Some aspects of sucrose transport and utilization in latex producing bark ofHevea brasiliensis Muel. Arg

The radiotracer technique is described for transport and metabolic studies in latex producing bark. This technique provided further evidence that carbohydrates are supplied to latex vessels as sucrose and that sucrose is readily used for rubber synthesis. Bark treatment with ethylene resulting in latex flow stimulation is shown to activate within a few hours sucrose and water transport into the latex vessels. The available knowledge suggests an implication of an energy-dependent process of proton excretion which could explain the earlier observed ethylene induced alkalization of latex cytosol enhancing invertase and sucrose utilization. An increase of tapping intensity decreases the level of latex sucrose and changes its distribution pattern in drained bark. There is a clonal difference in the early response to exploitation. Bark excision by conventional tapping inhibits downward sucrose translocation, the inhibition being proportional to the surface of consumed bark above the tapping cut. Seasonal variation in the level of latex sucrose is related to variations in sunshine duration and sucrose concentration rises during the process of refoliation. The results stress the importance of sucrose supply to latex vessels for latex producing potentialities and indicate an association of low sucrose availability in latex vessels with premature latex vessel degeneration and bark dryness.     

Control of carbohydrate metabolism by ethylene in latex vessels ofHevea brasiliensis MUEL. ARC in relation to rubber production

The treatment of the bark of regularly tapped trees with 2-ohloroethylphosphonio acid (CEPA) in the long run brings down sucrose level in the latex even when latex production is not increased. The sucrose deficit thus caused persists for a number of months after the treatment probably due to the depletion of carbohydrate reserves by the stimulation. The loss of carbohydrates not related to an increase of rubber produotion grows as the concentrations of applied CEPA are raised. The treatment increases the activity of phosphoenolpyruvate (PEP) carboxylase and decreases pyruvate deoarboxylation in mitochondria-free oytoplasmic latex serum. The pyruvate deoarboxylase has a sharp optimum at pH 6.5 and is almost inactive at pH 7.5 while PEP oarboxylase, like invertase, has the optimum at weakly alcaline pH and is only slightly active at pH 6.5. The CEPA-induced changes in the activity of these enzymes are related to an increase of latex pH after the treatment. Consistently with this the evolution of CO2 from sugars catabolised in the serum was found to decrease with increasing pH. It is concluded that the observed ethylene-induced changes in enzyme aotivities bring about in latex vessels a deviation of the glycolysis-fed metabolio flux from synthetic pathways to mitochondrial oxidations and consequently a decrease in the efficiency of sucrose conversion to rubber.     

Effect of strictly plant-related factors on the response ofHevea brasiliensis andTheobroma Cacao nodal explants culturedIn vitro

Summary Nodal cuttings (primary nodal explants) were used to initiate microcutting cultures ofHevea brasiliensis andTheobroma cacao. The physiological stage of growth reached by the branches from which the explants were taken had a considerable influence on culture initiation and response. In both species, the buds of the nodal explants taken from dormant branches (stages 11, 12 and 13 inTheobroma; stage D inHevea) revealed greater opening abilityin vitro and shoot elongation than buds taken from branches in a phase of active meristem growth inTheobroma (stages F1 and F2) or during a foliar growth phase inHevea (stage C). InHevea, the branch growth phase effect persisted into the multiplication phase when shoots developedin vitro were used as the source of secondary nodal explants. However, it disappeared during subcultures of primary nodal explants. In both species, the position of the nodes on the branch with respect to the apex also had an effect on culture initiation. The buds of nodes furthest from the apex were more likely to develop into shoots. InHevea, the position of secondary nodal explants obtained from the primary shoot was seen to vary in effect depending on growth regulator applications during primary culture. These results are analyzed in terms of variations in the balance between endogenous growth promoters/inhibitors in the buds, depending on morphogenic stages and the bud position on the branch.     

Stimulatory effects of 2,4-dichlorophenoxyacetic acid and of 1-naphthylacetic acid on sucrose level,invertase activity and sucrose utilization in the latex ofHevea brasiliensis

Summary Treatment of the bark ofHevea brasiliensis with 2,4-dichlorophenoxyacetic acid (2,4-D) or l-naphthylacetic acid (NAA) greatly increases sucrose level, invertase activity and sucrose utilization in the latex; the efficacy of 2,4-D is considerably greater than that of NAA. The greater sucrose utilization is the consequence of increased invertase activity. The changes occur as soon as the first tapping following bark treatment. It is suggested that the rise in both sucrose level and utilization in the latex serum mediate the effect of auxins on latex production. This is most likely related to a faciliation of latex outflow resulting from an increase in the osmotic and turgor pressure in the laticiferous tissue, as well as to enhanced regeneration of latex.The latex invertase has been found to be of a weakly alkaline type, with a sharp pH optimum at 7.15–7.20 in citrate-phosphate buffer. Its activity falls of rapidly on the acid side, being almost zero at pH 6.4. Since the natural pH of latex generally varies between pH 6.5 and 7.0, it is suggested that pH is an important factor in the regulation of invertase activity in the latex, and that the limiting nature of invertase-mediated sucrose hydrolysis in latex serum is caused by unfavourable conditions for invertase activity rather than by a scarcity of this enzyme.Expert of the International Atomic Energy Agency.     

The distribution of plasmodesmata in the phloem ofHevea brasiliensis in relation to laticifer loading

Summary Cell to cell connections, including plasmodesmata and perforations, were examined in the non-conducting secondary phloem ofHevea brasiliensis. Samples were taken from trunks of numerous trees, from several clones, and prepared for thin sectioning and transmission or scanning electron microscopy and as optical sections for fluorescence microscopy. Numerous plasmodesmata were found clustered in primary pit-fields between the ray and axial parenchyma cells. Between the laticifers and adjacent parenchyma sheath cells, structures corresponding to functional plasmodesmata were not observed. But some unusual structural features were occasionally seen in these walls. These observations are discussed in relation to the possible function of the cell types, and to the loss of latex on the tapping ofHevea. It is suggested that the loading of the laticifer might first require a symplastic pathway for the transport of metabolites, at the end of which the assimilates must enter the apoplast. A transmembrane active transport system then transfers the metabolites in the laticifer. The presumable role of parenchyma cells in the loading of laticifers is emphasized.     

Studies in the GenusHevea. VIII. Notes on infraspecific variants ofHevea brasiliensis (Euphorbiaceae)

Supplying 98% of the world's natural rubber,Hevea brasiliensis has transformed life around the world in only a century. This species-one of 10 in its genus-gives the best natural rubber, but much more remains to be studied concerning the numerous localised strains of the species. The British introduced to the Orient seeds from the lower Amazon-representatives of a not superior ecotype of the species but, at that time a century ago, the only available germ plasm. The time for study of the intraspecific variants ofH. brasiliensis and their use in programmes of genetic manipulation of the genus is long overdue. But basic to this desired programme is a clear taxonomic understanding of infraspecific variants, still awaiting evaluation. Although numerous taxonomic treatments of subspecific variants have been offered, there is little available to give genetic programmes concrete information.     

Ethylene stimulation of latex production in Hevea brasiliensis

Rubber tree (Hevea brasiliensis) is an important industrial crop for natural rubber production. Ethylene, as a stimulant of latex production in H. brasiliensis, has been widely used in commercial latex production. However, the mechanism of ethylene action are not completely elucidated, especially in molecular aspect. Here, we focus on the molecular biological progression of ethylene stimulation of latex production. Our data and all previous information showed ethylene had little direct effect on accelerating rubber biosynthesis. The prolonged latex flow and acceleration of sucrose metabolism by ethylene may be the main reasons for the stimulation of latex yield by ethylene.Key words: Hevea brasiliensis, ethylene, rubber production, gene, sucrose     

Base temperature and heat units for leaf flushing emission and growth ofHevea brasiliensis Muell. Arg.

In this study we determined the base temperature and Heat units for leaf flushing initiation and growth of RRIM-600 and GT1Hevea clones. The minimum base temperature was found to be approximately 16° C for leaf flushing emission and 19° C for shoot growth in terms of height. Approximately 420 degree days at 16° C as base temperature is required for successive leaf flushing initiation. Linear equations with correlation coefficients above 0.95 allow an estimation of the height increase from the accumulated degree days, corrected or otherwise for photoperiod. The highest correlation coefficients demonstrated a positive effect of the photoperiod among the factors influencing the shoot growth.     

Photosynthate allocation and productivity of latex vessels inHevea brasiliensis

Manipulations of production systems in rubber tree which were intended to improve sucrose translocation in tapped bark resulted in an increase of latex sucrose and of latex production and reduced the incidence of nonyielding laticiferous tissue. This was achieved by shortening the tapping cut from full to half spiral, by changing the descending direction into an ascending mode of tapping or by annual change-over of tapping panel allowing for a longer time the regeneration of bark removed above the location of the cut. The increase of latex yield did not result in a significant decrease in the growth of trees over a period of three years. Clonal differences in nonyielding bark appeared to be related to differences in sucrose depletion by tapping. In clone PB 235 which exhibited low latex sucrose, a reduction of tapping frequency resulted in an increase in sucrose level and in a decrease of bark “dryness” tending to an increase in total yield. The tapping manipulations examined did not affect latex flow characteristics such as the plugging index of latex vessels and the bursting index of lutoids. The results stress the importance of photosynthate allocation for the physiology of laticiferous system productivity and indicate the possibilities of improving assimilate economy in rubber trees. On mission as export of the International Atomic Energy Agency.     

The proton gradient across the vacuo-lysosomal membrane of lutoids from the latex ofHevea brasiliensis. I. Further evidence for a proton-translocating ATPase on the vacuo-lysosomal membrane of intact lutoids

Summary Lutoids (vacuo-lysosomal particles) were isolated from the latex ofHevea brasiliensis. Using flow dialysis with¹⁴C-methylamine uptake as a pH probe and⁸⁶Rb rubidium+valinomycin distribution for estimations of transmembrane electrical potential, intact lutoids exhibited a pH of 1 unit (interior more acid) and a of –70 mV (interior negative), when suspended in an isotonic medium at physiological concentration of potassium (30mm) and pH 7.0, in the absence of ATP. In most cases, the Donnan potential was shown to fully account for pH in nonenergized lutoids. The addition of Mg-ATP (5mm) resulted in a marked acidification of the lutoidic internal space (0.7 to 1 pH unit) depending on the composition of the medium, and in a membrane depolarization by 60 mV (interior becoming less negative). The resulting electrochemical potential of protons ( ) increased by a hundred millivolts when lutoids were energized by ATP. These data strongly support an inward electrogenic proton translocating function for the ATPase of the vacuo-lysosomal membrane of lutoids. Results are discussed in terms of thein vivo maintenance of large lutoids/cytoplasm proton gradients, and of the rôle of these vacuo-lysosomes in the homeostasis of the cytoplasmic metabolism.     

Anaerobic respiration and photoautotrophy in the evolution of prokaryotes

Anaerobic respiration preceded photosynthesis. Chemoautotrophy, photoautotrophy, and oxygen respiration were induced directly or indirectly from anaerobic respiration.     

Nutrients and limiting factors

The terms nutrient and limiting factor summarise the results of an experiment in which increase in supply results in an increased response. By extension they are often — perhaps usually — used when the user believes that were such an experiment made it would have this characteristic. If the supply is further increased the response diminishes and may, eventually, become negative. Nutrient and limiting factor therefore apply, strictly, only when the circumstances are specified: they cannot be attached to a particular substance without qualification. The claim that nitrogen is a nutrient (or limiting factor) is incomplete. All nutrients are limiting factors, but the reverse is not true. The widespread belief that only one factor can limit a complex process at one time is demonstrably false in general, though it may sometimes be true in particular cases.     

Polyphenol oxidases from latex of Hevea brasiliensis: purification and characterization

Polyphenol oxidase (PPO) was isolated from the B-serum obtained after repetitive freeze-thawing of the bottom fraction isolated from ultracentrifuged fresh latex. The B-serum was subjected to acetone precipitation and CM-Sepharose chromatography, affording two PPOs, PPO-I and PPO-II, which, upon SDS-PAGE, were 32 and 34 kDa, respectively. Both PPOs possessed the same pI (9.2), optimum pH (7) and optimum temperature (35-45 degrees C). They are stable up to 60 degrees C and active at broad pH ranges from 4-9. The K(m) values of PPO-I for dopamine, L-dopa and catechol as substrates are 2.08, 8.33 and 9.09 mM, while those for PPO-II are 2.12, 4.76 and 7.14 mM, respectively. Among various PPO inhibitors tested, 4-hexylresorcinol was the most potent. Anionic detergents were among the most effective activators of the enzymes, while cationic and nonionic detergents showed little and no effect on the PPO activities, respectively.     

Relationship between latex yield of Hevea brasiliensis and antecedent environmental parameters

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

Landscape genetics and limiting factors

Population connectivity is mediated by the movement of organisms or propagules through landscapes. However, little is known about how variation in the pattern of landscape mosaics affects the detectability of landscape genetic relationships. The goal of this paper is to explore the impacts of limiting factors on landscape genetic processes using simulation modeling. We used spatially explicit, individual-based simulation modeling to quantify the effects of habitat area, fragmentation and the contrast in resistance between habitat and non-habitat on the apparent strength and statistical detectability of landscape genetic relationships. We found that landscape genetic effects are often not detectable when habitat is highly connected. In such situations landscape structure does not limit gene flow. We also found that contrast in resistance values between habitat and non-habitat interacts with habitat extensiveness and fragmentation to affect detectability of landscape genetic relationships. Thus, the influence of landscape features critical to providing connectivity may not be detectable if gene flow is not limited by spatial patterns or resistance contrast of these features. We developed regression equations that reliably predict whether or not isolation by resistance will be detected independently of isolation by distance as a function of habitat fragmentation and contrast in resistance between habitat and non-habitat.     

Diffusion and the limiting substrate in two-substrate immobilized enzyme systems

The effects of mass transport resistances on two-substrate immobilized enzyme systems are investigated theoretically. It is shown that the effects of mass transport resistances on the overall reaction rate are related mainly to the transport of the limiting substrate. In the absence of external mass transport resistances, the limiting substrate can be identified by knowing only the ratio of the bulk substrate concentrations, the permeability of the support to the two substrates, and the stoichiometry of the reaction. However, a combination of internal and external mass transport resistances may result in the other substrate becoming limiting. These effects are most significant when the mass transport resistances are high. Applications in the design of enzyme electrodes and chemical reactors are discussed.     

Factors limiting mitochondrial respiration in media of high solute content

The state-3 rate of respiration of rat-liver mitochondria was depressed in media containing KCl, sucrose, or mannitol at concentrations in excess of 125 mM. At equivalent concentrations, glucose caused less inhibition than sucrose or mannitol, and no inhibition was observed with glycine. These observations establish that solute inhibition of respiration is not a consequence of the reduced chemical potential of water in the system. The accumulation of succinate by mitochondria was not reduced by high sucrose concentrations. Sonication only partially relieved inhibition by sucrose or mannitol, and not at all that by KCl, and the evidence indicates that solute inhibition is not primarily an inhibition of substrate entry into mitochondria. Sucrose in the assay media inhibited succinate dehydrogenase [succinate: PMS oxidoreductase (EC.1. 3. 91)] and malate dehydrogenase [l-malate: NAD oxidoreductase (EC.1.1.1.37)] activities, but these inhibitions were less than those of succinate-and malate-dependent oxygen uptake by mitochondria. Disruption of the mitochondrial membrane by detergent abolished the inhibition of respiration by sucrose, and the evidence indicates that solute inhibits the functional capacity of the membrane-associated respiratory system.     

Nippostrongylus brasiliensis and Ascaridia galli: mitochondrial respiration in free-living and parasitic stages

Aerobic respiratory pathways have been delineated and respiratory efficiency has been assessed in mitochondria isolated from embryonated eggs, infective larvae, and adult Nippostrongylus brasiliensis and Ascaridia galli. Mitochondrial respiration in free-living stages of N. brasiliensis is mediated mainly by a mammalian-like antimycin A- and cyanide-sensitive pathway; specific respiratory activity is high and oxidative phosphorylation efficient. In mitochondria of adult N. brasiliensis, antimycin A- and cyanide-sensitive respiration is decreased relative to respiration though an alternative pathway, and specific respiratory activity and mitochondrial efficiency are lower. Respiration in mitochondria from embryonated eggs and tissues of adult A. galli is comparable, and apparently mediated by an antimycin A- and cyanide-insensitive alternative respiratory pathway; no evidence for the presence of a mammalian-like respiratory pathway in embryonated eggs of A. galli was found. The results of this study are compared to mitochondrial respiration in eggs, larvae, and adult body wall muscle of Ascaris suum.