Exploration of plant genomes in the FLAGdb++ environment     

Sandra Dèrozier  Franck Samson  Jean-Philippe Tamby  Cécile Guichard  Véronique Brunaud  Philippe Grevet  Séverine Gagnot  Philippe Label  Jean-Charles Leplé  Alain Lecharny  Sébastien Aubourg 《Plant methods》2011,7(1):1-10

Background

Efficient high throughput screening systems of useful mutants are prerequisite for study of plant functional genomics and lots of application fields. Advance in such screening tools, thanks to the development of analytic instruments. Direct analysis in real-time (DART)-mass spectrometry (MS) by ionization of complex materials at atmospheric pressure is a rapid, simple, high-resolution analytical technique. Here we describe a rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic profiling by DART-MS.

Results

To determine whether this DART-MS combined by multivariate analysis can perform genetic discrimination based on global metabolic profiling, intact Arabidopsis thaliana mutant seeds were subjected to DART-MS without any sample preparation. Partial least squares-discriminant analysis (PLS-DA) of DART-MS spectral data from intact seeds classified 14 different lines of seeds into two distinct groups: Columbia (Col-0) and Landsberg erecta (Ler) ecotype backgrounds. A hierarchical dendrogram based on partial least squares-discriminant analysis (PLS-DA) subdivided the Col-0 ecotype into two groups: mutant lines harboring defects in the phenylpropanoid biosynthetic pathway and mutants without these defects. These results indicated that metabolic profiling with DART-MS could discriminate intact Arabidopsis seeds at least ecotype level and metabolic pathway level within same ecotype.

Conclusion

The described DART-MS combined by multivariate analysis allows for rapid screening and metabolic characterization of lots of Arabidopsis mutant seeds without complex metabolic preparation steps. Moreover, potential novel metabolic markers can be detected and used to clarify the genetic relationship between Arabidopsis cultivars. Furthermore this technique can be applied to predict the novel gene function of metabolic mutants regardless of morphological phenotypes.  相似文献   

Insertion sequence- and tandem repeat-based genotyping techniques for Xanthomonas citri pv. mangiferaeindicae     

Pruvost O  Vernière C  Vital K  Guérin F  Jouen E  Chiroleu F  Ah-You N  Gagnevin L 《Phytopathology》2011,101(7):887-893

Molecular fingerprinting techniques that have the potential to identify or subtype bacteria at the strain level are needed for improving diagnosis and understanding of the epidemiology of pathogens such as Xanthomonas citri pv. mangiferaeindicae, which causes mango bacterial canker disease. We developed a ligation-mediated polymerase chain reaction targeting the IS1595 insertion sequence as a means to differentiate pv. mangiferaeindicae from the closely related pv. anacardii (responsible for cashew bacterial spot), which has the potential to infect mango but not to cause significant disease. This technique produced weakly polymorphic fingerprints composed of ≈70 amplified fragments per strain for a worldwide collection of X. citri pv. mangiferaeindicae but produced no or very weak amplification for pv. anacardii strains. Together, 12 tandem repeat markers were able to subtype X. citri pv. mangiferaeindicae at the strain level, distinguishing 231 haplotypes from a worldwide collection of 299 strains. Multilocus variable number of tandem repeats analysis (MLVA), IS1595-ligation-mediated polymerase chain reaction, and amplified fragment length polymorphism showed differences in discriminatory power and were congruent in describing the diversity of this strain collection, suggesting low levels of recombination. The potential of the MLVA scheme for molecular epidemiology studies of X. citri pv. mangiferaeindicae is discussed.  相似文献   

Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa     

Yared Tesfai Tewoldemedhin  Mark Mazzola  Wilhelm J. Botha  Christoffel F. J. Spies  Adéle McLeod 《European journal of plant pathology / European Foundation for Plant Pathology》2011,130(2):215-229

Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South Africa, little is known about specific species in these genera and their pathogenicity toward apple. Therefore, these aspects were investigated along with the development and optimization of qPCR tests for detection and quantification of the most virulent oomycete species. In eight investigated orchards, the oomycete Phythophthora cactorum was widely distributed, while nine Pythium species were differentially distributed among the orchards. Pythium irregulare was the most widely distributed and the most virulent species along with P. sylvaticum, P. vexans and Ph. cactorum. Seven binucleate Rhizoctonia anastomosis groups (AGs) were also differentially distributed among the orchards, with the majority appearing to be non-pathogenic while certain AG-I and AG-F isolates exhibited low virulence on apple. In the genus Fusarium, F. oxysporum was widely distributed, but isolates were non-pathogenic. Fusarium solani and F. avenaceum were less frequently encountered, with only some isolates having low virulence. qPCR data obtained from seedling roots inoculated with the most virulent Pythium species (P. irregulare, P. sylvaticum and P. vexans) and the genus Phytophthora were not always reproducible between trials, or isolates of the same species. In general, seedling growth inhibition was associated with the presence of a low amount of pathogen DNA (±40 fg μl⁻¹ to 2 pg μl⁻¹) in roots. Pythium irregulare, although having the lowest DNA concentrations in roots, was the only species for which a significant negative correlation was found between seedling weight and pathogen DNA concentration.  相似文献   

<Emphasis Type="Italic">Cylindrocarpon</Emphasis> species associated with apple tree roots in South Africa and their quantification using real-time PCR     

Yared Tesfai Tewoldemedhin  Mark Mazzola  Lizel Mostert  Adéle McLeod 《European journal of plant pathology / European Foundation for Plant Pathology》2011,129(4):637-651

Cylindrocarpon species are known to be a component of the pathogen/pest complex that incites apple replant disease. In South Africa, no information is available on apple associated Cylindrocarpon species and their pathogenicity. Therefore, these aspects were investigated. Among the isolates recovered from apple roots in South Africa, four species (C. destructans, C. liriodendri, C. macrodidymum and C. pauciseptatum) were identified using β-tubulin gene sequencing and phylogenetic analysis. This is the first report of C. liriodendri, C. macrodidymum and C. pauciseptatum on apple trees. Cylindrocarpon macrodidymum was the most prevalent. Isolates within each of the four species were pathogenic towards apple seedlings, but varied in their virulence. With a single exception, all isolates were able to induce lesion development on seedling roots. Only 57% of the isolates, which represented all four species, were able to cause a significant reduction in seedling weight and/or height. The greatest seedling growth reductions were caused by two isolates of C. destructans, and one isolate each of C. liriodendri and C. macrodidymum. A quantitative real-time polymerase chain reaction (qPCR) method was developed for simultaneous detection of all four Cylindrocarpon species. qPCR analyses of Cylindrocarpon from the roots of inoculated seedlings showed that the amount of Cylindrocarpon DNA in roots was not correlated to seedling growth reductions (weight and height) or root rot. The qPCR method is, however, very useful for the rapid identification of apple associated Cylindrocarpon species in roots. The technique may also hold potential for being indicative of Cylindrocarpon disease potential if rhizosphere soil rather than roots are used.  相似文献   

Improving sooty blotch and flyspeck severity estimation on apple fruit with the aid of standard area diagrams     

Piérri Spolti  Luana Schneider  Rosa M. V. Sanhueza  Jean C. Batzer  Mark L. Gleason  Emerson Medeiros Del Ponte 《European journal of plant pathology / European Foundation for Plant Pathology》2011,129(1):21-29

Sooty blotch and flyspeck is caused by numerous species of fungi that colonize the surface of apple fruit and thereby lower its market value. Although this disease poses a substantial threat to apple growers’ profitability in some regions, reliable and cost-effective methods for epidemiological and disease control studies have not been validated, nor are they widely available. We modified a standard area diagram to aid sooty blotch and flyspeck severity assessments and quantified its impact on accuracy and precision of visual estimates. Samples of ‘Fuji’ and ‘Mutsu’ fruit were photographed both from the top and laterally. Severity was assessed from a sub-sample of 160 images using image analysis software. Validation of the diagram was performed by eight raters who independently assessed severity in two series of selected images representing the lateral view and the top view, initially unaided and subsequently with the aid of the scale. Severity estimates ranged from 0.4% to 98% (most fruit had <10% severity). Accuracy and precision of the estimates were significantly improved when using the diagrammatic scale; concordance correlation coefficient values increased from 0.81 to 0.95. A strong tendency to underestimate severity for the mid-range to high levels was minimized when using the aid, which also improved reproducibility of the estimates among raters. In addition to strengthening evidence that a standard area diagram can be used reliably in sooty blotch and flyspeck studies, we expanded its application to disease assessment in the peduncle region, which enhances the usefulness of the method for evaluating efficacy of management practices.  相似文献   

Diversity and fitness of <Emphasis Type="Italic">Plasmopara viticola</Emphasis> isolates resistant to QoI fungicides     

Marie-France Corio-Costet  Marie-Cécile Dufour  Jérémy Cigna  Pierre Abadie  Wei-Jen Chen 《European journal of plant pathology / European Foundation for Plant Pathology》2011,129(2):315-329

The effectiveness of Quinone outside Inhibitor (QoI) fungicides against grape downy mildew in European vineyards has significantly decreased in the last decade. One nucleotide polymorphism, G143A in the cytochrome b gene of Plasmopara viticola, is involved in resistance to QoIs. Previous genetic examination on the mitochondrial genomes showed four major haplotypes (IR, IS, IIR, IIS) coexisting in European vineyards. A resistant allele (G143A) was present in IR and IIR haplotypes. The purpose of the present study was to estimate the diversity of the different mitochondrial haplotypes and their distribution in QoI-resistant populations before evaluating the potential cost of the resistant mutation G143A in P. viticola population. From 2000 to 2004, the frequencies of resistant isolates ranged from 0% to 23.25% with an average of 4.64 % among the populations examined. To evaluate the fitness of sensitive and resistant isolates, a comparison of different biological parameters including latent period, spore production and infection frequency was performed, enabling a fitness index (F_I) to be determined. Resistant isolates exhibited greater infection frequency than sensitive isolates, whereas no significant difference was found in sporulation ability and latent period between sensitive and resistant isolates. To further investigate competitiveness among isolates, an assay including two resistant isolates in different proportion with a sensitive isolate was conducted on eight asexual growing cycles in the absence of a QoI fungicide. The competitiveness of resistant isolates varied according to their fitness parameters, suggesting that there is no noticeable cost of QoI resistance in controlled conditions in Plasmopara viticola.  相似文献   

Interactions between the fungicide benomyl and soil microorganisms     

L. Torstensson  B. Wessén 《Soil biology & biochemistry》1984,16(5):445-452

Decomposition of benomyl and carbendazim was studied in field experiments following repeated applications during autumn to winter cereals. Effects of the fungicides on straw decomposition, balance of straw fungal flora and mineralization of nitrogen in the soils were investigated in field and in laboratory experiments. Persistence in the field of the fungicides at doses of 0.1–0.2 kg ha^?1 was 9–12 months in clay soils and 12 months or longer in sand soil. Decomposition of straw in the field was not affected in clay soils by doses up to 2 kg ha^?1. In sand soil, doses up to 0.5 kg ha^?1 gave no effect but in one case at 2 kg ha^?1 the initial stages of straw decomposition were slightly inhibited. All doses tested in both clay and sand soils caused changes in composition of the straw fungal flora. In a laboratory experiment with benomyl in sand soil an increase in rate of nitrate accumulation was observed at a dose corresponding to 2 kg ha^?1.  相似文献   

Classification of wine and alcohol vinegar samples based on near-infrared spectroscopy. Feasibility study on the detection of adulterated vinegar samples     

Saiz-Abajo MJ  Gonzales-Saiz JM  Pizarro C 《Journal of agricultural and food chemistry》2004,52(25):7711-7719

Near-infrared (NIR) spectroscopy was used to discriminate between wine vinegar (red or white) and alcohol vinegar. One orthogonal signal correction method (OSC) was applied on a set of 73 vinegar NIR spectra from both origins and artificial blends made in the laboratory in order to remove information unrelated to a specific chemical response (tartaric acid), which was selected due to its high discriminant ability to differentiate between wine vinegar and alcohol vinegar samples. These corrected NIR spectra, as well as raw NIR spectra and 14 physicochemical variables, were used to develop separate classification models using the potential functions method as a class-modeling technique. The aforementioned models were compared to evaluate the suitability of NIR spectroscopy as a rapid method for discriminating between vinegar origins. The transformation of vinegar NIR spectra by means of an orthogonal signal correction method prompted a notable improvement in the specificity of the constructed classification models. The classification model developed was then applied to artificial vinegar blends made in the laboratory to test its capacity to recognize adulterated vinegar samples.  相似文献   

Compared effects of long-term pig slurry applications and mineral fertilization on soil denitrification and its end products (N2O, N2)     

Christophe Dambreville  Catherine Hénault  Florian Bizouard  Thierry Morvan  Rémi Chaussod  Jean-Claude Germon 《Biology and Fertility of Soils》2006,42(6):490-500

The long-term (9 years) effect of pig slurry applications vs mineral fertilization on denitrifying activity, N₂O production and soil organic carbon (C) (extractable C, microbial biomass C and total organic C) was compared at three soil depths of adjacent plots. The denitrifying activities were measured on undisturbed soil cores and on sieved soil samples with acetylene method to estimate denitrification rates under field or potential conditions. Pig slurry applications had a moderate impact on the C pools. Total organic C was increased by +6.5% and microbial biomass C by ≥25%. The potential denitrifying activity on soil suspension was stimulated (×1.8, P<0.05) 12 days after the last slurry application. This stimulation was still apparent, but not significant, 10 months later and, according to both methods of denitrifying activity measurement (r ²=0.916, P<0.01 on sieved soil; r ²=0.845, P<0.001 on soil cores), was associated with an increase in microbial biomass C above a threshold of about 105 mg kg⁻¹. The effect of pig slurry on denitrification and N₂O reduction rates was detected on the surface layer (0–20 cm) only. However, no pig slurry effect could be detected on soil cores at field conditions or after NO₃ ⁻ enrichments at 20°C. Although the potential denitrifying activity in sieved soil samples was stimulated, the N₂O production was lower (P<0.03) in the plot fertilized with pig slurry, indicating a lower N₂O/(N₂O + N₂) ratio of the released gases. The pig-slurry-fertilized plot also showed a higher N₂O reduction activity, which is coherent with the lower N₂O production in anaerobiosis.  相似文献   

Biochemical composition and kinetics of C and N mineralization of animal wastes: a typological approach     

Thierry Morvan  Bernard Nicolardot  Léonard Péan 《Biology and Fertility of Soils》2006,42(6):513-522

Forty-seven different animal wastes were characterized using chemical and organic matter fractionation methods (water extraction and Van Soest method) and 224-day incubation studies to assess their decomposition in soil. Simple correlation and multiple factor analysis were performed to establish relationships between the composition of these wastes and C and N mineralization. Carbon and N contents ranged from 101 to 469 mg C kg⁻¹ dry matter (d.m.) and from 4 to 39 mg N kg⁻¹ d.m. Soluble C and N represented less than 9% of organic C and 1.5% of total N at 20°C, respectively. The C fractions soluble at 100°C or in neutral detergent were larger and represented 14 and 32% of the organic C, respectively. The hemicellulose-like (HEM) and cellulose-like (CEL) fractions contained about 16.5 and 6% of the organic N, respectively. The C distribution in the lignin-like (LIG) and CEL fractions was comparable, but the former contained more N. Carbon mineralization varied from 5 to 62% of the organic C added during the 224-day incubation; 70% of the wastes induced net N mineralization at the end of incubation (from 3 to 51% of organic N). Other wastes induced net soil inorganic N immobilization, from −1 to −31% of the organic N added. Most highly significant correlations were established between the C mineralization and the C present in the water-soluble fraction at 20°C, and the HEM and LIG fractions. Relationships between N mineralization and biochemical characteristics were weak, except with the soluble Van Soest fraction, and highly significant correlations were observed between N mineralization rates calculated at 224 days of incubation and the organic N content or C/N ratio of wastes. Finally, an objective hierarchical classification based on composition criteria and C and N mineralization led to the definition of six different classes of wastes. It permitted differentiation between four composted wastes and intrinsically different wastes (i.e., cattle manures, pig manures, and poultry manures) which could not be objectively regrouped. It also placed some very different types of waste (solid phase from pig slurry separation, pig manures, and composted pig mixtures) in the same class.  相似文献