Effects of some non-pathogenic micro-organisms on the growth of gnotobiotic barley plants

Summary Mated cultures of Blakeslea trispora, which produce trisporic acid, inhibited root and shoot extension of barley plants grown in gnotobiotic cultures, where the carbon from roots was the sole source of carbon available to the microorganism. Unmated cultures of B. trispora and Mucor hiemalis, which do not synthesize trisporic acid, inhibited root extension only. Azotobacter chroococcum had a variable effect in different experiments on both root and shoot extension but could at least partially relieve the inhibitory effect of B. trispora on root growth. Plant growth regulators, applied to roots exogenously, produced effects on plants which, although comparable with some of the inhibitory effects of the micro-organisms, were not comparable with the stimulatory effects of A. chroococcum. re]19751127     

Aggressiveness of Hedera helix L. growing on monuments: Evaluation in Roman archaeological sites and guidelines for a general methodological approach

Ivy (Hedera helix L.) growing on monuments is a common issue in the study of nature’s relationship with artefacts in the Euro-Mediterranean area. This is due to its pervasiveness and its double role as protective and as deteriogen. We studied the effect of ivy colonization within the archaeological site of Villa of Massentius. We applied a quantitative method derived from monument mapping method by Fitzner, to evaluate deterioration due to ivy colonization on vertical surfaces. Instead, the capping ivy effects was investigated through a multi-parameter evaluation. We recognized three different damage categories: the superficial loss of surface (W), ivy residues on wall after the removal event (I), and the loss of compact stone fragments (P/O). Mean W corresponded to 58% and P/O to 14%. The total damage rate was positively correlated to the amount of ivy cover and to favourable shady conditions. For the capping ivy, we calculated an average loss of 4.34 cm³/m² and a high level of soil deposit, which led to a secondary ivy colonization process. This paper proposes a contribution to the identification and quantification of ivy deteriogen activity. The application of the proposed method also provides a useful tool for planning restoration activity.     

Microbial inoculation of plants

Summary Seed of maize, tomato, and wheat was inoculated with cultures of Azotobacter, Clostridium, and a nitrogen-fixing facultative Bacillus and grown in a nutrient-deficient sand and a highly fertile silt loam.In sand, wheat showed a significant positive response to inoculation with Azotobacter and Clostridium but maize and tomato were unaffected by inoculation.When inoculated seed was planted in Lima silt loam there were significant increases in the growth of maize, tomato, and wheat to treatment with Clostridium, inoculated maize and wheat responded to Azotobacter inoculation while only wheat responded to inoculation with the facultative Bacillus.In pure-culture studies of the ability of these cultures to establish upon plant roots it was shown that Azotobacter did not colonize the roots of lucerne, maize, tomato, or wheat to any great extent. Bacillus and Clostridium were moderate colonizers of plant roots reaching from 1 to 20 per cent the levels reached byPseudomonas fluorescens on the same plants.The author held a Fulbright Travel Grant for the 1961–1962 academic year.Agronomy Paper No. 595. Supported in part by funds provided by Regional Research Project NE 39.     

Nanoparticle biofabrication using English ivy (Hedera helix)

Background

English ivy (Hedera helix) is well known for its adhesive properties and climbing ability. Essential to its ability to adhere to vertical surfaces is the secretion of a nanocomposite adhesive containing spherical nanoparticles, 60?C85 nm in diameter, produced exclusively by root hairs present on adventitious roots. These organic nanoparticles have shown promise in biomedical and cosmetic applications, and represent a safer alternative to metal oxide nanoparticles currently available.

Results

It was discovered that the maximum adventitious root production was achieved by a 4 h application of 1 mg/ml indole-3 butyric acid (IBA) to juvenile English ivy shoot segments cultured in custom vessels. After incubation of the shoots under continuous light at 83 ??mol/m² s at 20°C for 2 weeks, the adventitious roots were harvested from the culture system and it was possible to isolate 90 mg of dry weight nanoparticles per 12 g of roots. The nanoparticle morphology was characterized by atomic force microscopy, and found to be similar to previous studies.

Conclusions

An enhanced system for the production of English ivy adventitious roots and their nanoparticles by modifying GA7 Magenta boxes and identifying the optimal concentration of IBA for adventitious root growth was developed. This system is the first such platform for growing and harvesting organic nanoparticles from plants, and represents an important step in the development of plant-based nanomanufacturing. It is a significant improvement on the exploitation of plant systems for the formation of metallic nanoparticles, and represents a pathway for the generation of bulk ivy nanoparticles for translation into biomedical applications.     

Seed inoculation withAzospirillum brasilense andAzotobacter chroococcum and the root biomass of rice (Oryza sativa L.)

Summary The root biomass of rice seedlings increased due to inoculation withAzospirillum brasilense andAzotobacter chroococcum singly or in combination. In general, the increase in the biomass of roots was better in unsterilized soil than in sterilized one with or without inorganic nitrogen in the form of urea.     

Responses of ivy (Hedera helix L.) to combinations of gibberellic acid,paclobutrazol and abscisic acid

A reduction in concentration of gibberellins has been implicated in the phase change from juvenile to mature forms of ivy (Hedera helix L.). Attempts were made to increase the effective internal concentration of gibberellins by exogenous application of GA₃, and to decrease them by various applications of abscisic acid (ABA) and paclobutrazol (PP333), alone or in combination with GA₃. ABA and GA₃ were fed directly into the xylem of ivy plants by a wick system (a less drastic procedure than the defoliation or decapitation used by earlier workers) whereas PP333 was applied as a soil drench.Mature ivy responded to the application of GA₃ by reversion to the juvenile form, although this reversion was incomplete with respect to leaf lobing and red (anthocyanin) pigmentation and could occur spontaneously without the application of GA₃. Contrary to expectation, applications of ABA and PP333 caused the stimulation of growth in juvenile ivy. No adult characteristics were induced. As similar concentrations of ABA and PP333 produced severe retardation of growth (which could be alleviated by the application of GA₃) in other species, it is suggested that ivy may be an unsuitable model system for the investigation of phase change in woody plants.     

Liberation of amino acids by heterotrophic nitrogen fixing bacteria

Summary. Large amounts of amino acids are produced by nitrogen-fixing bacteria such as Azotobacter, Azospirillum, Rhizobium, Mesorhizobium and Sinorhizobium when growing in culture media amended with different carbon and nitrogen sources. This kind of bacteria live in close association with plant roots enhanced plant growth mainly as a result of their ability to fix nitrogen, improving shoot and root development suppression of pathogenic bacteria and fungi, and increase of available P concentration. Also, it has been strongly evidenced that production of biologically substances such as amino acids by these rhizobacteria are involved in many of the processes that explain plant-grown promotion. This paper reviews literature concerning amino acids production by nitrogen-fixing bacteria. The role of amino acids in microbial interactions in the rhizosphere and establishment of plant bacterial association is also discussed.     

Factors concerned in the Rooting Responses of Isolated Leaves

The rooting responses of isolated leaves of the dwarf Frenchbean and the ivy were studied by applying solutions of growthsubstances to the petiole: ß-indolyl-butyric acid(I.B.A.) for the former and -naphthyl-acetic acid (N.A.A.) forthe latter. The following factors were investigated:

1. The variation in response of successive leaves from the apextowards the base of the plant.
2. The optimal concentrationsof the growth substances and therelation between duration ofapplication and concentration.
3. The relation of age of theleaf and optimal concentration(ivy).
4. The nutritive factorsconcerned in the response were studied,(a) by varying periodsof contact between lamina and petiole,(b) by varying durationof light, (c) by varying the amountof light, and (d) by feedingwith sucrose and asparagine.

The most responsive leaf in the bean was the 2nd from the apex,in the ivy the 9th leaf. Very young leaves were killed. Withpetioles immersed for 24 hours the optimal concentrations wereI.B.A. at 2·5 parts per million (p.p.m.) for the bean,and N.A.A. 100 p.p.m. for the ivy. It was shown that the activesubstance entered mainly through the cut end in the transpirationstream; entry through the cuticle was much slower. Starvation of the cuttings whether by low light intensity, darkness,or separation of the lamina from the petiole reduced root formation.Feeding with sucrose and asparagine increased the response. Analysis of the leaves was carried out at regular intervalsfor total sugar, total and soluble nitrogen. In the bean solublenitrogen increased slightly in the petiole during the first5 days after treatment, after which insoluble nitrogen continuedto accumulate until the 7 day, when the roots first emerged.After this the total nitrogen decreased. Sugar content of thepetiole at first increased but rapidly fell before the rootsemerged and then again increased. In the ivy petiole the increasein soluble and total nitrogen was very slow, the former beinggreatest on the 15th day and the latter on the 20th day aftertreatment. The total sugar increased to the 10th day, but thendecreased until the emergence of roots, after which it againincreased. Treated leaves of the bean and ivy of different agesand the leaves starved for varying periods were also analysedfor total sugars and total nitrogen. Treated bean leaves receivingvarying periods of daily illumination were also analysed toelucidate their rooting response. Histological examination showed that in the bean the roots arosein the rays between the vascular bundles and in the ivy externalto the vascular bundles opposite the phloem. The growth of the rooted leaves was followed for more than amonth in the bean and over one year in the ivy. Some growthwas found in the immature bean leaves which survived treatment,but the final size was much below that of the control leavesattached to the plant. In the ivy very little further growthoccurred although the leaves produced a large root system andafter 2 years are still alive: the original differences in sizeof the successive leaves at the time of removal from the plantstill remain.     

Micro-organisms in the rhizosphere of plants inoculated withAzotobacter chroococcum

Summary The hypothesis that inoculation withAzotobacter chroococcum affects the growth of plants indirectly through changing the rhizosphere microflora was investigated. Inoculated and uninoculated wheat and tomato plants were grown in the glasshouse in two different soils, and total bacteria, chitinolytic bacteria, actinomycetes, glucosefermenting bacteria, aerobic cellulose-decomposing bacteria, and anaerobes were determined in intervals in the rhizosphere and in the soil. Root-surface fungi were studied using the Harley and Waid's root-washing technique¹⁰. Azotobacter became established in the rhizosphere of wheat and tomato plants and stimulated their growth. All the bacterial groups examined were more abundant in the rhizosphere than in the soil. Inoculation with Azotobacter delayed the colonization of roots by bacteria, actinomycetes, and fungi in the rhizosphere, but had no effect on other organisms. Inoculation did not affect the dominant root-surface fungi, and minor changes were not consistent.Part of a thesis accepted by the University of London for the degree of Ph.D. in Microbiology.     

Biotoxic activity in the Mucorales

The toxigenicity of representatives of 15 species of Mucorales (Absidia glauca, Actinomucor elegans, Cunninghamella elegans, Helicostylum piriforme, Mortierella isabellina, Mortierella (Mucor) rammaniana, Mucor hiemalis, Mucor mucedo, Mucor spinosus, Phycomyces blakesleeanus, Rhizopus oligosporus, Rhizopus stolonifer, Syncephalastrum racemosum, Thamnidium elegans, Zygorhynchus moelleri) towards the larvae of brine shrimp (Artemia salina) and the growth of pea seedlings (Pisum sativum) and tobacco plants (Nicotiana tabacum) was evaluated. The fungi were cultivated on malt extract agar and aqueous solutions of the cultures were tested.Thamnidium elegans showed a marked toxic action towards brine shrimp (mortality: 74.1%) andPhycomyces, Actinomucor andSyncephalastrum were only weakly toxic. Length and weight of stems of pea seedlings were moderately reduced by extracts ofAbsidia, Cunninghamella, Zygorhynchus andThamnidium and to a lesser degree byMucor spinosus. Cunninghamella andMucor spinosus also inhibited the development of pea hypocotyls. The length of tobacco stems was reduced byMortierella ramanniana, Rhizopus stolonifer andCunninghamella elegans. Wilting or other toxic phenomena were never observed with both test plants. Considering the present results and data from literature it is suggested that species of Mucorales have only a weak toxigenicity.     

Mobilization of sulfane sulfur from cysteine desulfurases to the <Emphasis Type="Italic">Azotobacter vinelandii</Emphasis> sulfurtransferase RhdA

Mobilization of the l-cysteine sulfur for the persulfuration of the rhodanese of Azotobacter vinelandii, RhdA, can be mediated by the A. vinelandii cysteine desulfurases, IscS and NifS. The amount of cysteine was higher in mutant strains lacking rhdA (MV474) than in wild type. The diazotrophic growth of MV474 was impaired. Taking into account the functional results about rhodanese-like proteins and RhdA itself, it is suggested that RhdA-dependent modulation of l-cysteine levels must deal with a redox-related process.     

Associative effect of inoculation of different strains of Azotobacter and homologous rhizobium on the yield of moong (Vigna radiata), soybean (Glycine max) and pea (Pisum sativum)

Summary It has been observed that in the case ofVigna radiata andGlycine max incorporation of suitable strain of Azotobacter gave higher yield than obtained by the use of Rhizobium as inoculant. In the case ofVigna radiata even a strain of Azotobacter isolated from the rhizosphere of berseem gave similar yields as Rhizobium. In the case ofPisum sativum association of Rhizobium with a strain ofAzotobacter chroococcum isolated from the rhizosphere of pea gave numerically higher yield than Rhizobium alone. It may be possible that statistically higher yield may be obtained when a suitable strain of Azotobacter is used after screening a large number of strains of Azotobacter from the rhizosphere of pea.     

Structure, attachment properties, and ecological importance of the attachment system of English ivy (Hedera helix)

Root climbers such as English ivy (Hedera helix) rely on specialized adventitious roots for attachment, enabling the plants to climb on a wide range of natural and artificial substrates. Despite their importance for the climbing habit, the biomechanical properties of these specialized adventitious roots compared with standard roots and their performance in the attachment to different host species or inert substrates have not been studied. Here organs and tissues involved in the attachment are characterized and their significance in regard to a broader functional and ecological aspect is discussed. Depending on the substrate, the root clusters show different types of failure modes at various frequencies, demonstrating the close interaction between the climber and its substrates. With a Young's Modulus of 109.2 MPa, the attachment roots are relatively stiff for non-woody roots. The central cylinders of the attachment roots show a high tensile strength of 38 MPa and a very high extensibility of 34%. In host trees naturally co-distributed with English ivy, a 'balanced' occurrence of failure of the attachment system of the climber and the bark of the host is found, suggesting a co-evolution of climber and host. Maximum loads of root clusters normalized by the number of roots match those of individually tested attachment roots. In comparison with most subterranean roots the properties and structure of the attachment roots of English ivy show distinct differences. There exist similarities to the properties found for roots of Galium aparine, suggesting a trend in not fully self-supporting plants towards a higher extensibility.     

The Formation of Artificial Nitrogen-Fixing Symbioses with Rape (Brassica napusvar. napus) Plants in Nonsterile Soil

The treatment of rape plants grown in nonsterile soil with 2,4-dichlorophenoxyacetic acid (auxin-like growth-promoting substance) or their inoculation with the bacterial association Micrococcussp. + Rhodococcussp. and/or with the mixed nitrogen-fixing culture Azotobacter nigricans+ Bacillussp. led to the formation of paranodules on the rape roots. The introduced bacteria were detected both in the intercellular space and inside the cells of the paranodules and the rape roots. The nitrogen-fixing activity of the paranodulated plants was two times higher than that of the inoculated plants lacking paranodules and five times higher than that of the control (i.e., not inoculated) plants. The paranodulation led to a 40% increase in the crop yield of rape plants and provided for a statistically significant increase in the total nitrogen as well as protein nitrogen contents of the plants.     

Light intensity is a limiting factor to the inland expansion of Cape ivy (<Emphasis Type="Italic">Delairea odorata</Emphasis>)

Cape ivy (Delairea odorata) is a highly invasive climbing perennial vine that is primarily distributed in coastal communities of California and Oregon, with patchy infestations in some inland riparian areas. In this study, we evaluated light as a potential environmental limitation to the spread of Cape ivy into inland regions of the western United States. Cape ivy was collected from four locations representing the north to south range. Plants were grown for 9 to 11 weeks in full sunlight and under two shade regimes (20 and 6% of full sunlight). The experiment was conducted twice at two temperature regimes. Results show some within- and among-population variability, with the southernmost San Diego County population having the highest biomass under the warmer growing conditions and the three northern populations responding most favorably in the cooler growing conditions. Despite the minor differences within and between populations, Cape ivy grew very poorly in full sunlight in both experiments. Although plants growing under 6% light grew better than those in full sunlight, they were far less robust compared to plants growing at 20% light. Our results indicate that while Cape ivy will not persist in areas with prolonged high intensity sunlight, characterized by much of the interior regions of California and Oregon, it is expected to invade and spread in areas with reduced light, including coastal regions frequently exposed to fog or cloudy conditions, or sub-canopy layers of riparian forests or woodlands. These communities should be the target areas for early detection and rapid response programs to prevent further Cape ivy invasion.     

Prevalence of Different Horticultural Taxa of Ivy (Hederaspp., Araliaceae) in Invading Populations

‘English’ ivy (Hedera spp.) is a complex of invasive plant pests that are separated into several distinct taxa. To better understand the invasion by ivy of Pacific Northwest native forests, we investigated the taxonomic identity of 58 selected invasive populations in the Pacific Northwest. Random amplified polymorphic DNA (RAPD) markers revealed that 83% of the 119 samples from invading populations were derived form H. hibernica (Kirchner) Bean, which has been frequently sold as English ivy, although this apparently is an incorrect common name. It is used widely in urban landscapes in the Northwest. The remaining 20 samples were either H.helix ‘California,’ ‘Pittsburgh,’ ‘Star,’ other cultivars not investigated in the study or possible hybrids.     

Quantitative study and modelling of the litter decomposition in a European alluvial forest. Is there an influence of overstorey tree species on the decomposition of ivy litter (Hedera helix L.)?

The influence on the decomposition rate of ivy litter (Hedera helix L.) of three ligneous overstorey species (oak, Quercus robur L., white poplar, Populus alba and ash, Fraxinus excelsior L.) that support ivy was studied in an alluvial hardwood forest. The ivy provides an abundant litter at the end of spring. The decomposition of ivy litter and the nutrient release rate were analyzed over four months during the growing season of the canopy trees, the hypothesis being that throughfall could slow down the mass loss rate of ivy depending on the support species. Mathematical models for mass loss and nutrient (P, N, K and Mg) release rates were developed. Mass loss rate and the release rate of magnesium, nitrogen and phosphorus show significant differences depending on species whereas no influence of species was observed on the release rate of potassium. The results illustrate the significant effect of oak compared to ash and poplar in slowing down mass loss rate and nitrogen, phosphorus and magnesium release rates. The mass loss rate over time under the three species followed the proposed exponential model, whereas the release rates of phosphorus and magnesium did not follow this model. This model is fitted to the data in two cases out of three for potassium. Magnesium and potassium are released more rapidly when nitrogen accumulates and the phosphorus content does not change significantly. No species × date interaction was observed, except in the case of magnesium. The species effect is interpreted as an effect of composition of throughfalls and presence or absence of inhibitory substances such as phenolic compounds.     

Association of Azospirillum with Grass Roots

The association between grass roots and Azospirillum brasilense Sp 7 was investigated by the Fahraeus slide technique, using nitrogen-free medium. Young inoculated roots of pearl millet and guinea grass produced more mucilaginous sheath (mucigel), root hairs, and lateral roots than did uninoculated sterile controls. The bacteria were found within the mucigel that accumulated on the root cap and along the root axes. Adherent bacteria were associated with granular material on root hairs and fibrillar material on undifferentiated epidermal cells. Significantly fewer numbers of azospirilla attached to millet root hairs when the roots were grown in culture medium supplemented with 5 mM potassium nitrate. Under these growth conditions, bacterial attachment to undifferentiated epidermal cells was unaffected. Aseptically collected root exudate from pearl millet contained substances which bound to azospirilla and promoted their adsorption to the root hairs. This activity was associated with nondialyzable and proteasesensitive substances in root exudate. Millet root hairs adsorbed azospirilla in significantly higher numbers than cells of Rhizobium, Pseudomonas, Azotobacter, Klebsiella, or Escherichia. Pectolytic activities, including pectin transeliminase and endopolygalacturonase, were detected in pure cultures of A. brasilense when this species was grown in a medium containing pectin. These studies describe colonization of grass root surfaces by A. brasilense and provide a possible explanation for the limited colonization of intercellular spaces of the outer root cortex.     

Increased root exudation of14C-compounds by sorghum seedlings inoculated with nitrogen-fixing bacteria

Summary Organic components leaked fromSorghum bicolor seedlings (‘root exudates’) were examined by recovering¹⁴C labelled compounds from root solutions of seedlings inoculated withAzospirillum brasilense, Azotobacter vinelandii orKlebsiella pneumoniae nif-. Up to 3.5% of the total¹⁴C recovered from shoots, roots, and nutrient solutions was found in the root solutions. Inoculation with Azospirillum and Azotobacter increased the amounts of¹⁴C and decreased the amounts of carbohydrates in the root solutions. When sucrose was added as a carbon source for the bacteria, the increase of¹⁴C in the solutions did not occur. Quantities of¹⁴C found in the root solutions were proportional to amounts of mineral nitrogen supplied to the plants. Bacterial growth also was proportional to nitrogen levels. When sorghum plants were grown in soil and labelled with¹⁴CO₂, about 15% of the total¹⁴C recovered within 48 hours exposure was found in soil leachates.     

Indolyl-3-acetic acid formation byAzotobacter chroococcum

Summary Small amounts of indolyl-3-acetic acid (IAA) were detected in aerated cultures ofAzotobacter chroococcum grown with or withoutl-tryptophane in the medium, but IAA was detected in agar cultures only whenl-tryptophane was present. Most IAA was found in 7-day-old cultures and less in older cultures. Washed cells did not convert tryptophane enzymically to IAA. The time course of IAA formation byA. chroococcum strain A6 has been described and the effect of adding tryptophane to the medium has been studied. In contrast to results elsewhere strain A6 produced traces of IAA in aerated cultures with or without added tryptophane. IAA was detected only after the end of exponential growth when cells had begun to autolyse. The amount of IAA declined as cultures aged. The slight effect ofl- but not ofd-tryptophane in promoting IAA formation in ageing cultures suggests some kind of biological transformation but it seems unlikely that IAA formation is part of the normal metabolic processes of intact Azotobacter cells.