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PubMed1988

Nonlinear relationship between concentration and activity of a bacterial ice nucleation protein.

The expression level of an ice nucleation gene (inaZ) was varied in Escherichia coli to observe the relationship between activity and gene product. The ice nucleation activity increased as the 2nd to 3rd power of the membrane concentration of the inaZ gene product, implying that molecules of InaZ protein interact cooperatively in groups of two to three at the rate-limiting step of ice nucleus assembly. The 2nd to 3rd power relationship was independent of the threshold temperature at which ice nucleation was measured and was consistent over a 500-fold range of protein concentration. Such a relationship indicates that the same rate-limiting step must be common to the formation of ice nuclei displaying all the various threshold temperatures within a bacterial population. Observations of Pseudomonas syringae, expressing the inaZ gene at various levels, were consistent with a similar relationship and hence a similar mechanism of ice nucleus assembly in Pseudomonas.

Bacterial Proteins↗
PubMed1987

Copper resistance in bacteria.

Copper is a required trace element for many organisms, yet it can exert an inhibitory effect on bacterial growth at relatively low concentrations. However, there are some bacterial species that can tolerate high levels of copper. It also has been reported that copper resistance is plasmid-encoded in Escherichia coli, Proteus vulgaris and a Pseudomonas syringae isolate. It is not known if copper is effluxed from the cell, detoxified by binding to copper-binding proteins, or binds to cell-surface components. Actual data on copper transport in bacteria is also lacking, indicating that this area of research deserves serious attention.

Bacteria↗
PubMedDate not supplied

[Numerical systematics of bacteria of the genus Pseudomonas].

In 290 strains of bacteria belonging to the genus Pseudomonas, 120 morphological and physiologo-biochemical characters were studied and the results obtained thereby were analyzed by the methods of numerical taxonomy using computers. The majority of strains were subdivided into 11 clusters: Ps. aeruginosa (1), Ps. putida (2), Ps. rathonis (5), Ps. syringae (8), Ps. pseudoalcaligenes (9), Ps. maltophilia (10), Ps. acidovorans (11), Ps. testosteroni (12), Ps. mendocina (13), Ps. cepacia (14), Ps. fluorescens (3). The latter cluster included also the strains identified earlier as Ps. aurantiaca, Ps. lemonnieri, Ps. fluoro-violaceus, and Ps. aureofaciens. Three clusters contained strains which could not be identified and probably should be regarded as distinct species. The characteristics have been selected useful for diagnostics of the above Pseudomonas bacteria and the subgroups of Ps. fluorescens.

Computers↗
PubMed1997

[The experimental study of Pseudomonas contamination in soft drinks].

Bymeans of this study, we have found out about the situation of Pseudomons contamination in the soft drinks which are manufactured and sold at the area of Nanchang City. In the course of this study, we examined total 416 specimens, in which we found that 69 specimens were positive reaction. The positive rate made up 16.59% of the total specimens. From these 69 positive specimens we separate isolated andidentified the following Pseudomonas 111 strains including 16 different species: P. aeruginosa, 9; P. fluorescens biovars; 11; P. putida biovars, 14; P. syringae pathovars, 5; P. mendocina, 5; P. alcaligenes, 23; P. pseudoalcaligenes, 10; P. cepacia, 17; P. solanacearum, 1; P. testosteroni, 1; P. delafieldii, 3; P. facilis, 6; P. flava 1; P. psenudoflava, 2; P. palleronii, 1; and a new species of Pseudomonas--P. halosensibilis. The achievement of this study will provide scientific basis for working out the standards of food safety control and inspection, raising the level of food hygieneinspection, and directing the drinks menufacturers and selling units to strengthen the food safety control and inspection.

Carbonated Beverages↗
PubMed2003

Partial characterization and insecticidal properties of Ricinus communis L foliage flavonoids.

Aqueous leaf extract of Ricinus communis L (Euphorbiaceae), a cultivated plant in tropical countries, showed excellent insecticidal activity against Callosobruchus chinensis L (Coleoptera: Bruchidae). We have isolated and tested flavonoids as insecticidal and antimicrobial agents. The isolated flavonoids showed potential insecticidal, ovicidal and oviposition deterrent activities against C chinensis L. However, antimicrobial activity against the common microbial infestants of stored pulses, of which C chinensis is a major pest, was found to be insignificant. Two bands having Rf 0.63 and 0.69 were seen on HPTLC plates using mobile phase benzene + ethyl acetate + methanol + formic acid (12 + 4.5 + 2 + 1.5 by volume) as eluant. The Rf values and scanning of the spectrum in the ultraviolet region, showed identity with the flavonoids quercetin and kaempferol. This was further confirmed using HPLC and IR and UV spectrometry. HPLC and HPTLC chromatograms also suggested quercetin to be the major flavonoid present in the hydrolyzed aqueous leaf extract of R communis.

Animals↗
PubMed2003

Transmission of ice-nucleating active bacteria from a prey reduces cold hardiness of a predator (Araneae: Theridiidae).

The influence of ice-nucleating active (INA) bacteria on cold hardiness of the house spider, Achaearanea tepidariorum, was determined by measuring the supercooling point (SCP) of hatchlings given either INA-bacteria-fed or bacteria-free prey (Drosophila melanogaster). Spiders that had eaten INA-bacteria-fed flies showed higher SCPs than those fed on bacteria-free flies. Through feeding, INA bacteria in the prey reduce the cold hardiness of spiders. This fact should be taken into account before using INA agents as a means of pest management.

Animals↗
PubMed2006

Expression of the antimicrobial peptides in plants to control phytopathogenic bacteria and fungi.

Three antimicrobial peptides exhibiting in vitro antifungal activity were expressed in Arabidopsis to compare their in planta activity. Beta-Purothionin, cecropin B, and phor21 were expressed under an endogenous promoter with moderate-level activity and excreted extracellularly. Expression of beta-purothionin rendered the greatest antibacterial and antifungal resistance while cecropin B enhanced only antibacterial activity and phor21 did not improve antimicrobial resistance. The transgenic beta-purothionin arrested fungal growth on leaf surfaces and infection of stomata. Leaf extracts from plants producing beta-purothionin and cecropin B displayed membrane permeabilizing activity. The in planta antimicrobial activity of the tested peptides was consistent with previously reported in vitro experiments. The expression strategy allowed enhanced antifungal resistance without high-level transgene expression.

Antimicrobial Cationic Peptides↗
PubMed2005

Analysis of Arabidopsis growth factor gene 1 (GFG1) encoding a nudix hydrolase during oxidative signaling.

Maintenance of pyridine nucleotide homeostasis is vital for normal growth and development of plants and animals. We demonstrate that Arabidopsis Growth Factor Gene 1 (GFG1; At4g12720) encoding a nudix hydrolase, is an NADH pyrophosphatase and ADP-ribose pyrophosphatase. The affinity for NADH and ADP-ribose indicates that this enzyme could serve as a connection between sensing cellular redox changes and downstream signaling. GFG1 transcript levels were rapidly and transiently induced during both biotic stresses imposed by avirulent pathogens and abiotic stresses like ozone and osmoticum. T-DNA knock out plants of GFG1 gene, gfg1-1, exhibit pleiotropic phenotypes such as reduced size, increased levels of reactive oxygen species and NADH, microscopic cell death, constitutive expression of pathogenesis-related genes and enhanced resistance to bacterial pathogens. The recombinant protein failed to complement the mutator deficiency in SBMutT- strain of Escherichia coli, suggesting this protein may not play a role in sanitizing the nucleotide pool. Based on rapid transcriptional changes in response to various stresses, substrate specificity of the enzyme, and analysis of the knock out mutant, we propose that GFG1 is a key gene linking cellular metabolism and oxidative signaling.

Adenosine Diphosphate Ribose↗
PubMed2004

Activation of hypersensitive cell death by pathogen-induced receptor-like protein kinases from Arabidopsis.

In Arabidopsis, there is a family of receptor-like protein kinases (RLKs) containing novel cysteine-rich repeats in their extracellular domains. Genes encoding many of these cysteine-rich RLKs (CRKs) are induced by pathogen infection, suggesting a possible role in plant defense responses. We have previously generated Arabidopsis plants expressing four pathogen-regulated CRK genes (CRK5, 6, 10 and 11) under control of a steroid-inducible promoter and found that induced expression of CRK5, but not the other three CRK genes, triggered hypersensitive response-like cell death in transgenic plants. In the present study, we have analyzed the structural relationship of the CRK family and identified three CRKs (CRK4, 19 and 20) that are structurally closely related to CRK5. Genes encoding these three CRKs are all induced by salicylic acid and pathogen infection. Furthermore, induced expression of CRK4, 19 and 20 all activates rapid cell death in transgenic plants. Thus, the activity of inducing rapid cell death is shared by these structurally closely related CRKs. We have also performed yeast two-hybrid screens and identified proteins that interact with the kinase domains of CRKs. One of the identified CRK-interacting proteins is the kinase-associated type 2C protein phospohatase known to interact with a number of other RLKs through its kinase-interacting FHA domain. Other CRK-interacting proteins include a second protein with a FHA domain and another type 2C protein phosphatase. Interactions of CRKs with these three proteins in vivo were demonstrated through co-immunoprecipitation. These CRK-interacting proteins may play roles in the regulation and signaling of CRKs.

Amino Acid Sequence↗
PubMed2006

Plant stomata function in innate immunity against bacterial invasion.

Microbial entry into host tissue is a critical first step in causing infection in animals and plants. In plants, it has been assumed that microscopic surface openings, such as stomata, serve as passive ports of bacterial entry during infection. Surprisingly, we found that stomatal closure is part of a plant innate immune response to restrict bacterial invasion. Stomatal guard cells of Arabidopsis perceive bacterial surface molecules, which requires the FLS2 receptor, production of nitric oxide, and the guard-cell-specific OST1 kinase. To circumvent this innate immune response, plant pathogenic bacteria have evolved specific virulence factors to effectively cause stomatal reopening as an important pathogenesis strategy. We provide evidence that supports a model in which stomata, as part of an integral innate immune system, act as a barrier against bacterial infection.

Abscisic Acid↗
PubMed2006

Plant pathogens trick guard cells into opening the gates.

The stomata of plants regulate gas exchange and water transpiration in response to changing environmental conditions. New work reveals that stomata also have an important role in host defense. In this issue of Cell, Melotto et al. (2006) show that stomata close upon detection of potential microbial pathogens to prevent the infection of the leaf interior. Moreover, pathogenic bacteria have evolved strategies to suppress the closure of stomata.

Amino Acids↗
PubMed2006

Linalool and lilac aldehyde/alcohol in flower scents. Electrophysiological detection of lilac aldehyde stereoisomers by a moth.

The stereoisomers of linalool and lilac aldehyde/alcohol were determined in the flower scent of 15 plant species using enantioselective multidimensional gas chromatography/mass spectrometry (enantio-MDGC/MS). Both linalool and all 8 stereoisomers of lilac alcohol and lilac aldehyde were detected, and there was a species-specific pattern. Single stereoisomers were collected by micropreparative-enantio-MDGC and were electrophysiologically tested on antennae of the noctuid moth Hadena bicruris, a species known to rely on lilac aldehyde for finding its host plant. The moth responded to all 8 stereoisomers, though only four stereoisomers were found in the scent of its host plant. The moth was less sensitive to some isomers than to others.

Acyclic Monoterpenes↗
PubMed2006

Extra- and intra-cellular ice formation in Stage I and II Xenopus laevis oocytes.

We are currently investigating factors that influence intracellular ice formation (IIF) in mouse oocytes and oocytes of the frog Xenopus. A major reason for choosing these two species is that while their eggs normally do not possess aquaporin channels in their plasma membranes, these channels can be made to express. We wish to see whether IIF is affected by the presence of these channels. The present Xenopus study deals with control eggs not expressing aquaporins. The main factor studied has been the effect of a cryoprotective agent [ethylene glycol (EG) or glycerol] and its concentration. The general procedure was to (a) cool the oocytes on a cryostage to slightly below the temperatures at which extracellular ice formation occurs, (b) warm them to just below the melting point, and (c) then re-cool them to -50 degrees C at 10 degrees C/min. In the majority of cases, IIF occurs well into step (c), but a sizeable minority undergo IIF in steps (a) or (b). The former group we refer to as low-temperature flashers; the latter as high-temperature flashers. IIF is manifested as abrupt blackening of the egg, which we refer to as "flashing." Observations on the Linkam cryostage are restricted to Stage I and II oocytes, which have diameters of 200 300 microm. In the absence of a cryoprotective agent, that is in frog Ringers, the mean flash temperature for the low-temperature freezers is -11.4 degrees C, although a sizeable percentage flash at temperatures much closer to that of the EIF (-3.9 degrees C). When EG is present, the flash temperature for the low-temperatures freezers drops significantly to approximately -20 degrees C for EG concentrations ranging from 0.5 to 1.5 M. The presence of 1.5 M glycerol also substantially reduces the IIF temperature of the low-temperature freezers; namely, to -29 degrees C, but 0.5 and 1 M glycerol exert little or no effect. The IIF temperatures observed using the Linkam cryostage agree well with those estimated by calorimetry [F.W. Kleinhans, J.F. Guenther, D.M. Roberts, P. Mazur, Analysis of intracellular ice nucleation in Xenopus oocytes by differential scanning calorimetry, Cryobiology 52 (2006) 128-138]. The IIF temperatures in Xenopus are substantially higher than those observed in mouse oocytes [P. Mazur, S. Seki, I.L. Pinn, F.W. Kleinhans, K. Edashige, Extra- and intracellular ice formation in mouse oocytes, Cryobiology 51 (2005) 29-53]. Perhaps that is a reflection of their much larger size.

Animals↗
PubMed2005

An importin alpha homolog, MOS6, plays an important role in plant innate immunity.

Plant disease resistance is the consequence of an innate defense mechanism mediated by Resistance (R) genes [1]. The conserved structure of one class of R protein is reminiscent of Toll-like receptors (TLRs) and Nucleotide binding oligomerization domain (NOD) proteins-immune-response perception modules in animal cells [2, 3, and 4]. The Arabidopsis snc1 (suppressor of npr1-1, constitutive, 1) mutant contains a mutation in a TIR-NBS-LRR-type of R gene that renders resistance responses constitutively active without interaction with pathogens [5]. Few components of the downstream signaling network activated by snc1 are known. To search for regulators of R-gene-mediated resistance, we screened for genetic suppressors of snc1. Three alleles of the mutant mos6 (modifier of snc1, 6) partially suppressed constitutive-resistance responses and immunity to virulent pathogens in snc1. Furthermore, the mos6-1 single mutant exhibited enhanced disease susceptibility to a virulent oomycete pathogen. MOS6, identified by positional cloning, encodes importin alpha3, one of eight alpha importins in Arabidopsis [6]. alpha importins mediate the import of specific proteins across the nuclear envelope. We previously reported that MOS3, a protein homologous to human nucleoporin 96, is required for constitutive resistance in snc1 [7]. Our data highlight an essential role for nucleo-cytoplasmic trafficking, especially protein import, in plant innate immunity.

Active Transport, Cell Nucleus↗
PubMed2005

MOS2, a protein containing G-patch and KOW motifs, is essential for innate immunity in Arabidopsis thaliana.

Innate immunity is critical for sensing and defending against microbial infections in multicellular organisms. In plants, disease resistance genes (R genes) play central roles in recognizing pathogens and initiating downstream defense cascades. Arabidopsis SNC1 encodes a TIR-NBS-LRR-type R protein with a similar structure to nucleotide binding oligomerization domain (Nod) proteins in animals. A point mutation in the region between the NBS and LRR of SNC1 results in constitutive activation of defense responses in the snc1 mutant. Here, we report the identification and characterization of mos2-1, a mutant suppressing the constitutive defense responses in snc1. Analysis of mos2 single mutants indicated that it is not only required for resistance specified by multiple R genes, but also for basal resistance. Map-based cloning of MOS2 revealed that it encodes a novel nuclear protein that contains one G-patch and two KOW domains and has homologs across the animal kingdom. The presence of both G-patch and KOW domains in the MOS2 protein suggests that it probably functions as an RNA binding protein critical for plant innate immunity. Our discovery on the biological functions of MOS2 will shed light on functions of the MOS2 homologs in animals, where they may also play important roles in innate immunity.

Arabidopsis↗
PubMed2004

NO-degradation by alfalfa class 1 hemoglobin (Mhb1): a possible link to PR-1a gene expression in Mhb1-overproducing tobacco plants.

Tobacco plants overproducing alfalfa class 1 hemoglobin (HOT plants) have been shown to have reduced necrotic symptom development. Here, we show that this altered pathogenic response is linked to a significant increase in the nitric oxide (NO)-affected pathogenesis-related (PR-1a) transcript accumulation in the transgenic plants. Homogenates of HOT transgenic seedlings were also found to have higher NO-scavenging activity than non-transformed ones. The NO-scavenging properties of recombinant alfalfa class1 hemoglobin have been examined. Recombinant Mhb1 (rMhb1) was produced in bacteria and purified using polyethylene glycol (10-25%) fractionation, chromatography on DEAE-Sephacel, and Phenyl Superose columns. After the final purification step, the obtained preparations were near homogeneous and had a molecular weight of 44 kDa determined by size-exclusion chromatography and 23 kDa by SDS-PAGE, indicating that rMhb1 is a dimer. The protein participated in NO-degradation activity with NAD(P)H as a cofactor. After ion-exchange columns, addition of FAD was necessary for exhibiting maximal NO-degradation activity. The NAD(P)H-dependent NO-scavenging activity of rMhb1, which is similar to that of barley hemoglobin, supports a conclusion that both monocot and dicot class 1 hemoglobins can affect cellular NO levels by scavenging NO formed during hypoxia, pathogen attack and other stresses.

Base Sequence↗
PubMed2005

A RT/PCR-partial restriction enzymatic mapping (PREM) method for the molecular characterisation of the large satellite RNAs of Arabis mosaic virus isolates.

The satellite RNA of the grapevine isolate NW of Arabis mosaic virus (ArMV) was cloned and sequenced, and showed 75% identity at the nucleotide level to the satellite RNA of the lilac isolate of ArMV. In order to survey ArMV isolates from various geographical origins and natural hosts for the presence of large satellite RNAs and analyse their degree of variability, a RT/PCR-partial restriction enzymatic mapping (PREM) method was developed. The method is based on the incorporation of 5-methyl-dCTP in the RT/PCR reaction, and the subsequent digestion of the RT/PCR products by methyl-sensitive restriction enzymes. Satellites RNAs were detected by RT/PCR in eight isolates out of 47, six of them originating from grapevine, one from hop and one from lilac. The partial restriction digestion patterns allowed to distinguish six different types of satellites. Cloning and sequencing of the different satellites confirmed these results, the PREM proving able to discriminate sequences with 96% identity. The sizes of the different satellites varied between 1092 and 1139 nucleotides, their encoded proteins between 338 and 360 amino acids. Conserved domains were found in the amino and carboxy-termini between the sequences of the proteins encoded by the satellites of the different isolates of ArMV.

Amino Acid Sequence↗