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Gramene, a tool for grass genomics.

Gramene (http://www.gramene.org) is a comparative genome mapping database for grasses and a community resource for rice (Oryza sativa). It combines a semi-automatically generated database of cereal genomic and expressed sequence tag sequences, genetic maps, map relations, and publications, with a curated database of rice mutants (genes and alleles), molecular markers, and proteins. Gramene curators read and extract detailed information from published sources, summarize that information in a structured format, and establish links to related objects both inside and outside the database, providing seamless connections between independent sources of information. Genetic, physical, and sequence-based maps of rice serve as the fundamental organizing units and provide a common denominator for moving across species and genera within the grass family. Comparative maps of rice, maize (Zea mays), sorghum (Sorghum bicolor), barley (Hordeum vulgare), wheat (Triticum aestivum), and oat (Avena sativa) are anchored by a set of curated correspondences. In addition to sequence-based mappings found in comparative maps and rice genome displays, Gramene makes extensive use of controlled vocabularies to describe specific biological attributes in ways that permit users to query those domains and make comparisons across taxonomic groups. Proteins are annotated for functional significance using gene ontology terms that have been adopted by numerous model species databases. Genetic variants including phenotypes are annotated using plant ontology terms common to all plants and trait ontology terms that are specific to rice. In this paper, we present a brief overview of the search tools available to the plant research community in Gramene.

Avena↗

Genotype x environment interactions for postweaning performance in crossbred calves grazing winter wheat pasture or dormant native prairie.

Data from 403 calves from Angus, Brahman, and reciprocal-cross cows sired by Polled Hereford bulls were used to evaluate the impact of postweaning backgrounding forages on postweaning BW, gains, and carcass traits. Calves were born (spring of 1991 through 1994) and reared on either endophyte-infected tall fescue or common bermudagrass pastures. After weaning, calves were transported 360 km to the Grazinglands Research Laboratory, west of El Reno, OK, and, within breed and preweaning forage, were assigned to one of the following winter stocker treatments: 1) winter wheat pasture or 2) dormant native prairie plus supplemental CP. In March, winter stocker treatments were ended and calves were grazed as a single group on cool-season grasses until early July (1992, 1993, and 1994) or late May (1995), when the feedlot phase began. In the feedlot, calves were fed a high concentrate diet for an average of 120 d until a backfat thickness of > 10 mm was reached. Calves were shipped in truck load lots to Amarillo, TX (350 km), for processing and collection of carcass data. Averaged over calf breed group, calves wintered on wheat pasture gained faster (P < 0.01) during the stocker phase (0.71 vs 0.43 kg); had heavier (P < 0.01) final feedlot weights (535 vs 512 kg); lower feedlot (P < 0.01) ADG (1.37 vs 1.53 kg); heavier (P < 0.01) carcass weights (337 vs 315 kg); larger (P < 0.01) longissimus muscle (84.9 vs 81.8 cm2); higher percentage (P < 0.01) of kidney, heart, and pelvic fat (2.32 vs 2.26); and higher (P < 0.01) dressing percentage (62.2 vs 61.3) than calves backgrounded on native prairie. Maternal heterosis for stocker ADG was evident in calves backgrounded on native prairie but not on winter wheat (P < 0.10), but the two environments were similar in maternal heterosis for feedlot ADG and carcass traits. Calves wintered on native prairie were restricted in growth and expressed compensatory gain during the feedlot phase but not during the spring stocker phase. Dormant native grasses can be used to winter stocker calves excess to the winter wheat pasture needs, but ownership of these calves would have to be retained through the feedlot phase to realize any advantage of built-in compensatory gain. Finally, these data suggest that expression of maternal heterosis for weight gain is more likely in calves backgrounded on native prairie than in calves grazed on winter wheat.

Animal Feed↗

Bacillus species associated with wheat and sorghum dusts from combine harvester.

14 species and one species variety belonging to genus Bacillus were collected from wheat and sorghum dusts of combine harvester on either nutrient agar (NA) or sheep blood agar (SBA) at 30 degrees C (14 spp.), 45 degrees C (13 spp.) and 55 degrees C (5 spp.). The flora of Bacillus on both substrates and on the two isolation media were basically similar and the most common species were B. brevis, B. cereus, B. circulans, B. coagulans, B. licheniformis, B. stearothermophilus and B. subtilis. The total viable count of airborne bacteria in the two atmospheres were lower in wheat than in sorghum hay sites. The percentage count of haemolytic bacteria on SBA comprised 10% and 15% of the total viable count, respectively.

Air Microbiology↗

CPRF4a, a novel plant bZIP protein of the CPRF family: comparative analyses of light-dependent expression, post-transcriptional regulation, nuclear import and heterodimerisation.

Several DNA-binding proteins with conserved basic region/leucine zipper domains (bZIP) have been isolated from parsley. They all recognise defined ACGT-containing elements (ACEs), including ACE(PcCHSII) in the Light Regulatory Unit LRU1 of the CHS promoter which confers light responsiveness. A new member of this Common Plant Regulatory Factor (CPRF) family, designated CPRF4a, has been cloned, which displays sequence similarity to HBP-1a from wheat, as well as to other plant bZIP proteins. CPRF4a specifically binds as a homodimer to ACE(PcCHSII) and forms heterodimers with CPRF1 but not with CPRF2. In adult parsley plants, CPRF2 and CPRF4a mRNAs are found in all tissues and organs in which the chalcone synthase gene CHS is expressed. In protoplasts from suspension cultured cells, UV irradiation (290-350 nm) did not cause an increase in levels of CPRF1, CPRF2, or CPRF4a mRNA, whereas the corresponding CPRF proteins accumulated within 15 min of light treatment. Furthermore, the rapid light-mediated increase of CPRF proteins was insensitive to transcriptional inhibitors, suggesting that a post-transcriptional mechanism controls CPRF accumulation. CPRFs as well as Arabidopsis thaliana G-box binding factors (GBFs) are selectively transported from the cytosol into the nucleus, as shown in an in vitro nuclear transport system prepared from evacuolated parsley protoplasts, indicating that cytosolic compounds are involved in regulated nuclear targeting of plant bZIP factors.

Acyltransferases↗

The relative infectivities and genomic characterisation of three distinct mastreviruses from South Africa.

The genomic nucleotide sequences of the cloned agroinfectious genomes of three South African mastreviruses obtained from Zea mays, a Setaria sp., and Panicum maximum (designated MSV-Kom, MSV-Set, and PanSV-Kar respectively), were determined. Additionally, their relative infectivities and virulence were analysed in a range of differentially susceptible wheat, maize, and barley genotypes. MSV-Kom produced moderate to severe streak symptoms in all maize genotypes tested, but only moderate to very mild symptoms in the wheat and barley genotypes. MSV-Set infected only the susceptible to tolerant maize genotypes, but was generally more severe in the barley and wheat genotypes than MSV-Kom. PanSV-Kar was incapable of infecting any of the wheat and barley genotypes and only produced very mild symptoms on the three most sensitive maize genotypes. Genomic characteristics in common with related mastreviruses were identified. Phylogenetic analysis indicated that while MSV-Kom was closely related to previously sequenced MSV isolates, MSV-Set and PanSV-Kar represented distinctly novel strains of MSV and PanSV respectively. In the case of MSV-Set, this is the most distantly related MSV strain yet characterised.

Animals↗

Workshop on trichothecenes with a focus on DON: summary report.

A number of mycotoxins of the class of trichothecenes are produced by a variety of Fusarium fungi commonly found on cereals. Unfavourable weather conditions may lead to a high level of Fusarium infections in crops such as wheat and correspondingly high trichothecene contents. The ILSI Europe Natural Toxin Task Force therefore organised a workshop on trichothecenes with a special focus on deoxynivalenol (DON). A number of experts reviewed the current knowledge on trichothecenes with respect to occurrence, including aspects of mould growth, toxin formation, storage and effects of processing; prevention; analytical methodologies, including sampling; surveillance and exposure assessments; and toxicology and risk assessment. A number of recommendations were given under the headings: prevention, sampling and analytical methods, exposure assessment, and toxicology. Gaps in knowledge were also identified.

Animals↗

A lipid transfer protein binds to a receptor involved in the control of plant defence responses.

Lipid transfer proteins (LTPs) and elicitins are both able to load and transfer lipidic molecules and share some structural and functional properties. While elicitins are known as elicitors of plant defence mechanisms, the biological function of LTP is still an enigma. We show that a wheat LTP1 binds with high affinity sites. Binding and in vivo competition experiments point out that these binding sites are common to LTP1 and elicitins and confirm that they are the biological receptors of elicitins. A mathematical analysis suggests that these receptors could be represented by an allosteric model corresponding to an oligomeric structure with four identical subunits.

Algal Proteins↗

Mite-contaminated foods as a cause of anaphylaxis.

BACKGROUND: Although insect and arthropod contamination of certain foods has been recognized for many years, allergic manifestations caused by ingestion of mite allergens have only rarely been reported. OBJECTIVE: The purpose of this study is to present clinical observations in patients who experienced acute anaphylaxis after eating mite-contaminated foods. METHODS: Thirty atopic subjects who were first seen with systemic anaphylaxis precipitated by the ingestion of wheat-containing foods underwent skin prick tests with inhalant and food extracts, as well as with uncontaminated and mite-contaminated wheat flour. Flour samples were examined microscopically for identification and counting of mites. Der p 1 and Der f 1 levels were quantitated by using immunochemical methods. RESULTS: The most common symptoms were breathlessness, angioedema, wheezing, and rhinorrhea, which started between 10 and 240 minutes after eating. Abundant mites were present in the flour obtained from 28 patients; Suidasia spp. mites were found in grated bread from the other two patients. Positive prick test responses to Dermatophagoides farinae-and mite-contaminated flour and negative skin test responses to wheat extract, other food extracts, and uncontaminated wheat flour were found in all patients. Skin test responses were positive in volunteers with mite allergy even after heating the mite-contaminated flour at 100 degrees C. Screening of 35 unselected flour samples demonstrated the presence of mites in 13 of them (37.1%). CONCLUSIONS: Systemic anaphylaxis can occur after the ingestion of heated or unheated mite-contaminated foods. This problem may be more prevalent in tropical and subtropical countries than previously recognized.

Adolescent↗

Sudden infant death syndrome: a search for allergen hypersensitivity.

BACKGROUND: Sudden infant death syndrome (SIDS) remains a diagnosis by exclusion which leaves few if any pathologic clues to its etiology. Previous evaluations for anaphylaxis in SIDS have been few and limited. OBJECTIVE: To analyze forensic blood specimens for evidence of anaphylaxis in 51 (43 boys and 8 girls) children dying of SIDS and 13 (9 boys and 4 girls) age-matched controls who died from defined, nonanaphylactic causes. METHODS: Specimens collected over a 5-year period were assayed for (1) total IgE (IU/mL) by immunoenzymatic assay; (2) latex, cat, dust mite (Dermatophagoides farinae and Dermatophagoides pteronyssinus), milk, soy, wheat, peanuts, egg, and tomato specific-IgE by RAST; and (3) serum tryptase levels (U/L) by radioimmunoassay. RESULTS: The 51 SIDS cases (median age 3 months; range 1 to 9 months) and 13 control cases (median age 4 months; range 1 to 11 months) demonstrated similar total IgE of 9.8 +/- 1.1 IU/mL (mean +/- SEM) and 10.9 +/- 2.8 IU/mL (P = .59). The frequency of detectable (> 0.5 U/L) serum tryptase levels among SIDS cases (10/51) was similar to controls (3/13, P = .72). The frequency of positive RAST tests was 39% (20/51) in SIDS and 38% (5/13) in control subjects (P = .99). Differences in frequencies of positive RAST tests in SIDS and control cases were not statistically significant for any allergen tested. The most frequently detected allergen-specific IgE, to milk, was similar in SIDS (22%) and controls (31%, P = .48). CONCLUSIONS: Elevated tryptase levels and allergen-specific IgE (milk, soy, wheat, peanuts, egg, tomato, dust mites, cat, and latex) were demonstrated in some infant SIDS deaths but were no more common than in controls. We conclude that anaphylaxis is probably an uncommon etiology for SIDS.

Allergens↗

Accumulation of HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine) in indigenous and agricultural plants grown in HMX-contaminated anti-tank firing-range soil.

To investigate their potential for phytoremediation, selected agricultural and indigenous terrestrial plants were examined fortheir capacity to accumulate and degrade the explosive octahydro-1 ,3,5,7-tetra nitro-1,3,5,7-tetrazocine (HMX). Plant tissue and soil extracts were analyzed for the presence of HMX and possible degradative metabolites using high-performance liquid chromatography with diode-array UV detection (HPLC-UV), micellar electrokinetic chromatography with diode-array UV detection (MEKC-UV), and HPLC with electrospray ionization mass spectrometry (LC-MS). The pattern of HMX accumulation for alfalfa (Medicago sativa), bush bean (Phaseolus vulgaris), canola (Brassica rapa), wheat (Triticum aestivum), and perennial ryegrass (Loliumperenne) grown in a controlled environment on contaminated soil from an anti-tank firing range was similar to that observed for plants (wild bergamot (Monarda fistulosa), western wheat grass (Agropyron smithii), brome grass (Bromus sitchensis), koeleria (Koeleria gracilis), goldenrod (Solidago sp.), blueberry (Vaccinium sp.), anemone (Anemone sp.), common thistle (Circium vulgare), wax-berry (Symphoricarpos albus), western sage (Artemisia gnaphalodes), and Drummond's milk vetch (Astragalus drummondii)) collected from the range. No direct evidence of plant-mediated HMX (bio)chemical transformation was provided by the available analytical methods. Traces of mononitroso-HMX were found in contaminated soil extracts and were also observed in leaf extracts. The dominant mechanism for HMX translocation and accumulation in foliar tissue was concluded to be aqueous transpirational flux and evaporation. The accumulation of HMX in the leaves of most of the selected species to levels significantly above soil concentration is relevant to the assessment of both phytoremediation potential and environmental risks.

Azocines↗

Identification of hypophysiotropic luteinizing hormone-releasing hormone (LHRH) neurons by combined retrograde labeling and immunocytochemistry.

The decapeptide luteinizing hormone-releasing hormone (LHRH) is produced by telencephalic and diencephalic neurons and transported to the median eminence (ME). After having been released from nerve terminals, it is carried by the hypophysial portal vessels to the anterior pituitary, where it stimulates the production and release of luteinizing hormone (LH) and follicle stimulating hormone (FSH). Those LHRH neurons which project to the ME represent the final common pathway for the regulation of the pituitary/gonadal axis. We identified these neurons by injecting a retrograde tracer, the lectin wheat germ agglutinin (WGA), into the external zone of the ME. Eight to 24 hours later colchicine was given into the lateral ventricle and 24-48 hours after the WGA injection the animals were sacrificed. Vibratome sections of the brains were stained simultaneously for WGA and LHRH with a dual immunocytochemical technique. Approximately 70% of the LHRH neurons in the septum and the anterior hypothalamus were double-labeled, indicating that they projected to the ME. Double labeled LHRH cells were either smooth, fusiform or "spiny". WGA-accumulating LHRH perikarya were intermixed with single-labeled LHRH cells. The remaining 30% of the LHRH neurons which were not labeled with WGA appeared to project to different hypothalamic and extrahypothalamic areas of the brain. Our results suggest that there are at least two populations of LHRH neurons, one with access to the portal capillaries of the ME and functionally related to the regulation of the pituitary, and one without access to capillaries of the ME, functionally probably related to intracerebral neurotransmission or modulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Detection and identification of minor nucleotides in intact deoxyribonucleic acids by mass spectrometry.

A mass spectral method is described for the detection and identification of unusual nucleotide residues present in DNAs. Analysis by this method of intact, underivatized DNA from salmon sperm, calf thymus, mouse L-cells, wheat germ, M. lysodeikticus, E. Coli, and the bacteriophages 0X-174, fd, and lamda, yields diagnostic ions for the four common components of DNA as well as characteristic ions for 5-methyldeoxycytidine residues. The spectrum from T2 DNA contains ions indicative of 5-hydroxymethyldeoxycytidine and 5-methyldoxycytidine components but no ions corresponding to deoxycytidine residues. The DNAs of phages fd and 0X-174 also display ion products indicative of N6-methyldeoxyadenosine residues. Additional series of ions in the spectra of all four bacteriophage DNAs suggest the presence of 5-substituted deoxyuridine residues. The detection method exhibits considerable sensitivity in that amounts of DNA as low as 0.01 A260nm units can be used in the analysis, and thus, the procedure should prove of some value in the detection and location of modified components in specific regions of the various genomes by analysis of the appropriate endonuclease restriction fragments.

DNA↗

Distinct patterns of expression but similar biochemical properties of protein L-isoaspartyl methyltransferase in higher plants.

Protein L-isoaspartyl methyltransferase is a widely distributed repair enzyme that initiates the conversion of abnormal L-isoaspartyl residues to their normal L-aspartyl forms. Here we show that this activity is expressed in developing corn (Zea mays) and carrot (Daucus carota var. Danvers Half Long) plants in patterns distinct from those previously seen in winter wheat (Triticum aestivum cv Augusta) and thale cress (Arabidopsis thaliana), whereas the pattern of expression observed in rice (Oryza sativa) is similar to that of winter wheat. Although high levels of activity are found in the seeds of all of these plants, relatively high levels of activity in vegetative tissues are only found in corn and carrot. The activity in leaves was found to decrease with aging, an unexpected finding given the postulated role of this enzyme in repairing age-damaged proteins. In contrast with the situation in wheat and Arabidopsis, we found that osmotic or salt stress could increase the methyltransferase activity in newly germinated seeds (but not in seeds or seedlings), whereas abscisic acid had no effect. We found that the corn, rice, and carrot enzymes have comparable affinity for methyl-accepting substrates and similar optimal temperatures for activity of 45 degrees C to 55 degrees C as the wheat and Arabidopsis enzymes. These experiments suggest that this enzyme may have specific roles in different plant tissues despite a common catalytic function.

Daucus carota↗

The translational cis-regulatory element of mammalian ribosomal protein mRNAs is recognized by the plant translational apparatus.

The translational efficiency of mammalian ribosomal protein mRNAs correlates with the growth status of the cells and its control is mediated through a 5' terminal oligopyrimidine tract (5' TOP) common to all these mRNAs. In the present study, we demonstrate that the plant translational apparatus, as represented by wheat-germ extract, discriminates against mammalian mRNAs containing this motif to the same extent as do quiescent mammalian cells. Moreover, mutations in the 5' TOP, which abolish the growth-dependent translational control of the respective mRNAs in mammalian cells, render these mRNAs refractory to discrimination in the plant cell-free system. This selective discrimination reflects neither the specific instability of 5' TOP-containing mRNAs during the incubation in vitro nor a lower competitive potential for the cap-binding protein. The lower in vitro translational efficiency of these mRNAs is an inherent feature which is independent of whether they were derived from polysomes or messenger ribonucleoprotein particles of the transfected mammalian cells. The conservation of the discriminatory property of the translational apparatus between the animal and plant kingdoms is discussed from mechanistic and evolutionary points of view.

3T3 Cells↗

Rapid classification of partial waxy wheats using PCR-based markers.

Mutations in the three homeologous waxy loci Wx-A1, Wx-B1, and Wx-D1 of a waxy wheat line have previously been characterized at the molecular level. Using combinations of these mutations, six types of partial waxy wheat plus wild type and waxy wheat (types 1-8) can be produced. Here, we describe primer sets for all three loci that can be used under a single set of conditions, allowing 32 lines to be characterized as types 1-8 in a single PCR run using a 96-well plate. Using multiplex PCR, mutations at the Wx-B1 and Wx-D1 loci can be identified in a single PCR, reducing the number of reactions necessary to identify and select the desired partial waxy wheat line. A single multiplex PCR can be used to detect all three mutations when products are analyzed using capillary electrophoresis on a microchip device. The PCR conditions and primers are effective with a number of cultivars from other countries, indicating that the mutations found at the Wx-A1 and Wx-B1 loci of these cultivars likely have the same origins as the mutations in the corresponding loci of the waxy wheat line used in this study. The PCR selection method described here is an easy and effective alternative to the commonly used SDS-PAGE methods for identification of null alleles.

Amylose↗

Determination of selenium in Libyan food items using pseudocyclic instrumental neutron activation analysis.

Cyclic and pseudocyclic instrumental neutron activation analysis (INAA) has been used to determine the Se content of 40 Libyan food items. The selected samples include different varieties of local and imported foods such as wheat and barley products (bran and flours), rice, bread, almond, peanuts, vegetables as bean and peas, tea, coffee, sugar, and commonly used spices such as red and black paper, curry, cumin, mixture of spices, thyme, coriander, and fenugreek. Both conventional and anticoincidence gamma-ray spectrometry techniques have been employed. Pseudocyclic INAA in conjunction with anticoincidence counting has been found to provide the most reliable results. The precision of the method has been significantly improved by recycling the samples up to three times. The accuracy has been evaluated by analyzing a number of certified reference materials of varied Se levels. The detection limit has been found to vary between 26 and 90 ppb Se depending on the sample composition. The range of daily dietary intake has been calculated as 13-44 microg of Se per day.

Adult↗

Agrobacterium-mediated transformation of polyploid cereals. The efficiency of selection and transgene expression in wheat.

Three combinations of Agrobacterium tumefaciens strains and vectors were used in the transformation of selected Polish wheat cultivars. The combinations were: two hypervirulent strains, AGL1, containing the pDM805 binary plasmid, and EHA101, containing pGAH; and the common Agro strain LBA4404, harboring the super-binary pTOK233 vector. pDM805 contained bar under the control of Ubi1 promoter, pGAH had nptII under nos, and pTOK233 had hpt under 35S. Additionally, pDM805 and pTOK233 carried the gus reporter gene under the Act1 promoter or 35S promoter, respectively. The highest selection rate was 12.6% and was obtained with EHA101(pGAH) on a kanamycin-containing medium. Sixty-five of the plants grown on that medium were PCR positive. The second best combination was LBA4404(pTOK233) and kanamycin selection, which gave an average transformation rate of 2.3%. Phosphinothricin selection gave 1.0% transformation efficiency, while hygromycin, depending on the strain/vector used, gave from 0.2 to 0.4%. PCR tests in T1 revealed that 67% of the lines showed a 3:1 segregation ratio, and 11% a 15:1 ratio, while in 22%, segregation was non-Mendelian. The high number of T0 transgenic plants containing one copy of the transgene was confirmed via Southern blot analysis. Kanamycin resistance in the T1 generation was very low; in some lines, all the progeny were kanamycin sensitive. GUS expression, only tested in young T1 plants, was in agreement with Mendelian segregation in three out of the twelve tested. The factors influencing the efficiency of selection and transgene expression are discussed in this paper.

Agrobacterium tumefaciens↗

[Two cases of food-dependent exercise-induced anaphylaxis difficult to evoke symptoms by provocation test].

We report two cases of food-dependent exercise-induced anaphylaxis (FDEIA), which were hardly induced by provocation test in the hospital. Case 1: A 28-years-old Japanese female suffers repeated episodes of sternutation, nasal discharge and edema of eyelids after wheat ingestion of wheat followed by exercise. Case 2: A 14-years-old Japanese male suffers repeated episodes of wheal formation on whole body and dyspnea after lunch containing apple followed by exercise. Both of them had never developed symptoms by either ingestion or exercise alone. Provocation tests were performed on admission by combinations of the ingestion of suspected foods, exercise, and aspirin, but no symptoms were reproduced by any combination of them. After discharge, case 1 reproduced symptoms during exercise after the ingestion of wheat under prostration and cold climate. Case 2 reproduced symptoms during exercise after ingestion of apple when he suffered from common cold. Warm and comfortable condition in admission may make it harder to evoke symptoms by the provocation test. Frigidity, cold, prostration, and stress should be reckoned with in the provocation test to improve the accuracy of diagnosis for FDEIA.

Adolescent↗