Multi-Locus Genome-Wide Association Study and Genomic Selection of Kernel Moisture Content at the Harvest Stage in Maize.
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ABSTRACT: Kernel moisture content at the harvest stage (KMC) is an important trait that affects the mechanical harvesting of maize grain, and the identification of genetic loci for KMC is beneficial for maize molecular breeding. In this study, we performed a multi-locus genome-wide association study (ML-GWAS) to identify quantitative trait nucleotides (QTNs) for KMC using an association mapping panel of 251 maize inbred lines that were genotyped with an Affymetrix CGMB56K SNP Array and phenotypically evaluated in three environments. Ninety-eight QTNs for KMC were detected using six ML-GWAS models (mrMLM, FASTmrMLM, FASTmrEMMA, PLARmEB, PKWmEB, and ISIS EM-BLASSO). Eleven of these QTNs were considered to be stable, as they were detected by at least four ML-GWAS models under a uniformed environment or in at least two environments and BLUP using the same ML-GWAS model. With qKMC5.6 removed, the remaining 10 stable QTNs explained <10% of the phenotypic variation, suggesting that KMC is mainly controlled by multiple minor-effect genetic loci. A total of 63 candidate genes were predicted from the 11 stable QTNs, and 10 candidate genes were highly expressed in the kernel at different time points after pollination. High prediction accuracy was achieved when the KMC-associated QTNs were included as fixed effects in genomic selection, and the best strategy was to integrate all KMC QTNs identified by all six ML-GWAS models. These results further our understanding of the genetic architecture of KMC and highlight the potential of genomic selection for KMC in maize breeding.
Project description:Kernel moisture content at harvest stage (KMC) is an important factor affecting maize production, especially for mechanical harvesting. We investigated the genetic basis of KMC using an association panel comprising of 144 maize inbred lines that were phenotypically evaluated at two field trial locations. Significant positive or negative correlations were identified between KMC and a series of other agronomic traits, indicating that KMC is associated with other such traits. Combining phenotypic values and the Maize SNP3K Beadchip to perform a genome-wide association study revealed eight single nucleotide polymorphisms (SNPs) associated with KMC at P ? 0.001 using a mixed linear model (PCA+K). These significant SNPs could be converted into five quantitative trait loci (QTLs) distributed on chromosomes 1, 5, 8, and 9. Of these QTLs, three were colocalized with genomic regions previously reported. Based on the phenotypic values of the alleles corresponding to significant SNPs, the favorable alleles were mined. Eight maize inbred lines with low KMC and harboring favorable alleles were identified. These QTLs and elite maize inbred lines with low KMC will be useful in maize breeding.
Project description:Maize is China's largest grain crop. Mechanical grain harvesting is the key technology in maize production, and the kernel moisture concentration (KMC) is the main controlling factor in mechanical maize harvesting in China. The kernel dehydration rate (KDR) is closely related to the KMC. Thus, it is important to conduct genome-wide association studies (GWAS) of the KMC and KDR in maize, detect relevant quantitative trait nucleotides (QTNs), and mine relevant candidate genes. Here, 132 maize inbred lines were used to measure the KMC every 5 days from 10 to 40 days after pollination (DAP) in order to calculate the KDR. These lines were genotyped using a maize 55K single-nucleotide polymorphism array. QTNs for the KMC and KDR were detected based on five methods (mrMLM, FASTmrMLM, FASTmrEMMA, pLARmEB, and ISIS EM-BLASSO) in the package mrMLM. A total of 334 significant QTNs were found for both the KMC and KDR, including 175 QTNs unique to the KMC and 178 QTNs unique to the KDR; 116 and 58 QTNs were detected among the 334 QTNs by two and more than two methods, respectively; and 9 and 5 QTNs among 58 QTNs were detected in 2 and 3 years, respectively. A significant enrichment in cellular component was revealed by Gene Ontology enrichment analysis of candidate genes in the intervals adjacent to the 14 QTNs and this category contained five genes. The information provided in this study may be useful for further mining of genes associated with the KMC and KDR in maize.
Project description:Starch is the most abundant storage carbohydrate in maize kernels and provides calories for humans and other animals as well as raw materials for various industrial applications. Decoding the genetic basis of natural variation in kernel starch content is needed to manipulate starch quantity and quality via molecular breeding to meet future needs. Here, we identified 50 unique single quantitative trait loci (QTLs) for starch content with 18 novel QTLs via single linkage mapping, joint linkage mapping and a genome-wide association study in a multi-parent population containing six recombinant inbred line populations. Only five QTLs explained over 10% of phenotypic variation in single populations. In addition to a few large-effect and many small-effect additive QTLs, limited pairs of epistatic QTLs also contributed to the genetic basis of the variation in kernel starch content. A regional association study identified five non-starch-pathway genes that were the causal candidate genes underlying the identified QTLs for starch content. The pathway-driven analysis identified ZmTPS9, which encodes a trehalose-6-phosphate synthase in the trehalose pathway, as the causal gene for the QTL qSTA4-2, which was detected by all three statistical analyses. Knockout of ZmTPS9 increased kernel starch content and, in turn, kernel weight in maize, suggesting potential applications for ZmTPS9 in maize starch and yield improvement. These findings extend our knowledge about the genetic basis of starch content in maize kernels and provide valuable information for maize genetic improvement of starch quantity and quality.
Project description:Glyphosate (GP)-based herbicides have been widely applied to crops for weed control and pre-harvest desiccation. The objective of this research was to evaluate the effects of pre-harvest GP application on maize or how it physiologically alters this crop. Here, we applied four GP treatment (Control, GP150, GP200, and GP250) on maize lines of Z58 and PH6WC belonging to different maturity groups at grain-filling stages form DAP30 to DAP45. GP application significantly decreased the grain moisture content at harvest by 22-35% for Z58 and by 15-41% for PH6WC. However, the responses of grain weight to glyphosate vary with inbred lines and application time. A high concentration of glyphosate (GP250) reduced the grain weight of Z58 and low concentrations (GP150 and GP200) did not affect, while the grain weight of PH6WC significantly decreased under glyphosate treatment. In summary, our results revealed that timely and appropriate GP application lowers grain moisture content without causing seed yield and quality loss. GP application adversely affected photosynthesis by promoting maturation and leaf senescence. Meanwhile, it also enhanced non-structural carbohydrate (soluble sugars and starch) remobilization from the vegetative organs to the grains. Hence, GP treatment coordinates plant senescence and assimilate remobilization. RNA sequencing revealed that glyphosate regulated the transcript levels of sugar signaling-related genes and induced assimilate repartitioning in grains. This work indicates the practical significance of GP application for maize seed production and harvest, which highlights the contributions of source-sink communication to maize yield in response to external stress or pre-harvest desiccant application.
Project description:Unpredictable weather vagaries in the Asian tropics often increase the risk of a series of abiotic stresses in maize-growing areas, hindering the efforts to reach the projected demands. Breeding climate-resilient maize hybrids with a cross-tolerance to drought and waterlogging is necessary yet challenging because of the presence of genotype-by-environment interaction (GEI) and the lack of an efficient multi-trait-based selection technique. The present study aimed at estimating the variance components, genetic parameters, inter-trait relations, and expected selection gains (SGs) across the soil moisture regimes through genotype selection obtained based on the novel multi-trait genotype-ideotype distance index (MGIDI) for a set of 75 tropical pre-released maize hybrids. Twelve traits including grain yield and other secondary characteristics for experimental maize hybrids were studied at two locations. Positive and negative SGs were estimated across moisture regimes, including drought, waterlogging, and optimal moisture conditions. Hybrid, moisture condition, and hybrid-by-moisture condition interaction effects were significant (p ≤ 0.001) for most of the traits studied. Eleven genotypes were selected in each moisture condition through MGIDI by assuming 15% selection intensity where two hybrids, viz., ZH161289 and ZH161303, were found to be common across all the moisture regimes, indicating their moisture stress resilience, a unique potential for broader adaptation in rainfed stress-vulnerable ecologies. The selected hybrids showed desired genetic gains such as positive gains for grain yield (almost 11% in optimal and drought; 22% in waterlogging) and negative gains in flowering traits. The view on strengths and weaknesses as depicted by the MGIDI assists the breeders to develop maize hybrids with desired traits, such as grain yield and other yield contributors under specific stress conditions. The MGIDI would be a robust and easy-to-handle multi-trait selection process under various test environments with minimal multicollinearity issues. It was found to be a powerful tool in developing better selection strategies and optimizing the breeding scheme, thus contributing to the development of climate-resilient maize hybrids.
Project description:BackgroundThe chlorophyll content (CC) is a key factor affecting maize photosynthetic efficiency and the final yield. However, its genetic basis remains unclear. The development of statistical methods has enabled researchers to design and apply various GWAS models, including MLM, MLMM, SUPER, FarmCPU, BLINK and 3VmrMLM. Comparative analysis of their results can lead to more effective mining of key genes.ResultsThe heritability of CC was 0.86. Six statistical models (MLM, BLINK, MLMM, FarmCPU, SUPER, and 3VmrMLM) and 1.25 million SNPs were used for the GWAS. A total of 140 quantitative trait nucleotides (QTNs) were detected, with 3VmrMLM and MLM detecting the most (118) and fewest (3) QTNs, respectively. The QTNs were associated with 481 genes and explained 0.29-10.28% of the phenotypic variation. Additionally, 10 co-located QTNs were detected by at least two different models or methods, three co-located QTNs were identified in at least two different environments, and six co-located QTNs were detected by different models or methods in different environments. Moreover, 69 candidate genes within or near these stable QTNs were screened based on the B73 (RefGen_v2) genome. GRMZM2G110408 (ZmCCS3) was identified by multiple models and in multiple environments. The functional characterization of this gene indicated the encoded protein likely contributes to chlorophyll biosynthesis. In addition, the CC differed significantly between the haplotypes of the significant QTN in this gene, and CC was higher for haplotype 1.ConclusionThis study's results broaden our understanding of the genetic basis of CC, mining key genes related to CC and may be relevant for the ideotype-based breeding of new maize varieties with high photosynthetic efficiency.
Project description:In maize, doubled haploid (DH) line production capacity of large-sized maize breeding programs often exceeds the capacity to phenotypically evaluate the complete set of testcross candidates in multi-location trials. The ability to partially select DH lines based on genotypic data while maintaining or improving genetic gains for key traits using phenotypic selection can result in significant resource savings. The present study aimed to evaluate genomic selection (GS) prediction scenarios for grain yield and agronomic traits of one of the tropical maize breeding pipelines of CIMMYT in eastern Africa, based on multi-year empirical data for designing a GS-based strategy at the early stages of the pipeline. We used field data from 3,068 tropical maize DH lines genotyped using rAmpSeq markers and evaluated as test crosses in well-watered (WW) and water-stress (WS) environments in Kenya from 2017 to 2019. Three prediction schemes were compared: (1) 1 year of performance data to predict a second year; (2) 2 years of pooled data to predict performance in the third year, and (3) using individual or pooled data plus converting a certain proportion of individuals from the testing set (TST) to the training set (TRN) to predict the next year's data. Employing five-fold cross-validation, the mean prediction accuracies for grain yield (GY) varied from 0.19 to 0.29 under WW and 0.22 to 0.31 under WS, when the 1-year datasets were used training set to predict a second year's data as a testing set. The mean prediction accuracies increased to 0.32 under WW and 0.31 under WS when the 2-year datasets were used as a training set to predict the third-year data set. In a forward prediction scenario, good predictive abilities (0.53 to 0.71) were found when the training set consisted of the previous year's breeding data and converting 30% of the next year's data from the testing set to the training set. The prediction accuracy for anthesis date and plant height across WW and WS environments obtained using 1-year data and integrating 10, 30, 50, 70, and 90% of the TST set to TRN set was much higher than those trained in individual years. We demonstrate that by increasing the TRN set to include genotypic and phenotypic data from the previous year and combining only 10-30% of the lines from the year of testing, the predicting accuracy can be increased, which in turn could be used to replace the first stage of field-based screening partially, thus saving significant costs associated with the testcross formation and multi-location testcross evaluation.
Project description:Kernel water content (KWC) and kernel dehydration rate (KDR) are two main factors affecting maize seed quality and have a decisive influence on the mechanical harvest. It is of great importance to map and mine candidate genes related to KWCs and KDRs before physiological maturity in maize. 120 double-haploid (DH) lines constructed from Si287 with low KWC and JiA512 with high KWC were used as the mapping population. KWCs were measured every 5 days from 10 to 40 days after pollination, and KDRs were calculated. A total of 1702 SNP markers were used to construct a linkage map, with a total length of 1,309.02 cM and an average map distance of 0.77 cM. 10 quantitative trait loci (QTLs) and 27 quantitative trait nucleotides (QTNs) were detected by genome-wide composite interval mapping (GCIM) and multi-locus random-SNP-effect mixed linear model (mrMLM), respectively. One and two QTL hotspot regions were found on Chromosome 3 and 7, respectively. Analysis of the Gene Ontology showed that 2 GO terms of biological processes (BP) were significantly enriched (P ≤ 0.05) and 6 candidate genes were obtained. This study provides theoretical support for marker-assisted breeding of mechanical harvest variety in maize.
Project description:Genomic prediction models are often calibrated using multi-generation data. Over time, as data accumulates, training data sets become increasingly heterogeneous. Differences in allele frequency and linkage disequilibrium patterns between the training and prediction genotypes may limit prediction accuracy. This leads to the question of whether all available data or a subset of it should be used to calibrate genomic prediction models. Previous research on training set optimization has focused on identifying a subset of the available data that is optimal for a given prediction set. However, this approach does not contemplate the possibility that different training sets may be optimal for different prediction genotypes. To address this problem, we recently introduced a sparse selection index (SSI) that identifies an optimal training set for each individual in a prediction set. Using additive genomic relationships, the SSI can provide increased accuracy relative to genomic-BLUP (GBLUP). Non-parametric genomic models using Gaussian kernels (KBLUP) have, in some cases, yielded higher prediction accuracies than standard additive models. Therefore, here we studied whether combining SSIs and kernel methods could further improve prediction accuracy when training genomic models using multi-generation data. Using four years of doubled haploid maize data from the International Maize and Wheat Improvement Center (CIMMYT), we found that when predicting grain yield the KBLUP outperformed the GBLUP, and that using SSI with additive relationships (GSSI) lead to 5-17% increases in accuracy, relative to the GBLUP. However, differences in prediction accuracy between the KBLUP and the kernel-based SSI were smaller and not always significant.
Project description:For mechanized maize production, a low grain water content (GWC) at harvest is necessary. However, as a complex quantitative trait, understand the genetic mechanism of GWC remains a large gap, especially in hybrids. In this study, a hybrid population through two environments including 442 F1 was used for genome-wide association analysis of GWC and the grain dehydration rate (GDR), using the area under the dry down curve (AUDDC) as the index. Then, we identified 19 and 17 associated SNPs for GWC and AUDDC, including 10 co-localized SNPs, along with 64 and 77 pairs of epistatic SNPs for GWC and AUDDC, respectively. These loci could explain 11.39-68.2% of the total phenotypic variation for GWC and 41.07-67.02% for AUDDC at different stages, whose major effect was the additive and epistatic effect. By exploring the candidate genes around the significant sites, a total of 398 and 457 possible protein-coding genes were screened, including autophagy pathway and auxin regulation-related genes, and five inbred lines with the potential to reduce GWC in the combined F1 hybrid were identified. Our research not only provides a certain reference for the genetic mechanism analysis of GWC in hybrids but also provides an added reference for breeding low-GWC materials.Supplementary informationThe online version contains supplementary material available at 10.1007/s11032-022-01349-x.