Role of bacterial cyanide production in differential reaction of plant cultivars to deleterious rhizosphere pseudomonads

Lettuce and wheat cultivars, differing in reaction to root inoculation with plant growth-inhibitory bacteria, were tested for sensitivity to (i) gaseous metabolites produced by deleterious, cyanogenic isolates of Pseudomonas fluorescens, and to (ii) pure cyanide. Reactions were read as shoot and/or root elongation after exposure of seedlings to the volatiles in vitro. Lettuce cultivar Salad Bowl was significantly less sensitive than cv. Montana, both to bacterial volatiles and to pure cyanide, and a similar difference between these cultivars was also obtained in greenhouse experiments where bacteria were inoculated directly on the roots. Cultivar differences were, however, not recorded, either in vitro or in the greenhouse, when the bacteria were grown on a medium which did not support cyanide production. In wheat, a difference in sensitivity to bacteria-produced volatiles was recorded between two cultivars (Drabant and Besso) differing in reaction to bacterial inoculation, but in contrast to lettuce cultivars, the wheat cultivars tested did not react differentially to pure cyanide. The results suggest that in lettuce differential sensitivity to cyanide is one factor behind cultivar differences in reaction to the bacteria tested, even though bacterial metabolites other than cyanide may contribute to the plant growth inhibition recorded. In wheat, however, differential cultivar responses to these bacteria could not be shown to be related to cyanide.     

Construction of a rhizosphere pseudomonad with potential to degrade polychlorinated biphenyls and detection of bph gene expression in the rhizosphere.

The genetically engineered transposon TnPCB, contains genes (bph) encoding the biphenyl degradative pathway. TnPCB was stably inserted into the chromosome of two different rhizosphere pseudomonads. One genetically modified strain, Pseudomonas fluorescens F113pcb, was characterized in detail and found to be unaltered in important parameters such as growth rate and production of secondary metabolites. The expression of the heterologous bph genes in F113pcb was confirmed by the ability of the genetically modified microorganism to utilize biphenyl as a sole carbon source. The introduced trait remained stable in laboratory experiments, and no bph-negative isolates were found after extensive subculture in nonselective media. The bph trait was also stable in nonselective rhizosphere microcosms. Rhizosphere competence of the modified F113pcb was assessed in colonization experiments in nonsterile soil microcosms on sugar beet seedling roots. F113pcb was able to colonize as efficiently as a marked wild-type strain, and no decrease in competitiveness was observed. In situ expression of the bph genes in F113pcb was found when F113pcb bearing a bph'lacZ reporter fusion was inoculated onto sugar beet seeds. This indicates that the bph gene products may also be present under in situ conditions. These experiments demonstrated that rhizosphere-adapted microbes can be genetically manipulated to metabolize novel compounds without affecting their ecological competence. Expression of the introduced genes can be detected in the rhizosphere, indicating considerable potential for the manipulation of the rhizosphere as a self-sustaining biofilm for the bioremediation of pollutants in soil. Rhizosphere bacteria such as fluorescent Pseudomonas spp. are ecologically adapted to colonize and compete in the rhizosphere environment. Expanding the metabolic functions of such pseudomonads to degrade pollutants may prove to be a useful strategy for bioremediation.     

Interactive regulation of root exudation and rhizosphere denitrification by plant metabolite content and soil properties

Plant and Soil - Cyperaceae are common on nutrient-poor soils, including in western Japan. We examined the ability of the native Cyperaceae from western Japan to form dauciform roots and assessed...     

Beneficial and deleterious effects of HCN-producing pseudomonads on rhizosphere interactions

Rhizobacteria live around roots but also inside the cortical root tissues by utilizing organic substances released from root cells into the intercellular spaces and the root environment. The effects of metabolites of these rhizosphere-inhabiting bacteria on root physiology and plant development have hardly been studied. However, recent studies indicate that, depending on environmental factors and plant species, certain strains of rhizosphere Pseudomonas spp. and some of their metabolites such as HCN may inhibit or enhance plant establishment or inhibit development of plant disease. Cultural practices such as cropping frequency, no tillage, and soilless cultivation, as well as edaphic factors seem to determine these rhizosphere interactions.     

Peroxidase-catalyzed oxidation of eugenol: formation of a cytotoxic metabolite(s)

The oxidation of eugenol (4-allyl-2-methoxyphenol) by horseradish peroxidase was studied. Following the initiation of the reaction with hydrogen peroxide, eugenol was oxidized via a one-electron pathway to a phenoxyl radical which subsequently formed a transient, yellow-colored intermediate which was identified as a quinone methide. The eugenol phenoxyl radical was detected using fast-flow electron spin resonance. The radicals and/or quinone methide further reacted to form an insoluble complex polymeric material. The stoichiometry of the disappearance of eugenol versus hydrogen peroxide was approximately 2:1. The addition of glutathione or ascorbate prevented the appearance of the quinone methide and also prevented the disappearance of the parent compound. In the presence of glutathione, a thiyl radical was detected, and increases in oxygen consumption and in the formation of oxidized glutathione were also observed. These results suggested that glutathione reacted with the eugenol phenoxyl radical and reduced it back to the parent compound. Glutathione also reacted directly with the quinone methide resulting in the formation of a eugenol-glutathione conjugate(s). Using 3H-labeled eugenol, extensive covalent binding to protein was observed. Finally, the oxidation products of eugenol/peroxidase were observed to be highly cytotoxic using isolated rat hepatocytes as target cells.     

Bacterial diversity in tea plant (Camellia sinensis) rhizosphere soil from Qinling Mountains and its relationship with environmental elements

Plant and Soil - The bacterial diversity in rhizosphere soil is closely associated with its environmental factors, such as, pH, moisture and organic contents. The aims of this study were to...     

Lactobacillus plantarum; a deleterious contaminant of plant tissue cultures

Lactobacillus plantarum was isolated from in vitro plant cultures of Hemerocallis Stella d'ora, Catherine Woodbury and Stafford. Infected cultures deteriorated rapidly during three subcultures (15 weeks) when grown in vitro , showing chlorotic white shoots and grey calli. Weaned plants developed normally when transferred to the soil. The drop in multiplication rate of plant cultures coincided with a decrease in pH of the growth media. Uninfected plants of Hemerocallis Stella d'ora showed the same symptoms after inoculation with L. plantarum. Lactobacillus plantarum was reisolated from inoculated plant cultures that showed symptoms, thus fulfilling Koch's postulates. In plants inoculated with L. plantarum both the amount of dl-lactic acid formed and the number of plants showing symptoms increased with increasing numbers of bacteria in the inoculum, wheras plant multiplication rate and pH decreased. These effects could be reproduced by adding dl-lactic acid to the multiplication medium of plants free from L. plantarum , suggesting that the bacterial production of lactic acid was responsible for the changes in infected plants.     

The rhizosphere and plant nutrition: a quantitative approach

The role of the rhizosphere in relation to mineral nutrition is discussed within a quantitative framework using the Barber-Cushman model as a starting point. The uptake or release of nutrients by roots growing in soil leads to concentration gradients forming in the soil: the zone so affected is termed the rhizosphere. The nature of these gradients depends on three factors: the rate of uptake/release; the mobility of the nutrient in soil; and the rate of conversion between available and unavailable forms. The interplay between these factors determines the amount of mineral nutrients acquired by the plant and it is the complexity of the interplay which demands the use of mathematical models in order to understand which factors most limit uptake. Despite extensive experimental evidence of root-mediated changes to the physical, chemical and biological status of rhizosphere soil, the quantitative significance of these changes for mineral nutrition has not been assessed. The problems of making this quantitative transition are reviewed.     

Symbolic plant(s) of the Olympic Games

The victors of the Olympic Games in ancient Greece were awarded crowns made of olive branches. In Antiquity, the symbolism of plants was related to myths, properties, aesthetic values, and civilization. Theophrastus first classifies and identifies plants, and gathers information about them, in his classic books (4th century BC). Symbolic plants are native to the Mediterranean region and they exhibit some convergent behaviour with respect to their functional characteristics. These plants were collected (among other species) by Professor J. Sibthorp and his partners in two botanical journeys in the Levant during the 18th century, and they have been illustrated for Flora Graeca Sibthorpiana.     

Immunological interference of the synthetic aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD) and its metabolite(s) in the radioimmunoassay for testosterone

Radioimmunoassay (RIA) for testosterone (T) in unchromatographed plasma extracts from ATD-treated rats gave spuriously high values for T. Cross-reaction and Chromatographic studies subsequently showed that ATD and, to a much greater extent, its metabolite(s) were responsible for this overestimation. Celite column chromatography proved to be an effective way of separating T from ATD and its product(s)of metabolism.     

Studies on the rhizosphere microflora of a desert plant

The response of the bacterial population to edaphic drought of the Egyptian desert was investigated. The influence of the rhizosphere on the microbial population as compared with soil apart was studied. The total microbial flora, total simple nutritional microflora, spores of simple and complex nutritional requirements, Actinomycetes and Fungi were estimated. Some groups of organisms which carry out certain important biochemical processes in the soil were also investigated, namely, cellulose decomposers, nitrifiers, Azotobacter and Clostridia. It was found that significatnly higher donsities of microbial population existed in the rhizosphere than in soil apart. This was attributed to organic matter and root secretion furnished by the growing roots of the desert plant, in addition to the presence of relatively moister conditions around the root than in soil apart. The high densities of microflora present in the rhizosphere indicate that the desert soil of this locality is suitable for reclamation since the only limiting factor in the proliferation of soil microorganisms are soil organic matter and moisture. Cellulose decomposers and nitrogen fixers, were found in high densities in the rhizosphere but not detected in the soil apart. The significance of the rhizosphere as constituting a microhabitat is discussed. The probable contribution of the periotrophic mantle of bacteria for protecting root cells against edaphic drought is also noted. Mineralization of the high microbial protein in the rhizosphere supplies the plant with the necessary nutrients.

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Изучение микрофлоры ризосферы растений в пустыне

Изучалась реакция популяций бактерий на засуху в египетской пустыне. Исследовалось влияние ризосферы (по сравнению со свободной почвой) на популяцию микробов. Было определено общее количество микроорганизмов, количество микроорганизмов со скромными требованиями к питанию, количество спорообразующих микроорганизмов е простыми и комплексными требованиями к питанию и количество актиномицетов и грибов. Изучались также некоторые группы микроорганизмов, осуществляющих важные биологические превращения в почве, а именно микроорганизмов, разлагающих целлюлозу, нитрификаторов, Azotobacter и Clostridia. Было установлено, что в ризосфере микробы гораздо более многочисленны, чем в свободной почве. Кроме стносительно большей влажности среды вблизи корней это обусловливается и влиянием органического вещества и корневых выделений, попадающих в почву пустыни. Обильная микрофлора в ризосфере растений в прстине свидетельствует о том, что почва пустыни в исследуемых местах пригодна для обработки, так как лимитирующими факторами в развитии микроорганизмов являются только влажность и органическое вещество почвы.

     

Production of macrophage migration inhibitory factor(s) (MIF) by virus-transformed cells

Rodent cell lines transformed by SV40, polyoma virus and Rous sarcoma virus cultured in vitro release material into the culture medium which inhibits the migration of guinea pig macrophages. Similar macrophage migration inhibitory factors (MIF) were not detected in cell-free supernatants harvested from untransformed cell cultures. Comparison of the MIF produced by virus-transformed cells with MIF derived from peripheral lymphocytes stimulated in vitro by phytohemagglutinin (PHA) revealed that they had similar molecular weights (25 000), heat stability and were both inhibited by α-fucose and lotus agglutinin. Incubation of MIF-containing cell-free supernatants from transformed cells with pancreatic trypsin inhibitor, soybean trypsin inhibitor and diisopropylfluorophosphate eliminated the MIF activity. The possible identity of the MIF released by transformed cells as a protease is discussed with reference to a potential role in modifying the surface properties of transformed cells.     

New synthesis of (+/-)-alpha-CMBHC and its confirmation as a metabolite of alpha-tocopherol (vitamin E)

There is currently interest in the metabolism of the various compounds which make up the vitamin E family, especially with regards to the possible use of vitamin E metabolites as markers of oxidative stress and adequate vitamin E supply. A number of vitamin E metabolites have been described to date and we have recently developed a method to extract and quantitate a range of vitamin E metabolites in human urine. During the development of this method a new metabolite of alpha-tocopherol was identified, which we tentatively characterised as 5-(6-hydroxy-2,5,7,8-tetramethyl-chroman-2-yl)-2-methyl-pentanoic acid (alpha-CMBHC).(1) Here we describe the synthesis of alpha-CMBHC as a standard and confirm that it is a metabolite of alpha-tocopherol.     

Magnitude and sign epistasis among deleterious mutations in a positive-sense plant RNA virus

How epistatic interactions between mutations determine the genetic architecture of fitness is of central importance in evolution. The study of epistasis is particularly interesting for RNA viruses because of their genomic compactness, lack of genetic redundancy, and apparent low complexity. Moreover, interactions between mutations in viral genomes determine traits such as resistance to antiviral drugs, virulence and host range. In this study we generated 53 Tobacco etch potyvirus genotypes carrying pairs of single-nucleotide substitutions and measured their separated and combined deleterious fitness effects. We found that up to 38% of pairs had significant epistasis for fitness, including both positive and negative deviations from the null hypothesis of multiplicative effects. Interestingly, the sign of epistasis was correlated with viral protein-protein interactions in a model network, being predominantly positive between linked pairs of proteins and negative between unlinked ones. Furthermore, 55% of significant interactions were cases of reciprocal sign epistasis (RSE), indicating that adaptive landscapes for RNA viruses maybe highly rugged. Finally, we found that the magnitude of epistasis correlated negatively with the average effect of mutations. Overall, our results are in good agreement to those previously reported for other viruses and further consolidate the view that positive epistasis is the norm for small and compact genomes that lack genetic robustness.     

Characterization of antifungal metabolite phenazine from rice rhizosphere fluorescent pseudomonads (FPs) and their effect on sheath blight of rice

We have shown, the outcome of antifungal activity of phenazine derivatives which is produced by fluorescent pseudomonads (FPs) for the control of sheath blight of rice. A total of 50 fluorescent pseudomonads (FPs) were isolated from rice rhizosphere. Off which, 36 FPs exhibited antagonistic activity against Rhizoctonia solani, Macrophomina phaseolina, Fusarium oxysporum, Alternaria alternata and Sclerotium rolfsii up to 70–80% compared to control by dual culture method. BOX-PCR analyses of antagonistic isolates indicated that two phylogenetic group, where group I consisted of 28 isolates and eight isolates belongs to group II. Among 36 FPs, a total of 10 FPs revealed that the presence of phenazine derivatives on thin layer chromatography (TLC), which is coincided with that of authentic phenazine with R_f value 0.57. Similar to TLC analysis, antibiotic encoding gene phenazine-1-carboxamide (PCN) was detected in 10 FPs by PCR analysis with respective primer. Among, PCN detected isolates of FPs, a significant biocontrol potential possessing isolate designated as VSMKU1 and it was showed prominent antifungal activity against R. solani and other tested fungal pathogens. Hence, the isolate VSMKU1 was selected for further studies. The selected isolate VSMKU1 was identified as Pseudomonas aeruginosa by 16S rDNA sequence analysis. The antifungal metabolite phenazine like compound produced by VSMKU1 was confirmed by UV, FT-IR and HPLC analysis. The phenazine compound from VSMKU1 significantly arrest the growth of R. solani compared to carbendazim by well diffusion method. The detached leaf assay showed remarkable inhibition of lesion height 80 to 85% by the treatments of culture (VSMKU1), cell free culure filtrate and phenazine like compound compared to control and other treatments was observed in detached leaves of rice. These results emphasized that VSMKU1 isolate can be used as an alternative potential biocontrol agent against sheath blight of rice, instead of using commercial fungicide such as validamycin and carbendazim which cause environmental pollution and health hazards.     

Yeast flora of plant rhizosphere and phyllosphere]

The species belonging to the genera Cryptococcus and Hansenula with saturnian spores predominate in the rhizosphere of agricultural plants. The sporiferous strains of Debaryomyces, Hanseniaspora apiculata, Metschnikowia pulcherrima, and asporogenic Candida krusei and Trichosporon cutaneum prevail in the rhizosphere of wild plants. Candida krusei and Trichosporon cutaneum prevail in the rhizosphere of wild plants. The cultures of Rhodotorula, Candida krusei and Metschnikowia pulcherrima are typical of the phyllosphere of both cultural and wild plants. The phyllosphere of cultural plants contains also the asporogenic strains of Cryptococcus, Candida tropicalis, Trichosporon pullulans, Tr. cutaneum, and Hansenula, while Hanseniaspora apiculata and Saccharomyces cerevisiae predominate in the phyllosphere of wild plants. The yeast flora of the majority of studied plants is diverse and comprises 10--20 species (in cabbage, potato, linden, aspen, and pear trees). The rhizophere and phyllosphere of some plants contain only 2 to 4 yeast species (onion, hop, wild apple).