Xiao-Qi Zhang

1. Barley PanTs Supplementary Data1-8_10-14.xlsx
Ten Datasets referred to in the main text (Supplementary Data 1-10), presented in tabular format as separate sheets in a joint Excel formatted file.
Supplementary Data 1: Basic statistics of the RTDs
Supplementary Data 2: Ordering of genotypes incorporated into the linear pan-genome.
Supplementary Data 3: Gene categories of GsRTD
Supplementary Data 4: Alternative splicing events for highly expressed transcripts from core-single-copy genes (average TPM > 10)
Supplementary Data 5: Gene copy number variation cluster significantly correlated with the gene expression.
Supplementary Data 6: C-repeat/DRE-Binding Factor (CBF) genes identifier in GsRTD and their location in the genome
Supplementary Data 7: Genotypes with the 141Mb 7H inversion and non-inversion
Supplementary Data 8: Differentially expressed genes in the 7H inversion.
Supplementary Data 10: Detailed example of a split pattern in Golden Promise
Supplementary Data 11: Barley cv. Morex Expression Atlas Metadata
Supplementary Data 12: GA-Pathway gene expression in PanTs experiment
Supplementary Data 13: Yield and agro data for Ga2ox3-7
Supplementary Data 14: Statistics GA2ox and 7

2. Supplementary Data9.csv A csv file containing genotype-specific co-expression network results (modules and community assignments) with annotation and MorexV3 gene IDs.

3. Supplementary Data15.txt This tab delimited text file is too large for inclusion in the main Supplementary Data file and contains the details of how genes/transcripts from the genotype specific RTDs map onto genes in PanBaRT20.

Oat grain is a traditional human food that is rich in dietary fibre and contributes to improved human health1,2. Interest in the crop has surged in recent years owing to its use as the basis for plant-based milk analogues3. Oat is an allohexaploid with a large, repeat-rich genome that was shaped by subgenome exchanges over evolutionary timescales4. In contrast to many other cereal species, genomic research in oat is still at an early stage, and surveys of structural genome diversity and gene expression variability are scarce. Here we present annotated chromosome-scale sequence assemblies of 33 wild and domesticated oat lines, along with an atlas of gene expression across 6 tissues of different developmental stages in 23 of these lines. We construct an atlas of gene-expression diversity across subgenomes, accessions and tissues. Gene loss in the hexaploid is accompanied by compensatory upregulation of the remaining homeologues, but this process is constrained by subgenome divergence. Chromosomal rearrangements have substantially affected recent oat breeding. A large pericentric inversion associated with early flowering explains distorted segregation on chromosome 7D and a homeologous sequence exchange between chromosomes 2A and 2C in a semi-dwarf mutant has risen to prominence in Australian elite varieties. The oat pangenome will promote the adoption of genomic approaches to understanding the evolution and adaptation of domesticated oats and will accelerate their improvement.Oat grain is a traditional human food that is rich in dietary fibre and contributes to improved human health1,2. Interest in the crop has surged in recent years owing to its use as the basis for plant-based milk analogues3. Oat is an allohexaploid with a large, repeat-rich genome that was shaped by subgenome exchanges over evolutionary timescales4. In contrast to many other cereal species, genomic research in oat is still at an early stage, and surveys of structural genome diversity and gene expression variability are scarce. Here we present annotated chromosome-scale sequence assemblies of 33 wild and domesticated oat lines, along with an atlas of gene expression across 6 tissues of different developmental stages in 23 of these lines. We construct an atlas of gene-expression diversity across subgenomes, accessions and tissues. Gene loss in the hexaploid is accompanied by compensatory upregulation of the remaining homeologues, but this process is constrained by subgenome divergence. Chromosomal rearrangements have substantially affected recent oat breeding. A large pericentric inversion associated with early flowering explains distorted segregation on chromosome 7D and a homeologous sequence exchange between chromosomes 2A and 2C in a semi-dwarf mutant has risen to prominence in Australian elite varieties. The oat pangenome will promote the adoption of genomic approaches to understanding the evolution and adaptation of domesticated oats and will accelerate their improvement.

A pan-transcriptome describes the transcriptional and post-transcriptional consequences of genome diversity from multiple individuals within a species. We developed a barley pan-transcriptome using 20 inbred genotypes representing domesticated barley diversity by generating and analyzing short- and long-read RNA-sequencing datasets from multiple tissues. To overcome single reference bias in transcript quantification, we constructed genotype-specific reference transcript datasets (RTDs) and integrated these into a linear pan-genome framework to create a pan-RTD, allowing transcript categorization as core, shell or cloud. Focusing on the core (expressed in all genotypes), we observed significant transcript abundance variation among tissues and between genotypes driven partly by RNA processing, gene copy number, structural rearrangements and conservation of promotor motifs. Network analyses revealed conserved co-expression module::tissue correlations and frequent functional diversification. To complement the pan-transcriptome, we constructed a comprehensive cultivar (cv.) Morex gene-expression atlas and illustrate how these combined datasets can be used to guide biological inquiry.

Pangenomes are collections of annotated genome sequences of multiple individuals of a species1. The structural variants uncovered by these datasets are a major asset to genetic analysis in crop plants2. Here we report a pangenome of barley comprising long-read sequence assemblies of 76 wild and domesticated genomes and short-read sequence data of 1,315 genotypes. An expanded catalogue of sequence variation in the crop includes structurally complex loci that are rich in gene copy number variation. To demonstrate the utility of the pangenome, we focus on four loci involved in disease resistance, plant architecture, nutrient release and trichome development. Novel allelic variation at a powdery mildew resistance locus and population-specific copy number gains in a regulator of vegetative branching were found. Expansion of a family of starch-cleaving enzymes in elite malting barleys was linked to shifts in enzymatic activity in micro-malting trials. Deletion of an enhancer motif is likely to change the developmental trajectory of the hairy appendages on barley grains. Our findings indicate that allelic diversity at structurally complex loci may have helped crop plants to adapt to new selective regimes in agricultural ecosystems.

Seed dormancy is an important trait associated with pre-sprouting and malting quality in barley. Genome-wide association studies (GWASs) have been used to detect quantitative trait loci (QTLs) underlying complex traits in major crops. In the present study, we collected 295 barley (Hordeum vulgare L.) accessions from Australia, Europe, Canada and China. A total of 25,179 single nucleotide polymorphism (SNP)/diversity arrays technology sequence markers were used for population structure, linkage disequilibrium and GWAS analysis. Candidate genes within QTL regions were investigated, and their expression levels were analysed using RNAseq data. Five QTLs for seed dormancy were identified. One QTL was mapped on chromosome 1H, and one QTL was mapped on chromosome 4H, while three QTLs were located on chromosome 5H. This is the first report of a QTL on the short arm of chromosome 5H in barley. Molecular markers linked to the QTL can be used for marker-assisted selection in barley breeding programmes.

Sixteen green asparagus cultivars from 6 countries were planted in a greenhouse in Hangzhou, China (120.2°E, 30.3°N) in 2016. The annual cultivation system, consisting of one spring harvest using the conventional method, along with two summer and autumn harvests using the mother-stem method, was applied. The seasonal and annual total yields were determined in the first and second years of harvest, i.e., in 2017 and 2018. In every harvest season, spears were cut at the length of 37 cm and their quality and total yield were evaluated. ‘Pacific Challenger 1’, ‘Sunlim’, ‘Pacific Challenger 2’, ‘Pacific Endeavor’ and ‘H2-4’ were the top 5 yielders during the first two years of harvest. The average spear weight of ‘Italo’, ‘Sunlim’ and ‘Early California’ was over 20 g in all harvest seasons. In the second harvest year, the yield of ‘Pacific Challenger 1’, ‘Pacific Challenger 2’ and ‘Grande’ produced a relatively higher total yield than other cultivars in spring. The yield of ‘Pacific Challenger 2’ was the highest in summer, while ‘Italo’, ‘Pacific Challenger 1’, and ‘Pacific Challenger 2’ showed higher yield in autumn. Among three harvest seasons, the yield of all cultivars in autumn did not reach the expected level, which may have been caused by the typhoons, Capricorn and Winbia, in August 2018, as well as the deficiency of necessary fertilization. Overall, ‘Pacific Challenger 2’ seemed to be a good yielder in all harvest seasons during the second year of harvest and looked to be a promising cultivar to be cultivated in the greenhouse conditions in Hangzhou, China.

Divergent selection of populations in contrasting environments leads to functional genomic divergence. However, the genomic architecture underlying heterogeneous genomic differentiation remains poorly understood. Here, we de novo assembled two high-quality wild barley (Hordeum spontaneum K. Koch) genomes and examined genomic differentiation and gene expression patterns under abiotic stress in two populations. These two populations had a shared ancestry and originated in close geographic proximity but experienced different selective pressures due to their contrasting micro-environments. We identified structural variants that may have played significant roles in affecting genes potentially associated with well-differentiated phenotypes such as flowering time and drought response between two wild barley genomes. Among them, a 29-bp insertion into the promoter region formed a cis-regulatory element in the HvWRKY45 gene, which may contribute to enhanced tolerance to drought. A single SNP mutation in the promoter region may influence HvCO5 expression and be putatively linked to local flowering time adaptation. We also revealed significant genomic differentiation between the two populations with ongoing gene flow. Our results indicate that SNPs and small SVs link to genetic differentiation at the gene level through local adaptation and are maintained through divergent selection. In contrast, large chromosome inversions may have shaped the heterogeneous pattern of genomic differentiation along the chromosomes by suppressing chromosome recombination and gene flow. Our research offers novel insights into the genomic basis underlying local adaptation and provides valuable resources for the genetic improvement of cultivated barley.

Climate changes threaten global sustainable food supply by reducing crop yield. Estimates of future crop production under climate change have rarely considered the capacity of genetic improvement in breeding high-yielding and stress-tolerant crop varieties. We believe that technological advancements and developing climate-resilient crop varieties may offset the adverse effects of climate change. In this study, we examined the historical record of barley breeding and yield, and the trends of climate changes over the past 70 years in Australia. We related the selection of fast development varieties to yield improvement, and revealed the genetic connections of fast development and yield potential through genome-wide association studies. Historical records show that Australia's barley yield has experienced a steady growth despite that the seasonal production window has been shortened due to increased risk of frost damage at flowering stage and terminal heat during maturity since the 1970s. The increase in yield is largely the result of higher yield capacity of the more recently developed varieties that develop faster to counteract the impact of increased terminal heat. We also show that the changing temperature may soon reach a critical point that dramatically changes the barley flowering behaviour to impact yield by pushing its growth beyond the seasonal production window to face increasing frost damage. For the first time, we provide evidence that the effects of climate change on crop production might be less severe than what is currently believed because the advancement of technologies and development of climate-resilient crop varieties may mitigate the adverse effect of climate change to some extent. The greater use of genetic techniques in crop breeding will play a vital role in sustainable global food production in the era of climate change.