Root apical diameter and root elongation rate of rubber seedlings (Hevea brasiliensis) show parallel responses to photoassimilate availability

The hypothesis that root apical diameter may be used to evaluate root growth potential was tested. Temporal variations in the apical diameter of individual roots of rubber seedlings ( Hevea brasiliensis ) were studied together with their elongation patterns, using root observation boxes under controlled conditions. This study confirmed the overall positive correlation between apical diameter and growth rale. Moreover, the two parameters, varied in the same way during the life of a given root. For roots with short growth duration, there was a parallel quick decrease in both apical diameter and elongation rate, whereas roots that grew for longer periods showed synchronous fluctuations for both parameters. Since the mean values for the secondary roots within a root system exhibited the same trends, variations in apical diameter and elongation rates should depend on factors influencing the whole root system. When related to shoot rhythmic growth, both apical diameter and elongation rates were depressed during the periods of leaf growth. These effects were enhanced and/or prolonged by shading, hence reinforcing the hypothesis that this development depends on assimilate availability. Such results can be interpreted in terms of a source-sink relationship within the whole plant by considering the apical diameter, representing the size of the meristem related to the number of rneristematic cells, as an indicator of each root's growth potential.     

In-depth proteome analysis of the rubber particle of Hevea brasiliensis (para rubber tree)

The rubber particle is a special organelle in which natural rubber is synthesised and stored in the laticifers of Hevea brasiliensis. To better understand the biological functions of rubber particles and to identify the candidate rubber biosynthesis-related proteins, a comprehensive proteome analysis was performed on H. brasiliensis rubber particles using shotgun tandem mass spectrometry profiling approaches—resulting in a thorough report on the rubber particle proteins. A total of 186 rubber particle proteins were identified, with a range in relative molecular mass of 3.9–194.2 kDa and in isoelectric point values of 4.0–11.2. The rubber particle proteins were analysed for gene ontology and could be categorised into eight major groups according to their functions: including rubber biosynthesis, stress- or defence-related responses, protein processing and folding, signal transduction and cellular transport. In addition to well-known rubber biosynthesis-related proteins such as rubber elongation factor (REF), small rubber particle protein (SRPP) and cis-prenyl transferase (CPT), many proteins were firstly identified to be on the rubber particles, including cyclophilin, phospholipase D, cytochrome P450, small GTP-binding protein, clathrin, eukaryotic translation initiation factor, annexin, ABC transporter, translationally controlled tumour protein, ubiquitin-conjugating enzymes, and several homologues of REF, SRPP and CPT. A procedure of multiple reaction monitoring was established for further protein validation. This comprehensive proteome data of rubber particles would facilitate investigation into molecular mechanisms of biogenesis, self-homeostasis and rubber biosynthesis of the rubber particle, and might serve as valuable biomarkers in molecular breeding studies of H. brasiliensis and other alternative rubber-producing species.     

巴西橡胶树的分子生物学研究进展

巴西橡胶树是一种重要的热带经济作物,由于橡胶树体内橡胶含量多,且容易采收,所以橡胶树一直是天然橡胶的商业来源。相比于模式植物和粮食等经济作物来说,分子生物学研究显滞后。,而且橡胶树是多年生乔木,经济性状多集中于胶乳,因此研究难度大,研究也不多,本就巴西橡胶的分子生物学方面的研究进行综述。     

Methane fermentation of rubber (Hevea brasiliensis) latex effluent.

Four species of bacteria capable of CH4 fermentation of rubber latex effluent were isolated and identified as a Methanococcus, a strain of M. vannielii, a Methanobacterium and a strain of M. omelianskii. Auxanographic tests using the four strains showed growth and CH4 formation on a basal medium containing mineral salts or added H2 and Co2. Varied response was obtained when the basal medium was added to formate, acetate, butyrate, methanol, ethanol, and glucose. Previous work has established acid fermentation of Hevea latex arising from bacterial contamination and decomposition of the non-rubber constituents which consist of N-compounds, 2% quebrachitol, and smaller concentration of carbohydrates. This suggests that reduction of CO2 and fermentation of acids formed during metabolism of Hevea latex are possible pathways of CH4 production.     

不同树龄橡胶林土壤水分和细根生物量

采用根钻法,分析了橡胶幼树期(5 a)、初产期(9 a)和旺产期(16 a)林下土壤水分特征及橡胶树细根生物量.结果表明:橡胶树龄越大,土壤含水量越高,而橡胶树细根生物量越少;不同树龄橡胶林0～60 cm土壤含水量随着土层深度的增加而升高,年内变化则呈"双峰"型;不同树龄橡胶树细根生物量最大值均出现在10 cm土层,且随着土层深度的增加而减少,细根生物量年内变化同样呈"双峰"型,但不同树龄细根生物量峰值的出现时间不一致.土壤含水量和土层深度是橡胶树细根生物量的主要影响因子.     

西双版纳三叶橡胶林树干呼吸特征

采用红外气体分析法(IRGA)为期1a原位监测西双版纳三叶橡胶(Hevea brasiliensis)4个年龄段(7、15、27、40a)的树干呼吸情况,同时对每个年龄段树干监测2种高度(1.3 m-割胶部位、2.0 m-不割胶部位)和2个方向(南、北面)以及林内空气和树干1cm深温度.结果表明,4个年龄的树干呼吸有相同的季节规律,都是在雨季大于干季.林龄是影响橡胶树树干呼吸的一个重要因素,15、27a树干呼吸速率最大,分别为(4.989±0.278), (4.678±0.268) μmol·m-2·s-1,显著高于40a和7a树,40a树((3.753±0.205) μmol·m-2·s-1)显著大于7a树((2.299±0.129) μmol·m-2·s-1).所研究的高度和方向上树干呼吸速率无差异,割胶对树干表层破坏愈合后并不影响树干呼吸.树干呼吸与树干温度呈显著相关性,有良好的自然指数回归关系,Q10值为1.966～3.127,南北面Q10差异不明显,4个年龄段树干呼吸Q10值平均为2.452,大于已监测的热带树种.各年龄段橡胶林的主干(一级分枝以下部分树干)呼吸初步估算表明, 7、15、27a和40a橡胶树主干呼吸分别为1.74, 5.54, 7.53, 7.59 t C·hm-2·a-1.     

Synchronous flowering of the rubber tree (Hevea brasiliensis) induced by high solar radiation intensity

How tropical trees flower synchronously near the equator in the absence of significant day length variation or other meteorological cues has long been a puzzle. The rubber tree (Hevea brasiliensis) is used as a model to investigate this phenomenon. The annual cycle of solar radiation intensity is shown to correspond closely with the flowering of the rubber tree planted near the equator and in the subtropics. Unlike in temperate regions, where incoming solar radiation (insolation) is dependent on both day length and radiation intensity, insolation at the equator is due entirely to the latter. Insolation at the upper atmosphere peaks twice a year during the spring and autumn equinoxes, but the actual solar radiation that reaches the ground is attenuated to varying extents in different localities. The rubber tree shows one or two flowering seasons a year (with major and minor seasons in the latter) in accordance with the solar radiation intensity received. High solar radiation intensity, and in particular bright sunshine (as distinct from prolonged diffuse radiation), induces synchronous anthesis and blooming in Hevea around the time of the equinoxes. The same mechanism may be operational in other tropical tree species.     

Molecular cloning,characterization and expression of Mn-superoxide dismutase from the rubber tree (Hevea brasiliensis)

The gene and the RNA from Arabidopsis thaliana for the plastid-located glycerol-3-phosphate acyltransferase (GPAT; EC 2.3.1.15) and their encoded product have been studied. The gene (designated ATS1) was isolated by screening a DASH genomic library for cross-hybridization with a radiolabeled probe prepared from cDNA for GPAT from squash. cDNA clones representing the mRNA were isolated by screening a ZAPII cDNA library for hybridization with a radiolabeled probe prepared from a DNA fragment of ATS1. The nucleotide sequences of the gene and the cDNA were determined, and the 5 end of the RNA was mapped by primer extension. Sequences similar to the TATA box, polyadenylation sequences and intron-splicing sequences were found at the expected locations. The pre-mRNA was 3288 nucleotides long and contained 5 and 3-untranslated sequences of 57 and 442 nucleotides, respectively. The coding sequence of 1377 nucleotides was interrupted by 11 introns of 1412 nucleotides in total and the 3-untranslated sequence contained another intron of 94 nucleotides. The open-reading frame encoded a polypeptide of 459 amino acid residues, the amino acid sequence of which was highly homologous to those of precursors to plastid-located GPATs from squash and pea. The enzymatic activity of a gene product that was over-produced in Escherichia coli confirmed the indentity of the gene.Abbreviations ACP acyl carrier protein - GPAT glycerol-3-phosphate acyltransferase - IPTG isopropyl--thiogalactopyranoside.     

Co- and post-translational processing of the hevein preproprotein of latex of the rubber tree (Hevea brasiliensis)

Hevein is a chitin-binding protein of 43 amino acids found in the lutoid body-enriched fraction of rubber tree latex. A hevein cDNA clone (HEV1) (Broekaert, W., Lee, H.-i., Kush, A., Nam, C.-H., and Raikhel, N. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 7633-7637) encodes a putative signal sequence of 17 amino acids followed by a polypeptide of 187 amino acids. Interestingly, this polypeptide has two distinct domains: an amino-terminal domain of 43 amino acids, corresponding to mature hevein, and a carboxyl-terminal domain of 144 amino acids. To investigate the mechanisms involved in processing of the protein encoded by HEV1, three domain-specific antisera were raised against fusion proteins harboring the amino-terminal domain (N domain), carboxyl-terminal domain (C domain), and both domains (NC domain). Translocation experiments using an in vitro translation system show that the first 17-amino acid sequence encoded by the cDNA functions as a signal peptide. Immunoblot analysis of proteins extracted from lutoid bodies demonstrates that a 5-kDa protein comigrated with purified mature hevein and cross-reacted with N domain- and NC domain-specific antibodies. A 14-kDa protein was recognized by C domain- and NC domain-specific antibodies. A 20-kDa protein was cross-reactive with all three antibodies. Microsequencing data further suggest that the 5-kDa (amino-terminal domain) and 14-kDa (carboxyl-terminal domain) proteins are post-translational cleavage products of the 20-kDa polypeptide (both domains) which corresponds to the proprotein encoded by HEV1. In addition, it was found that the amino-terminal domain could provide chitin-binding properties to a fusion protein bearing it either amino terminally or carboxyl terminally.     

Compared root system architectures in seedlings and in vitro plantlets of Hevea brasiliensis,in the initial years of growth in the field

In vitro culture of Hevea was undertaken to propagate selected clones on their own roots. The challenge was to overcome the failure of cuttings due to the poor conformity of regenerated root systems. Trees of several juvenile or mature genotypes were propagated either by in vitro microcutting, or by somatic embryogenesis, and planted in the field. Certain static and dynamic components of the root system were observed at different growth stages, from 0 to 3 years, and compared to those of seedlings of the same age used in the trial as a reference. A simple method was designed for measuring the vigour and balance of the root system. The in vitro plantlets had a well-developed taproot and lateral root system, with an architecture similar to that of plants obtained from seed. Moreover, clear differences occurred between selected clones for the relative vigour of the tap roots, lateral roots and trunk. This revised version was published online in June 2006 with corrections to the Cover Date.     

Cloning and characterization of cDNA encoding farnesyl diphosphate synthase from rubber tree (Hevea brasiliensis)

Commercially used natural rubber (cis-1,4-polyisoprene) is a secondary metabolite of the rubber tree (Hevea brasiliensis). Previous studies have shown the involvement of a prenyl transferase in the final steps of natural rubber biosynthesis which includes polymerization of isopentenyl pyrophosphate into rubber. Using synthetic oligonucleotides corresponding to the partial amino acid sequences of this protein as probes to screen a laticifer-specific cDNA library, we have isolated a full-length cDNA which encodes a 47 kDa protein with strong homology to farnesyl diphosphate synthases from many species. The catalytic activity of this protein was confirmed by complementing the deletion yeast mutant. In Hevea, this gene is expressed in latex producing cells and in the epidermal region of the rubber plant suggesting a dual role for the protein in the biosyntheses of rubber and other isoprenoids. Although the expression level of this gene is not significantly affected by hormone treatment (e.g. ethylene), regeneration of latex due to tapping increases its expression level.     

Ribosomal DNA variations in wild and cultivated rubber tree (Hevea brasiliensis).

Ribosomal DNA variations were surveyed to assess the genetic variability among Hevea brasiliensis genetic resources. One hundred and sixty-eight individuals, including 73 cultivated Wickham clones and 95 wild clones from a prospection, were analyzed. Restriction mapping of rDNA units showed that RFLP variations are the result of both length and site (EcoRI) variations in the intergenic spacer (IGS). These variations can be revealed between as well as within individuals. A total of 12 spacer length variants is scored in the whole population, as well as two different ribosomal units (refered as type I and type II), defined by the presence or absence of an EcoRI site. Particular associations between spacer length variants and unit types can be revealed, leading to complex RFLP patterns. Cultivated clones appear to be less variable than prospections but show, however, a relatively high level of variability despite their narrow genetic base. Furthermore, IGS variations allowed a structuring within wild clones to be drawn, based mainly on their geographical origin. Some interesting discrepancies with previous work on isozyme variations are discussed and show the interest of surveying different genetic markers for diversity studies.     

Mites (Acari) from rubber trees (Hevea brasiliensis Muell. Arg., Euphorbiaceae) and spontaneous euphorbiaceous in rubber trees cultivation

Quarterly samples were done in 2001 on three rubber tree plantation in the northwest of the state of S?o Paulo. Three rubber trees of each locality were sampled. Between the rows of rubber tree four species of spontaneous euphorbiaceous were collected: Chamaesyce hirta, C. hyssopifolia, Euphorbia heterophylla and Phyllanthus tenellus. A total of 8.954 mites of 38 species, belonging to 31 genera of 11 families were collected. Tydeidae and Phytoseiidae had the highest diversity of species, 9 and 7, respectively. The most abundant families were Eriophyidae (3.594), Tydeidae (2.825) and Tenuipalpidae (1.027). The most abundant species on the rubber trees were: phytophagous - Calacarus heveae Feres, Tenuipalpus heveae Baker, Lorryia sp.2, Lorryia formosa Cooreman and Lorryia sp.1; predators - Zetzellia quasagistemas Hernandes & Feres, Pronematus sp., Iphiseiodes zuluagai Denmark & Muma and Euseius citrifolius Denmark & Muma. Among the spontaneous euphorbiaceous, predatory mites were abundantly found on C. hirta and E. heterophylla, mainly Pronematus sp. and E. citrifolius, suggesting that these plants could be important in the maintenance of these predators in the rubber tree cultivation areas. However, plants that can shelter predators and at the same time exert strong competition (nutrients, water etc) to rubber trees, can not be recommended for pest management programs. Studies about competition between rubber trees and spontaneous plants need to be conducted for feasible efficient programs of environmental management, aiming at the control of pest mites of rubber tree.     

Periodicity in the development of the root system of young rubber trees (Hevea brasiliensis Müell. Arg.): relationship with shoot development

The growth pattern of the root system of young rubber trees (Hevea brasiliensis) was studied in relation to shoot development over a period of 3 months. Temporal and spatial variations in elongation and branching processes were examined for the different root types, by means of root observation boxes. Shoot growth was typically rhythmic. Root development was periodic and related to leaf expansion. Root elongation was depressed during leaf growth, whereas branching was enhanced. Consequently, highly branched areas with vigorous secondary roots alternated along the taproot with poorly branched areas with shorter roots. Root types were not affected to the same degree by shoot competition: during leaf expansion, taproot growth was just depressed but remained continuous, the emergence and elongation rates of secondary roots were significantly affected and the elongation rates of tertiary roots fell to zero. These results were consistent with the hypothesis that root growth is related to competition for assimilates and to the sink strength of the different root types, whereas root branching appeared to be promoted by leaf development.     

巴西橡胶树SSR遗传图谱的构建

以热研88-13×IAN873的94个F1群体为试材, 利用简单序列重复(Simple sequence repeat, SSR)标记, 采用FsLinkageMAP 1.0软件, 构建了巴西橡胶树热研88-13×IAN873的遗传连锁图谱。从441对SSR引物中筛选出160对具有多态信息的引物, 在分离群体中共检测到206个多态性位点, 176个位点用于遗传图谱的构建; χ²检验结果显示, 有147个位点符合1:1分离比例, 有12个符合1:2:1分离比例, 有17个符合1:1:1:1的分离比例, 共有13个偏分离位点, 偏分离率低(7.38%); 91个SSR位点被分为18个连锁群, 覆盖橡胶树基因组1 937.06 cM, 每个连锁群包含2～16个位点, 标记间的平均距离为21.29 cM。     

Regulation of rubber biosynthetic rate and molecular weight in Hevea brasiliensis by metal cofactor

Metal ion cofactors are necessary for prenyltransferase enzymes. Magnesium and manganese can be used as metal ion cofactor by rubber transferase (a cis-prenyltransferase) associated with purified rubber particles. The rubber initiation rate, biosynthetic rate, and molecular weight produced in vitro from Hevea brasiliensis rubber transferase is regulated by metal ion concentration. In addition, varies significantly with [Mg(2+)]. decreases from 8000 +/- 600 microM at [Mg(2+)] = 4 mM to 68 +/- 10 microM at [Mg(2+)] = 8 mM and increases back to 970 +/- 70 microM at [Mg(2+)] = 30 mM. The highest affinity of rubber transferase for IPP.Mg occurred when [Mg(2+)] = A(max) (metal concentration that gives highest IPP incorporation rate). A metal ion is required for rubber biosynthesis, but an excess of metal ions interacts with the rubber transferase inhibiting its activity. The results suggest that H. brasiliensis could use [Mg(2+)] as a regulatory mechanism for rubber biosynthesis and molecular weight in vivo.