Barley
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The production of doubled haploid (DH) barley plants through anther culture is a very useful yet simple in vitro technique. DH plants derive from divisions of haploid microspores that have undergone a developmental switch under the appropriate conditions. The successive divisions lead to the formation of an embryo or callus rather than the formation of mature pollen grains. Plants that regenerate from these embryos are often either haploid, in which case their chromosome set can be doubled by treatment with colchicine, or spontaneous double haploids. The efficiency of DH plant production is highly variable depending on the genotype of the source material. Despite this limitation, DH plants have been widely used in breeding and research programs. Compared to conventional approaches, breeding strategies that makes use of DH plants achieve a homozygous state, allowing transgene or mutation stabilization in the genome, within a considerably shorter time, thus accelerating workflow or reducing work volume.
- MeSH
- barvení a značení MeSH
- DNA rostlinná genetika MeSH
- haploidie MeSH
- ječmen (rod) růst a vývoj MeSH
- kultivační média MeSH
- květy růst a vývoj MeSH
- pyl růst a vývoj MeSH
- regenerace MeSH
- somatická embryogeneze rostlin MeSH
- sterilizace MeSH
- techniky tkáňových kultur metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Understanding the mechanisms regulating the development of cereal seeds is essential for plant breeding and increasing yield. However, the analysis of cereal seeds is challenging owing to the minute size, the liquid character of some tissues, and the tight inter-tissue connections. Here, we demonstrate a detailed protocol for dissection of the embryo, endosperm, and seed maternal tissues at early, middle, and late stages of barley seed development. The protocol is based on a manual tissue dissection using fine-pointed tools and a binocular microscope, followed by ploidy analysis-based purity control. Seed maternal tissues and embryos are diploid, while the endosperm is triploid tissue. This allows the monitoring of sample purity using flow cytometry. Additional measurements revealed the high quality of RNA isolated from such samples and their usability for high-sensitivity analysis. In conclusion, this protocol describes how to practically dissect pure tissues from developing grains of cultivated barley and potentially also other cereals.
The discovery of radioactivity at the end of the nineteenth century played a key role in a series of historical landmarks that would lead to contemporary mutation breeding in agricultural crops. The aim of the earliest experiments was to test the effects of radiation on living organisms beginning with fruit flies. Exposure of plants to X-rays provided the first incontrovertible proof that phenotypic changes could be induced. Chemicals were a second type of mutagen tested from the 1940s and both forms are used today. This chapter is an overview of some of the historical developments that led to the use of mutagenesis in plants, with a focus on barley, a model species for mutation genetics and breeding as well as a major cereal crop. Perhaps the most well-known examples of mutant barley cultivars are Diamant, Golden Promise, and their hybrids.
- MeSH
- dějiny 19. století MeSH
- dějiny 20. století MeSH
- ječmen (rod) genetika růst a vývoj MeSH
- mutace genetika MeSH
- mutageneze genetika MeSH
- šlechtění rostlin ekonomika dějiny metody MeSH
- Check Tag
- dějiny 19. století MeSH
- dějiny 20. století MeSH
- Publikační typ
- časopisecké články MeSH
- historické články MeSH
- práce podpořená grantem MeSH
Wild barley is abundant, occupying large diversity of sites, ranging from the northern mesic Mediterranean meadows to the southern xeric deserts in Israel. This is also reflected in its wide phenotypic heterogeneity. We investigated the dynamics of DNA content changes in seed tissues in ten wild barley accessions that originated from an environmental gradient in Israel. The flow cytometric measurements were done from the time shortly after pollination up to the dry seeds. We show variation in mitotic cell cycle and endoreduplication dynamics in both diploid seed tissues (represented by seed maternal tissues and embryo) and in the triploid endosperm. We found that wild barley accessions collected at harsher xeric environmental conditions produce higher proportion of endoreduplicated nuclei in endosperm tissues. Also, a comparison of wild and cultivated barley strains revealed a higher endopolyploidy level in the endosperm of wild barley, that is accompanied by temporal changes in the timing of the major developmental phases. In summary, we present a new direction of research focusing on connecting spatiotemporal patterns of endoreduplication in barley seeds and possibly buffering for stress conditions.
- MeSH
- DNA rostlinná genetika MeSH
- endosperm genetika MeSH
- genetická variace genetika MeSH
- ječmen (rod) genetika MeSH
- polyploidie MeSH
- populační genetika metody MeSH
- semena rostlinná genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Izrael MeSH
Ramularia collo-cygni is the biotic factor responsible for the disease Ramularia leaf spot (RLS) of barley (Hordeum vulgare). Despite having been described over 100 years ago and being considered a minor disease in some countries, the fungus is attracting interest in the scientific community as a result of the increasing number of recorded economically damaging disease epidemics. New reports of disease spread and fungal identification using molecular diagnostics have helped redefine RLS as a global disease. This review describes recent developments in our understanding of the biology and epidemiology of the fungus, outlines advances made in the field of the genetics of both the fungus and host, and summarizes the control strategies currently available.
The bird cherry-oat aphid (Rhopalosiphum padi (L.)) is one of the most detrimental pests of cereals, causing harm mainly by transmitting Barley yellow dwarf virus (BYDV). R. padi migration has been monitored since 1992 using suction traps at five sites in the Czech Republic. The count data were subjected to the following different analyses: (i) the minimum temperature thresholds for the aphids to take off were determined; (ii) a partial redundancy analysis using the minimum, average, and maximum temperatures, as well as the wind speed, the precipitation total, and past aphid migration descriptors was performed to explain the relationship between aphid occurrences and weather patterns; and (iii) three types of models from the field of machine learning were used to predict aphid occurrences. According to our findings, (i) in Central Europe, 8°C is the temperature threshold for R. padi migration unless insufficient daylight delays the take-off; (ii) weather conditions occurring roughly 9 months before R. padi migration influence the migration size; (iii) the duration of the summer migration influences the autumn migration size; and (iv) the daytime and nighttime temperatures in the autumn determine the summer peak, whereas winter frosts and precipitation influence the autumn peak.
- MeSH
- ječmen (rod) * virologie MeSH
- Luteovirus * fyziologie MeSH
- migrace zvířat * MeSH
- mšice * fyziologie virologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Unraveling and exploiting mechanisms of disease resistance in cereal crops is currently limited by their large repeat-rich genomes and the lack of genetic recombination or cultivar (cv)-specific sequence information. We cloned the first leaf rust resistance gene Rph1 (Rph1a) from cultivated barley (Hordeum vulgare) using "MutChromSeq," a recently developed molecular genomics tool for the rapid cloning of genes in plants. Marker-trait association in the CI 9214/Stirling doubled haploid population mapped Rph1 to the short arm of chromosome 2H in a physical region of 1.3 megabases relative to the barley cv Morex reference assembly. A sodium azide mutant population in cv Sudan was generated and 10 mutants were confirmed by progeny-testing. Flow-sorted 2H chromosomes from Sudan (wild type) and six of the mutants were sequenced and compared to identify candidate genes for the Rph1 locus. MutChromSeq identified a single gene candidate encoding a coiled-coil nucleotide binding site Leucine-rich repeat (NLR) receptor protein that was altered in three different mutants. Further Sanger sequencing confirmed all three mutations and identified an additional two independent mutations within the same candidate gene. Phylogenetic analysis determined that Rph1 clustered separately from all previously cloned NLRs from the Triticeae and displayed highest sequence similarity (89%) with a homolog of the Arabidopsis (Arabidopsis thaliana) disease resistance protein 1 protein in Triticum urartu In this study we determined the molecular basis for Rph1-mediated resistance in cultivated barley enabling varietal improvement through diagnostic marker design, gene editing, and gene stacking technologies.
The detection of meiotic crossovers in crop plants currently relies on scoring DNA markers in a segregating population or cytological visualization. We investigated the feasibility of using flow-sorted haploid nuclei, Phi29 DNA polymerase-based whole-genome-amplification (WGA) and multi-locus KASP-genotyping to measure meiotic crossovers in individual barley pollen grains. To demonstrate the proof of concept, we used 24 gene-based physically mapped single nucleotide polymorphisms to genotype the WGA products of 50 single pollen nuclei. The number of crossovers per chromosome, recombination frequencies along chromosome 3H and segregation distortion were analysed and compared to a doubled haploid (DH) population of the same genotype. The number of crossovers and chromosome wide recombination frequencies show that this approach is able to produce results that resemble those obtained from other methods in a biologically meaningful way. Only the segregation distortion was found to be lower in the pollen population than in DH plants.
- MeSH
- chromozomy rostlin MeSH
- crossing over (genetika) * MeSH
- DNA rostlinná MeSH
- genom rostlinný MeSH
- haploidie MeSH
- ječmen (rod) genetika MeSH
- lokus kvantitativního znaku MeSH
- meióza genetika MeSH
- multilokusová sekvenční typizace * MeSH
- pyl genetika MeSH
- rekombinace genetická MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The effect of dietary administered young barley containing a mixture of phytochemicals to female rats for the prevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis was evaluated. After carcinogen administration (14 wk), mammary tumors were removed and prepared for histopathological and immunohistochemical analysis. Moreover, in vitro evaluation of possible mechanisms in MCF-7 breast cancer cell line was performed. Barley (0.3%) demonstrated mild antitumor effect in mammary carcinogenesis, yet 3% barley did not further improve this effect. Immunohistochemical analysis of rat tumor cells in treated groups showed significant increase in caspase-3 expression and significant reduction in Ki67 expression. In addition, 3% barley significantly decreased dityrosine levels versus control. Barley in higher dose significantly decreased serum low-density lipoprotein-cholesterol in rats. In vitro studies showed that barley significantly decreased survival of MCF-7 cells in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and significantly decreased 5-bromo-20-deoxyuridine incorporation versus control. Barley prevented cell cycle progression and extended incubation with barley showed significant increase in the percentage of annexin V/propidium iodide-positive MCF-7 cells. Our results propose an antitumor effect for the mixture of phytochemicals present in young barley in a breast cancer model.
- MeSH
- antikarcinogenní látky farmakologie MeSH
- apoptóza MeSH
- experimentální nádory mléčných žláz chemicky indukované patologie prevence a kontrola MeSH
- flavonoidy analýza MeSH
- ječmen (rod) * chemie MeSH
- lidé MeSH
- metabolismus lipidů MeSH
- methylnitrosomočovina toxicita MeSH
- MFC-7 buňky MeSH
- nádory prsu MeSH
- potkani Sprague-Dawley MeSH
- proliferace buněk MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
