Root growth profiles of Aveneae and Poeae plants in optically active α-methylbenzyl-3-p-tolylurea media

Using a root growth inhibition assay, we studied the diverse chiral responses of plants in the tribes, Aveneae and Poeae, to the optically active compounds, R- and S- 1-α-methylbenzyl-3- p -tolylurea (MBTU). We specifically examined the responses of grasses belonging to the Poeae tribe ( Lolium , Briza , Poa , Dactylis , and Festuca ) and the Aveneae tribe ( Avena , Holcus , Agrostis , Alopecurus , Beckmannia , and Phleum ). These plants include companion weeds of wheat and barley, and turf grass. The companion weeds of cereal crops, such as Poa annua , Poa pratensis , Dactylis glomerata , Avena fatua , Avena sativa , Holcus lanatus , Agrostis stolonifera , Alopecurus myosuroides , Al. aequalis , and Beckmannia syzigachne , showed significantly inhibited root growth in response to 20 µmol L⁻¹ R- MBTU, whereas the root growth of Triticum aestivum was not inhibited at this concentration. Like Oryza sativa , almost all the Poeae and Aveneae plants tested here preferentially responded to R -MBTU, but the four grasses, Lolium multiflorum , D. glomerata , Alopecurus species, and Phleum pratense , preferentially responded to S -MBTU. Among them, the Agrostis species were highly sensitive to R -MBTU and the Alopecurus species were highly sensitive to S -MBTU. All the plants among the genera, Poa , Avena , and Alopecurus , showed a homogeneous chiral preference.     

Bacillus cereus from the environment is genetically related to the highly pathogenic B. cereus in Zambia

To follow-up anthrax in Zambia since the outbreak in 2011, we have collected samples from the environment and the carcasses of anthrax-suspected animals, and have tried to isolate Bacillus anthracis. In the process of identification of B. anthracis, we collected two isolates, of which colonies were similar to B. anthracis; however, from the results of identification using the molecular-based methods, two isolates were genetically related to the highly pathogenic B. cereus, of which clinical manifestation is severe and fatal (e.g., pneumonia). In this study, we showed the existence of bacteria suspected to be highly pathogenic B. cereus in Zambia, indicating the possibility of an outbreak caused by highly pathogenic B. cereus.     

Design and storage stability of reference materials for microfluidic quantitative PCR-based equine gene doping tests

One method of gene doping in horseracing is administering of exogenous genetic materials, known as transgenes. Several polymerase chain reaction (PCR)-based methods have been developed for detecting transgenes with high sensitivity and specificity. However, novel designs for reference materials (RMs) and/or positive template controls (PTCs) are necessary for simultaneous analysis of multiple transgene targets. In this study, we designed and developed a novel RM for simultaneously detecting multiple targets via microfluidic quantitative PCR (MFQPCR). Twelve equine genes were selected as targets in this study. A sequence region including primers and probes for quantitative PCR was designed, and a 10 bp sequence was inserted to allow the RM to be distinguished from the original transgene sequences. The sequences of individual detection sites were then connected for 12 genes and cloned into a single plasmid vector. We performed fragment size analysis to distinguish between the PCR products of the original transgene sequence and those of the RM, enabling identification of RM contamination. PTCs diluted to 10,000, 1,000, 100, and 10 copies/µl with horse genomic DNA from RM were stably stored at 4°C for 1 year. As digital PCR enabled absolute quantification, the designed substances can serve as an RM. These findings indicate that the RM design and storage conditions were suitable for gene doping tests using MFQPCR.