Fine mapping and validation of a stable QTL for thousand-kernel weight in wheat(Triticum aestivum L.)

Thousand-kernel weight(TKW)is a measure of grain weight,a target of wheat breeding.The object of this study was to fine-map a stable quantitative trait loci(QTL)for TKW and identify its candidate gene in a recombinant inbred line(RIL)population derived from the cross of Kenong 9204(KN9204)and Jing411(J411).On a high-density genetic linkage map,24,26 and 25 QTL were associated with TKW,kernel length(KL),and kernel width(KW),respectively.A major and stable QTL,QTkw-2D,was mapped to an8.3 cM interval on chromosome arm 2DL.By saturation of polymorphic markers in its target region,QTkw-2D was confined to a 9.13 Mb physical interval using a secondary mapping population derived from a residually heterozygous line(F6:7).This interval was further narrowed to 2.52 Mb using QTkw-2D near-isogenic lines(NILs).NILs~(KN9204)had higher fresh and dry weights than NILsJ411at various grain-filling stages.The TKW and KW of NILs~(KN9204)were much higher than those of NILsJ411in field trials.By comparison of both DNA sequence and expression between KN9204 and J411,TraesCS2D02G460300.1(TraesKN2D01HG49350)was assigned as a candidate gene for QTkw-2D.This was confirmed by RNA sequencing(RNA-seq)of QTkw-2D NILs.These results provide the basis of map-based cloning of QTkw-2D,and DNA markers linked to the candidate gene may be used in marker-assisted selection.

Boosting wheat functional genomics via an indexed EMS mutant library of KN9204

A better understanding of wheat functional genomics can improve targeted breeding for better agronomic traits and environmental adaptation.However,the lack of gene-indexed mutants and the low transformation efficiency of wheat limit in-depth gene functional studies and genetic manipulation for breeding.In this study,we created a library for KN9204,a popular wheat variety in northern China,with a reference genome,transcriptome,and epigenome of different tissues,using ethyl methyl sulfonate(EMS)mutagenesis.This library contains a vast developmental diversity of critical tissues and transition stages.Exome capture sequencing of 2090 mutant lines using KN9204 genome-designed probes revealed that 98.79%of coding genes had mutations,and each line had an average of 1383 EMS-type SNPs.We identified new allelic variations for crucial agronomic trait-related genes such as Rht-D1,Q,TaTB1,and WFZP.We tested 100 lines with severemutations in 80 NAC transcription factors(TFs)under drought and salinity stress and identified 13 lines with altered sensitivity.Further analysis of three lines using transcriptome and chromatin accessibility data revealed hundreds of direct NAC targets with altered transcription patterns under salt or drought stress,including SNAC1,DREB2B,CML16,and ZFP182,factors known to respond to abiotic stress.Thus,we have generated and indexed a KN9204 EMS mutant library that can facilitate functional genomics research and offer resources for genetic manipulation of wheat.

BZR1 Family Transcription Factors Function Redundantly and Indispensably in BR Signaling but Exhibit BRI1-Independent Function in Regulating Anther Development in Arabidopsis

BRASSINAZOLE-RESISTANT 1 family proteins(BZRs)are central transcription factors that govern brassinosteroid(BR)-regulated gene expression and plant growth.However,it is unclear whether there exists a BZR-independent pathway that mediates BR signaling.In this study,we found that disruption of all BZRs in Arabidopsis generated a hextuple mutant(bzr-h)displaying vegetative growth phenotypes that were almost identical to those of the null mutant of three BR receptors,bri1brl1brl3(bri-t).By RNA sequencing,we found that global gene expression in bzr-h was unaffected by 2 h of BR treatment.The anthers of bzr-h plants were loculeless,but a similar phenotype was not observed in bri-t,suggesting that BZRs have a BR signaling-independent regulatory role in anther development.By real-time PCR and in situ hybridization,we found that the expression of SPOROCYTELESS(SPL),which encodes a transcription factor essential for anther locule development,was barely detectable in bzr-h,suggesting that BZRs regulate locule development by affecting SPL expression.Our findings reveal that BZRs are indispensable transcription factors required for both BR signaling and anther locule development,providing new insight into the molecular mechanisms underlying the microsporogenesis in Arabidopsis.