Enhancement in androgenesis efficiency in barley (Hordeum vulgare L.) and bread wheat (Triticum aestivum L.) by the addition of dimethyl sulfoxide to the mannitol pretreatment medium

24 Pags.- 7 Tabls. The definitive version is available at: http://link.springer.com/journal/11240Dimethyl sulfoxide (DMSO) is a well-known solvent widely used in cell biology owing to its specific physicochemical properties, which allow it to decrease the lipid bilayer’s thickness and membrane fluidity while increasing membrane permeability. To improve doubled haploid production using an anther culture method in barley and bread wheat, the addition of DMSO to the pretreatment medium was tested. The first experiment was carried out on four barley cultivars with varying degrees of androgenetic ability by exposing them to four DMSO concentrations (0, 0.2, 1 and 2 % v/v). The medium with 1 % DMSO was the most effective in increasing the numbers of embryos and plants. The highest concentration tested (2 %) negatively impacted all of the measured variables when compared with the results of the 1 % DMSO addition, probably owing to a toxic effect. The effects caused by this solvent were more remarkable on the most recalcitrant cultivars, in which there was a threefold increase in the number of green plants. Furthermore, a downward trend in the albinism rate was observed as the concentration of DMSO increased. In a second experiment, we compared a 1 % DMSO supplement with the control to determine whether its addition was effective in several cultivars and F1 crosses of bread wheat. As in barley, there was a marked increase in the number of green plants, leading to a twofold to fourfold increase in both cultivars and F1 crosses.This work has been supported by Projects AGL 2011-30529-C0207 and RTC-2014-2902-2 from the Spanish Ministry of Economy and Competitiveness.Peer reviewe

Identification and validation of QTLs for green plant percentage in barley (Hordeum vulgare L.) anther culture

The original version is available at: http://www.springerlink.com/content/688481281g18035q/fulltext.pdfIn cereals, albinism is a major obstacle to produce doubled haploids (DH) for breeding programs. In order to identify QTLs for green plant percentage in barley anther culture, a specific population was developed. This population, consisting of 100 DH lines, was generated by crossing the model cultivar for anther culture “Igri” with an albino-producing DH line (DH46) selected from Igri x Dobla, in search of a maximum segregation for the trait and minimum for the other anther culture variables. A combination of bulked segregant analysis and AFLP methodology was used to identify markers linked to the trait. A linkage map was constructed using these AFLPs, together with RAPD, STS and SSR markers. This study identified a new QTL for green plant percentage on chromosome 3H and confirmed the previously reported one on chromosome 5H. Up to 65.2% of the phenotypic variance for this trait was explained by the additive effects of these two QTLs. Thirty elite cultivars of barley from different origin, row type, growth habit and end use, were selected to validate these QTLs. Since two of the markers linked to the QTLs were AFLPs, we successfully converted them into simple PCR-based SCAR markers. Only the SSR HVM60, on chromosome 3H, was significantly associated with the trait, explaining near 20% of the phenotypic variance. Among the allelic variants identified for this marker, HVM60-120bp was associated with the highest values of green plant percentage.M. Muñoz-Amatriaín and X-W Chen were recipients of a fellowship from Ministry of Education and Science of Spain. The research was supported by Projects AGL2001-1631 and AGL2004-03396 from Plan Nacional de Recursos y Tecnologías Agroalimentarias of Spain.Peer reviewe

Transcriptome analysis of barley anthers: effect of mannitol treatment on microspore embryogenesis

The definitive version is available at: http://www.blackwell-synergy.com/doi/full/10.1111/j.1399-3054.2006.00729.xCarbohydrate starvation is an efficient stress treatment for induction of microspore embryogenesis. Transcriptome analysis of anthers response to mannitol treatment using the 22k Barley1 GeneChip revealed large changes in gene expression. Statistical analysis and filtering for 4-fold or greater changes resulted in 2,673 genes, of which 887 were up-regulated and 1,786 down-regulated. Great differences in some metabolic pathways, accompanied by a multi-dimensional stress response were found. Analysis of transcription factors showed that most of the down-regulated transcription factors were related to growth and development, and the up-regulated with abiotic and biotic stress responses and changes in developmental programs. Interestingly, the expression of most cell cycle related genes did not change significantly. Transcriptome analysis provided a successful approach to identify genes involved in mannitol treatment, essential for triggering microspore embryogenesis.María Muñoz-Amatriaín is recipient of a FPI fellowship and a Short-term Scientific Mission to UCR, from Ministerio de Educación y Ciencia of Spain. The research was supported by Projects AGL-2001-1631 and AGL-2004-03396, from Plan Nacional I+D+I, Ministerio de Educación y Ciencia of Spain.Peer reviewe

Comparative mapping of the Oregon Wolfe Barley using doubled haploid lines derived from female and male gametes

28 Pag., 2 Tabl. 4 Fig. The definitive version is available at: http://www.springerlink.com/content/0040-5752/The Oregon Wolfe Barley mapping population is a resource for genetics research and instruction. Prior reports are based on a population of doubled haploid (DH) lines developed by the Hordeum bulbosum (H.b.) method, which samples female gametes. We developed new DH lines from the same cross using anther culture (A.C.), which samples male gametes. Linkage maps were generated in each of the two subpopulations using the same 1,328 single nucleotide polymorphism markers. The linkage maps based on DH lines derived from the products of megasporogeneis and microsporogenesis revealed minor differences in terms of estimated recombination rates. There were no differences in locus ordering. There was greater segregation distortion in the A.C.-derived subpopulation than in the H.b.-derived subpopulation, but in the region showing the greatest distortion, the cause was more likely allelic variation at the ZEO1 plant height locus rather than to DH production method. The effects of segregation distortion and pleiotropy had greater impacts on estimates of quantitative trait locus effect than population size for reproductive fitness traits assayed under greenhouse conditions. The Oregon Wolfe Barley (OWB) population and data are community resources. Seed is available from three distribution centers located in North America, Europe, and Asia. Details on ordering seed sets, as well as complete genotype and phenotype data files, are available at http://wheat.pw.usda.gov/ggpages/maps/OWB/.L. Cistué was recipient of a Senior Research Fellowship from the Spanish Ministry of Science and Innovation during his time with the Oregon State University Barley Project and his research was financed by the Spanish Ministry of Science and Innovation thought the National Plan Projects AGL2005-07195-C02-01 and AGL2008-05541-C02-01. Alfonso Cuesta-Marcos was supported by a postdoctoral fellowship from the Spanish Ministerio de Ciencia e Innovación (MICINN). The BOPA SNPs were developed under the auspices of USDA-CSREES-NRI Grant No 2006- 55606-16722 “Barley Coordinated Agricultural Project: Leveraging Genomics, Genetics, and Breeding for Gene Discovery and Barley Improvement”.Peer reviewe

Genetic basis of barley contributions to beer flavor

13 Pags.- 1 Fig.- 3 Tabls. © 2022 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license.Barley malt is critical for the malting, brewing, and distilling industries, as it is one of the main ingredients of beer and some types of spirits. There is growing evidence that barley genotype - via malt - can impact the flavors of beers and spirits. However, information on the barley genes involved in these flavors is lacking. Therefore, we used quantitative trait locus (QTL) mapping of malt quality traits, beer sensory descriptors and metabolic compounds on a biparental population of doubled haploids derived from the cross of the cultivars Golden Promise and Full Pint. Putative candidate genes for QTLs were identified by alignment with the reference barley genome sequence. There were thirty-seven QTLs across all chromosomes except 4H, with three QTL clusters located on 3H (1 cluster) and 5H (2 clusters: mid-5H and end-5H). Those “hotspots” contained QTLs for multiple phenotypes. Several candidate genes that regulate plant metabolism were identified within the QTLs and included HvAlaAT, HvDep1, HvMKK3, HvGA20ox1 and HvGA20ox2. These genes are involved in seed dormancy and plant height. Alleles at these loci, and perhaps at physically linked loci, can have key downstream effects on malting quality, beer flavor, and abundance of volatile metabolites.Research at Oregon State University was supported by the Agricultural Research Foundation Barley Progress Fund. At Colorado State University, research was supported by CSU's College of Agricultural Sciences, with partial support from the American Malting Barley Association.Peer reviewe

The Oregon Promise Barley Population: A tool for understanding the genetic basis of traits fundamental for barley production, malting, brewing, and distilling

The simultaneous availability of unique germplasm resources and cost-effective high-throughput genotyping allows for accelerated genome exploration and gene discovery. Our germplasm -the Oregon Promise population- is an array of 200 barley doubled haploids developed from the cross of Full Pint x Golden Promise. The spring 2-row parents have contrasting alleles at two of the dwarfing genes deployed in current varieties. The four homozygous combinations of these plant height alleles lead to contrasting phenotypes and each allele has pleiotropic effects on a range of other traits. Golden Promise is an iconic variety for malting, brewing, and distilling; Full Pint is a contributor to the craft brew Renaissance. Accordingly, the Oregon Promise will provide a valuable resource for extending current knowledge of malting and brewing genes to the frontiers of sensory assessment. The population shows transgressive segregation for adult plant resistance to stripe rust. As this disease is likely to become increasingly prevalent as a consequence of climate change, expanding the catalog of genes conferring durable resistance to this pathogen is an essential defensive breeding step. The availability of a quick-turnaround and cost effective SNP genotyping service (400+ markers) at Eureka Genomics (developed in collaboration with the James Hutton Institute) allows accelerated linkage map construction, QTL detection, and unraveling of gene interactions and pleiotropic effects based on the multi-environment, multi-trait phenotyping of the Oregon Promise population. This project is possible thanks to the tools and knowledge generated by the USDA-NIFA T-CAP project.Peer Reviewe

Estudios citogenéticos en Beta vulgaris L.

206 Pag.Los objetivos de la presente tesis doctoral pueden dividirse en dos grandes apartados: a. Desarrollo y aplicación de técnicas citológicas que permiten la identificación positiva de los distintos componentes del complemento cromosómico de Beta vuigaris L. Este objetivo se subdivide en: 1. Obtención del cariotipo metafásico en diploides. 2. Obtención de haploides como situación óptima para estudiar diferencias entre los nueve tipos cromosómicos. 3. Obtención del cariotipo prometafásico en dipicides. 4. Obtención del cariotipo profásico. Posibilidad y utilidad en diploides. 5. Diferencias entre metafases, prometafases y profases. 6. Estudio de la contracción del cromosoma. b. Identificación positiva de cada uno de los nueve componentes de la serie trisómica primaria de Beta vulgaris L. a través de técnicas citológicas. Este objetivo puede subdividirse en: 1. Obtención de placas cariotipicas de plantas trisómicas. 2. Determinar que cromosoma está triplicado en cada uno de los nueve grupos morfológicos de la línea homocigótica trisómica de I. Romagosa. 3. Comprobar si existe alguna anormalidad cromosómíca en la línea trisómica de Butterfass..Peer reviewe

Wheat doubled haploids: Production to sequencing. What makes them so appealing?

35 Pags.- Chapter Book of the Authors into vol. 3 of the World Wheat Book. This volume is devoted to a updating of the first two volumes by adding to the countries discussed, giving an update on associated wheat technologies and giving an insight into the means to enhance production in a changing environment.Peer reviewe

Gametic embryogenesis in Triticum: A study of some critical factors in haploid (microspore) embryogenesis

This review of embryogenesis from microspores in two types of wheat examines the influences of various procedures used to induce and produce embryoids from the male gametes. The various factors that influence the quality of the microspores are critical and include growth of donor plants, pretreatments and culture and regeneration media. There are also many genetic and genotype factors, with a major problem being production of albino plants. Genotype differences in response and gametic selection during culture may be overcome through improved response in culture such as has been achieved in barley. Chromosome doubling to obtain uniform doubled haploid lines is being continually improved through pretreatments to cause nuclear fusion and through anti-microtubule agents in the induction media. Finally, the interesting aspects of cell and tissue changes that occur during the induction and development of the embryoids are briefly described and illustrated.Peer Reviewe