Corrélations de croissance entre le mycélium et les rhizomorphes chez le Sphaerostilbe repens

Correlation of Growth between Mycelium and Rhizomorphs in Sphaerostilbe repens In Sphaerostilbe repens Berkeley and Broome, cultured on a solid medium in Petri dishes, growth of mycelium and chizomorphs depends on the thickness of the culture medium underlying the thallus. On shallow layers of agar medium, aggregated units were fewer and rhizomorph growth reduced. Vegetative mycelium which starts growth earlier than rhizomorphs contributed to inhibit the development of these latter. On deep layers of substrate, rhizomorphs were numerous, elongated rapidly, and eventually inhibited undifferentiated mycelium. – There was no evidence for the involvement of any inhibitor substance diffusing into culture medium or located in the thallus. Development of aerial mycelium and rhizomorphs, could, on the contrary, be interpreted as nutritional competition between these two parts of the colony.     

Armillaria luteobubalina mycelium develops air pores that conduct oxygen to rhizomorph clusters

Armillaria luteobubalina produces air pores in culture. They consist of two parts: a basal region of tissue elevated to form a mound covered with a rind continuous with that of the colony, but perforated; and an apical region of long parallel hyphae, cemented together by scattered patches of extracellular material. This forms a hydrophobic structure that is elevated above the general level of the mycelial crust and does not easily become waterlogged. Air pores develop near the inoculum plug shortly after inoculation, arising directly from the mycelium, and rhizomorphs are initiated from them. The air pore contains a complex system of gas space connecting the atmosphere with the central canal of each rhizomorph. The tissue beneath the melanised colony crust also contains gas space, especially near air pores. This is also connected with the gas space of each rhizomorph and of each air pore. Measurements with oxygen electrodes show that air pores and their associated rhizomorphs conduct oxygen. The average oxygen conductance of a group of air pores with associated rhizomorphs, within agar blocks, but with rhizomorph apices cut off, was about 700 × 10⁻¹² m³ s⁻¹, equivalent to about 200 × 10⁻¹² m³ s⁻¹ for each air-pore. We conclude that the air pores conduct oxygen into the gas space below the pigmented mycelium of the colony, where the rhizomorphs - which also conduct oxygen - originate. A. luteobubalina thus has a complex aerating system which allows efficient diffusion of oxygen into rhizomorphs, and this is likely to facilitate extension of inoculum into low-oxygen environments.     

Polar Transport of Auxin across Gravistimulated Roots of Maize and Its Enhancement by Calcium

The effect of Ca on the polar movement of [³H]indoleacetic acid ([³H] IAA) in gravistimulated roots was examined using 3-day-old seedlings of maize (Zea mays L.). Transport of label was measured by placing an agar donor block containing [³H]IAA on one side of the elongation zone and measuring movement of label across the root into an agar receiver block on the opposite side. In vertically oriented roots, movement of label across the elongation zone into the receiver was slight and was not enhanced by incorporating 10 millimolar CaCl₂ into the receiver block. In horizontally oriented roots, movement of label across the root was readily detectable and movement to a receiver on the bottom was about 3-fold greater than movement in the opposite direction. This polarity was abolished in roots from which the caps were removed prior to gravistimulation. When CaCl₂ was incorporated into the receivers, movement of label across horizontally oriented intact roots was increased about 3-fold in both the downward and upward direction. The ability of Ca to enhance the movement of label from [³H]IAA increased with increasing Ca concentration in the receiver up to 5 to 10 millimolar CaCl₂. With the inclusion of CaCl₂ in the receiver blocks, gravity-induced polar movement of label into receiver blocks from applied [³H]IAA was detectable within 30 minutes, and asymmetric distribution of label within the tissue was detectable within 20 minutes. The results indicate that gravistimulation induces a physiological asymmetry in the auxin transport system of maize roots and that Ca increases the total transport of auxin across the root.     

Régénération des boutures d’apex de rhizomorphes chez lesphaerostilbe repens B. et Br.

The tips of the rhizomorphs ofSphaerostilbe repens transferred on agar medium regenerate some aggregated organs without an intermediate mycelium stage. Each extremity of the drawn rhizomorphs can give rise to eoremia on its air face and to rhizomorphs on the contact with tho culture medium. These heterogeneous environmental factors impose polarized regeneration, easily inverted, upon the rhizomorphs. Nevertheless, a residual and stable polarity remains; it is attributed to the apex which is more capable of bearing new organs than the other extremity. Tho efficiency to regenerate depends on the ago of collulos and on tho size of tho rhizomorph tip; howover it is very difficult to provent the formation of new aggregated organs.     

Biomechanics of invasive growth by Armillaria rhizomorphs

Rhizomorphs of wood-decay basidiomycetes are root-like structures produced by the coordinated growth of thousands of hyphae. Very little is known about their development nor the way that they penetrate soils and rotting wood. In this study, we applied techniques used in previous studies on hyphae to explore the mechanics of the invasive growth process in Armillaria gallica. Growth rate measurements were made in media with different gel strengths. The osmolyte composition of rhizomorph sap was determined spectroscopically and the forces exerted by growing tips were measured using a force transducer. Cultured rhizomorphs extended at much faster rates than unbundled hyphae (3.5 mm d⁻¹ versus 1.5 mm d⁻¹) and their growth accelerated in response to increased medium gel strength (to 7.4 mm d⁻¹). Measurements of rhizomorph osmolality provided a turgor pressure estimate of 760 kPa (7.5 atm.), and spectroscopic analysis showed that this pressure was generated by the accumulation of erythritol, mannitol, and KCl. Forces exerted by growing tips ranged from 1 to 6 mN, corresponding to pressures of 40–300 kPa (0.4–3.0 atm.). Pressures exerted by extending rhizomorphs are comparable to those produced by individual vegetative hyphae. This suggests that the mechanical behavior of hyphae is similar whether they grow as unbundled cells or aggregate to form macroscopic rhizomorphs.     

Temporal changes in the elemental composition of Rhizopogon rhizomorphs during colonization of patches with fresh organic matter or acid-washed sand

A laboratory experiment was performed to estimate the elemental composition of rhizomorphs of an ectomycorrhizal (EM) fungus growing in a patchy environment. Successive samples of Rhizopogon rhizomorphs, located adjacent to patches with organic matter or patches with acid-washed sand, were taken over a period of 45 d. The mass per unit area was analyzed with scanning transmission ion microscopy (STIM), and the elemental content of elements heavier than Mg were analyzed with particle induced X-ray emission (PIXE). Rhizomorphs associated with the organic matter on average were three times heavier per unit area than rhizomorphs associated with sand. The Ca concentration (mg g(-1)) increased in rhizomorphs adjacent to patches with sand, while it decreased in rhizomorphs adjacent to patches with organic matter. Fe concentration was higher in rhizomorphs adjacent to patches with sand. We concluded that the EM fungus responded to the organic matter by producing larger rhizomorphs, rather than increasing the concentration of elements in the rhizomorphs, to improve the transport of elements to the roots. The elemental composition of rhizomorphs varied considerably over time, and the accumulation of Ca in rhizomorphs in the sand-filled compartments could be the effect of acropetal flow of solutes from the plant roots toward the mycelial front.     

Studies on ectomycorrhiza

Summary Plants ofPicea abies (L.) Karst were grown in mycorrhizal association withTelephora terrestris (Pers. ex Fr.) andPisolithus tinctorius (Mich. ex Pers.) Coker and Couch on sphagnum peat in petri dishes or Perspex chambers. After 1 yearT. terrestris had formed prominent rhizomorphs which were characterized by light microscopy and investigated for³²P-orthophosphate uptake. The absorbed phosphate was transported to sinks throughout the rhizomorphal system as well as into the plant. The calculated translocation velocity and flux rate in the rhizomorph were in the range of 1–3 cm/h and 0.5–4.0 × 10^-10 mol cm^-2 s^-1, respectively. Label was observed to accumulate in the needles 2–3 days after application. Feeding a non-mycorrhized root with³²P-orthophosphate led to an accumulation of label in needles within 1 h, but no radioactivity appeared in the associatedT. terrestris rhizomorphs. The rhizomorphs ofP. tinctorius revealed a higher structural differentiation than those ofT. terrestris. Translocation of labelled phosphorus through rhizomorphs ofP. tinctorius into spruce needles was also demonstrated.     

The accumulation of mercury by the thornback ray, Raja clavata L.

The accumulation of inorganic and organic mercury by the thornback ray has been studied using ²⁰³HgCl₂ and CH₃²⁰³HgCl. Observations have been made on the rates of intake and loss of ²⁰³Hg, from both labelled food and sea water, and the internal distributions of the isotope compared with that of total mercury. Both forms were readily absorbed from sea water. Retention of the two forms from food was, however, dissimilar in that, in contrast to inorganic mercury, methylmercury was readily absorbed and only slowly eliminated. The results are compared with data on the accumulation of mercury by the plaice.     

Internal recycling of respired CO2 may be important for plant functioning under changing climate regimes

Recent studies have provided evidence of a large flux of root-respired CO₂ in the transpiration stream of trees. In our study, we investigated the potential impact of this internal CO₂ transport on aboveground carbon assimilation and CO₂ efflux. To trace the transport of root-respired CO₂, we infused a ¹³C label at the stem base of field-grown Populus deltoides Bartr. ex. Marsh trees. The ¹³C label was transported to the top of the stem and throughout the crown via the transpiration stream. Up to 17% of the ¹³C label was assimilated by chlorophyll-containing tissues. Our results provide evidence of a mechanism for recycling respired CO₂ within trees. Such a mechanism may have important implications for how plants cope with predicted increases in intensity and frequency of droughts. Here, we speculate on the potential significance of this recycling mechanism within the context of plant responses to climate change and plants currently inhabiting arid environments.     

Foliar nutrition of <Emphasis Type="Italic">in vitro</Emphasis>-cultured <Emphasis Type="Italic">Prosopis chilensis</Emphasis> (Molina) Stuntz shoots

Summary Foliar nutrition has been conceived as a possible means of overcoming the recalcitrance of Prosopis chilensis (Molina) Stuntz explants to standard in vitro culture. The foliar uptake of cations (K from 20 gl⁻¹ KNO₃ and Ca from 50 gl⁻¹ CaCl₂), anions (NO₃ from 50 gl⁻¹ KNO₃ and PO₄ from 50 gl⁻¹ NaH₂PO₄), and glucose from a 100 mg l⁻¹ solution studied. All of the nutrients examined were absorbed. The efficacy of foliar nutrition in prolonging the vigor of micropropagated P. chilensis shoot tips was compared with nutrients supplied as a liquid to the base of the stem (liquid) or as an agar-solidified medium (agar). A foliar-feeding apparatus was constructed that employed pressurization of the medium reservoir to drive the medium into the culture vessel with a passive return by a siphoning effect. The medium used was Murashige and Skoog with 30 gl⁻¹ sucrose, 0.1 mgl⁻¹ benzylaminopurine, and 1 mgl⁻¹ indole-3-butyric acid. Over a 9-wk test period it was found that explants cultured by foliar nutrition performed significantly better than those grown on agar for shoot length, nodal production, and leaf retention; and better than liquid MS for node production. There was no significant difference among the three treatments in percentage survival, percentage rooting, or the mean number of roots.     

Characterization of Japanese Lactarius section deliciosi ectomycorrhizae: Toward the use of mycorrhizae for taxonomy and expansion of available cultures

We described the ectomycorrhizae (ECM) of the Japanese Lactarius section Deliciosi species L. akahatsu, L. hatsudake, L. laeticolor, and L. subindigo with the aim of promoting the use of mycorrhizae for taxonomy and expansion of available cultures. Lactarius tottoriensis (sect. Lactarius) was also included as a comparator. The ECM of the four species of sect. Deliciosi showed orange laticifers and plectenchymatous mantles, whereas those of L. tottoriensis showed white laticifers and pseudoparenchymatous mantles. The ECM of L. hatsudake and L. subindigo showed complex rhizomorphs with thicker walls and more developed vessel-like hyphae compared to the other three species. Cystidium-like cells were rarely found on the mantles of L. laeticolor and L. subindigo. Fungal cultures of the five Lactarius species from their ECM were morphologically characterized. Cultures produced laticifers on agar medium, but the five species showed different growth rates. Rhizomorphs of L. hatsudake and L. subindigo in cultures showed a similar morphology to their ECM. Molecular phylogenetic analyses of cultures from the ECM demonstrated that they were of the targeted Lactarius species and suggested the need for taxonomic studies of L. hatsudake, L. subindigo, and L. tottoriensis at the species and section levels.     

Correlations between Gravitropic Curvature and Auxin Movement across Gravistimulated Roots of Zea mays

We compared the kinetics of auxin redistribution across the caps of primary roots of 2-day-old maize (Zea mays, cv Merit) seedlings with the time course of gravitropic curvature. [³H] indoleacetic acid was applied to one side of the cap in an agar donor and radioactivity moving across the cap was collected in an agar receiver applied to the opposite side. Upon gravistimulation the roots first curved upward slightly, then returned to the horizontal and began curving downward, reaching a final angle of about 67°. Movement of label across the caps of gravistimulated roots was asymmetric with preferential downward movement (ratio downward/upward = ca. 1.6, radioactivity collected during the 90 min following beginning of gravistimulation). There was a close correlation between the development of asymmetric auxin movement across the root cap and the rate of curvature, with both values increasing to a maximum and then declining as the roots approached the final angle of curvature. In roots preadapted to gravity (alternate brief stimulation on opposite flanks over a period of 1 hour) the initial phase of upward curvature was eliminated and downward bending began earlier than for controls. The correlation between asymmetric auxin movement and the kinetics of curvature also held in comparisons between control and preadapted roots. Both downward auxin transport asymmetry and downward curvature occurred earlier in preadapted roots than in controls. These findings are consistent with suggestions that the root cap is not only the site of perception but also the location of the initial redistribution of effectors that ultimately leads to curvature.     

Nitrogen isotopes in ectomycorrhizal sporocarps correspond to belowground exploration types

Nitrogen isotope values (δ¹⁵N) are higher in ectomycorrhizal fungi than in their plant hosts but the wide variability in δ¹⁵N among sporocarps of different fungal taxa is unexplained. We propose that fungal δ¹⁵N reflects sequestration of fungal nitrogen to build fungal biomass, and should accordingly reflect fungal exploration strategies and hyphal properties. To test this, we compared δ¹⁵N to exploration types, hyphal hydrophobicity, and the presence of rhizomorphs in ectomycorrhizal species from surveys at four sites in temperate and boreal coniferous forests. Fungi with exploration types of high biomass, such as long-distance (e.g., Suillus), medium-distance mat (e.g., Hydnellum), and medium-distance fringe (e.g., Cortinarius) were 4–7‰ more enriched in ¹⁵N than fungi with exploration types of low biomass [medium-distance smooth (e.g., Amanita), short-distance (e.g., Inocybe), and contact (e.g., Hygrophorus)]. High biomass types comprised 79% (Åheden, northern Sweden), 65% (Deer Park, Pacific Northwest, USA), 45% (Stadsskogen, central Sweden), and 39% (Hoh, Pacific Northwest, USA) of ectomycorrhizal species, with these types more prevalent at sites of lower nitrogen availability. Species with hydrophobic hyphae or with rhizomorphs were 3–4‰ more enriched in ¹⁵N than taxa with hydrophilic hyphae or without rhizomorphs. The consistency of these patterns suggest that δ¹⁵N measurements could provide insights into belowground functioning of poorly known taxa of ectomycorrhizal fungi and into relative fungal biomass across ectomycorrhizal communities.     

Uptake of HCO3? and CO2 in Cells and Chloroplasts from the Microalgae Chlamydomonas reinhardtii and Dunaliella tertiolecta

Mass-spectrometric disequilibrium analysis was applied to investigate CO₂ uptake and HCO₃⁻ transport in cells and chloroplasts of the microalgae Dunaliella tertiolecta and Chlamydomonas reinhardtii, which were grown in air enriched with 5% (v/v) CO₂ (high-Ci cells) or in ambient air (low-Ci cells). High- and low-Ci cells of both species had the capacity to transport CO₂ and HCO₃⁻, with maximum rates being largely unaffected by the growth conditions. In high- and low-Ci cells of D. tertiolecta, HCO₃⁻ was the dominant inorganic C species taken up, whereas HCO₃⁻ and CO₂ were used at similar rates by C. reinhardtii. The apparent affinities of HCO₃⁻ transport and CO₂ uptake increased 3- to 9-fold in both species upon acclimation to air. Photosynthetically active chloroplasts isolated from both species were able to transport CO₂ and HCO₃⁻. For chloroplasts from C. reinhardtii, the concentrations of HCO₃⁻ and CO₂ required for half-maximal activity declined from 446 to 33 μm and 6.8 to 0.6 μm, respectively, after acclimation of the parent cells to air; the corresponding values for chloroplasts from D. tertiolecta decreased from 203 to 58 μm and 5.8 to 0.5 μm, respectively. These results indicate the presence of inducible high-affinity HCO₃⁻ and CO₂ transporters at the chloroplast envelope membrane.     

Differential regulation of the N-methyl transferases responsible for phosphatidylcholine synthesis in Saccharomyces cerevisiae

The enzymes catalyzing the conversion of phosphatidylethanolamine to phosphatidylcholine were assayed by measuring the incorporation of label from [¹⁴C-CH₃]-S-adenosyl-methionine into the endogenous phospholipids of particulate, cell-free preparations from S. cerevisiae grown in the presence of N-methylethanolamine, N,N-dimethylethanolamine, or choline. The results indicate that each base in the growth medium results in reduced levels of all the N-methyltransferase activity involved in the formation of the phosphatidyl ester of the given base. By following the conversion of exogenous [³²P]-phosphatidyldimethylethanolamine to [³²P]-phosphatidylcholine it has been shown that the activity of the third methyl transfer is 90% lower in particles prepared from choline grown cells than in particles prepared from cells grown without choline. The results suggest that there are at least two enzymes involved in the conversion of phosphatidylethanolamine to phosphatidylcholine and that their levels can be regulated individually.Supplementing the growth medium with any of the three methylated aminoethanols results in markedly increased cellular levels of their corresponding phosphatidyl esters and decreased levels of the precursor phosphatidyl esters. The fatty acid composition of phosphatidylcholine also changes when the medium is supplemented with choline suggesting that the proportions of the molecular species of this phosphatide depends on whether synthesis is via methylation of phosphatidylethanolamino or from the supplemented aminoethanol.     

Absorption and transport of dietary lipid in Pieris brassicae

The digestion and absorption of dietary glycerol tri(1-¹⁴C)oleate and oleic acid-1-¹⁴C and subsequent transport of the label was followed during the fifth instar in Pieris brassicae. The rate of incorporation of the label in tissue lipid was similar in both diets. Triolein was hydrolysed to free fatty acids (FFA), diglycerides (DGL), and monoglycerides (MGL). DGL were rapidly absorbed. FFA were less readily absorbed and some were excreted. In the gut wall the label was found in phospholipids (PL), triglycerides (TGL), DGL, and FFA. In haemolymph most of the label was in DGL and PL, but later appeared also in TGL and sterol esters (SE). The results suggest that DGL are released from the gut wall and carried in haemolymph into the fat body. In the fat body lipid is stored mainly as TGL, and released as DGL, TGL, and SE. The turnover of oleate in haemolymph DGL is rapid in comparison to haemolymph SE or TGL. Synthesis of PL in gut lumen is apparent. Much of this PL is excreted but some may be absorbed.     

Effects of 6-benzylaminopurine on photosystem II functionality and leaf anatomy of <Emphasis Type="Italic">in vitro</Emphasis> cultivated <Emphasis Type="Italic">Aechmea blanchetiana</Emphasis>

Cytokinins (CKs) are often used during the in vitro cultivation of plant species. However, it is not clear how CKs, such as 6-benzylaminopurine (BAP), affect photosystem PS) II functionality and leaf anatomy over a long period of in vitro plant propagation. The aim of this study was to analyze the residual effects of BAP on the photosynthetic performance and leaf anatomy of Aechmea blanchetiana after 120 d without exposure to CKs. Aechmea blanchetiana plants previously grown in vitro were transferred to Murashige and Skoog (MS) culture media containing 0, 5, 10, 15, or 20 μM BAP. After 60 d on the MS medium with BAP, explants were subcultivated twice on the MS medium without growth regulators, first in a stationary liquid medium for 60 d and then in a solidified medium with 6 g dm^-3 agar for 60 d. Leaf anatomy, pigment content, and chlorophyll a fluorescence were assessed for plants from each treatment after 120 d on the CK-free medium. Stomatal density presented a negative linear correlation with BAP concentration. Pigment content decreased in plants subjected to previous BAP exposure. An increase in absorbed energy flux per reaction center (ABS/RC) and a sharp decrease in energy transport flux (ETo/RC) followed by an increase in energy dissipation flux (DIo/RC) also occurred. Furthermore, maximum quantum yield (F_V/F_M) decreased as a function of BAP concentration. Thus, the use of BAP during in vitro propagation of A. blanchetiana induced long-term physiological defects.     

Detection of Lactobacillus acidophilus in Feces of Humans, Pigs, and Chickens

Lactobacilli in fecal material from humans, pigs, and chickens were enumerated on lactobacillus selective agar (LBS). In all samples, higher numbers of lactobacilli were detected when plates were incubated in a system flushed with CO₂ rather than in air. Much higher numbers of bacteria from human feces were detected when the LBS agar plates were incubated anaerobically in a hydrogen-carbon dioxide atmosphere (GasPak) than when incubated in CO₂. The bacteria from human feces isolated on LBS agar incubated anaerobically were predominately bifidobacteria. Cultures from all three sources isolated on LBS agar incubated under CO₂ were lactobacilli, including Lactobacillus acidophilus. Differences were observed in biochemical characteristics of some of the L. acidophilus isolated from all three sources. Guanine plus cytosine base ratios of deoxyribonucleic acid isolated from L. acidophilus cultures from humans were lower, in most cases, than those from pigs and chickens.     

Ammonium uptake by nitrogen fixing bacteria

Both the changes in the activities of nitrogenase, glutamine synthetase and glutamate dehydrogenase and in the extracellular and intracellular NH₄ ⁺ concentrations were investigated during the transition from an NH₄ ⁺ free medium to one containing NH₄ ⁺ ions for a continuous culture of Azotobacter vinelandii. If added in amounts causing 80–100% repression of nitrogenase, ammonium acetate, lactate and phosphate are absorbed completely, whereas chloride, sulfate and citrate are only taken up to about 80%. After about 1–2 hrs the NH₄ ⁺ remaining in the medium is absorbed too, indicating the induction or activation of a new NH₄ ⁺ transport system. One of the new permeases allows the uptake of citrate in the presence of sucrose. Addition of inorganic NH₄ ⁺ salts leads to acidification of the culture. Anaerobiosis suppresses NH₄ ⁺ transport. A rise in the extracellular NH₄ ⁺ level leads to a reversible rise in the glutamine synthetase activity, which is not prevented by chloramphenicol, and to a reversible decrease in nitrogenase activity. During these measurements glutamate dehydrogenase activity remains close to zero. The intracellular NH₄ ⁺ level of about 0.6 mM does not change when extracellular NH₄ ⁺ is taken up and repression of nitrogenase starts.     

Activation of a Na+-dependent amino acid transport system in preimplantation mouse embryos

The uptake of l-methionine-methyl-³H and l-leucine-³H from completely defined medium into acid-soluble fractions of preimplantation mouse embryos has been studied. Late four-cell embryos and early blastocysts raised in vitro can concentrate both amino acids by processes which exhibit saturable, Michaelis-Menten type kinetics, characteristic of carrier-mediated active transport systems. This uptake is temperature-sensitive and inhibited by certain amino acids which compete for the same uptake sites. Methionine uptake seems to be mediated by a single transport system (K_m = 6.25 × 10^?5M) at the four-cell stage. Complex kinetics suggest that two distinct transport systems exist at the early blastocyst stage (K_m = 6.25 × 10^?5M; 8.9 × 10^?4M). V_max values (mg/embryo/15 min) for methionine and leucine transport increase significantly from the late four-cell stage to the blastocyst stage, suggesting that additional carriers are produced or activated during development.Most importantly, leucine and methionine transport is Na⁺-independent at the four-cell stage, methionine transport is partially dependent at the morula stage, and both amino acids are completely Na⁺-dependent at the blastocyst stage. The cumulative results suggest that preimplantation embryos accumulate leucine and methionine by specific, chemically mediated, active transport systems. The qualitative and quantitative developmental changes in cell membrane function may represent preparatory steps for subsequent growth of embryonic and/or trophoblastic cells.