Differential expression of molecular rust resistance components have distinctive profiles in <Emphasis Type="Italic">Coffea arabica</Emphasis> - <Emphasis Type="Italic">Hemileia vastatrix</Emphasis> interactions

Countering the economic hurdle caused by coffee leaf rust disease is most appealing at this time as it has posed a major threat to coffee production around the world. Establishing differential expression profiles at different times following pathogen invasion in both innate and acquired immunities unlocks the molecular components of resistance and susceptibility. Suppression subtractive hybridization (SSH) was used to identify genes differentially over-expressed and repressed during incompatible and compatible interactions between Coffea arabica and Hemileia vastatrix. From 433 clones of expressed sequence tags (ESTs) sequenced, 352 were annotated and categorized of which the proportion of genes expressed during compatible interaction were relatively smaller. The result showed upregulation and downregulation of various genes at 12 and 24 h after pathogen inoculation in both interactions. The use of four different databases in searching for gene homology resulted in different number of similar sequences. BLASTx against EMBL-EBI (European Molecular Biology Laboratory-European Bioinformatics Institute) database being with the maximum (100%) hits for all the annotated sequences. RT-qPCR analysis of seven resistance-signaling genes showed similar expression patterns for most of the genes in both interactions, indicating these genes are involved in basal (non-specific) defense during which immune reactions are similar. Using SSH, we identified different types of resistance related genes that could be used for further studies towards resistant cultivar development. The potential role of some of the resistance related proteins found were also discussed.     

Multilocus sequence typing as a tool for studying the molecular epidemiology and population structure of Brachyspira hyodysenteriae

The purpose of this study was to develop and apply a multilocus sequence typing (MLST) scheme to study the molecular epidemiology of Brachyspira hyodysenteriae, the aetiological agent of swine dysentery. Sequences of seven conserved genomic loci were examined in 111 B. hyodysenteriae strains. Fifty-eight of these previously had been analysed by multilocus enzyme electrophoresis (MLEE), and for some the results of pulsed field gel electrophoresis (PFGE), restriction endonuclease analysis (REA) and/or serotyping also were available. The discriminatory power of these methods was compared. The strains were divided into 67 sequence types (STs) and 46 amino acid types (AATs) by MLST. The Index of Association value was significantly different from zero, indication that the population was clonal. Eleven clonal complexes (Cc) comprising between 2 and 10 STs were recognised. A population snapshot based on AATs placed 77.5% of the isolates from 30 of the AATs into one major cluster. The founder type AAT9 included 13 strains from nine STs that were isolated in Australia, Sweden, Germany and Belgium, including one from a mallard. The MLST results were generally comparable to those produced by MLEE. The MLST system had a similar discriminatory power to PFGE, but was more discriminatory than REA, MLEE or serotyping. MLST data provided evidence for likely transmission of strains between farms, but also for the occurrence of temporal “micro-evolution” of strains on individual farms. Overall, the MLST system proved to be a useful new tool for investigating the molecular epidemiology and diversity of B. hyodysenteriae.     

Antibody Formation in Embryos

The production of agglutinins to Salmonella typhi was studied in the opossum embryo in the period immediately before and after the development of thymic and lymph node lymphoid tissue. Antibody was found only in embryos older than 8 days which corresponds to an 8- to 10-week human embryo in terms of organogenesis and is the earliest stage at which antibody production has been recorded.     

Release of atrazine and alachlor from clay-oxamide controlled-release formulations

Controlled-release herbicide formulations have been shown to decrease the leaching potential of several herbicides under laboratory and field conditions. The utility and efficacy of these formulations may be improved by combining several herbicides and a fertilizer source in a single formulation. The objective of these studies was to develop granular alginate formulations that were composed of a combination of the herbicides atrazine and alachlor with the slow-release nitrogen source oxamide (ethanediamide). Controlled release of the herbicides was obtained by addition of selected minerals, including calcium bentonite, fine-grind bentonite, montmorillonite K10, kaolinite and iron (III) oxide. A formulation without clay was used as a comparison. The formulations tested had herbicide active ingredient contents ranging from ∽0·02 to 0·54% and a nitrogen content of 21%. Release of the herbicides was studied by equilibrating the formulations with deionized water on a rotary shaker at 200 rev min^-1 and sampling at regular time intervals up to 104 hours. The minerals used in the different formulations influenced the herbicide active ingredient composition, as well as the release properties of the individual formulations. The atrazine content of the formulations decreased in the order calcium bentonite>fine-grind bentonite>kaolinite>montmorillonite=iron oxide>no clay. For alachlor the content decreased in the order of calcium bentonite>fine-grind bentonite>montmorillonite>iron oxide>kaolinite>no clay. Controlled release of atrazine (i.e. reduction in release rate) varied in the order calcium bentonite>iron oxide>montmorillonite>fine-grind bentonite= kaolinite>no clay, and for alachlor fine-grind bentonite>calcium bentonite>montmorillonite>no clay=kaolinite=iron oxide. A certain percentage of the applied active ingredient of both alachlor and atrazine was not recovered. From 5 to 27% of the active ingredient was not released, with the greatest retention by the bentonite formulations. Release of nitrogen was not strongly influenced by mineral type, although a trend indicated greater release with formulations containing kaolinite. © 1998 SCI     

Identification and evaluation of chloroplast uni- and trinucleotide sequence repeats in citrus

A cpDNA fragment of 34 genotypes belonging to Citrus and four related genera was amplified and sequenced. Chloroplast microsatellites were revealed with the length of repeats ranging from 25 to 44 bases. Other than the normal uninucleotide poly(A) repeats, a trinucleotide poly(TAA) motif was also found, the first report of such repeats in a plant chloroplast genome. According to SSR structure variations, 18 Chloroplast SSR Types (CST) were identified. The CST sequences were informative for better understanding the genetic relationships of chloroplast genomes among the analyzed genotypes and confirmed some previous hypotheses about the female parent of several hybrid accessions.