The Andean black cherry (
P. serotina
) is an underutilized fruit species that could contribute to the development of sustainable food systems in the Andean region. The species displays gametophytic ...self-incompatibility (GSI), a mechanism controlled by the multiallelic
S-locus
which prevents crossbreeding between genetically related individuals and hinders breeding efforts. To design effective crosses, breeders require accurate knowledge of the
S-
haplotypes of parental lines. However,
S-
haplotype diversity is commonly evaluated using PCR-based methods that fail to accurately discriminate alleles. To address this limitation, we developed a new method to identify
S-
alleles in
P. serotina
using nanopore sequencing technology. Our method uses the Native Barcoding protocol and MinION sequencer from Oxford Nanopore Technologies to enable scalable, multiplex amplicon sequencing. For sequence analysis, we developed a bioinformatic pipeline that uses Porechop for sample demultiplexing, MeshClust for sequence alignment and clustering, and the Ugene Consensus algorithm to determine allelic variants. In this study, we evaluated the
S-RNase
gene of 24
P. serotina
accessions using our nanopore sequencing and bioinformatic workflow. Among these accessions, we identified 12 previously reported and 6 putative new
S-
alleles that could not be identified with existing
S-
genotyping methods. Five accessions were classified as homozygous, while the other 19 were heterozygous with two or three alleles. Our results demonstrate that nanopore sequencing provides a cost-effective alternative for
S-
allele profiling that improves on the accuracy of existing PCR-based methods. Because of the versatility of MinION sequencing, the reported workflow can be used to characterize the diversity of other useful genes in the species, which are of relevance for conservation and breeding efforts.
SUMMARY
Loss‐of‐function alleles of plant MLO genes confer broad‐spectrum resistance to powdery mildews in many eudicot and monocot species. Although barley (Hordeum vulgare) mlo mutants have been ...used in agriculture for more than 40 years, understanding of the molecular principles underlying this type of disease resistance remains fragmentary. Forward genetic screens in barley have revealed mutations in two Required for mlo resistance (Ror) genes that partially impair immunity conferred by mlo mutants. While Ror2 encodes a soluble N‐ethylmaleimide‐sensitive factor‐attached protein receptor (SNARE), the identity of Ror1, located at the pericentromeric region of barley chromosome 1H, remained elusive. We report the identification of Ror1 based on combined barley genomic sequence information and transcriptomic data from ror1 mutant plants. Ror1 encodes the barley class XI myosin Myo11A (HORVU.MOREX.r3.1HG0046420). Single amino acid substitutions of this myosin, deduced from non‐functional ror1 mutant alleles, map to the nucleotide‐binding region and the interface between the relay‐helix and the converter domain of the motor protein. Ror1 myosin accumulates transiently in the course of powdery mildew infection. Functional fluorophore‐labeled Ror1 variants associate with mobile intracellular compartments that partially colocalize with peroxisomes. Single‐cell expression of the Ror1 tail region causes a dominant‐negative effect that phenocopies ror1 loss‐of‐function mutants. We define a myosin motor for the establishment of mlo‐mediated resistance, suggesting that motor protein‐driven intracellular transport processes are critical for extracellular immunity, possibly through the targeted transfer of antifungal and/or cell wall cargoes to pathogen contact sites.
Significance Statement
We report the identification of a barley (Hordeum vulgare) gene that is required for the agriculturally relevant broad‐spectrum powdery mildew resistance conditioned by mlo mutants. The gene encodes a class XI myosin motor protein that partly associates with peroxisomes and might be needed for the timely intracellular transport of defense‐related cargoes in the context of extracellular immunity.
The Pierre Auger Cosmic Ray Observatory Aab, A.; Abreu, P.; Aglietta, M. ...
Nuclear instruments & methods in physics research. Section A, Accelerators, spectrometers, detectors and associated equipment,
10/2015, Letnik:
798, Številka:
C
Journal Article
Recenzirano
Odprti dostop
The Pierre Auger Observatory, located on a vast, high plain in western Argentina, is the world׳s largest cosmic ray observatory. The objectives of the Observatory are to probe the origin and ...characteristics of cosmic rays above 1017eV and to study the interactions of these, the most energetic particles observed in nature. The Auger design features an array of 1660 water Cherenkov particle detector stations spread over 3000km2 overlooked by 24 air fluorescence telescopes. In addition, three high elevation fluorescence telescopes overlook a 23.5km2, 61-detector infilled array with 750m spacing. The Observatory has been in successful operation since completion in 2008 and has recorded data from an exposure exceeding 40,000km2sryr. This paper describes the design and performance of the detectors, related subsystems and infrastructure that make up the Observatory.
Methyl tert-butylether (MTBE) used as fuel oxygenate poses problems for water suppliers since it is persistent in the aquatic environment and the removal efficiency by conventional water treatment ...methods (aeration or activated carbon filtration) is rather low. Substitution by other ether compounds such as ethyl tert-butylether (ETBE), tert-amylmethylether (TAME) or di-isopropylether (DIPE) is discussed, however, their environmental behaviour is similar to that of MTBE. Experiments investigating the elimination efficiency of AOP were carried out in tap water and water from Lake Constance. The elimination efficiency for all treatment processes was found to follow the order: MTBE << TAME approximately equal ETBE < DIPE For all compounds under investigation, neither pure ozonation nor UV irradiation yield a considerable concentration decline. Only the formation of highly reactive OH radicals shows a potential for removing the ethers from water. Therefore the addition of H2O2 in equimolar ratio prior to ozone admixing proved to be quite efficient. The application of combined UV/H2O2 showed good results in all cases; the best concentration decline was achieved with UV/ozone. The rate of elimination of the three substitutes for MTBE (ETBE, TAME and DIPE) is higher in all processes; nevertheless, no complete removal could be achieved. Therefore, from the point of view of water suppliers, the use of other ethers as substitute for MTBE is posing the same problems as MTBE.
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