Hordeum bulbosum L., a wild relative of barley （Hordeum vulgare L.）, has been considered as a valuable source of genetic diversity for barley improvement. Since the 1990s, a considerable number of barley/H, bulbosum...Hordeum bulbosum L., a wild relative of barley （Hordeum vulgare L.）, has been considered as a valuable source of genetic diversity for barley improvement. Since the 1990s, a considerable number of barley/H, bulbosum introgression lines （IL）s has been generated, with segments introgressed from H. bulbosum harboring a diverse set of desirable traits. However, the efficient utilization of these ILs has been hampered, largely due to the lack of suitable molecular tools for their genetic characterization and highly reduced interspecific recombination frequencies in the region of the introgression. In the present study, we utilized genotyping-by-sequencing for the detailed molecular characterization of 145 ILs. Genotypic information allows the genetic diversity within the set of ILs to be determined and a strategy was outlined to tackle the obstacle of reduced recombination frequencies. Furthermore, we compiled exome capture re-sequencing information of barley and H. bulbosum and designed an integrated barley/ H. bulbosum sequence resource with polymorphism information on interspecific and intraspecific sequence variations of both species. The integrated sequence will be valuable for marker development in barley/H, bulbosum ILs derived from any barley and H. bulbosum donors. This study provides the tools for the widespread utilization of barley/H, bulbosum ILs in applied barley breeding and academic research.展开更多
Barley is a diploid species with a genome smaller than those of other members of the Triticeae tribe,making it an attractive model for genetic studies in Triticeae crops.The recent development of barley genomics has c...Barley is a diploid species with a genome smaller than those of other members of the Triticeae tribe,making it an attractive model for genetic studies in Triticeae crops.The recent development of barley genomics has created a need for a high-throughput platform to identify genetically uniform mutants for gene function investigations.In this study,we report an ethyl methanesulfonate(EMS)-mutagenized population consisting of 8525M_(3) lines in the barley landrace“Hatiexi”(HTX),which we complement with a high-quality de novo assembly of a reference genome for this genotype.The mutation rate within the population ranged from 1.51 to 4.09 mutations per megabase,depending on the treatment dosage of EMS and the mutation discrimination platform used for genotype analysis.We implemented a three-dimensional DNA pooling strategy combined with multiplexed amplicon sequencing to create a highly efficient and cost-effective TILLING(targeting induced locus lesion in genomes)platform in barley.Mutations were successfully identified from 72 mixed amplicons within a DNA pool containing 64 individual mutants and from 56 mixed amplicons within a pool containing 144 individuals.We discovered abundant allelic mutants for dozens of genes,including the barley Green Revolution contributor gene Brassinosteroid insensitive 1(BRI1).As a proof of concept,we rapidly determined the causal gene responsible for a chlorotic mutant by following the MutMap strategy,demonstrating the value of this resource to support forward and reverse genetic studies in barley.展开更多
文摘Hordeum bulbosum L., a wild relative of barley （Hordeum vulgare L.）, has been considered as a valuable source of genetic diversity for barley improvement. Since the 1990s, a considerable number of barley/H, bulbosum introgression lines （IL）s has been generated, with segments introgressed from H. bulbosum harboring a diverse set of desirable traits. However, the efficient utilization of these ILs has been hampered, largely due to the lack of suitable molecular tools for their genetic characterization and highly reduced interspecific recombination frequencies in the region of the introgression. In the present study, we utilized genotyping-by-sequencing for the detailed molecular characterization of 145 ILs. Genotypic information allows the genetic diversity within the set of ILs to be determined and a strategy was outlined to tackle the obstacle of reduced recombination frequencies. Furthermore, we compiled exome capture re-sequencing information of barley and H. bulbosum and designed an integrated barley/ H. bulbosum sequence resource with polymorphism information on interspecific and intraspecific sequence variations of both species. The integrated sequence will be valuable for marker development in barley/H, bulbosum ILs derived from any barley and H. bulbosum donors. This study provides the tools for the widespread utilization of barley/H, bulbosum ILs in applied barley breeding and academic research.
基金funded by grants from the National Key Research and Development Program of China(2018YFD1000702/2018YFD1000700)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS),China.
文摘Barley is a diploid species with a genome smaller than those of other members of the Triticeae tribe,making it an attractive model for genetic studies in Triticeae crops.The recent development of barley genomics has created a need for a high-throughput platform to identify genetically uniform mutants for gene function investigations.In this study,we report an ethyl methanesulfonate(EMS)-mutagenized population consisting of 8525M_(3) lines in the barley landrace“Hatiexi”(HTX),which we complement with a high-quality de novo assembly of a reference genome for this genotype.The mutation rate within the population ranged from 1.51 to 4.09 mutations per megabase,depending on the treatment dosage of EMS and the mutation discrimination platform used for genotype analysis.We implemented a three-dimensional DNA pooling strategy combined with multiplexed amplicon sequencing to create a highly efficient and cost-effective TILLING(targeting induced locus lesion in genomes)platform in barley.Mutations were successfully identified from 72 mixed amplicons within a DNA pool containing 64 individual mutants and from 56 mixed amplicons within a pool containing 144 individuals.We discovered abundant allelic mutants for dozens of genes,including the barley Green Revolution contributor gene Brassinosteroid insensitive 1(BRI1).As a proof of concept,we rapidly determined the causal gene responsible for a chlorotic mutant by following the MutMap strategy,demonstrating the value of this resource to support forward and reverse genetic studies in barley.
