Ethephon Stimulation Research Articles

Genetic analysis of agronomic and physiological traits associated with latex yield revealed complex genetic bases in Hevea brasiliensis

Hevea brasiliensis, a natural rubber producing species, is widely cultivated due to its high rubber yield potential. Natural rubber is synthesised in the rubber particles of laticifers. Latex diagnosis (LD) was established to characterise the physiological state of the laticiferous system by measuring its physiological parameters, i.e., sucrose, inorganic phosphorous (Pi), thiols and total solid content (TSC). Rubber clones are often classified in three groups i.e., quick starters, medium starters and slow starters. To better understand the genetic bases of latex yield, a biparental population was generated from a cross between the quick-starter clone PB 260 and the medium-starter clone SP 217. LD was performed during the peak latex production season and used to calculate sucrose loading. The agronomic and physiological parameters associated with latex yield led to the classification of genotypes according to the rubber clonal typology and to the identification of quantitative trait loci (QTL) using a high-density map. Inorganic phosphorous content was positively associated with yield during the first year of production thus enabling identification of quick-starter clones. In addition, the LD-based clonal typology led to determine the long-term yield potential and the use of appropriate ethephon stimulation. QTL analysis successfully identified several QTLs related to yield, sucrose, Pi and TSC. One QTL related to sucrose loading was identified in the same position as the QTL for sucrose on linkage group 1. To our knowledge, this is the first study to report QTL analysis for this trait. The use of a high-density map enables the identification of genes underlying QTLs. Several putative genes underlying QTLs related to yield, sucrose and TSC were identified.

Integrated physiology, transcriptome and proteome analyses highlight the potential roles of multiple hormone-mediated signaling pathways involved in tapping panel dryness in rubber tree

Currently, one of the most serious threats to rubber tree is the tapping panel dryness (TPD) that greatly restricts natural rubber production. Over-tapping or excessive ethephon stimulation is regarded as the main cause of TPD occurrence. Although extensive studies have been carried out, the molecular mechanism underlying TPD remains puzzled. An attempt was made to compare the levels of endogenous hormones and the profiles of transcriptome and proteome between healthy and TPD trees. Results showed that most of endogenous hormones such as jasmonic acid (JA), 1-aminocyclopropanecarboxylic acid (ACC), indole-3-acetic acid (IAA), trans-zeatin (tZ) and salicylic acid (SA) in the barks were significantly altered in TPD-affected rubber trees. Accordingly, multiple hormone-mediated signaling pathways were changed. In total, 731 differentially expressed genes (DEGs) and 671 differentially expressed proteins (DEPs) were identified, of which 80 DEGs were identified as putative transcription factors (TFs). Further analysis revealed that 12 DEGs and five DEPs regulated plant hormone synthesis, and that 16 DEGs and six DEPs were involved in plant hormone signal transduction pathway. Nine DEGs and four DEPs participated in rubber biosynthesis and most DEGs and all the four DEPs were repressed in TPD trees. All these results highlight the potential roles of endogenous hormones, signaling pathways mediated by these hormones and rubber biosynthesis pathway in the defense response of rubber trees to TPD. The present study extends our understanding of the nature and mechanism underlying TPD and provides some candidate genes and proteins related to TPD for further research in the future.

Analysis of reduced and oxidized antioxidants in Hevea brasiliensis latex reveals new insights into the regulation of antioxidants in response to harvesting stress and tapping panel dryness

Latex diagnosis (LD) is applied to optimize the natural rubber production and prevent tapping panel dryness (TPD), a physiological syndrome affecting latex production in Hevea brasiliensis. The reduced thiol content (RSH) is one of the biochemical parameters associated with the risk of TPD. However, RSH is difficult to interpret because of the influence of the environment. In order to better understand the regulation of antioxidants and to better interpret RSH, a key parameter of LD, this study analysed in latex both oxidised and reduced forms of ascorbic acid (AsA) and glutathione, and their cofactors as well as other latex diagnosis parameters in response to harvesting stress (tapping and ethephon stimulation) and TPD occurrence. The content of antioxidants in latex had a high variability among five rubber clones. The concentration in AsA was about ten times higher than GSH in laticifer, GSH accounting for about 50% of RSH. For short-term harvesting stress, RSH increased with tapping frequency and ethephon stimulation. TPD is associated with high latex viscosity and bursting of lysosomal particles called lutoids, as well as for several rubber clones with lower RSH and GSH contents. These results suggest that a high level of RSH shows the capacity of laticifer metabolism to cope with harvesting stress, while a drop in RSH is the sign of long stress related to lower metabolic activity and TPD occurrence. RSH remains an essential physiological parameter to prevent TPD when associated with reference data under low and high harvesting stress. This study paves the way to understand the role of AsA and GSH, and carry out genetic studies of antioxidants.

Effects of Ethephon Stimulation and Cut Length by Vertically Tapping on Rubber Yield and Latex Quality

Effects of Ethephon Stimulation and Cut Length by Vertically Tapping on Rubber Yield and Latex Quality

Effect of ethephon stimulation on natural rubber latex properties; new insight into ethephon stimulation

Stimulation based low intensity harvesting (LIH) systems are accepted all over the world as an agronomic tool to overcome issues such as high cost of production, worker scarcity and low economic lifespan of trees in rubber plantation industries. When developing a protocol for ethephon stimulation, yield levels were concerned regardless of the effect on associated latex quality parameters. Therefore, this study elucidated the effect of ethephon on dry rubber yield and associated latex quality parameters together with latex rheological behaviour and physiological parameters. Five stimulation levels of ethephon (1 − 5 %) were investigated with RRIC 121 genotype under S/2 d3 (tapping half of the spiral once in three days frequency) harvesting system with unstimulated latex as the control. The results revealed that all the properties studied were significantly affected by the ethephon concentration and maximum yield with optimum properties could be achieved with the application of 3 % ethephon under S/2 d3 harvesting system.

Transcriptome analysis in Hevea brasiliensis latex revealed changes in hormone signalling pathways during ethephon stimulation and consequent Tapping Panel Dryness

Tapping Panel Dryness (TPD) affects latex production in Hevea brasiliensis. This physiological syndrome involves the agglutination of rubber particles, which leads to partial or complete cessation of latex flow. Latex harvesting consists in tapping soft bark. Ethephon can be applied to stimulate latex flow and its regeneration in laticifers. Several studies have reported transcriptome changes in bark tissues. This study is the first report on deep RNA sequencing of latex to compare the effect of ethephon stimulation and TPD severity. Trees were carefully selected for paired-end sequencing using an Illumina HiSeq 2000. In all, 43 to 60 million reads were sequenced for each treatment in three biological replicates (slight TPD trees without ethephon stimulation, and slight and severe TPD trees with ethephon treatment). Differentially expressed genes were identified and annotated, giving 8,111 and 728 in response to ethephon in slight TPD trees and in ethephon-induced severe TPD trees, respectively. A biological network of responses to ethephon and TPD highlighted the major influence of metabolic processes and the response to stimulus, especially wounding and jasmonate depression in TPD-affected trees induced by ethephon stimulation.

Curative Activity of Watery Fermented Compost Extract as a Bark Treatment against Tapping Panel Dryness

Background: Tapping panel dryness (TPD) is a stress-related disorder that afflicts rubber trees, contributing to yield losses in nearly every rubber-growing region. Method: We demonstrated the curative effects of biostimulants containing a fermented watery extract of shrimp waste-enriched compost (SWCE) on TPD in field trials. Undiluted SWCE was applied to lightly scraped bark in the first, third, and fourth trials, and applied directly without bark scraping in the second trial. Results: Bark treatment significantly (p < 0.05) reduced tapping cut dryness and increased latex yield, suggesting recovery from the disorder. When SWCE was applied to pre-scraped bark, 80% and 30% of trees with partial and complete TPD, respectively, recovered from tapping dryness within 2 months. The latex dry weight of treated trees with partial and complete TPD was 77.5% and 21.1% that of healthy trees, respectively. We observed slight recovery from TPD in trees treated without bark scraping and in trees with a history of ethephon stimulation. No curative effect of SWCE was demonstrated in treated trees without a tapping rest period. These findings suggest that compost extract could be a useful treatment for partial TPD.

Yield-Growth Dynamics of Hevea Hybrid Clones in Tripura

Evaluation of growth and yield for twelve selected hybrids from two recombination breeding programmes was conducted in a clonal nursery trial at Tripura (North East India). Girth, increment in girth and yield based on test tapping as well as under ethephon stimulation were evaluated. Based on mean girth and girth increment, 99/5/9, 99/3/61, 99/1/15 and 98/46 were identified as vigorous clones. Test tap yield in the peak season was highest for reference clone RRIM 600 followed by 98/46 Hybrid clones 99/5/9, 99/3/61 and 99/5/35 also had test tap yield above the grand mean in both peak season and summer season. Assessment of intra-clonal variability through boxplot analysis revealed that among the promising clones 98/46, 99/3/61 and 99/5/9 had a lower range in girth, while 98/46 and 99/5/9 had better uniformity in yield. Significant yield-girth correlation was observed within clones in the second year of test tapping. Hybrid clones were classified based on mean and proportion of trees in the above average category. Clones 99/3/61, 99/5/9 and 98/46, belonging to the high girth and high yield categories, recorded higher mean yield than the grand mean over three years of test tapping, were comparable with RRIM 600 and identified as potential clones for further evaluation.

Establishment of Hevea brasiliensis lines overexpressing genes involved in ethylene signalling pathway

The gaseous plant hormone ethylene has a wide variety of applications in agriculture and horticulture. Ethylene Response Factors (ERF) are the last transcription factors of the ethylene signalling pathway and control a large number of ethylene-responsive genes. Two Hevea brasiliensis ERF, HbERF-IXc4 and HbERF-IXc5, are orthologs to ERF1 a key regulator at the crosstalk of ethylene and jasmonate signalling pathways. These genes were suggested to play an important role in regulating latex cell metabolism in response to tapping and ethephon stimulation. In this study, transgenic lines overexpressing HbERF-IXc4 and HbERF-IXc5 under control of 35S CaMV and HEV2.1 promoter have been conducted. Transgenic Hevea lines were obtained by Agrobacterium tumefaciens-mediated genetic transformation. The somatic embryogenesis process was affected by these modifications. Agrobacterium tumefaciens genetic transformation procedure has been developed from friable callus line for clone PB260. Hevea callus was sub-cultured as small aggregates on paromomycin selection medium. Transgenic callus lines were established from sub-aggregates showing full GFP activity. Ten transgenic lines were confirmed as transgenic by Southern blot hybridization. This result showed successfully establishment of H. brasiliensis transgenic lines. Further plant regeneration and characterization were necessary to understand the function HbERF-IXc4 and HbERF-IXc5 in latex.

Establishment of Hevea brasiliensis lines overexpressing genes involved in ethylene signalling pathway

The gaseous plant hormone ethylene has a wide variety of applications in agriculture and horticulture. Ethylene Response Factors (ERF) are the last transcription factors of the ethylene signalling pathway and control a large number of ethylene-responsive genes. Two Hevea brasiliensis ERF, HbERF-IXc4 and HbERF-IXc5, are orthologs to ERF1 a key regulator at the crosstalk of ethylene and jasmonate signalling pathways. These genes were suggested to play an important role in regulating latex cell metabolism in response to tapping and ethephon stimulation. In this study, transgenic lines overexpressing HbERF-IXc4 and HbERF-IXc5 under control of 35S CaMV and HEV2.1 promoter have been conducted. Transgenic Hevea lines were obtained by Agrobacterium tumefaciens-mediated genetic transformation. The somatic embryogenesis process was affected by these modifications. Agrobacterium tumefaciens genetic transformation procedure has been developed from friable callus line for clone PB260. Hevea callus was sub-cultured as small aggregates on paromomycin selection medium. Transgenic callus lines were established from sub-aggregates showing full GFP activity. Ten transgenic lines were confirmed as transgenic by Southern blot hybridization. This result showed successfully establishment of H. brasiliensis transgenic lines. Further plant regeneration and characterization were necessary to understand the function HbERF-IXc4 and HbERF-IXc5 in latex.

Ethephon Increases Rubber Tree Latex Yield by Regulating Aquaporins and Alleviating the Tapping-Induced Local Increase in Latex Total Solid Content

The concentration of phloem solute generally falls from leaves to roots. However, a local increase in latex total solid content (LILTSC) was identified near the tapping cut of rubber trees. To understand the mechanism of ethephon-stimulated latex yield, the formation and ethephon (an ethylene releaser) alleviation of the LILTSC near the tapping cut were examined. It was found that the LILTSC near the tapping cut of a tapped rubber tree was caused by the tapping-accelerated rubber biosynthesis which began following the first tapping and became significant after the fourth tapping. Ethephon stimulation markedly reduced the LILTSC. The latex yield change pattern upon ethephon stimulation was associated with the kinetic change of LILTSC and the decomposition dynamic of ethephon into ethylene. Once the LILTSC was reduced by ethylene release upon ethephon stimulation, the latex yield increased; however, when the ethylene release upon ethephon stimulation receded, the LILTSC was restored and the effect of ethephon stimulation dissipated. The reduction of LILTSC by ethephon stimulation could be ascribed to the translocation property of ethylene in plants and its regulation of aquaporins. Because maximum ethylene release upon tapping-cut-ethephon-application occured close to the tapping cut, the aquaporins were more up-regulated in this region, leading to a reduction of the LILTSC and an increase in latex yield. All these results suggest that the LILTSC near the tapping cut was caused by tapping; the ethephon-induced aquaporin up-regulation and LILTSC reduction are involved in the mechanism of ethephon-promoted latex yield.

Insight into prevention of enzymatic browning and increase of antioxidant potential of crepe rubber with ethephon stimulation

The effect of different ethephon concentrations 1%-5% (w/v) on colour of crepe rubber and plasticity retention index (PRI) was studied. The study was conducted with the RRISL 121 clone using S/2, d3 harvesting system. The results revealed that ethephon stimulation affects strongly the antioxidant and polyphenol oxidase (PPO) inhibitory potential. Further, thiol content and PRI values were increased while giving low Lovibond colour index upto 3% ethephon concentration and the properties were found to be inferior above 3% concentration. A probable reaction mechanism is proposed for colour improvement with optimum concentration of ethephon.

Genome-wide identification of rubber tree (Hevea brasiliensis Muell. Arg.) aquaporin genes and their response to ethephon stimulation in the laticifer, a rubber-producing tissue.

BackgroundNatural rubber, an important industrial raw material, is specifically synthesized in laticifers located inside the rubber tree (Hevea brasiliensis Muell. Arg.) trunk. Due to the absence of plasmodesmata, the laticifer water balance is mediated by aquaporins (AQPs). However, to date, the characterization of H. brasiliensis AQPs (HbAQPs) is still in its infancy.ResultsIn this study, 51 full-length AQP genes were identified from the rubber tree genome. The phylogenetic analysis assigned these AQPs to five subfamilies, including 15 plasma membrane intrinsic proteins (PIPs), 17 tonoplast intrinsic proteins (TIPs), 9 NOD26-like intrinsic proteins (NIPs), 4 small basic intrinsic proteins (SIPs) and 6 X intrinsic proteins (XIPs). Functional prediction based on the analysis of the aromatic/arginine (ar/R) selectivity filter, Froger’s positions and specificity-determining positions (SDPs) showed a remarkable difference in substrate specificity among subfamilies. Homology analysis supported the expression of 44 HbAQP genes in at least one of the examined tissues. Furthermore, deep sequencing of the laticifer transcriptome in the form of latex revealed a key role of several PIP subfamily members in the laticifer water balance, and qRT-PCR analysis showed diverse expression patterns of laticifer-expressed HbAQP genes upon ethephon treatment, a widely-used practice for the stimulation of latex yield.ConclusionsThis study provides an important genetic resource of HbAQP genes, which will be useful to improve the water use efficiency and latex yield of Hevea.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-2152-6) contains supplementary material, which is available to authorized users.

Involvement of Ethylene in the Latex Metabolism and Tapping Panel Dryness of Hevea brasiliensis.

Ethephon, an ethylene releaser, is used to stimulate latex production in Hevea brasiliensis. Ethylene induces many functions in latex cells including the production of reactive oxygen species (ROS). The accumulation of ROS is responsible for the coagulation of rubber particles in latex cells, resulting in the partial or complete stoppage of latex flow. This study set out to assess biochemical and histological changes as well as changes in gene expression in latex and phloem tissues from trees grown under various harvesting systems. The Tapping Panel Dryness (TPD) susceptibility of Hevea clones was found to be related to some biochemical parameters, such as low sucrose and high inorganic phosphorus contents. A high tapping frequency and ethephon stimulation induced early TPD occurrence in a high latex metabolism clone and late occurrence in a low latex metabolism clone. TPD-affected trees had smaller number of laticifer vessels compared to healthy trees, suggesting a modification of cambial activity. The differential transcript abundance was observed for twenty-seven candidate genes related to TPD occurrence in latex and phloem tissues for ROS-scavenging, ethylene biosynthesis and signalling genes. The predicted function for some Ethylene Response Factor genes suggested that these candidate genes should play an important role in regulating susceptibility to TPD.

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.

Effect of low frequency harvesting in <i>Hevea brasiliensis</i> on major raw rubber properties

Harvesting latex, a key activity in the production of natural rubber (NR), involves repeated tapping of the bark on the main trunk of the trees. Low frequency harvesting (LFH) systems are only economically feasible with the use of Ethephon (ET) which stimulates the tree to provide high yield per tap as a compensation for the yield loss due to lowering the tapping frequency. Ethephon concentration and its frequency of application may vary with the frequency of harvesting used. Latex samples were collected from genotypes; RRIC 100, RRIC 102, RRIC 121 and RRIC 130 harvested under S/2 d2, S/2 d3, S/2 d4 and S/2 d6 (i.e. tapping half of the spiral of tree once in two, three, four and six days, respectively) systems were processed into lace crepe and assessed for raw rubber properties viz. initial plasticity (Po), plasticity retention index (PRI), Mooney viscosity (VR), colour, nitrogen and ash content. No significant differences were observed among different LFH systems with respect to each raw rubber property tested. However, clonal differences in those raw rubber properties were prominent. Therefore, the study revealed that the low frequency harvesting systems tested with Ethephon stimulation do not have any effect on the quality of raw rubber produced.

Ethephon Stimulation and Yield Response of SomeHeveaClones in the Humid Forests of South West Cameroon

Several exploitation systems are being used today to sustainably improve dry rubber production by the rubber tree (Hevea brasiliensis). These involve different combinations of tapping frequencies, cut lengths, stimulation frequencies, and stimulant concentrations. Such combinations are much easier to ascertain for confirmed clones as opposed to new introductions, for which extensive testing is required. A study was therefore conducted in the South West region of Cameroon characterized by a monomodal rainfall regime (one dry and one rainy season) to evaluate yield response to Ethephon stimulation (conc. 2.5%) of some newly introduced rubber clones (IRCA 18, IRCA 19, RRIC 100, and RRIC 110) for large-scale planting in Cameroon. Generally, annual yields and tree productivity ranged between the referenced clones tested: some closer to the intermediate yielding GT 1 (IRCA 18 and IRCA 19) and others to the high yielding PB clones (RRIC 100 and RRIC 110) indicating thereby the possible convenient adoption of some established exploitation regimes for these new introductions. Climatic factors like cumulative rainfall and relative humidity conditioned rubber yields of clones tested and considerably accounted for yield variations. These results could be used as a first step towards deriving regional climate models for predicting rubber yields, especially in an era of global climate change.

HbMT2, an ethephon-induced metallothionein gene from Hevea brasiliensis responds to H2O2 stress

Metallothioneins (MTs) are the cysteine-rich proteins with low molecular weight, which play important roles in maintaining intracellular ion homeostasis, detoxification of heavy metal ions and protecting against intracellular oxidative damages. In this study a novel ethephon-induced metallothionein gene, designated as HbMT2, was isolated and characterized from Hevea brasiliensis. The HbMT2 cDNA contained a 237 bp open reading frame encoding 78 amino acids and the deduced protein showed high similarity to the type 2 MTs from other plant species. Expression analysis revealed more significant accumulation of HbMT2 transcripts in leaves and latex than in roots and barks. The transcription of HbMT2 in latex was strongly induced by ethephon and hydrogen peroxide (H(2)O(2)) stress. Overproduction of recombinant HbMT2 protein gave the Escherichia coli cells more tolerance on Cu(2+) and Zn(2+), and the recombinant HbMT2 could scavenge the reactive oxidant species (ROS) in vitro. All these results indicated that HbMT2 could respond to ethephon stimulation and H(2)O(2) stress as a ROS scavenger in H. brasiliensis. It is also suggested that HbMT2 function in improving the tolerance of rubber trees to heavy metal ions, and repressing the ethephon-induced senilism and tapping panel dryness (TPD) development by ROS scavenge system in H. brasiliensis.

Carbohydrate reserves as a competing sink: evidence from tapping rubber trees

Carbohydrate reserve storage in trees is usually considered a passive function, essentially buffering temporary discrepancies between carbon availability and demand in the annual cycle. Recently, however, the concept has emerged that storage might be a process that competes with other active sinks for assimilate. We tested the validity of this concept in Hevea brasiliensis Müll. Arg. (rubber) trees, a species in which carbon availability can be manipulated by tapping, which induces latex regeneration, a high carbon-cost activity. The annual dynamics of carbohydrate reserves were followed during three situations of decreasing carbon availability: control (no tapping), tapped and tapped with Ethephon stimulation. In untapped control trees, starch and sucrose were the main carbohydrate compounds. Total nonstructural carbohydrates (TNC), particularly starch, were depleted following bud break and re-foliation, resulting in an acropetal gradient of decreasing starch concentration in the stem wood. During the vegetative season, TNC concentration increased. At the end of the vegetative season, there were almost no differences in TNC concentration along the trunk. In tapped trees, the vertical gradient of starch concentration was locally disturbed by the presence of the tapping cut. However, the main effect of tapping was a dramatic increase in TNC concentration, particularly starch, throughout the trunk and in the root. The difference in TNC concentration between tapped and untapped trees was highest when latex production was highest (October); the difference was noticeable even in areas of the trees that are unlikely to be directly involved in latex regeneration, and it was enhanced by Ethephon stimulation, which is known to increase latex metabolism and flow duration. Thus, contrary to what could be expected if reserves serve as a passive buffer, a decrease in carbohydrate availability resulted in a net increase in carbohydrate reserves at the trunk scale. Such behavior supports the view that trees tend to adjust the amount of carbohydrate reserves stored to the level of metabolic demand, at the possible expense of growth.

Effects of Ethephon Stimulation on Clone PB 260 Tapped on Panel BO-1 : Preliminary Results

Tapping of Hevea brasiliensis for latex is usually accompanied by stimulation. In this study two methods of stimulating clone PB 260 on panel BO-1 tapped on half spiral cut downward on d/4 frequency were evaluated. The preliminary results show that stimulation from eleven rounds with 1.25 per cent ethephon, per year gave yields that were higher than that of the unstimulated control and similar to yields from six rounds of stimulation per year with 2.5 per cent ethephon per year, during the first year of recording. However a sharp decline in yield responses was noted for both stimulated treatments in the second year of recording. The percentage dry cut length was significantly lower during the second year for trees stimulated with 1.25 per cent ethephon (eleven rounds per year) than trees stimulated with 2.5 per cent ethephon six rounds per year. Control and stimulated trees showed no differences with regard to the physiological parameters monitored. Keywords: Clone, drycut length, Hevea brasiliensis, production, stimulation.