Rooting Substances in Water Extracts of Castanea sativa and Salix viminalis

The rooting activity of water extracts of cuttings of chestnut (Castanea sativa Mill.) and willow (Salix viminalis L.) was studied throughout the year by the bean rooting test. The chestnut extract nullified the rooting activity of 10^?5M IAA, but on the whole it did not modify the rooting pattern of the bean cuttings. The willow extract increased rooting and showed a rhythm in the content of endogenous growth substances. IAA was found at the time when the extract showed the strongest rooting activity. The extract from willow contained two synergistic root-promoting fractions with the R_f's of 0.0–0.2 and 0.6–0.8 in isopropanol: ammonia: water. The latter one promoted rooting without added IAA. In this fraction a high amount of catechol was detected. No clear correlation was found between the ability to induce roots and the total phenol content in the two species.     

Effect of Reciprocal Bud-Graft Transfers Between Old Home and Bartlett Pears and Centrifugatiou on Trans-location of Endogenous Growth Substances in Hardwood Cuttings

An investigation was made of the effect of bud-graft exchanges between two pear varieties—the easily-rooted Old Home and the hard-to-root Bartlett—on the root-promoting activity of extracts obtained from basal segments of hardwood cuttings of each variety. The effect of a centrifugal force obtained from 2000 rpm for 30 minutes on the root-promoting activity of diffusates and extracts from bases of centrifuged cuttings of these two varieties was also studied. Bud exchange treatments showed no appreciable influence on the root-promoting or inhibiting activity of basal extracts unless the transbudded shoots (used for making cuttings) were girdled one month prior to the sampling date; in such cases an influence did appear. The centrifugation treatment increased the number of buds sprouting on cuttings. It also increased the root-promoting activity of extracts from the basal parts of cuttings and of their growth-active components (separated by paper chromalography) when bioassayed by the mung bean rooting test. Apparently some inhibitory materials from the buds were removed through the bases of the cuttings by centrifuging.     

The Nature of Root-Promoting Substances in Lycopersicon esculentum Seedlings

Determination of the organ sites and chemical nature of root-promoting substances in Lycopersicon esculentum seedlings was made. Results of organ excision indicated that the young cotyledons were the primary site of root-promoting substances. As the cotyledon matured, its root-promoting activity decreased while that of the developing plumular leaves increased. Application of 2,3,5-triiodobenzoic acid (TIBA) at 10^?3 M in lanolin to the hypocotyl below the cotyledonary node significantly prevented root formation. Bioassay, using a new tomato rooting test, of the basic, acidic and bound fractions of the cotyledon extracts after paper chromatography showed biological activities in all three fractions. Gas-liquid chromatography of selected active regions of the paper chromatogram resulted in a partial tentative identification of indolepropionic acid, 5-methylindole, 5-hydroxy-indole and 3-indoleacetonitrile. The combined evidence indicated that the root-promoting substances in tomato seedlings resemble auxin compounds.     

THE EFFECT OF CHARCOAL ON THE GROWTH OF LEGUMINOUS PLANTS IN SAND CULTURE

The addition of small proportions (0.5-2.0 %) of activated charcoal to the rooting medium of inoculated peas in nitrogen-free sand culture resulted in marked increases in dry weight of the plants and in nitrogen fixation. Wood charcoal in larger proportions had a similar effect, while animal charcoal severely inhibited growth. The number of nodules was greatly reduced in the presence of activated charcoal, but such nodules as formed were much larger and the nodule tissues per unit weight were more active in nitrogen fixation. Activated charcoal also led to an increase in dry weight of non-inoculated peas supplied with inorganic combined nitrogen. It is tentatively suggested that these favourable effects arise from the adsorption by the charcoal of harmful excretions from roots or micro-organisms or of excess nutrients, and from the maintenance of a more favourable pH in the rooting medium. The examination of barley intersown with the peas, and the results of Kjeldahl analyses on the rooting media, provided no evidence that the enhanced fixation in the presence of activated charcoal was attended by any considerable excretion of fixed nitrogen.     

Initiation and development of wound tissue and roots on hypocotyl cuttings of Pinus sylvestris in vitro

The rooting of hypocotyl cuttings from 20-day-old seedlings of Pinus sylvestris L. cultured in vitro is discussed. About 40% of the cuttings cultured on medium lacking activated charcoal produced roots during the first two months. When activated charcoal was added to the medium, either root formation (75% formed roots) or wound tissue growth (95% formed large wound tissues) was stimulated in different experiments. These large wound tissues did not develop any roots. The anatomical changes in the basal part of the cuttings were similar during the first two weeks in all the cuttings studied. A vascular cylinder composed of short tracheids with many pores developed. Thereafter the differentiation process became varied. The amount of wound tissue produced and the time for rooting differed among the cuttings. Tracheid nests which were in contact with the vascular system in the hypocotyl via short tracheids were observed after three weeks. Subsequently, roots developed from the tracheid nests. The longer root formation was delayed, the larger the wound tissue became.
Short tracheids were found close to the wound tissue surface. Their ability to adsorb nutrients and water is discussed.     

Root-promoting Substances in Salix alba

Root-promoting substances were extracted from softwood cuttings of Salix alba L. by centrifuging them with water or by shaking the ground freeze-dried stems with water. Rooting substances were partitioned by paper chromatography or chemical fractionation and their rooting activity was tested by mung bean cuttings. Both extracts indicated three major root -promoting fractions at Rf 0-0.1, 0.7-0.8, and 0.3-0.4 in a decreasing order of their activities when paper chromatographed with isopropanol:ammonia:water 8:1:1 v/v. The strongest one indicated an apparent synergistic rooting effect with indol-3yl-acetic acid (IAA) regardless of the extraction method. These results indicate that water can extract from freeze -dried sample the similar rooting substances found in the centrifugal diffusates. The Rf 0–0.1 fraction consisted of at least four fractions and the strongest one did not move from the starting line on the chromatogram when isopropanol:ammonia:water 8:1:1 was used. This starting line fraction was extremely strong in rooting activity and its highest concentration resulted in 8.7 times as many roots as controls. More thain additive rooting effect between IAA and the fraction was found only at the highest concentration. The fraction was very soluble in water but insoluble in chloroform or ethyl ether and only stimulated rooting of mung bean cuttings when it was applied within 3 days after cuttings were made. It had no effect in lengthening roots. The starting line fraction was further found to have four root-promoting subfractions at Rf 0.05, 0.35, 0.65, and 0.85 when it was chromatographed in 60 % isopropanol. Among these four, the subfractions at Rf 0.65 and 0.35 were strongly root promotive and displayed more than additive root promotion with IAA at the highest concentrations studied.     

Isolation, Purification, and Characterization of an Endogenous Root-promoting Factor Obtained from Basal Sections of Pear Hardwood Cuttings

Basal segments taken from Old Home and Bartlett pear hardwood cuttings collected at intervals during the rooting period in September were extracted with ethanol and fractionated by paper chromatography in different solvent systems. Different zones on the chromatograms were bioassayed by the mung bean rooting test, which showed high levels of promotion in Old Home basal extracts when the cuttings were obtained during the period of maximum rooting. Extracts from Bartlett cuttings, however, showed considerably less promotion activity in the bioassay but did show high levels of inhibitory activity.

After the easily-rooted Old Home cuttings had been in the rooting medium for 10 days, a highly active endogenous root-promoting material was found in extracts from basal segments of cuttings having buds and which had been treated with indolebutyric acid. Similar extracts obtained from disbudded cuttings, or from cuttings with buds but not treated with indolebutyric acid, lacked this rooting-factor. Extracts obtained from all types of the difficult-to-root Bartlett cuttings also lacked this rooting-factor. The latter is believed to be produced by physiologically active Old Home buds, and is very effective in the mung bean bioassay, even at extremely low concentrations.

From paper chromatographic studies, tests with spray reagents, solubility determinations, biological tests, UV spectrum analysis, and infrared spectroscopy, it is believed that this rooting factor could be a condensation product between exogenous auxin (indolebutyric acid) and a phenolic compound produced by physiologically active Old Home pear buds.

     

Aerobic Heterotrophic Bacterial Populations of Sewage and Activated Sludge: I. Enumeration1

Agar plating media containing solely activated sludge extracts yielded, in general, higher viable counts of activated sludge bacteria than any other culture medium tested. Activated sludge extracts made from different treatment plants varied in efficacy in evoking maximal viable counts. Frequently, homologous plating, i.e., plating inocula of activated sludges on extracts made from the same activated sludges, tended to yield lower counts than the heterologous platings tried in this investigation. The counts obtained by homologous plating of activated sludge were not significantly lower and sometimes were even significantly higher than the counts obtained on standard Nutrient Agar, which had been found by previous workers to be a good medium for counting activated sludge bacteria. The higher counts obtained with activated sludge extracts set objectives for formulating reproducible or defined culture media for the enumeration of activated sludge bacteria.     

Effect of Sucrose Concentration, Charcoal, and Indole-3-Butyric Acid on Germination of Abies Numidica Somatic Embryos

The dependency of radicle elongation in Abies numidica somatic embryos on germination media has been studied. No significant differences were detected between the Murashige and Skoog (MS) and Schenk and Hildebrandt (SH) medium. The addition of 10 g dm^–3 activated charcoal or 0.05 mg dm^–3 indole-3-butyric acid (IBA) into both media had positive influence on embryo germination. Difference between activated charcoal and IBA effects were significant. The high rooting percentage (85 %) was recorded on half SH medium with 10 g dm^–3 sucrose and activated charcoal. After IBA addition rooting percentage was increased to 95 %. During 7 months 73 % of plantlets survived transfer to soil and in 54 % of plantlets shoot growth was observed.     

In vitro regeneration of Semecarpus anacardium L. from axenic seedling-derived nodal explants

Semecarpus anacardium (Anacardiaceae), a deciduous forest tree, is a potent source of medicinal compounds. Poor seed viability of this species limits the conventional propagation practice. Proliferation of shoots from axillary meristem was achieved in semisolid WPM medium supplemented with BAP 4.44 μM and KN 4.64 μM. Factors including culture vessels, gelling agents and antioxidants were identified and optimized for proliferation and growth of shoots in vitro. Cotton-plugged culture vessels were more favorable. Phytagel 0.2% as gelling agent and activated charcoal 0.2% as antioxidant were superior to other agents and antioxidants tested. All the shoots rooted in half-strength WPM liquid medium with IBA 2.46 μM. Rooted shoots survived (91%) in the soil–sand 1:1 mixture. Ex vitro rooting of shoots and hardening of plants were achieved in 80% of the explants in the soil–sand mixture. Hardened plants were maintained in a greenhouse. This is the first report on in vitro regeneration of Semecarpus anacardium.     

Charcoal and activated carbon as adsorbate of phytotoxic compounds – a comparative study

This study compares the potential of natural charcoal from Scots pine ( Pinus sylvestris L.) and activated carbon to improve germination under the hypothesis that natural charcoal adsorbs phytotoxins produced by dwarf-shrubs, but due to it's chemical properties to a lesser extent than activated carbon. Activated carbon has been used in many bioassays as an adsorbate to clean aqueous solutions.
We used aqueous extracts from young leaves of Calluna vulgaris (L.) Hull and Vaccinium myrtillus (L.) as phytotoxin sources in two different concentrations (10 and 14 gr. of dried leaves in 100 ml distilled water). Germination of pine seeds was prevented by the higher concentration of both species, while the lower ones did not show significantly reduced germination. Both ericaceous species showed a very similar potential to prevent germination of Scots pine seeds.
Supplemented carbon (activated carbon, powdered or granulated pine charcoal) restored germination in strong extracts. Adding activated carbon resulted in germination of almost 100%. With pine charcoals added, lower germination percentages were observed. The charcoal powder was more effective (60% for C. vulgaris ; 28% for V. myrtillus ) than the charcoal granulate (30% and 16%, respectively) in restoring germination.
Chemical and surface analysis of the three carbon supplements revealed that activated carbon had by far the biggest active surface area (641 m² g⁻¹), and thus many more cavities to bind phytotoxins than natural charcoal (total surface area of 142 m² g⁻¹).
We conclude, that charcoal produced by forest fires can have a positive effect on seed germination, but to a much lesser extent than activated carbon. Previous studies, which used activated carbon as an equivalent for charcoal, overestimated the effect of charcoal on germination.     

In vitro rooting of micropropagated shoots from juvenile and mature Pinus pinaster explants: influence of activated charcoal

The ability for adventitious rooting of micropropagated shoots from juvenile and mature Pinus pinaster Ait. explants was assessed in vitro on a rooting expression medium. The different rooting traits observed, namely the rooting rate, the number and the length of the adventitious roots, and the root score, were greatly influenced by the age of the donor plant: 98% of juvenile explants rooted, while only 49% of mature explants did. Addition of activated charcoal in the rooting expression medium improved the overall rooting capacity of the mature explants to an average of 78%. Whatever the plant material, the number and the length of the adventitious roots, as well as the root score, fluctuated according to the sampling date.Abbreviations BA 6-benzyladenine - NAA naphthaleneacetic acid - REM rooting expression medium - RIM rooting induction medium     

In vitro propagation of Vicia faba L. by micro-cutting and multiple shoot induction

The influences of nitrogen sources, culture temperature and activated charcoal supplements were studied in relation to the rooting ability of V. faba cuttings. The interaction of these factors led to quantitative and qualitative modifications of the culture responses. Low temperatures (14–18°C) were suitable for in vitro culture, limiting the formation of phenolics in plant material and making activated charcoal supplement unnecessary. Nitrogen supplements contributed in modifying the different plant responses, in accordance with temperature. Multiple shoot formation was obtained from the cotyledonary node and from the stem nodes cultivated in the presence of 6-benzylaminopurine (BAP). BAP at 4 mg l^-1 was the most effective concentration in promoting high rates of shoot development. The original position of stem nodes was found to determine the explant response to plant growth regulator treatments, possibly due to the effect of residual apical dominance.     

Partial purification of a legume nodulation factor present in coconut water

The nodulation of adventitious roots growing from segments of bean hypocotyl tissue was used as a bioassay for the material present in coconut water which stimulated nodulation. The active material in coconut water is acidic, but it was not possible to extract it from an acid solution with organic solvents. A purification of approximately 70-fold (on a dry wt basis) was obtained using activated charcoal, but at least 10 different compounds were present in the active fractions. A purified fraction of coconut water, which is stimulatory to the growth of carrot root explants, was active in the nodulation assay at a concentration of 2 μg/ml. This represents a 4000-fold purification of the diffusible fraction of coconut water. The charcoal fractionation procedure can be applied to the active material present in extracts of bean leaves.     

Effect of particle size of activated charcoal on separation of Triton X-100 from protein, liver cytosol, and lipoxygenase extracts

Triton X-100 was almost completely removed from bovine serum albumin solutions, BALB/c mouse liver extracts, and avocado peel lipoxygenase extracts by stirring the samples for 30 min in the presence of 250-350 mesh activated charcoal. The procedure did not remove protein significantly and did not reduce enzyme activity. At higher charcoal particle sizes, the efficiency of Triton adsorption was decreased and protein adsorption was increased. ElevateD temperatures enhanced Triton and protein adsorption. Adsorption on activated charcoal of 250-350 mesh is a simple and rapid procedure for Triton removal at a ratio of 0.23 g Triton X-100 per gram of activated charcoal.     

金线莲组织培养(简报)

以金线莲茎段为外植体进行离体快速繁殖体系研究。结果表明：芽诱导最佳培养基为1/2MS+6-BA 1.5 mg·L^-1+NAA 0.2 mg·L^-1;增殖培养基为1/2MS +6-BA 2.0 mg·L^-1+NAA 0.5 mg·L^-1+香蕉100 g·L^-1;增殖系数达6～8倍;生根培养基为1/2MS + IBA 1.0 mg·L^-1+NAA 0.5 mg·L^-1+活性炭0.2 g·L^-1,生根率可达95%。将生根苗移栽入滤水性好的细兰石与炭渣混合基质中,盖膜保湿。成活率可达90%。     

Factors affecting induction and development of in vitro rooting in apple rootstocks

Shoots of apple rootstocks raised in vitro were transferred to various rooting media to study the effect of different factors on root initiation and development. Various concentrations of indole-3-butyric acid (IBA) initiated rooting but maximum rooting percentage was found with 2.0 and 2.5 mg l(-1) of IBA in M7 and with 1.0 mg l(-1) of IBA in MM106. The drawback was that the roots were thick, short and with profuse callus. The presence of activated charcoal (AC) in the rooting medium improved the rooting quality but reduced the rooting percentage in both the rootstocks. In high auxin dip of 70, 80 and 90 mg l(-1) IBA for 2, 2 and 1 hr showed 75-85 per cent rooting in M7, but lacked reproducibility of the results. Whereas in MM106, 66 - 70 % rooting was achieved with 70 mg l(-1) of IBA dip for 3 h. Root induction in shoots in IBA containing liquid medium (LM) in dark for few days and root elongation in IBA--free medium in light proved most effective. On the other hand, continuous light treatment showed reduced rooting. Reduction of MS salts and sucrose in root elongation medium showed decreased rooting. Plantlets from two--stage rooting procedure showed more rapid growth and satisfactory survival during hardening of plants and on transfer to field.     

Microbial utilization of levoglucosan in wood pyrolysate as a carbon and energy source

The Waterloo Fast Pyrolysis Process (WFPP) can produce an organic liquid high in levoglucosan (1, 6-anhydro-beta-D-glucopyranose) content from suitably pretreated lignocellulosics. A variety of fungi and yeasts were screened for their ability to utilize and ferment this organic liquid. To enhance its fermentability, the pyrolysis tar was posttreated in three different ways: (1) an aqueous extract (lignin removed); (2) activated charcoal treated (lignin and aromatics removed); and (3) acid hydrolysate (lignin and aromatics removed with the levoglucosan hydrolyzed to glucose). Four fungal strains were examined. None grew in the aqueous extract, but all grew equally well in both the activated charcoal treated and the acid hydrolysate, suggesting that the aromatic species were inhibitory to growth. Seven yeast species were examined, two of which did not grow on any of the extracts. Five of the yeast strains grew well on both the aqueous extract as well as the activated charcoal extract. The hydrolysate was optimal in terms of biomass yield and ethanol production. Ethanol yields on the hydrolysate were comparable or better than those on glucose. Ethanol was also produced in the aqueous extract and activated charcoal-treated substrate, but yields were considerably lower than on the hydrolysate or glucose. It is apparent that a wood pyrolysate maximized for levoglucosan can serve as a fermentable substrate, although postpyrolysis clean-up appears necessary. (c) 1993 John Wiley & Sons, Inc.     

N-phenyl indolyl-3-biityraitiide and phenyl indole-3-thiolobutyrate enhance adventitious root primordium development

N-phenyl indoryl-3-butyramide (NP-IBA) and phenyl indole-3-thiolobutyrate (P-ITB) are new synthetic auxins that enhance rooting of bean ( Phaseolus vulgaris cv. Top Crop) hypocotyl cuttings and jack pine ( Pinus banksiana Lamb.) seedling cuttings. In comparison with an equal concentration of indole-3-butyric acid (IBA), NP-IBA treatment resulted in greater rooting of bean and jack pine seedling cuttings. IBA and P-ITB treatment at equal concentration were equally effective in inducing rooting of bean cuttings. However, in comparison with an equal concentration of IBA, P-ITB treatment promoted greater rooting of jack pine seedling cuttings, but P-ITB treatment was less effective than NP-IBA treatment. Application of crystalline chemical to jack pine seedling cuttings indicated lesser toxic effects and earlier and greater root-promoting effects of P-ITB and NP-IBA treatments, in comparison with IBA treatment. The earlier rooting of NP-IBA-treated jack pine seedling cuttings was positively related to increase in fresh weight of basal 1 cm stem before primordia were initiated. Weight of basal steins of NP-IBA-treated cuttings initially increased during propagation at day 2 in comparison with the control, and at day 3 in comparison with IBA-treated cuttings. Results that were obtained with seedling jack pine cuttings were generally confirmed for cuttings from 7-year-old trees.     

Improvements in rooting regenerated safflower (Carthamus tinctorius L.) shoots

A continuing obstacle for regenerating safflower (Carthamus tinctorius L.) plants from cultured explants or callus has been a reliable method for rooting shoots. For shoots directly regenerated from primary explants, 76% of shoots rooted after a 7-d exposure to 10 mg/1 indole-3-butyric acid. Auxin source, concentration or exposure time did not greatly affect root formation or morphology, but strongly affected callus production. Shoots infected with Agrobacterium rhizogenes produced massive numbers of fibrous roots, but shoots did not elongate or survive transfer to soil. Shoot hyperhydricity symptoms were reduced by including 1 g/1 activated charcoal in rooting media. The optimal protocol for inducing root formation consisted of a 7-d exposure to 10 mg/l indole-3-butyric acid in root induction media, followed by incubation in media containing 15 g/l sucrose and 1 g/1 activated charcoal for 21 d.Abbreviations IBA indole-3-butyric acid - MS Murashige and Skoog (1962) medium - NAA anaphthalene acetic acid - POP 2,3,5-trichloro--phenoxypropionic acid