Synthesis of Polygalacturonate Trans-eliminase and Polygalacturonase by a Strain of Enterobacter cloacae Isolated from Ponded Sitka Spruce

During the ponding of Sitka spruce in lake water there was a change from a diverse, aerobic flora (16 species) to a restricted, facultatively anaerobic flora (1 or 2 species). This change corresponded with a marked increase in the degradation of pectin and a concomitant increase in the permeability of the wood to preservatives. A strain of Enterobacter cloacae (NCPPB 2909) isolated during ponding, synthesized an extracellular and intracellular polygalacturonase (PG) and an intracellular polygalacturonate trans-eliminase (PGTE). Both PG and PGTE were growth–linked; extracellular PG was produced initially and then replaced by intracellular PG and PGTE. A change in the pH value of the medium did not alter the relative synthesis of enzyme. Reduced oxygen tension retarded growth but had no effect on enzyme activities. PG and PGTE of E. cloacae were shown to have specific ion requirements and when tap-water was used in the preparation of a medium growth did not occur. The results are discussed in relation to an artificial system of sprinkling water, seeded with known species of bacteria, on to spruce wood in order to control the rate of pectin degradation and thus the permeability of the wood to preservatives.     

Pectinase Activity of Anaerobic and Facultatively Anaerobic Bacteria Associated with Soft Rot of Yam (Diascorea rotundata)

Anaerobic and facultatively anaerobic bacteria associated with soft rot of yam (Diascorea rotundata) were isolated by the looping-out method and found to consist of Clostridium (three isolates), Corynebacterium (three isolates), Vibrio (one isolate), and Bacillus lentus (one isolate). Enzyme assay for hydrolase, lyase, and pectinesterase activities by the cup-plate method showed that except for Vibrio sp., B. lentus, and two isolates of Corynebacterium no pectinase activity could be detected for organisms cultured on pectin medium. Most of the cultures on yam tissue, however, showed activities for the three enzymes. The viscometric assay for hydrolase and lyase enzymes indicated a significant level of hydrolase activity (a 40.90% decrease in viscosity for Vibrio sp. and Corynebacterium spp.), but no lyase activity for most of the isolates. Two isolates of Corynebacterium and B. lentus caused changes in fresh yams suggestive of soft rot.     

Pectinase Activity of Anaerobic and Facultatively Anaerobic Bacteria Associated with Soft Rot of Yam (Diascorea rotundata)

[This corrects the article on p. 564 in vol. 41.].     

Characteristics of Polygalacturonate Lyase C from Bacillus subtilis 7-3-3 and Its Synergistic Action with PelA in Enzymatic Degumming

An alkaline polygalacturonate lyase (PGL) from Bacillus subtilis 7-3-3, PelC, with diverse depolymerization abilities for different pectin substrates was found. The PGL activity of PelC decreased with increasing degree of methyl esterification of the substrate. PelA and PelC displayed notable synergistic effects in the enzymatic degumming of ramie fibers. Gum loss rates increased by 62% when PelC was used to replace up to three-eighths of the PelA dose (PelC, 60 U g⁻¹ ramie fibers). To the best of our knowledge, this study is the first to report the synergistic action of members of polysaccharide lyase families 1 and 3, represented by PelA and PelC, respectively. The present paper provides new insights into the improvement and production of enzymes used in enzymatic degumming.     

Ultraviolet-B-Responsive Anthocyanin Production in a Rice Cultivar Is Associated with a Specific Phase of Phenylalanine Ammonia Lyase Biosynthesis

Seedlings of 17 rice (Oryza sativa L.) cultivars were classified on the basis of anthocyanin pigmentation into three groups: an acyanic group with 9 cultivars, a moderately cyanic group with 5 cultivars, and a cyanic group with 3 cultivars. Seedlings of the cyanic group were deep purple in color, possessing copious amounts of anthocyanin in shoots. Sunlight (SL)-mediated anthocyanin and phenylalanine ammonia lyase (PAL) induction in a cyanic cultivar, purple puttu, was compared with an acyanic cultivar, black puttu. A brief exposure of dark-grown purple puttu seedlings to SL induced anthocyanin formation during a subsequent dark period with a peak at 24 h. The magnitude of SL-mediated anthocyanin induction is age dependent, the 4-d-old seedlings being the most responsive to SL. The anthocyanin induction in purple puttu seedlings is mediated exclusively by the ultraviolet-B (UV-B) component of SL. The SL-triggered anthocyanin induction was reduced by about 30% by a terminal far-red light pulse and was restored by a red light pulse, indicating the role of phytochrome in modulation of anthocyanin level. The SL-mediated induction of PAL showed two peaks, one at 4 h and the other at 12 h. Whereas the first PAL peak (4 h) was induced by phytochrome and was seen in both cultivars, the second PAL peak (12 h) was inducible by UV-B only in the cyanic purple puttu cultivar.     

Reevaluation of Production of Paralytic Shellfish Toxin by Bacteria Associated with Dinoflagellates of the Portuguese Coast

Paralytic shellfish toxins (PSTs) are potent neurotoxins produced by certain dinoflagellate and cyanobacterial species. The autonomous production of PSTs by bacteria remains controversial. In this study, PST production by two bacterial strains, isolated previously from toxic dinoflagellates, was evaluated using biological and analytical methods. Analyses were performed under conditions determined previously to be optimal for toxin production and detection. Our data are inconsistent with autonomous bacterial PST production under these conditions, thereby challenging previous findings for the same strains.     

Bacteria Associated with Benthic Diatoms from Lake Constance: Phylogeny and Influences on Diatom Growth and Secretion of Extracellular Polymeric Substances

The composition of diatom-associated bacterial communities was studied with 14 different unialgal xenic diatom cultures isolated from freshwater epilithic biofilms of Lake Constance, Germany. A clear dominance of Alphaproteobacteria was observed, followed by Betaproteobacteria, Gammaproteobacteria, Bacteroidetes, and Verrucomicrobia. Pure cultures of the diatom Cymbella microcephala, which was found to be dominant in epilithic biofilms in Lake Constance, were cocultivated with six associated bacterial strains. All these bacterial strains were able to grow in C. microcephala cultures in the absence of organic cosubstrates. Diatom growth was generally enhanced in the presence of bacteria, and polysaccharide secretion was generally increased in the presence of Proteobacteria. The monomer composition of extracellular polysaccharides of C. microcephala changed in relation to the presence of different bacteria, but the dominant monomers were less affected. Our results indicate that these changes were caused by the diatom itself rather than by specific bacterial degradation. One Bacteroidetes strain strongly influenced carbohydrate secretion by the alga via extracellular soluble compounds. Biofilms were formed only in the presence of bacteria. Phylogenetic analysis and coculture studies indicate an adaptation of Proteobacteria and Bacteroidetes to the microenvironment created by the diatom biofilm.     

Drought-Tolerance of Wheat Improved by Rhizosphere Bacteria from Harsh Environments: Enhanced Biomass Production and Reduced Emissions of Stress Volatiles

Water is the key resource limiting world agricultural production. Although an impressive number of research reports have been published on plant drought tolerance enhancement via genetic modifications during the last few years, progress has been slower than expected. We suggest a feasible alternative strategy by application of rhizospheric bacteria coevolved with plant roots in harsh environments over millions of years, and harboring adaptive traits improving plant fitness under biotic and abiotic stresses. We show the effect of bacterial priming on wheat drought stress tolerance enhancement, resulting in up to 78% greater plant biomass and five-fold higher survivorship under severe drought. We monitored emissions of seven stress-related volatiles from bacterially-primed drought-stressed wheat seedlings, and demonstrated that three of these volatiles are likely promising candidates for a rapid non-invasive technique to assess crop drought stress and its mitigation in early phases of stress development. We conclude that gauging stress by elicited volatiles provides an effectual platform for rapid screening of potent bacterial strains and that priming with isolates of rhizospheric bacteria from harsh environments is a promising, novel way to improve plant water use efficiency. These new advancements importantly contribute towards solving food security issues in changing climates.     

Synthesis and Secretion of Hypoxyproline-containing Macromolecules in Carrots: III. Metabolic Requirements for Secretion

Using pulse-chase experiments with radioactive proline, it is possible to study the rapid transfer from the cytoplasm to the cell wall of the hydroxyproline-rich protein found in the cell walls of higher plants. The secretion of this protein is not obligatorily coupled to protein synthesis. Secretion is completely inhibited by uncouplers of oxidative phosphorylation and strongly inhibited by the inhibitors of electron transport, cyanide and azide. It is concluded that the transfer of proteins from the cytoplasm to the cell wall is an energy-requiring step.     

Ferric Iron Reduction by Bacteria Associated with the Roots of Freshwater and Marine Macrophytes

In vitro assays of washed, excised roots revealed maximum potential ferric iron reduction rates of >100 μmol g (dry weight)⁻¹ day⁻¹ for three freshwater macrophytes and rates between 15 and 83 μmol (dry weight)⁻¹ day⁻¹ for two marine species. The rates varied with root morphology but not consistently (fine root activity exceeded smooth root activity in some but not all cases). Sodium molybdate added at final concentrations of 0.2 to 20 mM did not inhibit iron reduction by roots of marine macrophytes (Spartina alterniflora and Zostera marina). Roots of a freshwater macrophyte, Sparganium eurycarpum, that were incubated with an analog of humic acid precursors, anthroquinone disulfate (AQDS), reduced freshly precipitated iron oxyhydroxide contained in dialysis bags that excluded solutes with molecular weights of >1,000; no reduction occurred in the absence of AQDS. Bacterial enrichment cultures and isolates from freshwater and marine roots used a variety of carbon and energy sources (e.g., acetate, ethanol, succinate, toluene, and yeast extract) and ferric oxyhydroxide, ferric citrate, uranate, and AQDS as terminal electron acceptors. The temperature optima for a freshwater isolate and a marine isolate were equivalent (approximately 32°C). However, iron reduction by the freshwater isolate decreased with increasing salinity, while reduction by the marine isolate displayed a relatively broad optimum salinity between 20 and 35 ppt. Our results suggest that by participating in an active iron cycle and perhaps by reducing humic acids, iron reducers in the rhizoplane of aquatic macrophytes limit organic availability to other heterotrophs (including methanogens) in the rhizosphere and bulk sediments.     

Isolation and Characterization of Beneficial Bacteria Associated with Citrus Roots in Florida

Cultivable diversity of bacteria associated with citrus was investigated as part of a larger study to understand the roles of beneficial bacteria and utilize them to increase the productive capacity and sustainability of agro-ecosystems. Citrus roots from Huanglongbing (HLB) diseased symptomatic and asymptomatic citrus were used in this study. A total of 227 and 125 morphologically distinct colonies were isolated and characterized from HLB asymptomatic and symptomatic trees, respectively. We observed that the frequency of bacterial isolates possessing various plant beneficial properties was significantly higher in the asymptomatic samples. A total of 39 bacterial isolates showing a minimum of five beneficial traits related to mineral nutrition [phosphate (P) solubilization, siderophore production, nitrogen (N) fixation], development [indole acetic acid (IAA) synthesis], health [production of antibiotic and lytic enzymes (chitinase)], induction of systemic resistance [salicylic acid (SA) production], stress relief [production of 1-amino-cyclopropane-1-carboxylate deaminase] and production of quorum sensing [N-acyl homoserine lactones] signals were characterized. A bioassay using ethidium monoazide (EMA)-qPCR was developed to select bacteria antagonistic to Candidatus Liberibacter asiaticus. Using the modified EMA-qPCR assay, we found six bacterial isolates showing maximum similarity to Paenibacillus validus, Lysinibacillus fusiformis, Bacillus licheniformis, Pseudomonas putida, Microbacterium oleivorans, and Serratia plymutica could significantly reduce the population of viable Ca. L. asiaticus in HLB symptomatic leaf samples. In conclusion, we have isolated and characterized multiple beneficial bacterial strains from citrus roots which have the potential to enhance plant growth and suppress diseases.     

Production of Pectinase and Cellulase by Cladosporium cucumerinum with Dissolved Carbohydrates and Isolated Cell Walls of Cucumber as Carbon Sources

The scab fungus Cladosporium cucumerinum can use pectins and polygalacturonic acid as sole sources of carbon. Cellulose and Ca-polygalacturonate are not available carbon sources for the fungus. When growing on sucrose or pectin, pectinase is produced. In these cases the production of cellulase is insignificant. On a mixture of pectin and carboxymethylcellulose also cellulase is produced. Both pectinase and cellulase are released into the culture filtrate when the fungus grows on cell walls without ionic proteins, whereas only cellulase is released when cell walls with ionic proteins are the carbon source. Pectinase produced by the pathogen can bind to isolated cell walls. The bound pectinase can be extracted with 1 M NaCl from cell walls without ionic proteins, but not from cell walls with ionic proteins. A water-extract or 1 M NaCl-extract of cucumber hypocotyls with visible disease symptoms contains cellulase but no pectinase activity. Lack of pectinase activity in the 1 M NaCl-extract may be due to inhibition by a component that could be extracted by NaCl from the cucumber cell walls.     

兰科植物内生细菌物种多样性及其促生机理研究进展

内生细菌影响兰科植物菌根形成和共生关系的稳定性,在兰科植物的生活史中起着重要作用。内生细菌通过分泌植物激素、采用光合作用、生物固氮或促进矿质营养的循环以及产生铁载体、合成其他活性物质等途径来促进兰科植物生长发育。综述了兰科植物内生细菌物种多样性的研究方法及其对兰科植物的促生机理,基于兰科植物与共生微生物的密切关系,认为内生细菌间、内生细菌与兰科植物菌根真菌间的互作是揭示兰科植物与内生细菌互作机理的重要方向。     

Syndrome of Inappropriate Antidiuretic Hormone Secretion Associated with Coproporphyria: Case Report and Review of Literature

ObjectiveTo remind physicians to consider the hepatic porphyrias in the differential diagnosis of the syndrome of inappropriate antidiuretic hormone secretion.MethodsWe present a case report of a patient seen in the hospital for severe hyponatremia, who was discovered to have the syndrome of inappropriate antidiuretic hormone secretion attributable to coproporphyria. Results of laboratory tests of the patient and her family are presented.ResultsA 54-year-old woman was seen in the hospital because of severe hyponatremia accompanied by generalized seizures. Her serum sodium concentration was 112 mEq/L, with concomitant serum and urine osmolalities of 235 and 639 mOsm/kg, respectively. Renal, thyroid, and adrenal functions were normal. Brain, chest, abdominal, and pelvic imaging studies were negative for occult malignant disease. Urinary excretions of porphobilinogen and aminolevulinic acid were substantially elevated. Results of follow-up urine, plasma, and fecal porphyrin studies were consistent with coproporphyria. Results of porphyrin metabolic studies of the patient’s family showed normal findings in her parents and a minimally increased fecal coproporphyrin concentration and urinary uroporphyrin excretion in her sister.ConclusionAn endocrinology consultation is often requested for patients with hyponatremia. It is important to consider the acute hepatic porphyrias in the differential diagnosis, even though these are rare disorders and the family history may not always be helpful because of the high frequency of asymptomatic carriers. (Endocr Pract. 2007;13:164-168)     

Effects of Bacteria Associated with Pine Wood Nematode (Bursaphelenchus xylophilus) on Development and Egg Production of the Nematode

We tested the effects of four bacterial strains carried on the surface of the pine wood nematode (PWN), Bursaphelenchus xylophilus, on egg hatch, development rate and egg production. Strains GcM5‐1A (Pseudomonas fluorescens) and ZpB1‐2A (P. putida), were strong phytotoxin producers, while strains JnB1B (Pantoea sp.) and AcB1C (Peptostreptococcus asaccharalyticus) did not produce phytotoxins. None of the strains had any effect on egg hatch. GcM5‐1A and ZpB1‐2A promoted egg production, developmental rate, body length and diameter growth in both male and female PWN, whereas JnB1B and AcB1C had no such effects on the nematode. Indeed, the latter two strains completely inhibited egg production of the nematode. The results suggest that GcM5‐1A and ZpB1‐2A may provide PWN with food and/or essential nutrients for development and egg production. These results provide further evidence for our previous finding of a mutualistic symbiosis between the PWN and certain strains of bacteria carried by this nematode ( Zhao et al., 2003, 2005 ).     

The Kinetics of Isocitrate Lyase Formation in Chlorella: Evidence for the Promotion of Enzyme Synthesis by Photophosphorylation

The addition of acetate to aerobic Chlorella pyrenoidosa indarkness was followed by the formations of isocitrate lyaseactiity. After a lag period of 40 minutes the formation proceededat a constant rate. By use of actylamide gel electrophoresisit was shown that the increase in enzyme activity was accompaniedby the formation of a new protein which, after separation byelectrophoresis, contained isocitrate lyase activity. The formationof isocitrate lyase was repressed by glucose; it was repressedby light in the presence of carbon dioxide, but not when DCMUwas added. In light, plus DCMU, isocitrate lyase was formedanaerobically and the capacity for photo-formation of isocitratelyase was saturated at 500 ergs/cm²/sec. In this respect theprocess resembled the photo-conversion of glucose to polysaccharidebut differed from the photo-assimilation of carbon dioxide whichbecame saturated at a heigher light intensity. Monochromaticlight of 706 mµ wavelength supported both isocitrate layseformation and the conversion of glucose to polysaccharide butnot carbon dioxide fixation. It is concluded that ATP generatedby cyclic photophosphorylatin can provide the energy for isocitratelyase synthesis in Chlorella.     

Inhibition of Protease Production of Various Bacteria by Ammonium Salts: Its Effect on Toxin Production and Virulence

Production of protease by many bacteria was found to be inhibited by ammonium salts, and the enzyme production was more sensitive to the salts than was growth of the organisms. Inhibition of protease production by some pathogenic bacteria may result in the recognition of an exotoxin which otherwise would have been digested by the protease. In the case of Pseudomonas aeruginosa, qualitatively different toxicities could be demonstrated in the culture fluids, depending on the presence or absence of protease in such a fluid. The toxicity of the culture in the presence of a high titer of protease may be due primarily to the protease, whereas the toxicity exhibited in the absence of protease could be due to proteinacious exotoxin. Producers of high titers of protease tended to be less virulent in vivo than producers of low titers of the enzyme, which exert their toxicities by a separate exotoxin.