Powdery mildew disease resistance and markerassisted screening at the Pm60 locus in wild diploid wheat Triticum urartu

Triticum urartu,a diploid wild wheat and progenitor of the A genome of bread wheat,is an important resource for resistance to powdery mildew fungus caused by Blumeria graminis f.sp.tritici(Bgt).In the present study we systematically characterized the interaction between the Bgt fungus and T.urartu at the microscopic level.We also tested 227 T.urartu accessions for reaction to Bgt isolate E09 and discovered previously uncategorized powdery mildew resistance in this collection.Pm60 is a CC-NB-LRR type powdery mildew resistance gene that has at least three functional alleles,Pm60,Pm60a,and Pm60b.A marker-assisted screen targeting the Pm60 locus identified a non-functional allele of Pm60a,designated as Pm60a′.A sequence comparison of Pm60a′and Pm60a revealed that they differed by 58 SNPs and one 3-nucleotide deletion.Based on the sequence variations two molecular markers were developed to differentiate the functional Pm60a allele from the non-functional Pm60a′.Our screen revealed the presence of a previously uncharacterized powdery mildew resistance in T.urartu and provides new insights into the Pm60 locus.We believe that the two molecular markers developed here and new T.urartu resistant accessions will facilitate further identification of novel powdery mildew resistance genes and benefit breeding for powdery mildew resistance.

Low-affinity SPL binding sites contribute to subgenome expression divergence in allohexaploid wheat

Expression divergence caused by genetic variation and crosstalks among subgenomes of the allohexaploid bread wheat(Triticum aestivum.L.,BBAADD)is hypothesized to increase its adaptability and/or plasticity.However,the molecular basis of expression divergence remains unclear.Squamosa promoter-binding protein-like(SPL)transcription factors are critical for a wide array of biological processes.In this study,we constructed expression regulatory networks by combining DAP-seq for 40 SPLs,ATACseq,and RNA-seq.Our findings indicate that a group of low-affinity SPL binding regions(SBRs)were targeted by diverse SPLs and caused different sequence preferences around the core GTAC motif.The SBRs including the low-affinity ones are evolutionarily conserved,enriched GWAS signals related to important agricultural traits.However,those SBRs are highly diversified among the cis-regulatory regions(CREs)of syntenic genes,with less than 8%SBRs coexisting in triad genes,suggesting that CRE variations are critical for subgenome differentiations.Knocking out of Ta SPL7A/B/D and Ta SPL15A/B/D subfamily further proved that both high-and low-affinity SBRs played critical roles in the differential expression of genes regulating tiller number and spike sizes.Our results have provided baseline data for downstream networks of SPLs and wheat improvements and revealed that CRE variations are critical sources for subgenome divergence in the allohexaploid wheat.

Chromatin accessibility landscapes revealed the subgenome-divergent regulation networks during wheat grain development

Development of wheat(Triticum aestivum L.)grain mainly depends on the processes of starch synthesis and storage protein accumulation,which are critical for grain yield and quality.However,the regulatory network underlying the transcriptional and physiological changes of grain development is still not clear.Here,we combined ATAC-seq and RNA-seq to discover the chromatin accessibility and gene expression dynamics during these processes.We found that the chromatin accessibility changes are tightly associated with differential transcriptomic expressions,and the proportion of distal ACRs was increased gradually during grain development.Specific transcription factor(TF)binding sites were enriched at different stages and were diversified among the 3 subgenomes.We further predicted the potential interactions between key TFs and genes related with starch and storage protein biosynthesis and found different copies of some key TFs played diversified roles.Overall,our findings have provided numerous resources and illustrated the regulatory network during wheat grain development,which would shed light on the improvement of wheat yields and qualities.