Plant AT-rich sequence and zinc-binding proteins (PLATZs) are a novel category of plant-specific transcription factors involved in growth, development, and abiotic stress responses. However, the PLATZ gene family has not been identified in barley. In this study, a total of 11 HvPLATZs were identified in barley, and they were unevenly distributed on five of the seven chromosomes. The phylogenetic tree, incorporating PLATZs from Arabidopsis, rice, maize, wheat, and barley, could be classified into six clusters, in which HvPLATZs are absent in Cluster VI. HvPLATZs exhibited conserved motif arrangements with a characteristic PLATZ domain. Two segmental duplication events were observed among HvPLATZs. All HvPLATZs were core genes present in 20 genotypes of the barley pan-genome. The HvPLATZ5 coding sequences were conserved among 20 barley genotypes, whereas HvPLATZ4/9/10 exhibited synonymous single nucleotide polymorphisms (SNPs); the remaining ones showed nonsynonymous variations. The expression of HvPLATZ2/3/8 was ubiquitous in various tissues, whereas HvPLATZ7 appeared transcriptionally silent; the remaining genes displayed tissue-specific expression. The expression of HvPLATZs was modulated by salt stress, potassium deficiency, and osmotic stress, with response patterns being time-, tissue-, and stress type-dependent. The heterologous expression of HvPLATZ3/5/6/8/9/10/11 in yeast enhanced tolerance to salt and osmotic stress, whereas the expression of HvPLATZ2 compromised tolerance. These results advance our comprehension and facilitate further functional characterization of HvPLATZs.

β-amylase activity is related to the polymorphisms of Bmy1 intron III; however, no attention has been given to such relationships under environmental stresses such as drought. In this study, 73 cultivated barley genotypes and 52 Tibetan wild barley accessions were used to test the association between Bmy1 gene intron III polymorphisms and β-amylase activity under drought stress. Our results showed that three alleles, Bmy1.a, Bmy1.b, and Bmy1.c, existed in the examined barley genotypes. Tibetan wild barley had a higher proportion of Bmy1.b, whereas cultivated barley showed a higher proportion of Bmy1.a. Impressively, barley genotypes with Bmy1.b showed a significant increase in β-amylase activity under drought stress, compared with those with Bmy1.a or Bmy1.c, indicating that the Bmy1.b allele might provide more chances for developing barley cultivars with higher β-amylase activity under water stress than both the Bmy1.a and Bmy1.c alleles. Furthermore, the Tibetan wild barley XZ147, belonging to the Bmy1.b allele type, showed significantly higher β-amylase activity than Triumph under drought stress. This might be the result of the unique amino acid substitution M527 or the amino acid composition of R115, D165, A233, S347, and M527 of XZ147.

Abscisic acid (ABA)-stress-ripening, or ABA-, stress-, and ripening-induced (ASR) proteins play an important role in responses to environmental stimuli. A total of ten barley HvASRs were identified in this study, which were unevenly distributed on three chromosomes. ASRs from barley, wheat, Brachypodium distachyon, rice, maize, foxtail millet, and tomato were classified into two distinct clusters based on phylogenetic analysis. Notably, ASRs from Poaceae were evenly distributed between these two clusters. HvASRs contained a typical ABA/WDS domain, and exhibited similar motif arrangements. Two gene pairs of tandem duplicates (HvASR4/5/6/7 and HvASR8/9) were identified among HvASRs. Cis-acting elements involved in hormone and stress responses, including ABRE, MYB, ARE, and STRE, were consistently identified in the promoters of HvASRs. The expression of HvASRs was substantially influenced by salt, osmotic, and ABA treatments in the roots and leaves of barley seedlings. HvASR2 acts as a transcriptional repressor, whereas HvASR3 serves as a transcriptional activator. These results enhance our understanding of the HvASR family and provide a foundation for further functional characterization.

The chloroplast is an important cellular organelle and metabolic hub, which is not only responsible for plant photosynthesis but is also involved in the de novo biosynthesis of pigments, fatty acids, and hormone metabolisms. Several genes that are responsible for rice leaf color variations have been reported to be directly or indirectly involved in chlorophyll biosynthesis and chloroplast development, whereas a few genes have been functionally confirmed to be responsible for leaf color changes in barley at the molecular level. In this study, we obtained a yellow leaf and dwarf ygl8 mutant from the progeny of Morex (a variety of barley) seeds treated with EMS. We performed bulked-segregant analysis (BSA) and RNA-seq analysis and targeted a UMP kinase encoding gene, YGL8, which generated a splicing site change between exon 5 and 6 of YGL8 due to a G to A single-nucleotide transition in the 5th exon/intron junction in the ygl8 mutant. The splicing site change between exon 5 and 6 of YGL8 had no effects on chloroplast subcellular localization but resulted in an additional loop in the UMP kinase domain, which might disturb the access of the substrates. On one hand, the splicing site change between exon 5 and 6 of YGL8 downregulated the transcriptional expression of chloroplast-encoded genes and chlorophyll-biosynthesis-related genes in a temperature-dependent manner in the ygl8 mutant. On the other hand, the downregulation of bioactive GA-biosynthesis-related GA20ox genes and cell-wall-cellulose-biosynthesis-related CesA genes was also observed in the ygl8 mutant, which led to a reduction in plant height. Our study will facilitate the understanding of the regulation of leaf color and plant height in barley.

Genomic structural variants (SVs) are major sources of genetic variation and have profound impacts on phenotypic traits. However, their functional effects remain largely unexplored in soybean. Here, we resequence 940 soybean accessions. Together with 44 publicly available datasets, we identify 602,281 SVs. Using a graph-based genome, we detect an additional 58,760 presence/absence variations (PAVs) that broadly affect gene expression. Population genomic analyses reveal that SVs serve as a core driving force for soybean domestication and improvement. Integrating SVs with QTLs for oil and protein content, and performing GWAS on 27 traits, we identify key functional SVs. These include transposable element insertions altering seed coat color, multiple insertions within a cytochrome P450 gene modifying flower and hypocotyl color, and a GmMATE1 deletion enhancing seed size. Together, our study establishes a comprehensive SV map of soybean, offering a valuable resource for dissecting the genetic basis of complex traits to accelerate molecular breeding.

Barley (Hordeum vulgare) is one of the earliest domesticated cereal crops and remains important for global feed, food, and malting industries. Although high-quality genomic resources are now available, its large repeat-rich genome has complicated genome-wide characterization of genomic features associated with barley evolution and domestication. Here, we integrated comparative genomics, methylomics, population genomics, and Hi-C analyses to investigate genome evolution and domestication in barley. Comparative analyses across representative grass species revealed that barley exhibits lineage-specific genome expansion associated with the extensive proliferation of long terminal repeat retrotransposons. Genome-wide DNA methylation analyses revealed distinct cytosine methylation landscapes in barley. Population genomic analyses of 291 wild and domesticated barley accessions revealed relatively slow linkage disequilibrium decay and identified 243 candidate domestication-associated genes. These candidate genes were enriched in starch and sucrose metabolism pathways. Comparative Hi-C analyses of three barley accessions further uncovered differences in A/B chromatin compartments and topologically associating domain (TAD) organization despite their largely conserved gene content. In addition, the enrichment of DNA transposons and Copia retrotransposons at TAD boundaries suggests potential associations between specific TE classes and higher-order chromatin organization. Together, these complementary analyses connect genome expansion, epigenetic regulation, domestication-associated selection, and chromatin organization into a unified framework for understanding barley genome evolution and domestication.