Long non-coding RNAs (lncRNAs) have emerged as important regulators in plant stress response. Here, we report a genome-wide lncRNA transcriptional analysis in response to drought stress using an expanded series of maize samples collected from three distinct tissues spanning four developmental stages. In total, 3488 high-confidence lncRNAs were identified, among which 1535 were characterized as drought responsive. By characterizing the genomic structure and expression pattern, we found that lncRNA structures were less complex than protein-coding genes, showing shorter transcripts and fewer exons. Moreover, drought-responsive lncRNAs exhibited higher tissue- and development-specificity than protein-coding genes. By exploring the temporal expression patterns of drought-responsive lncRNAs at different developmental stages, we discovered that the reproductive stage R1 was the most sensitive growth stage with more lncRNAs showing altered expression upon drought stress. Furthermore, lncRNA target prediction revealed 653 potential lncRNA-messenger RNA (mRNA) pairs, among which 124 pairs function in cis-acting mode and 529 in trans. Functional enrichment analysis showed that the targets were significantly enriched in molecular functions related to oxidoreductase activity, water binding, and electron carrier activity. Multiple promising targets of drought-responsive lncRNAs were discovered, including the V-ATPase encoding gene, vpp4. These findings extend our knowledge of lncRNAs as important regulators in maize drought response.

Summary: In maize, kernel traits strongly impact overall grain yields, and it is known that sophisticated spatiotemporal programs of gene expression coordinate kernel development, so advancing our knowledge of kernel development can help efforts to improve grain yields. Here, using phenotype, genotype and transcriptomics data of maize kernels at 5 and 15 days after pollination (DAP) for a large association mapping panel, we employed multiple quantitative genetics approaches—genome‐wide association studies (GWAS) as well as expression quantitative trait loci (eQTL) and quantitative trait transcript (QTT) analyses—to gain insights about molecular genetic basis of kernel development in maize. This resulted in the identification of 137 putative kernel length‐related genes at 5 DAP, of which 43 are located in previously reported QTL regions. Strikingly, we identified an eQTL that overlaps the locus encoding a maize homolog of the recently described m⁶A methylation reader protein ECT2 from Arabidopsis; this putative ^epieQTL is associated with 53 genes and may represent a master epi‐transcriptomic regulator of kernel development. Notably, among the genes associated with this ^epieQTL, 10 are for the main storage proteins in the maize endosperm (zeins) and two are known regulators of zein expression or endosperm development (Opaque2 and ZmICE1). Collectively, beyond cataloging and characterizing genomic attributes of a large number of eQTL associated with kernel development in maize, our study highlights how an eQTL approach can bolster the impact of both GWAS and QTT studies and can drive insights about the basic biology of plants.

Drought is one of the most critical environmental factors constraining maize production. When it occurs at the flowering stage, serious yield losses are caused, and often, the damage is irretrievable. In this study, anthesis to silk interval (ASI), plant height (PH), and ear biomass at the silking date (EBM) of 279 inbred lines were studied under both water-stress (WS) and well-water (WW) field conditions, for three consecutive years. Averagely, ASI was extended by 25.96%, EBM was decreased by 17.54%, and the PH was reduced by 12.47% under drought stress. Genome-wide association studies were carried out using phenotypic values under WS, WW, and drought-tolerance index (WS-WW or WS/WW) and applying a mixed linear model that controls both population structure and relative kinship. In total, 71, 159, and 21 SNPs, located in 32, 59, and 12 genes, were significantly (P < 10⁻⁵) associated with ASI, EBM, and PH, respectively. Only a few overlapped candidate genes were found to be associated with the same drought-related traits under different environments, for example, ARABIDILLO 1, glycoprotein, Tic22-like, and zinc-finger family protein for ASI; 26S proteasome non-ATPase and pyridoxal phosphate transferase for EBM; 11-ß-hydroxysteroid dehydrogenase, uncharacterised, Leu-rich repeat protein kinase, and SF16 protein for PH. Furthermore, most candidate genes were revealed to be drought-responsive in an association panel. Meanwhile, the favourable alleles/key variations were identified with a haplotype analysis. These candidate genes and their key variations provide insight into the genetic basis of drought tolerance, especially for the female inflorescence, and will facilitate drought-tolerant maize breeding.

Variation in flowering plays a major role in maize photoperiod adaptation during long-term domestication. It is of high value to investigate the genetic basis of maize flowering under a wide range of environmental conditions in order to overcome photoperiod sensitivity or enhance stress tolerance. A recombinant inbred line (RIL) population derived from a cross between Huangzaosi and Mo17, composed of 121 lines and genotyped by 8329 specifically developed markers, was field evaluated in two consecutive years under two planting densities (67,500 and 120,000 plants ha⁻¹) and two water treatments (normal irrigation and drought stress at the flowering stage). The days to silking (DTS), days to anthesis (DTA), and anthesis to silking interval (ASI) were all evaluated. Within the RIL population, DTS and DTA expanded as planting density and water deficit increased. For DTA, DTS, ASI, and ASI-delay, a total of 22, 17, 21, and 11 QTLs were identified, respectively. More than two significant QTLs were identified in each of the nine chromosomal intervals. Under diverse conditions and locations, six QTLs (quantitative trait locus) for DTS and DTA were discovered in Chr. 8: 118.13–125.31 Mb. Three chromosome regions, Chr. 3: 196.14–199.89 Mb, Chr. 8: 169.02–172.46 Mb, and Chr. 9: 128.12–137.26 Mb, all had QTLs for ASI-delay under normal and stress conditions, suggesting their possible roles in stress tolerance enhancement. These QTL hotspots will promote early-maturing or multiple abiotic stress-tolerant maize breeding, as well as shed light on the development of maize varieties with a broad range of adaptations.