Key message Combined linkage and association mapping analyses facilitate the emphasis on the candidate genes putatively involved in maize husk growth. The maize (Zea mays L.) husk consists of multiple leafy layers and plays important roles in protecting the ear from pathogen infection and in preventing grain dehydration. Although husk morphology varies widely among different maize inbred lines, the genetic basis of such variation is poorly understood. In this study, we used three maize recombinant inbred line (RIL) populations to dissect the genetic basis of three husk traits: i.e., husk length (HL), husk width (HW), and the number of husk layers (HN). Three husk traits in all three RIL populations showed wide phenotypic variation and high heritability. The HL showed stronger correlations with ear traits than did HW and HN. A total of 21 quantitative trait loci (QTL) were identified for the three traits in three RIL populations, and some of them were commonly observed for the same trait in different populations. The proportions of total phenotypic variation explained by QTL in three RIL populations were 31.8, 35.3, and 44.5% for HL, HW, and HN, respectively. The highest proportions of phenotypic variation explained by a single QTL were 14.7% for HL in the By815/K22 RIL population (BYK), 13.5% for HW in the By815/DE3 RIL population (BYD), and 19.4% for HN in the BYD population. A combined analysis of linkage mapping with a previous genome-wide association study revealed five candidate genes related to husk morphology situated within three QTL loci. These five genes were related to metabolism, gene expression regulation, and signal transduction.

DNA methylation plays vital roles in repressing transposable element (TE) activity and regulating gene expression. The chromatin-remodeling factor Decrease in DNA methylation 1 (DDM1) is crucial for maintaining DNA methylation across diverse plant species, and is required for RNA-directed DNA methylation (RdDM) to maintain mCHH islands in maize (Zea mays). However, the mechanisms by which DDM1 is involved in RdDM are not well understood. In this work, we used chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-seq) to ascertain the genome-wide occupancy of ZmDDM1 in the maize genome. The results revealed that ZmDDM1 recognized an 8-bp-long GC-rich degenerate DNA sequence motif, which is enriched in transcription start sites (TSSs) and other euchromatic regions. Meanwhile, 24-nucleotide siRNAs and CHH methylation were delineated at the edge of ZmDDM1-occupied sites. ZmDDM1 co-purified with Argonaute 4 (ZmAGO4) proteins, providing further evidence that ZmDDM1 is a component of RdDM complexes in planta. Consistent with this, the vast majority of ZmDDM1-targeted regions co-localized with ZmAGO4-bound genomic sites. Overall, our results suggest a model that ZmDDM1 may be recruited to euchromatic regions via recognition of a GC-rich motif, thereby remodeling chromatin to provide access for RdDM activities in maize.

Summary Plants absorb sulfur from the environment and assimilate it into suitable forms for the biosynthesis of a broad range of molecules. Although the biochemical pathway of sulfur assimilation is known, how genetic differences contribute to natural variation in sulfur assimilation remains poorly understood. Here, using a genome‐wide association study, we uncovered a single‐nucleotide polymorphism (SNP) variant in the sulfite reductase (SiR) gene that was significantly associated with SiR protein abundance in a maize natural association population. We also demonstrated that the synonymous C to G base change at SNP69 may repress translational activity by altering messenger RNA secondary structure, which leads to reduction in ZmSiR protein abundance and sulfur assimilation activity. Population genetic analyses showed that the SNP69C allele was likely a variant occurring after the initial maize domestication and accumulated with the spread of maize cultivation from tropical to temperate regions. This study provides the first evidence that genetic polymorphisms in the exon of ZmSiR could influence the protein abundance through a posttranscriptional mechanism and in part contribute to natural variation in sulfur assimilation. These findings provide a prospective target to improve maize varieties with proper sulfur nutrient levels assisted by molecular breeding and engineering.

Background The maize husk consists of numerous leafy layers and plays vital roles in protecting the ear from pathogen infection and dehydration. Teosinte, the wild ancestor of maize, has about three layers of small husk outer covering the ear. Although several quantitative trait loci (QTL) underlying husk morphology variation have been reported, the genetic basis of husk traits between teosinte and maize remains unclear. Results A linkage population including 191 BC2F8 inbred lines generated from the maize line Mo17 and the teosinte line X26–4 was used to identify QTL associated with three husk traits: i.e., husk length (HL), husk width (HW) and the number of husk layers (HN). The best linear unbiased predictor (BLUP) depicted wide phenotypic variation and high heritability of all three traits. The HL exhibited greater correlation with HW than HN. A total of 4 QTLs were identified including 1, 1, 2, which are associated with HL, HW and HN, respectively. The proportion of phenotypic variation explained by these QTLs was 9.6, 8.9 and 8.1% for HL, HN and HW, respectively. Conclusions The QTLs identified in this study will pave a path to explore candidate genes regulating husk growth and development, and benefit the molecular breeding program based on molecular marker-assisted selection to cultivate maize varieties with an ideal husk morphology.