Natural polymorphisms in ZMET2 encoding a DNA methyltransferase modulate the number of husk layers in maize.

DNA methylation affects agronomic traits and the environmental adaptability of crops, but the natural polymorphisms in DNA methylation-related genes and their contributions to phenotypic variation in maize (Zea mays) remain elusive. Here, we show that a polymorphic 10-bp insertion/deletion variant in the 3'UTR of Zea methyltransferase2 (ZMET2) alters its transcript level and accounts for variation in the number of maize husk layers. ZMET2 encodes a chromomethylase and is required for maintaining genome-wide DNA methylation in the CHG sequence context. Disruption of ZMET2 increased the number of husk layers and resulted in thousands of differentially methylated regions, a proportion of which were also distinguishable in natural ZMET2 alleles. Population genetic analyses indicated that ZMET2 was a target of selection and might play a role in the spread of maize from tropical to temperate regions. Our results provide important insights into the natural variation of ZMET2 that confers both global and locus-specific effects on DNA methylation, which contribute to phenotypic diversity in maize.

Maize decrease in DNA methylation 1 targets RNA-directed DNA methylation on active chromatin.

DNA methylation plays vital roles in repressing transposable element 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 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 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.

The RHW1-ZCN4 regulatory pathway confers natural variation of husk leaf width in maize.

Maize husk leaf - the outer leafy layers covering the ear - modulates kernel yield and quality. Despite its importance, however, the genetic controls underlying husk leaf development remain elusive. Our previous genome-wide association study identified a single nucleotide polymorphism located in the gene RHW1 (Regulator of Husk leaf Width) that is significantly associated with husk leaf-width diversity in maize. Here, we further demonstrate that a polymorphic 18-bp InDel (insertion/deletion) variant in the 3' untranslated region of RHW1 alters its protein abundance and accounts for husk leaf width variation. RHW1 encodes a putative MYB-like transcriptional repressor. Disruption of RHW1 altered cell proliferation and resulted in a narrower husk leaf, whereas RHW1 overexpression yielded a wider husk leaf. RHW1 positively regulated the expression of ZCN4, a well-known TFL1-like protein involved in maize ear development. Dysfunction of ZCN4 reduced husk leaf width even in the context of RHW1 overexpression. The InDel variant in RHW1 is subject to selection and is associated with maize husk leaf adaption from tropical to temperate regions. Overall, our results identify that RHW1-ZCN4 regulates a pathway conferring husk leaf width variation at a very early stage of husk leaf development in maize.

Genome-wide association study reveals the genetic architecture of root hair length in maize.

BACKGROUND: Root hair, a special type of tubular-shaped cell, outgrows from root epidermal cell and plays important roles in the acquisition of nutrients and water, as well as interactions with biotic and abiotic stress. Although many genes involved in root hair development have been identified, genetic basis of natural variation in root hair growth has never been explored. RESULTS: Here, we utilized a maize association panel including 281 inbred lines with tropical, subtropical, and temperate origins to decipher the phenotypic diversity and genetic basis of root hair length. We demonstrated significant associations of root hair length with many metabolic pathways and other agronomic traits. Combining root hair phenotypes with 1.25 million single nucleotide polymorphisms (SNPs) via genome-wide association study (GWAS) revealed several candidate genes implicated in cellular signaling, polar growth, disease resistance and various metabolic pathways. CONCLUSIONS: These results illustrate the genetic basis of root hair length in maize, offering a list of candidate genes predictably contributing to root hair growth, which are invaluable resource for the future functional investigation.

Genetic analysis of three maize husk traits by QTL mapping in a maize-teosinte population.

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.

A natural single-nucleotide polymorphism variant in sulfite reductase influences sulfur assimilation in maize.

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.

Comparison of Fractional Micro-Plasma Radiofrequency and Fractional Microneedle Radiofrequency for the Treatment of Atrophic Acne Scars: A Pilot Randomized Split-Face Clinical Study in China.

BACKGROUND AND OBJECTIVE: Both fractional micro-plasma radiofrequency (RF) and fractional microneedle RF are novel devices that can be applied for the treatment of atrophic acne scars, and they have both been proved to be effective. To compare the clinical effectiveness and adverse reactions of fractional micro-plasma RF and fractional microneedle RF for the therapy of facial atrophic acne scars in a randomized split-face study. STUDY DESIGN/MATERIALS AND METHODS: Sixty patients with facial atrophic acne scars received three applications at 2-month intervals in a randomized split-face study using fractional micro-plasma RF and fractional microneedle RF on different sides of the face. Three independent dermatologists evaluated the improvement in acne scars using the ECCA grading scale (Echelle d'Evaluation Clinique des Cicatrices d'Acné) by comparing the digital images and graded the improvement in the acne scars. Patients were asked to provide a self-evaluation of satisfaction for efficacy and safety. Adverse effects were also recorded after each treatment. RESULTS: In total sixty patients completed the entire study. A significant improvement was observed in the appearance of acne scars, and the mean ECCA scores improved significantly after both modalities. The mean decrease in ECCA scores from the baseline was significantly more pronounced in fractional micro-plasma RF as compared with fractional microneedle RF (41.33 ± 20.19 vs 32.17 ± 17.35; P < 0.05). The degree of clinical improvement was also significantly better for fractional micro-plasma RF. Pain, erythema, and swelling were observed in all patients after both treatments. The pain was more intense during micro-plasma RF treatment (P = 0.000), and the duration of pain and erythema were longer than with fractional microneedle RF (P = 0.000). Postinflammatory hyperpigmentation (PIH) was observed in one patient on the fractional micro-plasma RF side while no PIH was observed on the fractional microneedle RF side. No infections or worsening of scarring was observed with either treatment. No subject was dissatisfied with the efficacy of either device. Rolling scars tended to respond better to fractional micro-plasma RF treatment compared with fractional microneedle RF (P = 0.000). CONCLUSIONS: Both fractional micro-plasma RF and fractional microneedle RF are effective and safe methods for improving atrophic acne scars. Fractional micro-plasma RF is significantly more effective for atrophic acne scars, especially for rolling scars. However, fractional microneedle RF has fewer side effects plus shorter downtime, and patients are more comfortable after the treatment. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

Linkage mapping combined with association analysis reveals QTL and candidate genes for three husk traits in maize.

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.