Early detection of bovine respiratory disease in young bulls using reticulo-rumen temperature boluses

The use of reticulo-rumen temperature boluses to detect bovine respiratory disease (BRD) was investigated in young bulls following their entry into a fattening unit. Twenty-four bulls received a bolus at entry and were observed for 40 days. As soon as a reticulo-rumen hyperthermia (RH) episode was detected using the bolus, clinical examination was performed by a veterinarian and then repeated every 12-24h until the end of RH episode. Fifty-two RH episodes were detected in 22 animals. High rectal temperatures (40.1±0.6°C) were observed during these episodes. BRD was diagnosed on the basis of clinical examination during 38/52 RH episodes in 21 animals (positive predictive value 73%). The onset of BRD signs always occurred after the onset of RH episodes, with a time-lag from 12 to 136 h, depending on BRD signs. Monitoring reticulo-rumen temperature permits early detection of BRD; however, clinical examination is required to confirm BRD.     

A simple and efficient method for isolating small RNAs from different plant species

Background  

Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species.     

Soybean cultivar performance in the presence of soybean Asian rust,in relation to chemical control programs

Soybean rust is caused by an obligate parasite (Phakopsora pachyrhizi) which has spread in Brazil in each new season since 2001 and, despite the efforts to control the disease, losses have occurred every year. Its control demands several tactics amongst which chemical control with fungicides is the main method and remains indispensable. Control strategies such as the use of cultivars with partial resistance are desirable, but are not yet commercially available. The present study analyzed the existing differences in the reactions of short, medium and long cycle soybean cultivars against Asian rust and their responses to fungicide sprays. The experiment was conducted at Uberlandia-MG, Brazil, under field conditions from December 2007 to May 2008, in the Syngenta Seeds Experimental Station. The high pressure of the disease in the experiment simulated the natural pressure that the disease often reaches in Brazil. The studied variables were: visual severity (percentage of infected leaf area), percentage defoliation and productivity (kg ha⁻¹). Disease severity was expressed as AUDPC (area under disease progress curve). Variance analysis and comparison of means by the Tukey test (5% significance) were done for all variables studied. Significant differences were observed between cultivar effects and chemical control programs. The results obtained here indicate that the cultivars M-Soy 8199RR and Emgopa 315RR were less susceptible to disease, and that a control program termed “monitoring” (in which the appearance of new pustules of the pathogen were monitored to make the decision at each fungicide spray) was the most effective.     

Uncommon associations in target resistance among French populations of Myzus persicae from oilseed rape crops

Within the framework of a molecular exploration of target resistance in populations of Myzus persicae on oilseed rapes in France, (1) the S431F mutation (coding gene ace2), although previously reckoned to be rare, revealed to be frequent, (2) M918L (phenotypically characterised) and L932F (both on para) were found for the first time in M. persicae, and (3) a linkage was revealed between M918L and S431F. While until recently populations developing on French oilseed rapes were dominated by genotypes possessing pyrethroid target resistance and esterase overproduction, to date a different type of dominating genotype, equipped with carbamate and pyrethroid target resistance, seems to be invading such fields.     

Two new species of Clestobothrium (Cestoda: Bothriocephalidea), parasites of Merluccius australis and m. hubbsi (Gadiformes: Merlucciidae) from the Patagonian shelf of Argentina, with comments on Clestobothrium crassiceps (Rudolphi, 1819)

Abstract: Two new species of bothriocephalidean cestodes, Clestobothrium splendidum sp. n. from Merluccius australis (Hutton) and Clestobothrium cristinae sp. n. from Merluccius hubbsi Marini from the Patagonian shelf of Argentina, are described. Clestobothrium splendidum can be typified by the following characteristics: a medium-sized strobila composed of410-528 proglottides that are much wider than long; 49-90 testes per mature proglottis, partially surrounding the ovary posteriorly; a transversely elongated genital pore situated anterior to spurious articulations; presence of a genital atrium; a globular cirrus-sac occupying 4-6% of mature proglottis width; a vagina with sphincter and three pairs of osmoregulatory canals on each side of the proglottis. Clestobothrium cristinae is characterised by its small size; 71-219 proglottides; 39-64 testes per mature proglottis, usually surrounding completely the ovary posteriorly; a rounded genital pore situated at the same level of spurious articulations; an oval cirrus-sac occupying 8-16% of mature proglottis width; and three pairs of osmoregulatory canals on each side of the proglottis. Clestobothrium cristinae shares with C. splendidum the type and distribution of microtriches, except for the central surface delimited by two lips. Additionally, type and voucher materials of Clestobothrium crassiceps (Rudolphi, 1819) from Merluccius merluccius were studied. A key to species is provided.     

Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research

Background

The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways.

Results

We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system.

Conclusions

The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk.     

Exploration of plant genomes in the FLAGdb++ environment

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.     

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

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.     

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

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.     

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

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.