The leaf is not only the main site of photosynthesis, but also an important organ reflecting plant salt tolerance. Discovery of salt-stress-responding genes in the leaf is of great significance for the molecular improvement of salt tolerance in wheat varieties. In this study, transcriptome sequencing was conducted on the leaves of salt-tolerant wheat germplasm CH7034 seedlings at 0, 1, 6, 24, and 48 h after NaCl treatment. Based on weighted gene correlation network analysis of differentially expressed genes (DEGs) under salt stress, 12 co-expression modules were obtained, of which, 9 modules containing 4029 DEGs were related to the salt stress time-course. These DEGs were submitted to the Wheat Union database, and a total of 904,588 SNPs were retrieved from 114 wheat germplasms, distributed on 21 wheat chromosomes. Using the R language package and GAPIT program, association analysis was performed between 904,588 SNPs and leaf salt injury index of 114 wheat germplasms. The results showed that 30 single nucleotide polymorphisms (SNPs) from 15 DEGs were associated with salt tolerance. Then, nine candidate genes, including four genes (TaBAM, TaPGDH, TaGluTR, and TaAAP) encoding enzymes as well as five genes (TaB12D, TaS40, TaPPR, TaJAZ, and TaWRKY) encoding functional proteins, were identified by converting salt tolerance-related SNPs into Kompetitive Allele-Specifc PCR (KASP) markers for validation. Finally, interaction network prediction was performed on TaBAM and TaAAP, both belonging to the Turquoise module. Our results will contribute to a further understanding of the salt stress response mechanism in plant leaves and provide candidate genes and molecular markers for improving salt-tolerant wheat varieties.

The root is an important organ by which plants directly sense variation in soil moisture. The discovery of drought stress-responsive genes in roots is very important for the improvement of drought tolerance in wheat varieties via molecular approaches. In this study, transcriptome sequencing was conducted on the roots of drought-tolerant wheat cultivar YH1818 seedlings at 0, 2, and 7 days after treatment (DAT). Based on a weighted gene correlation network analysis of differentially expressed genes (DEGs), 14 coexpression modules were identified, of which five modules comprising 3107 DEGs were related to 2 or 7 DAT under drought stress conditions. A total of 223,357 single-nucleotide polymorphisms (SNPs) of these DEGs were retrieved from public databases. Using the R language package and GAPIT program, association analysis was performed between the 223,357 SNPs and the drought tolerance coefficient (DTC) values of six drought resistance-related traits in 114 wheat germplasms. The results revealed that 18 high-confidence SNPs of 10 DEGs, including TaPK, TaRFP, TaMCO, TaPOD, TaC3H-ZF, TaGRP, TaDHODH, TaPPDK, TaLectin, and TaARF7-A, were associated with drought tolerance. The RT–qPCR results confirmed that these genes were significantly upregulated by drought stress at 7 DAT. Among them, TaARF7-A contained three DTC-related SNPs, which presented two haplotypes in the tested wheat germplasms. YH1818 belongs to the Hap1 allele, which is involved in increased drought tolerance. This study revealed key modules and candidate genes for understanding the drought-stress response mechanism in wheat roots.

The stay-green (SG) trait enhances photosynthetic activity during the late grain-filling period, benefiting grain yield under drought and heat stresses. CH7034 is a wheat breeding line with SG. To clarify the SG loci carried by CH7034 and obtain linked molecular markers, in this study, a recombinant inbred line (RIL) population derived from the cross between CH7034 and non-SG SY95-71 was genotyped using the Wheat17K single-nucleotide polymorphism (SNP) array, and a high-density genetic map covering 21 chromosomes and consisting of 2159 SNP markers was constructed. Then, the chlorophyll content of flag leaf from each RIL was estimated for mapping, and one QTL for SG on chromosome 7D was identified, temporarily named QSg.sxau-7D, with the maximum phenotypic variance explained of 8.81~11.46%. A PCR-based diagnostic marker 7D-16 for QSg.sxau-7D was developed, and the CH7034 allele of 7D-16 corresponded to the higher flag leaf chlorophyll content, while the 7D-16 SY95-71 allele corresponded to the lower value, which confirmed the genetic effect on SG of QSg.sxau-7D. QSg.sxau-7D located in the 526.4~556.2 Mbp interval is different from all the known SG loci on chromosome 7D, and 69 high-confidence annotated genes within the interval expressed throughout the entire period of flag leaf senescence. Moreover, results of an association analysis based on the diagnostic marker showed that there is a positive correlation between QSg.sxau-7D and thousand-grain weight. Our results revealed a novel QTL QSg.sxau-7D whose CH7034 allele had a strong effect on SG, which can be applied in further wheat molecular breeding.

Yellow rust (YR), caused by Puccinia striiformis f. sp. tritici (Pst), is a global disease infecting wheat that seriously affects the yield and the quality of grains. Wheat breeding line C855 is immune to the mixed Pst isolates CYR32 + CYR33 + CYR34 under field conditions. To identify the Yr-loci carried by C855, in this study, an F₂ population derived from the crossing of C855 with Yannong 999, a YR-sensitive cultivar, was established, and the infection type (IT) of each F₂ individual was estimated. The correlation analysis results show that YR resistance was significantly positively correlated with grain weight and grain size. Using a 120K single-nucleotide polymorphism (SNP) array, the F₂ population was genotyped, and a high-density genetic map covering 21 wheat chromosomes and consisting of 5362 SNP markers was built. Then, five Yr-QTLs on chromosomes 1B, 2A, 2B, and 2D were identified. Of these, the QTL on chromosome 2A, temporarily named QYr.sxau-2A.1, is a major-effect QTL explaining 15.62% of the phenotypic variance. One PCR-based marker SSR2A-14 for QYr.sxau-2A.1 was developed, and the C855 allele of SSR2A-14 corresponded to the stronger Yr resistance. QYr.sxau-2A.1, located in the 228.02~241.58 Mbp physical interval, is different from all the known Yr loci on chromosomes 2A. Within the interval, there are 30 annotated genes, including a nucleotide-binding site and a leucine-rich repeat (NBS-LRR)-encoding gene with the linkage marker NRM2A-16 of QYr.sxau-2A.1. Our results reveal a novel major-effect QYr.sxau-2A.1, which provided resistance to YR and is a molecular marker for wheat breeding.

Wheat is among the most important staple crops worldwide; however, its yield and quality are severely threatened by stripe rust caused by Puccinia striiformis f. sp. tritici (Pst). CH806 is a Thinopyrum intermedium-derived resistant breeding line developed in our laboratory that is highly resistant to the prevalent Chinese Pst races CYR32, CYR33, and CYR34 in field trials. A genetic population was developed by crossing CH806 with the susceptible cultivar Chuanmai 24. Phenotypic evaluation of the progeny under field conditions revealed segregation for stripe rust resistance in the F₂ generation. On the basis of the resistance phenotypes of the F₂ and F_2:3 populations, homozygous resistant and homozygous susceptible F₂ individuals were selected to construct resistant and susceptible DNA bulks, respectively, for genotyping using the Wheat 120K SNP array. Bulked segregant analysis indicated that the most significant SNPs were predominantly clustered on chromosome 4A. Subsequently, publicly available simple sequence repeat (SSR) markers on chromosome 4A and newly developed SSR markers within the candidate region that were enriched for polymorphic SNPs were used for linkage analysis. The resistance locus, temporarily designated YrCH806, was mapped to an interval flanked by markers Xwmc48/Xwmc89 and SSR4A-60, with genetic distances of 4.4 cM and 2.5 cM, respectively, corresponding to a physical position of 515.8–574.7 Mb on the wheat reference genome. The closest flanking marker, SSR4A-60, was successfully converted into a Kompetitive Allele-Specific PCR (KASP) marker. This high-throughput marker was subsequently utilized to screen a panel of wheat germplasms for the distribution of YrCH806. This study provides a novel resistance source and associated molecular markers for improving stripe rust resistance in wheat breeding programs.