Saccharomyces cerevisiae wine strains differing in copper resistance exhibit different capability to reduce copper content in wine

Two wine strains of Saccharomyces cerevisiae, characterized by a different degree of copper resistance, were tested in grape must fermentation in the presence of different copper concentrations. The sensitive strain SN9 was strongly affected by copper concentration (32 ppm, (32 mg/l)), whereas the resistant strain SN41 exhibited a good growth activity in presence of 32 ppm of copper and only a reduced activity in presence of 320 ppm. The different strain fermentation performance in response to the copper addition corresponded to a different capability to accumulate copper inside the cells. Both strains exhibited the capacity to reduce the copper content in the final product, eventhough a significantly greater reducing activity was exerted by the resistant strain SN41, which was able to reduce by 90% the copper concentration in the final product and to accumulate the metal in great concentrations in the cells. As high concentrations of copper can be responsible for wine alterations, the selection of S. cerevisiae strains possessing high copper resistance and the ability to reduce the copper content of wine has a great technological interest, in particular for the fermentation of biological products. From the results obtained, the technique proposed is not only suitable for the assay of copper residues in must, wine and yeast cells, but it also offers the advantage of easy sample preparation and low detection limit in the ppb (g/l) range.     

Effects of continuous nitrogen application and nitrogen preconditioning on nodulation and growth ofCeanothus griseus varhorizontalis

Rooted cuttings ofCeanothus griseus varhorizontalis were irrigated with 0, 10, 20, 50, 75 or 100ppm nitrogen as NH₄NO₃ for eight weeks prior to inoculation with infectiveFrankia. After inoculation, half of the plants for each treatment nitrogen level continued to be irrigated with the preconditioning nitrogen level and half were given no more supplemental nitrogen. For plants continuously receiving nitrogen, nodule initiation (nodule number) was inversely correlated with increasing supplemental nitrogen levels, and suppressed above 50 ppm N. Leaf nitrogen above 2% in continuous-N plants correlated with greatly reduced or suppressed nodulation. Plants maintained after inoculation without supplemental nitrogen showed influence of the prior nitrogen treatment on nodulation. Preconditioning at 50 ppm and above greatly reduced the number of nodules formed. The evidence suggests that stored internal nitrogen can regulate nodulation.Plant biomass accumulated maximally when nodulation was suppressed, at 75 and 100 ppm supplemental N applied continuously. Internode elongation during the nodulation period occurred only on nodulated plants, or in the presence of supplemental N (10 ppm and above).     

Effect of different heavy-metal concentrations on Drosophila melanogaster larval growth and development

The survival percentage of Drosophila melanogaster larvae on synthetic media containing different concentrations of heavy metals, including Cd, Cu, Pb, and Zn, in the first generation indicated no significant reduction in their growth and development up to 500 ppm for all tested heavy-metal concentrations. At 500 ppm, results showed that there was a significant reduction in pupa and adult stages: 65% and 25% for Cd, 50% and 25% for Cu, 100% and 95% for Pb, and 85% and 75% for Zn, respectively. The survival percentages at 1000 ppm were further significantly reduced: 15% and 0% for Cd, 35% and 15% for Cu, 45% and 90% for Pb, and 65% and 35% for Zn, respectively for pupa and adult stages. For the second generation, there was no significant reduction in survival growth and development up to 100 ppm, but above 500 ppm, there was a significant reduction. For most of these heavy-metal concentrations, the survival percentages of the second generation at the pupa stage was higher than the first generation, whereas for the adult, there was a lower survival percentage, indicating some effect on metamorphosis of these heavy-metal concentrations on Drosophila melanogaster Comparing the survival percentage between first and second generations at 500 ppm for pupa indicate a significant increase on Cu only, and for the adult, there was a significant reduction for Cd.     

Toxicity and anticholinesterase activity of the fungal metabolites territrems to the corn earworm, Helicoverpa zea

The toxicity and anticholinesterase activity of tremorgenic fungal metabolites, territrems, to the corn earworm, Helioverpa zea (Boddie) (Lepidoptera, Noctuidae) were examined. In oral toxicity studies, territrem A significantly inhibited growth by 40% at 25 ppm and by 89% at 250 ppm. Territrem B and an epoxy-derivative significantly inhibited growth by ca. 45% at 250 ppm. Piperonyl butoxide administered orally synergized the toxicity of the territrems tested. In topical toxicity studies, the epoxy-derivative caused 26% mortality and 25% growth retardation at 10 mg/gm insects. Territrem A and B were not significantly lethal, but did reduce growth by 15–20% at 10 mg/gm insect. Paraoxon tested in the same way caused 100% mortality at 25 ppm orally and 10 mg/gm topically. However, all territrems tested in vitro as inhibitors of H. zea head acetylcholinesterase were at least comparable to or were more active than paraoxon. Topically administered epoxy-territrem B also inhibited H. zea head acetylcholinesterase. The potential for development of new insecticides from a territrem lead structure is discussed.     

Effect of delayed harvest on contamination of pearl millet grain with mycotoxin-producing fungi and mycotoxins

The response to delayed harvest of fungal and mycotoxin contamination of grain of the pearl millet hybrid HGM 100 was examined in 1992 and 1993. Samples of grain were assayed from seven plantings at locations near Tifton, Georgia, USA. Grain was harvested at 30, 40, and 50 days after anthesis and evaluated for infection byFusarium species andAspergillus flavus, and mycotoxin contamination. Mean isolation frequencies ofF. semitectum (35.6%) andF. chlamydosporum (17.2%) increased linearly with delayed harvest.Fusarium moniliforme andF. equisiti were infrequently isolated (<0.5%) and did not increase in the grain when harvest was delayed. Low mean concentrations of zearalenone (0.17 ppm), nivalenol (0.42 ppm), and deoxynivalenol (0.01 ppm) were detected but were not affected by delayed harvest. Isolation frequencies ofF. chlamydosporum andF. equiseti were correlated (P=0.07) with levels of nivalenol.Aspergillus flavus was not isolated from the grain, and aflatoxin concentrations averaged 1.9 ppb.     

Selenium-mediated differential response of β-glucosidase and β-galactosidase of germinatingTrigonella foenum-graecum

β-Glucosidase and β-galactosidase activity profile tested in different seeds during 24 h germination revealed reasonably high levels of activity inVigna radiata, Cicer arietinum, andTrigonella foenum-graecum. In all seeds tested, β-galactosidase activity was, in general, higher than that of β-glucosidase.T. foenum-graecum seedlings exhibited maximal total and specific activities for both the enzymes during 72 h germination. Se supplementation as Na₂SeO₃ up to 0.75 ppm was found to be beneficial to growth and revealed selective enhancement of β-galactosidase activity by 40% at 0.5 ppm Se. The activities of both the enzymes drastically decreased at 1.0 ppm level of Se supplementation. On the contrary, addition of Na₂SeO₃ in vitro up to 1 ppm to the enzyme extracts did not influence these activities. Hydrolytic rates of β-glucosidase in both control and Se-supplemented groups were enhanced by 20% with 0.05M glycerol in the medium and 30% at 0.1M glycerol. The rates were marginally higher in Se-supplemented seedlings than the controls, irrespective of added glycerol in the medium. In contrast, hydrolysis by β-galactosidase showed a trend of decrease in Se-supplemented seedlings compared to the control, when glycerol was present in the medium. Addition of Se in vitro in the assay medium showed no difference in the hydrolytic rate by β-galactosidase when compared to control, while the activity of β-glucosidase declined by 50%. Se-grown seedlings showed an enhancement of transglucosidation rate by 40% in the presence of 0.1M glycerol. The study reveals a differential response to Se among the β-galactosidase and β-glucosidase ofT. foenumgraecum with increase in the levels of β-galactosidase activity.     

Bacterial removal of chromium (VI) and (III) in a continuous system

The capacity of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans to reduce different concentrations of hexavalent chromium in shake flask cultures has been investigated. A. ferrooxidans reduces 100% of chromium (VI) at concentrations of 1, 2.5 and 5 ppm, but in the presence of 10 ppm only 42.9% of chromium (VI) was reduced after 11 days of incubation. A. thiooxidans showed a lower capacity to reduce this ion and total reduction of chromium (VI) was only obtained for concentrations of 1 and 2.5 ppm, whereas 64.7% and 30.5% was reached for 5 and 10 ppm, respectively, after 11 days. A continuous flow mode system was subsequently investigated, in which A. thiooxidans was immobilized on elemental sulphur and the acidic medium obtained was employed to solubilize chromium (III) and to reduce chromium (VI) present in a real electroplating waste [30% of chromium (III) and 0.1% of chromium (VI)]. The system enabled the reduction of 92.7% of hexavalent chromium and represents a promising way to treat this type of waste in the industry.     

Effect of brassicaceous plants on the survival and infectivity of insect pathogenic fungi

Studies were conducted to investigate potentialinteractions between brassicaceous plants, theisothiocyanates they produce and insectpathogenic fungi. Studies in vitro showedthat 100 ppm of 2-phenylethyl isothiocyanatecompletely inhibited growth of Metarhiziumanisopliae and Tolypocladiumcylindrosporum. T. cylindrosporum wassignificantly inhibited by 10 ppm 2-phenylethylisothiocyanate, but not by 1 ppm. M.anisopliae was not inhibited by 10 or 1 ppm2-phenylethyl isothiocyanate. The effect offreshly grated rutabaga on virulence of M.anisopliae to Galleria mellonella larvaewas also tested, resulting in reduced infectionby M. anisopliae. To study the effect ofplants in the Brassicaceae under morefield-like conditions fungi were added to soilwithout plants or at the bases of two differentplant species, Barbarea vulgaris and Eruca vesicaria sativa. Soil was sampled at 5,10 and 15 days to determine the concentrationof colony forming units (CFUs). Treatments withplants did not have fewer CFUs than thecontrol. Instead significantly fewer CFUsresulted when M. anisopliae was added topots with soil only than to pots with plants.There were no significant differences betweenthe two plant species in the number of CFUs atday 5. For days 10 and 15, however,significantly more M. anisopliae CFUswere found in pots containing E. vesicariasativa than B. vulgaris. T.cylindrosporum was less affected by thedifferent treatments than M. anisopliae.Our studies demonstrated that whileisothiocyanates can inhibit insect pathogenicfungi in Petri dishes, when using a morerealistic fungus/plant/soil microcosm no fungalinhibition was found.     

Influence of long-term hypoxia on the energy metabolism of the haemoglobin-containing bivalve Scapharca inaequivalvis: critical O2 levels for metabolic depression

Summary The oxygen consumption rate of Scapharca inaequivalvis measured under normoxic conditions over 48 h showed a significant daily cycle with lowest values occurring shortly after the dark period; all hypoxia exposure experiments were carried out during the declining part of the cycle. Animals were exposed to a constant level of hypoxia for a 12-h period in a series of 14 experiments, each at a different oxygen tension. The oxygen consumption was measured continuously, and the extent of accumulation of end-products (succinate and propionate), and the inhibitory effect of adenosine triphosphate on phosphofructokinase were determined at the end of exposures. All three parameters (oxygen consumption, end-product accumulation, phosphofructokinase inhibition) showed a remarkable correlation with major changes occurring between 2.5 and 1.5 ppm (7 and 4 kPa) O₂. The oxygen consumption rates showed a drop to 6% of the normoxic rate, but a consistent low consumption remained below 2 ppm (5.5 kPa) which partly recovered over the 12-h exposure period by about three-fold. Succinate and propionate accumulated progressively between 2.5 and 1.5 ppm (7 and 4 kPa); at [O₂]<1.5 ppm (4kPa) the concentration did not increase further, indicating that anaerobic metabolism had reached a maximum. Over the same range, phosphofructokinase showed an increased sensitivity for adenosine triphosphate, the lower inhibitor concentration at 50% V _max value pointing to depression of glycolytic rate. Despite the activation of anaerobic metabolism and the evident depression of aerobic metabolism, simple calculation demonstrates that Scapharca inaequivalvis relies mainly on aerobic metabolism even during severe hypoxia. It is assumed that the occurrence of haemoglobin in this species is essential for its capacity to survive long periods of hypoxia.Abbreviations ATP adenosine triphosphate - I₅₀ inhibitor concentration at 50% V _max - PFK phosphofructokinase - P _c critical PO₂ - SEM standard error of mean - VO₂ oxygen consumption rate - ww wet weight     

Suppression of spore germination and aflatoxin biosynthesis in Aspergillus parasiticus during and after exposure to high levels of phosphine

Agar cultures of toxigenic Aspergillus parasiticus NRRL 2999 were exposed to phosphine (PH3), in levels ranging from 0 to 2000 ppm (vol/vol). It was found that with PH3 concentrations of 400 ppm or higher the growth of the fungus was totally arrested. When PH3 was vented and the agar plates were exposed to open air, 100% of the initial CFU developed into fully grown colonies after PH3 levels below 300 ppm, but at higher PH3 concentrations only 50% of the colonies developed. The same strain of A. parasiticus was inoculated into high moisture corn under conditions highly favorable for aflatoxin production, and it was exposed to a range of PH3 levels. After exposure to 500 ppm PH3, both fungal growth and aflatoxin synthesis resumed shortly after elimination of the toxic gas, but after exposure to PH3 levels of 1000 ppm and higher, the physical appearance of the contaminated corn was remarkably changed, showing reduced mycelial growth and almost complete absence of green pigmentation. In addition, aflatoxin synthesis was totally absent for the remainder of the experiment (20 days). These results strongly suggest that exposure to PH3 levels of 1000 ppm or higher could bring about persistent metabolic changes in surviving Aspergillus organisms. This revised version was published online in June 2006 with corrections to the Cover Date.     

Selection of entomopathogenic fungi for use in combination with sub-lethal doses of imidacloprid: perspectives for the control of the leaf-cutting ant <Emphasis Type="Italic">Atta sexdens</Emphasis><Emphasis Type="Italic">rubropilosa</Emphasis> Forel (Hymenoptera: Formicidae)

In the first part of this study, four isolates of the fungus Beauveria bassiana (Bals.) Vuillemin (LPP1, LPP2, CG05 and CG24) and one isolate of Metarhizium anisopliae (Metsch.) Sorokin (CG46) were tested against adult foragers of Atta sexdens rubropilosa. Ants were allowed to walk on filter paper discs, inside Petri dishes, previously impregnated with 1 ml of a conidia suspension (2 × 10⁷ conidia ml⁻¹), maintained at 80% RH and 26°C for 24 h and subsequently, transferred to sterile Petri dishes, maintained at 23°C, 80% RH, 24 h dark. The mean values of LT₅₀ for LPP2, LPP1, CG46, CG24 and CG05 were 3.5, 3.7, 3.8, 4.2 and 4.4 days, respectively. Control insects for all tests in this study showed less than 10% mortality. Experiments were carried out to test the toxicity of imidacloprid (IMI) to A. sexdens rubropilosa. Mortality was evaluated 10 days following a 24 h exposure to the insecticide. Percent mortality caused by 500, 200, 100 and 10 ppm IMI was 77.8, 56.7, 45.5 and 5.5 respectively. Insects treated with 10 ppm IMI were observed to have reduced locomotor activity 24 h after exposure to the insecticide. The LC₅₀ of IMI was 154.3 ppm. Subsequent tests were carried out to evaluate the combination of a sub-lethal dose of IMI (10 ppm) and infection by CG24 (1 × 10⁷ conidia ml⁻¹). Mortality due to fungal infection alone was 43.3%. Mortality of insects treated with IMI followed by exposure to the fungus was 64.3%. These results indicate that IMI significantly increases the susceptibility of ants to infection by B. bassiana CG24.     

Studies on sulphur in vertisols

Summary Some soil and plant test methods were evaluated for predicting response of soybean crop (Glycine max (L.) Merr.) to S application in vertisols. Morgan's reagent, 500 ppm P containing Ca(H₂PO₄)₂.H₂O and KH₂PO₄ solutions, 0.5N NH₄OAc+0.25N HOAc and 0.15% CaCl₂ were found to be suitable extractants for measuring available soil S. The critical limits of extractable S were 9.0 ppm by Morgan's reagent, 10.0 ppm by phosphate solutions, 8.0 ppm by 0.5N NH₄OAc +0.25N HOAc and 14.0 ppm by 0.15% CaCl₂. Morgan's reagent was regarded as superior to other soil test methods in view of its high relationship with S uptake by plants, A values and relative yield. Critical S concentration in soybean plants varied with age. It was 0.15% and 0.185% for 36 and 60 days old plants, respectively. The critical N/S ratio on the other hand appeared to be constant at about 16.5 during vegetative growth period. Constancy of critical N/S ratio in plants was attributed to the near constancy of N/S ratio in plant proteins. There was highly significant relationship between response of soybean to S and to N, supporting the conclusion of some earlier workers that any soil showing large responses to N may not be supplying adequate S from the mineralization of soil organic matter.     

Detection of high-phosphorus tolerant VAM-fungi colonizing hops and peppermint

Summary In a continuing search for field sources of endomycorrhizal fungi that tolerate high levels of available phosphorus in soil, samples were obtained from long-established liberally fertilized hops (Humulus lupulus L.) and peppermint (Mentha piperita L.) fields. In hops fields, where extractable P (Bray) levels ranged from 21–196 ppm, spores of twoAcaulospora spp. and sixGlomus spp. were isolated. From peppermint fields with P levels of 44 to 244 ppm, spores of oneAcaulospora sp., fourGlomus spp. and oneGigaspora sp. were obtained. Vesicular-arbuscular mycorrhizal (VAM) fungi were found colonizing most roots to some extent, despite the very high fertility of several sites. Although percentages of colonization in hops tended to be low, ranging from 0 to 9.3%, colonizations in peppermint roots were considerably higher averaging 26.5% between the sampled fields, notwithstanding P levels as high as 244 ppm. Curiously, considerable external VAM hyphae were found adherent to or entangling roots in many cases, even where percentage of root length with internal VAM structures was low. The functional benefit, neutrality, or detriment to plants of such high-P tolerant species remains to be determined.Contribution from the USDA-ARS, Horticultural Crops Research Laboratory and the Dept. of Soil Science, Oregon State University, in cooperation with the Oregon State University Agricultural Experiment Station, Corvallis, OR 97331. Technical paper no. 6979 of the latter     

Chemical composition and larvicidal effects of essential oil of Dendropanax morbifera against Aedes aegypti L.

Essential oils obtained from the flowers of Dendropanax morbifera were extracted and the chemical composition and larvicidal effects were studied. The analyses were conducted by gas chromatography and mass spectroscopy (GC–MS) revealed that the essential oil of D. morbifera contained 27 compounds. The major chemical components identified were γ-elemene (18.59%), tetramethyltricyclohydrocarbon (10.82%), β-selinene (10.41%), α-zingibirene (10.52%), 2-isopropyl-5-methylbicylodecen (4.2%), β-cubebene (4.19), and 2,6-bis(1,1-Dimethylethyl)-4-phenol (4.01%). The essential oil had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L. with an LC₅₀ value of 62.32 ppm and an LC₉₀ value of 131.21 ppm. The results could be useful in search for newer, safer, and more effective natural larvicidal agents against A. aegypti.     

Effect of phosphinothricin (glufosinate) on photosynthesis and photorespiration of C3 and C4 plants

Phosphinothricin (glufosinate), an irreversible inhibitor of glutamine synthetase, causes an inhibition of photosynthesis in C₃ (Sinapis alba) and C₄ (Zea mays) plants under atmospheric conditions (400 ppm CO₂, 21% O₂). This photosynthesis inhibition is proceeding slower in C₄ leaves. Under non-photorespiratory conditions (1000 ppm CO₂, 2% O₂) there is no inhibition of photosynthesis. The inhibition of glutamine synthetase by phosphinothricin results in an accumulation of NH₄ ⁺. The NH₄ ⁺-accumulation is lower in C₄ plants than in C₃ plants. The inhibition of glutamine synthetase through phosphinothricin in mustard leaves results in a decrease in glutamine, glutamate, aspartate, asparagine, serine, and glycine. In contrast to this, a considerable increase in leucine and valine following phosphinothricin treatment is measured. With the addition of either glutamine, glutamate, aspartate, glycine or serine, photosynthesis inhibition by phosphinothricin can be reduced, although the NH₄ ⁺-accumulation is greatly increased. This indicates that NH₄ ⁺-accumulation cannot be the primary cause for photosynthesis inhibition by phosphinothricin. The investigations demonstrate the inhibition of transmination of glyoxylate to glycine in photorespiration through the total lack of amino donors. This could result in a glyoxylate accumulation inhibiting ribulose-1,5-bisphosphate-carboxylase and consequently CO₂-fixation.Abbreviations GOGAT glutamine-2-oxoglutarate-amidotransferase - GS glutamine synthetase - PPT phosphinothricin - MSO methionine sulfoximine - RuBP ribulose-1,5-bisphosphate     

Effects of soil phosphorus and Glomus intraradices on growth,nonstructural carbohydrates,and photosynthetic activity of Citrus aurantium

Sour orange (Citrus aurantium L.) grown in low-P (9–12 ppm) and high-P (420 ppm) soil inoculated with or without Glomus intraradices (G.i.), were evaluated for biomass, carbohydrates, ribulose bisphosphate carboxylase (RuBPCase), phosphoenolpyruvate carboxylase (PEPCase) activity, leaf ¹⁴CO₂ incorporation, and other physiological parameters. Growth of plants in the low-P, noninoculated soil was lowest, with total dry biomass reduced up to half of the low-P, inoculum treatment. Total nonstructural carbohydrates were 40% lower in leaves of plants in the low-P, noninoculated soil, compared with the other treatments. Inoculation of the low-P soil enhanced leaf ¹⁴CO₂ incorporation by 67%, total chlorophyll content by 28%, and RuBPCase activity by 42%, compared with low-P, noninoculated treatment. Improved P-use efficiency by G.i. in low-P soil was comparable to high-P nutrition in improving leaf ¹⁴CO₂ incorporation and concentration of major leaf photosynthetic products that include starch and sucrose. Leaf PEPCase activity in the low-P, noninoculated treatment, however, was at least threefold higher than the other treatments, suggesting a possible alteration in organic acid metabolism in sour orange leaves as a result of P deficiency.     

Nitrogen and carbon dynamics in C3 and C4 estuarine marsh plants grown under elevated CO2 in situ

Summary Carbon dioxide concentrations were elevated in three estuarine communities for an entire growing season. Open top chambers were used to raise CO₂ concentrations ca. 336 ppm above ambient in monospecific communities of Scirpus olneyi (C₃) and Spartina patens (C₄), and a mixed community of S. olneyi, S. patens and Distichlis spicata (C₄). Nitrogen and carbon concentration (% wt) of aboveground tissue was followed throughout growth and senescence. Green shoot %N was reduced and %C was unchanged under elevated CO₂ in S. olneyi. This resulted in a 20%–40% increase in tissue C/N ratio. There was no effect of CO₂ on either C₄ species. Maximum aboveground N (g/m²) was unchanged in S. olneyi, indicating that increased productivity under elevated CO₂ was dependent on reallocation of stored N. There was no change in the N recovery efficiency of S. olneyi in pure stand and a decrease in the mixed community. Litter C/N ratio was not affected by elevated CO₂ suggesting that decomposition and N mineralization rates will also remain unchanged. Continued growth responses to elevated CO₂ could, however, be limited by the ability of S. olneyi to increase the total aboveground N pool.     

Insect growth regulating effects of neem extract and azadirachtin on aphids

Neem,Azadirachta indica (A. Juss.), seed oil (NSO) applied to leaf discs at a concentration of 1.0% resulted in 94% to 100% mortality of second instar nymphs of currant-lettuce aphid,Nasonovia ribis-nigri (Mosley), and green peach aphid,Myzus persicae (Sulzer), after nine days. The equivalent amount of pure azadirachtin (AZA) (≈40 ppm), the principle active ingredient of neem, was as effective as NSO. The survival of adult aphids was unaffected by NSO or AZA, but the survival of offspring from treated adultM. persicae andN. ribis-nigri was reduced significantly. The lethal concentration of AZA resulting in 50% mortality of second instar nymphs of nine species of aphids ranged from 2.4 ppm forM. persicae on pepper to 635.0 ppm for the strawberry aphid,Chaetosiphon fragaefolii (Cockerell), on strawberry. ForM. persicae, the growth regulating effect of AZA was influenced by the host plant and the nymphal instar treated.     

Toxicity of cadmium to twoStreptomyces species as affected by clay minerals

Summary The effect of cadmium on the growth ofStreptomyces rimosus andS. bottropensis (both isolated from soil) was investigated. The modifying effect of the presence of the clay minerals kaolinite, bentonite and vermiculte on Cd toxicity was also included. After four days no growth was observed at 100 ppm CdCl₂ ofS. bottropensis and at 150 ppm in case ofS. rimosus. After six days some growth ofS. rimosus occurred at 150 ppm CdCl₂ and ofS. bottropensis at 100 ppm. Addition of the three clay minerals decreased the Cd toxicity considerably.     

INDUCTION OF WOUND-VESSEL DIFFERENTIATION IN ISOLATED COLEUS STEM SEGMENTS IN VITRO

Excised internodes and 2-mm-thick transverse stem segments of Coleus blumei were incubated 7 days on media containing 2% sucrose, 1% agar, and various growth substances. Wound-vessel members differentiated in the 2-mm-thick tissue slices incubated on medium containing no exogenous auxin (control). Compared to control slices, the addition to the medium of either IAA (50 or 5 ppm), 2, 4-D (10, 1, or 0.1 ppm), TIBA (50, 5, or 0.5 ppm), or kinetin (50, 5, 0.5 or 0.05 ppm) inhibited wound-vessel differentiation. Simultaneous treatment of tissue slices with IAA and kinetin inhibited wound-vessel differentiation, as did the incubation of tissue slices on medium containing no sucrose. Low concentrations of IAA (0.05 ppm) or 2, 4-D (0.01 ppm) resulted in over a 100% increase in the numbers of wound-vessel members differentiated. These results are interpreted as indicating auxin synthesis by the tissue slices and the participation of auxin as a limiting factor in xylogenesis. The inhibition of wound-vessel differentiation by relatively high concentrations of 2,4-D, TIBA, or kinetin is interpreted as a reflection of the inhibition of polar auxin transport by these substances, and an indication that polar auxin transport enhances xylogenesis.