bluuebunny/crossref_metadata_2025_split huggingface dataset:
Coxiella burnetii express type IV secretion system proteins that function similarly to components of the Legionella pneumophila Dot/Icm system
Authors: Zamboni Dario S. , McGrath Susan , Rabinovitch Michel , Roy Craig R. 
Published: July 2003

Summary Coxiella burnetii is an obligate intracellular pathogen that replicates in large endocytic vacuoles. Genomic sequence data indicate that 21 genes encoding products that are similar to components of the Legionella pneumophila Dot/Icm type IV secretion system are located on a contiguous 35 kb region of the Coxiella chromosome. It was found that several dot/icm genes were expressed by Coxiella during host cell infection and that dot/icm gene expression preceded the formation of large replicative vacuoles. To determine whether these genes encode a functional type IV secretion system, we have amplified the Coxiella dotB, icmQ, icmS and icmW genes and produced the encoded proteins in Legionella mutants in which the native copy of each gene had been deleted. The Coxiella dotB, icmS and icmW products restored dot/icm‐dependent growth of Legionella mutants in eukaryotic host cells. The Coxiella IcmQ protein and the Legionella IcmR protein did not interact, which could explain why the Coxiella icmQ gene was unable to restore growth to a Legionella icmQ mutant. Thus, Coxiella encodes functional components of a type IV secretion system expressed in vivo that is mechanistically related to the Legionella Dot/Icm apparatus. These studies suggest that a dot/icm‐related secretion system could play an important role in creating the specialized vacuole that supports Coxiella replication.
Multiple transporters associated with malaria parasite responses to chloroquine and quinine
Authors: Mu Jianbing , Ferdig Michael T. , Feng Xiaorong , Joy Deirdre A. , Duan Junhui , Furuya Tetsuya , Subramanian G. , Aravind L. , Cooper Roland A. , Wootton John C. , Xiong Momiao , Su Xin‐zhuan 
Published: July 2003

SummaryMutations and/or overexpression of various transporters are known to confer drug resistance in a variety of organisms. In the malaria parasite Plasmodium falciparum, a homologue of P‐glycoprotein, PfMDR1, has been implicated in responses to chloroquine (CQ), quinine (QN) and other drugs, and a putative transporter, PfCRT, was recently demonstrated to be the key molecule in CQ resistance. However, other unknown molecules are probably involved, as different parasite clones carrying the same pfcrt and pfmdr1 alleles show a wide range of quantitative responses to CQ and QN. Such molecules may contribute to increasing incidences of QN treatment failure, the molecular basis of which is not understood. To identify additional genes involved in parasite CQ and QN responses, we assayed the in vitro susceptibilities of 97 culture‐adapted cloned isolates to CQ and QN and searched for single nucleotide polymorphisms (SNPs) in DNA encoding 49 putative transporters (total 113 kb) and in 39 housekeeping genes that acted as negative controls. SNPs in 11 of the putative transporter genes, including pfcrt and pfmdr1, showed significant associations with decreased sensitivity to CQ and/or QN in P. falciparum. Significant linkage disequilibria within and between these genes were also detected, suggesting interactions among the transporter genes. This study provides specific leads for better understanding of complex drug resistances in malaria parasites.
The BceRS two‐component regulatory system induces expression of the bacitracin transporter, BceAB, in Bacillus subtilis
Authors: Ohki Reiko , Giyanto  , Tateno Kozue , Masuyama Waka , Moriya Shigeki , Kobayashi Kazuo , Ogasawara Naotake 
Published: July 2003

Summary BceA and bceB encode a nucleotide‐binding domain (NBD) and membrane‐spanning domain (MSD) subunit, respectively, of an ATP‐binding cassette (ABC) transporter in Bacillus subtilis. Disruption of these genes resulted in hypersensitivity to bacitracin, a peptide antibiotic that is non‐ribosomally synthesized in some strains of Bacillus. Northern hybridization analyses showed that expression of the bceAB operon is induced by bacitracin present in the growth medium. The bceRS genes encoding a two‐component regulatory system are located immediately upstream of bceAB. Deletion analyses of the bceAB promoter together with DNase I footprinting experiments revealed that a sensor kinase, BceS, responds to extracellular bacitracin either directly or indirectly and transmits a signal to a cognate response regulator, BceR. The regulator binds directly to the upstream region of the bceAB promoter and upregulates the expression of bceAB genes. The bcrC gene product is additionally involved in bacitracin resistance. The expression of bcrC is dependent on the ECF σ factors, σM and σX, but not on the BceRS two‐component system. In view of these results, possible roles of BceA, BceB and BcrC in bacitracin resistance of B. subtilis 168 are discussed.
An intramolecular disulphide bond reduces the efficacy of a lipoprotein plasma membrane sorting signal
Authors: Robichon Carine , Bonhivers Mélanie , Pugsley Anthony P. 
Published: July 2003

SummaryTo study lipoprotein sorting in Escherichia coli, we devised a novel screen in which sensitivity or resistance to bacteriophage T5 and colicin M reflects the membrane localization of the bacteriophage T5‐encoded lipoprotein Llp, which inactivates the outer membrane (OM) T5 receptor (FhuA). When processed by lipoprotein signal peptidase, Llp has a serine at position +2, immediately after the fatty acylated N‐terminal cysteine. As predicted by the ‘+2 lipoprotein sorting rule’ that determines the localization of lipoproteins in the cell envelope, Llp is located in the OM. However, contrary to expectations, when serine +2 was replaced by aspartate, the canonical plasma membrane lipoprotein retention signal, Llp was still ≥ 40% targeted to the OM and protected cells against colicin M and phage T5. OM association of this Llp derivative was abolished when a peptide spacer was inserted between the aspartate and the rest of Llp or when the formation of an intramolecular disulphide bond in Llp was prevented by substituting one or other of the cysteines involved. Furthermore, analysis of a MalE–Llp hybrid protein with or without a lipid moiety demonstrated that fatty acylation of Llp is essential for its OM association and for protection against colicin M and bacteriophage T5. These data suggest (i) that phage‐encoded Llp uses the endogenous E. coli Lol pathway for lipoprotein sorting to the OM and (ii) that the conformation of a lipoprotein can affect its sorting within the cell envelope.
In vivo magnetic resonance imaging of xylem vessel contents in woody lianas
Authors: CLEARWATER M. J. , CLARK C. J. 
Published: July 2003

ABSTRACTPrevious reports suggest that in some plant species the refilling of embolized xylem vessels can occur while negative pressure exists in the xylem. The aim of this experiment was to use non‐destructive nuclear magnetic resonance imaging (MRI) to study the dynamics of xylem cavitation and embolism repair in‐vivo. Serial 1H‐MRI was used to monitor the contents of xylem vessels in stems of two dicotyledonous (Actinidia deliciosa and Actinidia chinensis, kiwifruit) and one monocotyledonous (Ripogonum scandens, supplejack) species of woody liana. The configuration of the horizontal wide bore magnet and probe allowed the imaging of woody stems up to 20 mm in diameter. Tests using excised stems confirmed that the image resolution of 78 µm and digital image subtraction could be used to detect the emptying and refilling of individual vessels. Imaging was conducted on both intact plants and excised shoots connected to a water supply. In the case of Ripogonum the excised shoots were long enough to allow the distal end of the shoot, including all leaves, to be exposed to ambient conditions outside the building while the proximal end was inside the MRI magnet. In total, six stems were monitored for 240 h while the shoots were subjected to treatments that included light and dark periods, water stress followed by re‐watering, and the covering of all leaves to prevent transpiration. The sudden emptying of water‐filled vessels occurred frequently while xylem water potential was low (below −0.5 MPa for Actinidia, −1.0 MPa for Ripogonum), and less frequently after xylem water potential approached zero at the end of water‐stress treatments. No refilling of empty vessels was observed at any time in any of the species examined. It is concluded that embolism repair under negative pressure does not occur in the species examined here. Embolism repair may be more likely in species with narrower xylem vessels, but further experiments are required with other species before it can be concluded that repair during transpiration is a widespread phenomenon.
Transfer conductance in second growth Douglas‐fir (Pseudotsuga menziesii (Mirb.)Franco) canopies
Authors: WARREN C. R. , ETHIER G. J. , LIVINGSTON N. J. , GRANT N. J. , TURPIN D. H. , HARRISON D. L. , BLACK T. A. 
Published: July 2003

ABSTRACTThe internal conductance from intercellular spaces to the sites of carboxylation (gi) has only been measured in a few tree species and not in conifers, despite the fact it may impose a large limitation on photosynthesis. The present study provides the first estimates of gi for a coniferous species, and examines variation in gi with height and its relationships to anatomical, biochemical and physiological traits. Measurements were made on upper and lower canopy current‐year needles of 50‐year‐old Douglas‐fir (Pseudotsuga menziesii (Mirb.) Franco). Needle thickness and specific leaf area decreased by 30% from the top to bottom of the canopy. These anatomical/morphological changes were accompanied by modest variation in allocation of N to chlorophyll and the chlorophyll a/b ratio. Allocation of N to Rubisco did not vary with height, but the ratio of Rubisco to chlorophyll did owing to the aforementioned changes in allocation to chlorophyll. The value of gi was estimated in one tree from concurrent measurements of carbon isotope discrimination and net photosynthesis. To examine the variation in gi among trees a second independent method based on day respiration and the difference between the chloroplastic and intercellular photocompensation points (photocompensation point method) was used. Estimates of gi obtained by the two methods agreed well with values varying between 0.14 and 0.20 mol m−2 s−1. It is estimated that gi limits photosynthesis by approximately 20% as compared to an approximately 30% stomatal limitation (under well‐watered conditions). The value of gi scaled approximately with maximum rates of photosynthesis, which were significantly greater in upper canopy needles. Nevertheless, gi did not vary significantly with canopy height, owing to greater variability in gi than photosynthesis.
Nitrate photo‐assimilation in tomato leaves under short‐term exposure to elevated carbon dioxide and low oxygen
Authors: SEARLES P. S. , BLOOM A. J. 
Published: July 2003

ABSTRACTThe role of photorespiration in the foliar assimilation of nitrate (NO3–) and carbon dioxide (CO2) was investigated by measuring net CO2 assimilation, net oxygen (O2) evolution, and chlorophyll fluorescence in tomato leaves (Lycopersicon esculentum). The plants were grown under ambient CO2 with ammonium nitrate (NH4NO3) as the nitrogen source, and then exposed to a CO2 concentration of either 360 or 700 µmol mol−1, an O2 concentration of 21 or 2%, and either NO3– or NH4+ as the sole nitrogen source. The elevated CO2 concentration stimulated net CO2 assimilation under 21% O2 for both nitrogen treatments, but not under 2% O2. Under ambient CO2 and O2 conditions (i.e. 360 µmol mol−1 CO2, 21% O2), plants that received NO3– had 11–13% higher rates of net O2 evolution and electron transport rate (estimated from chlorophyll fluorescence) than plants that received NH4+. Differences in net O2 evolution and electron transport rate due to the nitrogen source were not observed at the elevated CO2 concentration for the 21% O2 treatment or at either CO2 level for the 2% O2 treatment. The assimilatory quotient (AQ) from gas exchange, the ratio of net CO2 assimilation to net O2 evolution, indicated more NO3– assimilation under ambient CO2 and O2 conditions than under the other treatments. When the AQ was derived from gross O2 evolution rates estimated from chlorophyll fluorescence, no differences could be detected between the nitrogen treatments. The results suggest that short‐term exposure to elevated atmospheric CO2 decreases NO3– assimilation in tomato, and that photorespiration may help to support NO3– assimilation.
Sugar transport together with environmental conditions controls time lags between xylem and stem diameter changes
Authors: SEVANTO S. , VESALA T. , PERÄMÄKI M. , NIKINMAA E. 
Published: July 2003

ABSTRACTIn the present study the seasonal patterns of time lags between diurnal xylem and whole stem diameter variations at the top and at the base of two Scots pine trees (Pinus sylvestris L.) were compared. The diameter variations were measured during the summers of 2001 and 2002. Time lags were determined using the cross‐correlation method. The lags were found to vary in time according to the different stages of growth. At the top the xylem lagged behind the whole stem between the beginning of stem growth and the end of shoot growth in both years. In 2001 the time lags at the base showed a similar behaviour during stem growth. That kind of seasonal pattern of the time lags would result from the changes in the sink strength due to changing growth rate at different parts of the tree and the differences in the annual rhythm of growth and water availability in the soil (based on precipitation measurements) between the years 2001 and 2002 were reflected in the patterns. The time lags of shrinking and swelling periods during high and low photosynthetic activity (measured using a shoot chamber) were also compared. It was found, for example, that in 2001 in the middle of the growing season at the top of the tree the whole stem lagged on average 15 min more behind the xylem on the days of high photosynthetic activity than on the days of low or moderate. These results show for the first time that the transportation of carbohydrates and variable sink activity could be detected during the growing season in field conditions using stem and xylem diameter variation measurements. Furthermore, these results provide evidence of the pressure gradient‐driven flow also in the phloem of gymnosperms.
Differences in hydraulic architecture account for near‐isohydric and anisohydric behaviour of two field‐grown Vitis vinifera L. cultivars during drought
Authors: SCHULTZ H. R. 
Published: July 2003

ABSTRACTA comparative study on stomatal control under water deficit was conducted on grapevines of the cultivars Grenache, of Mediterranean origin, and Syrah of mesic origin, grown near Montpellier, France and Geisenheim, Germany. Syrah maintained similar maximum stomatal conductance (gmax) and maximum leaf photosynthesis (Amax) values than Grenache at lower predawn leaf water potentials, Ψleaf, throughout the season. The Ψleaf of Syrah decreased strongly during the day and was lower in stressed than in watered plants, showing anisohydric stomatal behaviour. In contrast, Grenache showed isohydric stomatal behaviour in which Ψleaf did not drop significantly below the minimum Ψleaf of watered plants. When g was plotted versus leaf specific hydraulic conductance, Kl, incorporating leaf transpiration rate and whole‐plant water potential gradients, previous differences between varieties disappeared both on a seasonal and diurnal scale. This suggested that isohydric and anisohydric behaviour could be regulated by hydraulic conductance. Pressure‐flow measurements on excised organs from plants not previously stressed revealed that Grenache had a two‐ to three‐fold larger hydraulic conductance per unit path length (Kh) and a four‐ to six‐fold larger leaf area specific conductivity (LSC) in leaf petioles than Syrah. Differences between internodes were only apparent for LSC and were much smaller. Cavitation detected as ultrasound acoustic emissions on air‐dried shoots showed higher rates for Grenache than Syrah during the early phases of the dry‐down. It is hypothesized that the differences in water‐conducting capacity of stems and especially petioles may be at the origin of the near‐isohydric and anisohydric behaviour of g.
Expression of low molecular weight heat‐shock proteins and total antioxidant activity in the Mediterranean tree Quercus ilex L. in relation to seasonal and diurnal changes in physiological parameters
Authors: VERDAGUER D. , ARANDA X. , JOFRÉ A. , EL OMARI B. , MOLINAS M. , FLECK I. 
Published: July 2003

ABSTRACTThe total antioxidant activity (TAA) and the accumulation of small heat shock proteins (sHsps) were analysed under field conditions in Quercus ilex with regard to organ ontogeny and the physiological state of the plant. The results point toward the participation of sHsps and an increase of TAA in the general adaptation syndrome (GAS) of woody Mediterranean evergreens. In leaves and stems, there was a definite TAA seasonal pattern but no effect from diurnal variations or from the current stage of organ ontogeny. TAA was about twice as large in summer as in spring and winter and the hydrophilic antioxidant content was about 16 times greater than that of lipophilic antioxidants. The accumulation of sHsps in leaves also showed a seasonal pattern, but no effect from diurnal variations or from leaf ontogeny. In summer days, the sHsps content remained invariable even during the daylight hours in which the leaves were physiologically recovered. However, the accumulation of sHsps in stems did vary in relation to organ ontogeny. Old stems had a high accumulation of sHsps throughout the year, whereas in young stems, accumulation of sHsps was detected only in summer. This is probably due to a higher quantity of lignified and suberized tissues in the older stems. Using two‐dimensional immunodetection for leaves and stems, two sets of Hs protein species (17 and 10 kDa regions) were observed. In stems, there was an increase in 10 kDa proteins from winter to summer. These results are discussed and the possible role of the two types of polypeptides in the face of environmental and endogenous oxidative stress are considered.
Seasonal detection of pear decline phytoplasma by nested‐PCR in different pear cultivars
Authors: Garcia‐Chapa M. , Medina V. , Viruel M. A. , Laviña A. , Batlle A. 
Published: July 2003

Seasonal detection of pear decline phytoplasma was studied in three pear cultivars: Bartlett, Limonera and Blanquilla. Samples from 43 infected trees were collected monthly over 2 years and analysed by nested PCR. The three cultivars each showed a different pattern of phytoplasma detection. The maximum detection rate of pear decline phytoplasma occurred in December in the three orchards, and it remained high throughout the winter months. In spring, when new buds appeared and sap was produced, the detection rate decreased. Leaf midribs, buds and stems were compared to determine which sample was more reliable for phytoplasma detection. The best indicators were stems. The presence of phytoplasma in sieve tubes during the dormant season was determined by grafting. The results suggest that phytoplasmas could overwinter in shoots, with the implication that vegetative propagation during this period could also disseminate the disease.
Genetic structure of a population of Ganoderma boninense on oil palm
Authors: Pilotti C. A. , Sanderson F. R. , Aitken E. A. B. 
Published: July 2003

Ganoderma boninense (the causal agent of basal stem rot of oil palm in Papua New Guinea) has a tetrapolar mating system with multiple alleles. Investigations into the population structure of G. boninense, using interfertility between isolates as a marker, revealed that the population on oil palm was comprised predominantly of genetically distinct individuals, although a number of isolates were found to share single mating alleles. No direct hereditary relationship was found between isolates on neighbouring or spatially separated diseased palms, indicating that outcrossing had probably occurred over several generations in the founder population prior to colonization of oil palm. In this study, a total of 81 A and 83 B mating type alleles (factors) were detected with 18 allelic repeats at the A locus and 17 at the B locus. Alleles appeared to be randomly dispersed throughout the population in each study block, although there was a significantly (P < 0·01) higher proportion of allelic repeats between estates separated by several kilometres compared to within estates. This is the first study on the population structure of this species and confirms that sexual reproduction is a significant component of the epidemiology of this disease.
Evaluating Pochonia chlamydosporia in a double‐cropping system of lettuce and tomato in plastic houses infested with Meloidogyne javanica
Authors: Verdejo‐Lucas S. , Sorribas F. J. , Ornat C. , Galeano M. 
Published: July 2003

The effect of Pochonia chlamydosporia, a facultative fungal parasite of nematode eggs, alone or in combination with oxamyl was evaluated in a double‐cropping system of lettuce and tomato in unheated plastic houses infested with Meloidogyne javanica at two sites for two consecutive growing seasons. An additional treatment of methyl bromide fumigation was included to compare crop yield in nematode‐free vs. nematode‐infested soil. Final population densities, reproductive rate, root gall rating, and egg production were determined after each crop. Pochonia chlamydosporia was isolated from nematode eggs up to nine months after application to soil. The fungus survived in the rhizosphere for the entire growing season at one site, but only at low densities. Final population densities of M. javanica decreased after cultivation of lettuce and increased after tomato, and this pattern of population fluctuation was unaffected by treatment, experiment or site. The reproductive rate on lettuce was equal to or below 1, and it was similar among treatments in both experiments at both sites. Eggs were not found on lettuce roots. On tomato, the reproductive rate in the fungus + oxamyl treatment was significantly lower (P < 0·05) than other treatments in experiment 1 at both sites. Fungus + oxamyl consistently reduced root gall ratings on tomato in all cases, but numbers of eggs per g root varied depending on treatment. Methyl bromide‐treated plots remained free of M. javanica at the end of the 2‐year study.
Jurassic smectite and kaolinite trends of the East European Platform: implications for palaeobathymetry and palaeoclimate
Authors: Šimkevičius Petras , Ahlberg Anders , Grigelis Algimantas 
Published: July 2003

ABSTRACTClay minerals cannot only give information on sedimentary depositional environment but also the climate regime under which sediment was weathered. The successions of non‐marine to marine Middle and Late Jurassic strata of the East European Platform show upwards increasing contents of smectite and decreasing contents of kaolinite. There is also an increasing smectite vs. kaolinite ratio laterally, in a basinward direction within individual formations. Vertical smectite and kaolinite content curves coincide broadly with the Sahagian and Jones eustatic sea‐level curve. All this implies differential settling of smaller smectite particles offshore and larger kaolinite particles onshore. Smectite formation, preservation and supply were further enhanced as the eastwards expanding Late Jurassic arid zone of Laurasia influenced weathering conditions in the sediment source areas, to the north and east of the East European Platform.