bluuebunny/crossref_metadata_2025_split huggingface dataset:
Revising the Critical Care Family Needs Inventory for the Emergency Department
Authors: Redley Bernice , Beanland Christine 
Published: December 2003

Background.  The Critical Care Family Needs Inventory (CCFNI) has been used widely over the last two decades for analysing the needs of family members in the intensive care unit. However, it has significant limitations as a needs assessment tool for use with families in the Emergency Department (ED). This paper discusses the methodological challenges encountered during the process of reviewing and adapting this tool for use in the ED.Aims.  The purpose of this study was to revise and adapt the CCFNI for use with a population of family members of critically ill patients in an Australian Emergency Department.Instrument.  The process of tool revision, adaptation and reconstruction included: critique of the CCFNI; concept definition; item review; content and structure revision; scale revision; and testing with a sample of the target population.Methods.  Data collection methods were aimed at accessing a vulnerable population, while enhancing response rate and data quality. A sample of 84 relatives of critically ill patients from one Melbourne Metropolitan Emergency Department was used, 73% of whom returned questionnaires.Results.  Pilot data were examined with the specific purpose of identifying elements of the tool that required refinement or modification. Methods used for establishing reliability and validity of the revised tool provided satisfactory results.Limitations.  Limitations of this study include inadequate sample size for exploratory factor analysis, and an incomplete response set for some items, which influenced item analysis.Conclusion.  The process used for addressing the identified methodological issues in reviewing and adapting the CCFNI for use in the ED provides a framework for adapting an established tool for a specific purpose.
Potato virus Y NIa protease activity is not sufficient for elicitation of Ry‐mediated disease resistance in potato
Authors: Mestre Pere , Brigneti Gianinna , Durrant Marcus C. , Baulcombe David C. 
Published: December 2003

Summary Ry confers extreme resistance (ER) to all strains of potato virus Y (PVY). In previous work, we have shown that the protease domain of the nuclear inclusion a protease (NIaPro) from PVY is the elicitor of the Ry‐mediated resistance and that integrity of the protease active site is required for the elicitation of the resistance response. Two possibilities arise from these results: first, the structure of the active protease has elicitor activity; second, NIa‐mediated proteolysis is required to elicit the resistance response. To resolve these possibilities, the NIaPro from PVY was randomly mutagenised and the clones obtained were screened for elicitation of cell death as an indicator of resistance and proteolytic activity. We did not find any mutants that had retained the ability to elicit cell death but had lost protease activity, as measured by processing of the NIa cleavage site in the viral genome. This was consistent with the idea that protease activity is necessary for elicitor activity. However, protease activity was not sufficient because we found three elicitor‐defective mutants in which there was a high level of protease activity in this assay.
Anionic lipids are required for chloroplast structure and function in Arabidopsis
Authors: Yu Bin , Benning Christoph 
Published: December 2003

SummaryPhotosynthetic membranes of plants primarily contain non‐phosphorous glycolipids. The exception is phosphatidylglycerol (PG), which is an acidic/anionic phospholipid. A second major anionic lipid in chloroplasts is the sulfolipid sulfoquinovosyldiacylglycerol (SQDG). It is hypothesized that under severe phosphate limitation, SQDG substitutes for PG, ensuring a constant proportion of anionic lipids even under adverse conditions. A newly constructed SQDG and PG‐deficient double mutant supports this hypothesis. This mutant, sqd2 pgp1‐1, carries a T‐DNA insertion in the structural gene for SQDG synthase (SQD2) and a point mutation in the structural gene for phosphatidylglycerolphosphate synthase (PGP1). In the sqd2 pgp1‐1 double mutant, the fraction of total anionic lipids is reduced by approximately one‐third, resulting in pale yellow cotyledons and leaves with reduced chlorophyll content. Photoautotrophic growth of the double mutant is severely compromised, and its photosynthetic capacity is impaired. In particular, photosynthetic electron transfer at the level of photosystem II (PSII) is affected. Besides these physiological changes, the mutant shows altered leaf structure, a reduced number of mesophyll cells, and ultrastructural changes of the chloroplasts. All observations on the sqd2 pgp1‐1 mutant lead to the conclusion that the total content of anionic thylakoid lipids is limiting for chloroplast structure and function, and is critical for overall photoautotrophic growth and plant development.
Synthesis of ketocarotenoids in the seed of Arabidopsis thaliana
Authors: Stålberg Kjell , Lindgren Ove , Ek Bo , Höglund Anna‐Stina 
Published: December 2003

SummaryA cDNA coding for a gene necessary for synthesis of ketocarotenoids was cloned from the alga Haematococcus pluvialis and expressed in the seed of Arabidopsis thaliana. The expression of the algal β‐carotene‐oxygenase gene was directed to the seed by use of the 2S, seed storage protein promoter napA. Extracts from seeds of the transgenic plants were clearly red because of accumulation of ketocarotenoids, and free and esterified forms of ketocarotenoids were found in addition to the normal carotenoid composition in the seed. The major ketocarotenoids in the transgenic plants were: 4‐keto‐lutein (3,3′‐dihydroxy‐β‐,ε‐carotene‐4‐one), adonirubin (3‐hydroxy‐β‐,β′‐carotene‐4,4′‐dione) and canthaxanthin (β‐,β′‐carotene‐4,4′‐dione). 4‐Keto‐lutein differs from the more common adonixanthin only in the position of one double bond. To increase the substrate availability for the β‐carotene‐oxygenase, these transformants were crossed with transgenic plants overexpressing a construct of an endogenous phytoene synthase gene, also under the control of the napA promoter. The resulting crossings gave rise to seeds with a 4.6‐fold relative increase of the total pigment, and the three major ketocarotenoids were increased 13‐fold compared to seeds of transgenic plants carrying only the β‐carotene‐oxygenase construct.
Antisense LOX expression increases herbivore performance by decreasing defense responses and inhibiting growth‐related transcriptional reorganization in Nicotiana attenuata
Authors: Halitschke Rayko , Baldwin Ian T. 
Published: December 2003

SummaryInhibition of jasmonic acid (JA) signaling has been shown to decrease herbivore resistance, but the responsible mechanisms are largely unknown because insect resistance is poorly understood in most model plant systems. We characterize three members of the lipoxygenase (LOX) gene family in the native tobacco plant Nicotiana attenuata and manipulate, by antisense expression, a specific, wound‐ and herbivory‐induced isoform (LOX3) involved in JA biosynthesis. In three independent lines, antisense expression reduced wound‐induced JA accumulation but not the release of green leaf volatiles (GLVs). The impaired JA signaling reduced two herbivore‐induced direct defenses, nicotine and trypsin protease inhibitors (TPI), as well as the potent indirect defense, the release of volatile terpenes that attract generalist predators to feeding herbivores. All these defenses could be fully restored by methyl‐JA (MeJA) treatment, with the exception of the increase in TPI activity, which was partially restored, suggesting the involvement of additional signals. The impaired ability to produce chemical defenses resulted in lower resistance to Manduca sexta attack, which could also be restored by MeJA treatment. Expression analysis using a cDNA microarray, specifically designed to analyze M. sexta‐induced gene expression in N. attenuata, revealed a pivotal role for LOX3‐produced oxylipins in upregulating defense genes (protease inhibitor, PI; xyloglucan endotransglucosylase/hydrolase, XTH; threonine deaminase, TD; hydroperoxide lyase, HPL), suppressing both downregulated growth genes (RUBISCO and photosystem II, PSII) and upregulated oxylipin genes (α‐dioxygenase, α‐DOX). By genetically manipulating signaling in a plant with a well‐characterized ecology, we demonstrate that the complex phenotypic changes that mediate herbivore resistance are controlled by a specific part of the oxylipin cascade.
The ethanol switch: a tool for tissue‐specific gene induction during plant development
Authors: Deveaux Yves , Peaucelle Alexis , Roberts Gethin R. , Coen Enrico , Simon Rudiger , Mizukami Yukiko , Traas Jan , Murray James A. H. , Doonan John H. , Laufs Patrick 
Published: December 2003

SummaryControlled gene expression in time and space is a powerful tool for the analysis of gene function during plant development. Here, we report ethanol inducible gene expression in defined sub‐domains of the shoot apical and floral meristems. For this, expression of an ethanol‐regulated transcription factor, ALCR, is restricted to precise domains using specific promoters. Gene expression activation is followed using reporters under the control of the alcA promoter, which responds to ALCR only in the presence of the ethanol. We demonstrate that precise control of spatially limited gene expression can be achieved. The kinetics of reporter gene activation and inactivation following a pulse of ethanol induction shows that the system is dynamic and suitable for precise temporal control of expression. The system is both flexible and robust, permitting simultaneous expression of two genes in a given domain or, conversely, the expression of a gene in two separate domains. We also show that this strategy can be applied to mis‐express genes with developmental roles, by manipulating expression of the SHOOT MERISTEMLESS (STM) and CYCLIN D3;1 (CYCD3;1) genes during plant development.
The NOMEGA gene required for female gametophyte development encodes the putative APC6/CDC16 component of the Anaphase Promoting Complex in Arabidopsis
Authors: Kwee Hong‐Sien , Sundaresan Venkatesan 
Published: December 2003

SummaryDevelopment of the female gametophyte involves several rounds of nuclear divisions during which nuclei are rearranged and finally cellularized to form a mature seven‐celled embryo sac. During these nuclear divisions, key proteins involved in the cell cycle need to be degraded quickly in order to facilitate both the metaphase–anaphase transition stage and late anaphase. Here, we report the characterization of an Arabidopsis mutant nomega, which results in arrest of the embryo sac development at the two‐nucleate stage. The NOMEGA gene product shows high homology to the APC6/cell division cycle (CDC)16 subunit of the Anaphase Promoting Complex/Cyclosome (APC/C). The phenotype of the nomega mutant is quite different from that of the hobbit mutant, which had suggested a role for the plant APC/C in auxin signalling. We show that nomega mutant embryo sacs are unable to degrade Cyclin B, an important APC/C substrate, providing further evidence of a role for the NOMEGA gene product and the plant APC/C in cell cycle progression during gametophyte development.
(R,S)‐Reticuline 7‐O‐methyltransferase and (R,S)‐norcoclaurine 6‐O‐methyltransferase of Papaver somniferum – cDNA cloning and characterization of methyl transfer enzymes of alkaloid biosynthesis in opium poppy
Authors: Ounaroon Anan , Decker Gabriele , Schmidt Jürgen , Lottspeich Friedrich , Kutchan Toni M. 
Published: December 2003

Summary S‐Adenosyl‐l‐methionine:(R,S)‐reticuline 7‐O‐methyltransferase converts reticuline to laudanine in tetrahydrobenzylisoquinoline biosynthesis in the opium poppy Papaver somniferum. This enzyme activity has not yet been detected in plants. A proteomic analysis of P. somniferum latex identified a gel spot that contained a protein(s) whose partial amino acid sequences were homologous to those of plant O‐methyltransferases. cDNA was amplified from P. somniferum RNA by reverse transcription PCR using primers based on these internal amino acid sequences. Recombinant protein was then expressed in Spodoptera frugiperda Sf9 cells in a baculovirus expression vector. Steady‐state kinetic measurements with one heterologously expressed enzyme and mass spectrometric analysis of the enzymatic products suggested that this unusual enzyme is capable of carrying through sequential O‐methylations on the isoquinoline and on the benzyl moiety of several substrates. The tetrahydrobenzylisoquinolines (R)‐reticuline (4.2 sec−1 mm−1), (S)‐reticuline (4.5 sec−1 mm−1), (R)‐protosinomenine (1.7 sec−1 mm−1), and (R,S)‐isoorientaline (1.4 sec−1 mm−1) as well as guaiacol (5.9 sec−1 mm−1) and isovanillic acid (1.2 sec−1 mm−1) are O‐methylated by the enzyme with the ratio kcat/K m shown in parentheses. A P. somniferum cDNA encoding (R,S)‐norcoclaurine 6‐O‐methyltransferase was similarly isolated and characterized. This enzyme was less permissive, methylating only (R,S)‐norcoclaurine (7.4 sec−1 mm−1), (R)‐norprotosinomenine (4.1 sec−1 mm−1), (S)‐norprotosinomenine (4.0 sec−1 mm−1) and (R,S)‐isoorientaline (1.0 sec−1 mm−1). A phylogenetic comparison of the amino acid sequences of these O‐methyltransferases to those from 28 other plant species suggests that these enzymes group more closely to isoquinoline biosynthetic O‐methyltransferases from Coptis japonica than to those from Thalictrum tuberosum that can O‐methylate both alkaloid and phenylpropanoid substrates.
Enhancement of stress tolerance in transgenic tobacco plants overexpressing Chlamydomonas glutathione peroxidase in chloroplasts or cytosol
Authors: Yoshimura Kazuya , Miyao Kazuhiro , Gaber Ahmed , Takeda Toru , Kanaboshi Haruo , Miyasaka Hitoshi , Shigeoka Shigeru 
Published: December 2003

SummaryTo evaluate the physiological potential of the defense system against hydroperoxidation of membrane‐lipid components caused by environmental stresses in higher plants, we generated transgenic tobacco plants expressing a glutathione peroxidase (GPX)‐like protein in the cytosol (TcGPX) or chloroplasts (TpGPX). The activities toward α‐linolenic acid hydroperoxide in TcGPX and TpGPX plants were 47.5–75.3 and 32.7–42.1 nmol min−1 mg−1 protein, respectively, while no activity was detected in wild‐type plants. The transgenic plants showed increased tolerance to oxidative stress caused by application of methylviologen (MV: 50 µm) under moderate light intensity (200 µE m−2 sec−1), chilling stress under high light intensity (4°C, 1000 µE m−2 sec−1), or salt stress (250 mm NaCl). Under these stresses, the lipid hydroperoxidation (the production of malondialdehyde (MDA)) of the leaves of TcGPX and TpGPX plants was clearly suppressed compared with that of wild‐type plants. Furthermore, the capacity of the photosynthetic and antioxidative systems in the transgenic plants remained higher than those of wild‐type plants under chilling or salt stress. These results clearly indicate that a high level of GPX‐like protein in tobacco plants functions to remove unsaturated fatty acid hydroperoxides generated in cellular membranes under stress conditions, leading to the maintenance of membrane integrity and increased tolerance to oxidative stress caused by various stress conditions.
The tomato resistance protein Bs4 is a predicted non‐nuclear TIR‐NB‐LRR protein that mediates defense responses to severely truncated derivatives of AvrBs4 and overexpressed AvrBs3
Authors: Schornack Sebastian , Ballvora Agim , Gürlebeck Doreen , Peart Jack , Ganal Martin , Baker Barbara , Bonas Ulla , Lahaye Thomas 
Published: December 2003

SummaryThe Lycopersicon esculentum Bs4 resistance (R) gene specifies recognition of Xanthomonas campestris pv. vesicatoria (Xcv) strains that express the cognate AvrBs4 avirulence protein. Bs4 was isolated by positional cloning and is predicted to encode a nucleotide‐binding leucine‐rich repeat (NB‐LRR) protein that is homologous to tobacco N and potato Y‐1 resistance proteins. Xcv infection tests demonstrate that Bs4 confers perception of AvrBs4 but not the 97% identical AvrBs3 protein. However, when delivered via Agrobacterium T‐DNA transfer, both, avrBs4 and avrBs3 trigger a Bs4‐dependent hypersensitive response, indicating that naturally occurring AvrBs3‐homologues provide a unique experimental platform for molecular dissection of recognition specificity. Transcript studies revealed intron retention in Bs4 transcripts. Yet, an intron‐deprived Bs4 derivative still mediates AvrBs4 detection, suggesting that the identified splice variants are not crucial to resistance. The L. pennellii bs4 allele, which is >98% identical to L. esculentum Bs4, has a Bs4‐like exon‐intron structure with exception of a splice polymorphism in intron 2 that causes truncation of the predicted bs4 protein. To test if the receptor‐ligand model is a valid molecular description of Bs4‐mediated AvrBs4 perception, we conducted yeast two‐hybrid studies. However, a direct interaction was not observed. Defense signaling of the Bs4‐governed reaction was studied in Nicotiana benthamiana by virus‐induced gene silencing and showed that Bs4‐mediated resistance is EDS1‐ and SGT1‐dependent.
The FUS3 transcription factor functions through the epidermal regulator TTG1 during embryogenesis in Arabidopsis
Authors: Tsuchiya Yuichiro , Nambara Eiji , Naito Satoshi , McCourt Peter 
Published: December 2003

SummaryLoss‐of‐function mutations in the FUSCA3 (FUS3) gene of Arabidopsis result in alterations in cotyledon identity, inability to complete late seed maturation processes, and the premature activation of apical and root embryonic meristems, which indicates that this transcription factor is an essential regulator of embryogenesis. Although FUS3 shows a complex pattern of expression in the embryo, this gene is only required in the protoderm to carry out its functions. Moreover, the epidermal morphogenesis regulator TRANSPARENT TESTA GLABRA1 (TTG1) is negatively regulated by FUS3 in the embryo. When a loss‐of‐function ttg1 mutation is introduced into a fus3 mutant, a number of fus3‐related phenotypes are rescued, indicating a functional TTG1 gene is required to manifest the fus3 mutant phenotype. It therefore appears that one of the functions of FUS3 is to restrict the domain of expression of TTG1 during embryogenesis. The FUS3–TTG1 interaction is both maternal and zygotic, suggesting a complex relationship is required between these gene products to allow correct seed development.
TRANSPARENT TESTA 19 is involved in the accumulation of both anthocyanins and proanthocyanidins in Arabidopsis
Authors: Kitamura Satoshi , Shikazono Naoya , Tanaka Atsushi 
Published: December 2003

SummaryFlavonoid compounds such as anthocyanins and proanthocyanidins (PAs; so‐called condensed tannins) have a multitude of functions in plants. They must be transported from the site of synthesis in the cytosol to their final destination, the vacuoles. Three models have been proposed for sequestering anthocyanins in vacuoles, but the transport machinery for PAs is poorly understood. Novel Arabidopsis mutants, transparent testa 19 (tt19), which were induced by ion beam irradiation, showed a great reduction of anthocyanin pigments in the vegetative parts as well as brown pigments in the seed coat. The TT19 gene was isolated by chromosome walking and a candidate gene approach, and was shown to be a member of the Arabidopsis glutathione S‐transferase (GST) gene family. Heterologous expression of a putative ortholog, petunia anthocyanin 9 (AN9), in tt19 complemented the anthocyanin accumulation but not the brown pigmentation in the seed coat. This suggests that the TT19 gene is required for vacuolar uptake of anthocyanins into vacuoles, but that it has also a function different from that of AN9. The depositional pattern of PA precursors in the mutant was different from that in the wild type. These results indicate that TT19 participates in the PA pathway as well as the anthocyanin pathway of Arabidopsis. As involvement of GST in the PA pathway was previously considered unlikely, the function of TT19 in the PA pathway is also discussed in the context of the putative transporter for PA precursors.
Two MAPK cascades, NPR1, and TGA transcription factors play a role in Pto‐mediated disease resistance in tomato
Authors: Ekengren Sophia K. , Liu Yule , Schiff Michael , Dinesh‐Kumar S. P. , Martin Gregory B. 
Published: December 2003

SummaryThe tomato Pto kinase confers resistance to the causative agent of bacterial speck disease, Pseudomonas syringae pv. tomato, by recognizing the pathogen effector proteins AvrPto or AvrPtoB. Pto‐mediated resistance requires multiple signal transduction pathways and has been shown to activate many defense responses including an oxidative burst, rapid changes in the expression of over 400 genes, and localized cell death. We have tested the role in Pto‐mediated resistance in tomato of a set of 21 genes from other species known to be involved in defense‐related signaling. Expression of each gene was suppressed by virus‐induced gene silencing (VIGS) and the effect on disease symptoms and bacterial growth during the tomato–Pseudomonas incompatible interaction was determined. We found that Pto‐mediated resistance was compromised by silencing of genes encoding two mitogen‐activated protein (MAP) kinase kinases, MEK1 and MEK2, two MAP kinases, NTF6 and wound‐induced protein kinase (WIPK), a key regulator of systemic acquired resistance (SAR), NPR1, and two transcription factors, TGA1a and TGA2.2. A lesser impact on Pto‐mediated resistance was observed in plants silenced for RAR1 and COI1. The identification of nine genes that play a role in resistance to bacterial speck disease both advances our knowledge of Pto signal transduction and demonstrates the conservation of many defense signaling components among diverse plant species.
A new method for rapid visualization of defects in leaf cuticle reveals five intrinsic patterns of surface defects in Arabidopsis
Authors: Tanaka Toshihiro , Tanaka Hirokazu , Machida Chiyoko , Watanabe Masaru , Machida Yasunori 
Published: December 2003

SummaryThe epidermis of higher plants generates the cuticle layer that covers the outer surface of each plant. The cuticle plays a crucial role in plant development, and some mutants with defective cuticle exhibit morphological abnormalities, such as the fusion of organs. The way in which the cuticle forms and its contribution to morphogenesis are poorly understood. Conventional detection of the cuticle by transmission electron microscopy (TEM) requires laborious procedures, which include fixation, staining with osmium, and preparation of ultra‐thin sections. It is also difficult to survey entire surfaces of expanded leaves because of the limited size of specimens that can be examined. Thus, TEM is unsuitable for large‐scale screening for mutants with defective cuticle. We describe here a rapid and inexpensive method, designated the toluidine‐blue (TB) test, for detection of cuticular defects in whole leaves. We demonstrated the validity of the TB test using mutants of Arabidopsis thaliana, including abnormal leaf shape1 (ale1), fiddlehead (fdh), and five eceriferum (cer) mutants, in which the structure and/or function of the cuticle is abnormal. Genetic screening for mutants using the TB test allowed us to identify seven loci. The cuticle‐defective regions of leaves of the mutants revealed five intrinsic patterns of surface defects (classes I through V), suggesting that formation of functional cuticle on leaves involves various spatially regulated factors.