable. Total depigmentation was observed in presence of 80% or more of depigmenteted patches. Conclusions: The phytotherapeutic preparation proposed is effective on treatment of depigmenteted patches in vitiligo.

RNAi technology targeting PbGP43 and PbP27 in Paracoccidioides brasiliensis

Efficient technologies for gene silencing would be important to carry out functional analysis with P. brasiliensis genes, as well as for a better understanding of the biology and pathogenesis of this pathogenic fungus. Due to the fact that homologous recombination is unusual in P. brasiliensis, the development of knockout isolates is currently non-feasible. The goal of this work was to assess RNA interference (RNAi) technology as an alternative tool for gene silencing previously employed successfully in H. capsulatum. For this purpose, we built different inverted repeat transgenic hairpin constructs to down-regulate the PbGP43 and PbP27 genes known to codify for two fungal immunogenic proteins that elicit a strong immune response during experimental paracoccidioidomycosis. Using the RNAi strategy, a reduction in the mRNA levels of the PbGP43 and PbP27 genes was observed during the first 20 days after selection;however, in the transformed yeast cells, the gene silencing status proved non-stable through the assay. We demonstrated that electrotransformation was suitable to transform P. brasiliensis yeast cells and integrate the hairpin constructions;nonetheless, gene silencing was not stable along the experimental time. A detailed analysis of the underlying molecular RNAi machinery may provide further insights into the intracellular mechanism that governs this reverse genetic tool.

巴西橡胶树(Hevea brasiliensis)HbMlo2基因克隆与表达分析

Mlo (mildew resistance locus O)是一种植物防卫相关基因,对植物的抗病性有负向调控作用。本研究采用基因编辑技术等方法为培育橡胶树抗病品种作出铺垫,进一步填补Mlo家族成员。通过对拟南芥的AtMlo2基因在巴西橡胶树上进行比对,根据同源性克隆得到一个cDNA序列,将其命名为HbMlo2。根据生物信息学分析得出,该基因cDNA全长1 725 bp,编码574个氨基酸,该蛋白的分子量为65.78 kD,等电点为8.98,8个跨膜域,没有信号肽,HbMlo2基因与已知的木薯的Mlo基因相似性达到83%。白粉菌侵染橡胶树叶组织后,通过qRT-PCR发现HbMlo2基因显著上调。本研究结果初步表明,HbMlo2可能参与橡胶树的抗病反应机制,为橡胶树抗病育种的研究提供新思路。