Hybrid Maize Breeding Research Articles

Genetic diversity and population structure analyses of tropical maize inbred lines using Single Nucleotide Polymorphism markers.

Analyses of the genetic distance and composition of inbred lines are a prerequisite for parental selection and to exploit heterosis in plant breeding programs. The study aimed to assess genetic diversity and population structure of a maize germplasm panel comprising 182 founder lines and 866 derived inbred lines using Single Nucleotide Polymorphism (SNP) markers to identify genetically unique lines for hybrid breeding. The founder lines were genotyped with 1201 SNPs, and the derived lines with 1484 SNPs. Moderate genetic variation, with genetic diversity ranging from 0.004 to 0.44 with a mean of 0.25, was recorded for the founder lines, while corresponding values of 0.004 to 0.34 with a mean of 0.13 were recorded for the derived lines. Heterozygosity values ranging from 0.00 to 0.24 and a mean of 0.08 were recorded for both lines. Of the SNP markers used, 82% of the 1201 markers and 84% of the 1484 markers exhibited polymorphism information content ranging from 0.25 to 0.50. Analysis of molecular variance revealed significant genetic differences (P ≤ 0.001) among and within populations in the founder and derived lines. Most detected variations, i.e., 97% and 88.38%, were attributed to within populations in the founder and derived lines, respectively. Population structure analysis identified three distinct subpopulations among founder lines and two among derived lines. Cluster analysis supported the population structure The following genetically distant founder and derived inbred lines were selected: G15NL337 and G15NL312 (Cluster 1), 15ARG152 and RGS-PL44 (Cluster 2), RGS-PL44 and 15ARG149 (Cluster 2), and RGS-PL33 and RGS-PL44 (Cluster 2), respectively. The selected lines are genetically distinct and recommended for marker-assisted hybrid maize breeding to exploit the frequency of beneficial alleles. This study provides valuable insights for maize breeding programs, enabling the exploitation of beneficial alleles and contributing to improved crop yields and food security through hybrid breeding.

Genomic Prediction of Kernel Water Content in a Hybrid Population for Mechanized Harvesting in Maize in Northern China

Genomic prediction enables rapid selection of maize varieties with low kernel water content (KWC), facilitating the development of mechanized maize harvesting and reducing costs. This study evaluated and characterized the KWC and grain yield (GY) of hybrid maize in northern China and used genomic prediction to identify superior hybrid combinations with low kernel water content at maturity (MKWC) and high GY adapted to northern China. A total of 285 hybrids obtained from single crosses of 34 inbred lines from Stiff Stalk and Non-Stiff Stalk heterotic groups were used for genomic prediction of KWC and GY. We tested 20 different statistical prediction models considering additive effects and evaluating the impact of dominance and epistasis on prediction accuracy. Employing 10-fold cross-validation, it showed that the average prediction accuracy ranged drastically from 0.386 to 0.874 across traits and models. Eight linear statistical methods displayed a very similar prediction accuracy for each trait. The average prediction accuracy of machine learning methods was lower than that of linear statistical methods for KWC-related traits, but the random forest model had a high prediction accuracy of 0.510 for GY. When genetic effects were incorporated into the prediction model, the prediction accuracy for each trait was improved. Overall, the model with dominant and epistatic effects (G:AD(AA)) performed best. For the same number of markers, predictions using trait-specific markers resulted in higher prediction accuracy than randomly selected markers. When the number of trait-specific SNPs was set to 100, the prediction accuracy of GY increased by 33.27%, from 0.406 to 0.541. Out of all the 561 potential hybrids, the TOP 30 hybrids selected by genomic prediction would lead to a 1.44% decrease in MKWC compared with Xianyu335, a hybrid with a fast kernel water dry-down, and these hybrids also had higher GY simultaneously. Our results confirm the value of genomic prediction for hybrid breeding low MKWC suitable for maize mechanized harvesting in northern China. In conclusion, this study highlights the potential of genomic prediction to optimize maize hybrid breeding, enhancing efficiency and providing insights into genotype-accuracy relationships. The findings offer new strategies for hybrid design and advancing mechanized harvesting in northern China.

Assessment of genetic diversity and heterotic alignment of CIMMYT and IITA maize inbred lines adapted to sub‐Saharan Africa

AbstractDespite the breeding efforts by many institutions, maize (Zea mays L.) productivity in sub‐Saharan Africa is still low. A limited number of productive maize hybrids have been developed partly due to a lack of knowledge on the diversity and heterotic relationship of the germplasm, especially in public breeding programs. Understanding the extent of diversity, structure, and heterotic grouping of available maize germplasm originating from different breeding programs is important to enhance long‐term genetic gain in hybrid maize breeding programs by optimizing heterotic pools using modern breeding tools. Information about the genetic structure of the available germplasm could help breeders design effective breeding strategies to improve yield. This study was conducted to determine the genetic diversity, population structure, and heterotic alignment among 187 elite maize inbred lines from the IITA (International Institute of Tropical Agriculture) and CIMMYT (International Maize and Wheat Improvement Center) maize breeding programs. The inbred lines were genotyped with 9857 Diversity Array Technology sequencing based single nucleotide polymorphism markers. Hierarchical clustering revealed three major groups, with some subgroups consistent with the selection history, and pedigree of the inbred lines. Three broad groups were detected: two consisting of CIMMYT lines and a mixed group consisting of both CIMMYT and IITA inbred lines. The STRUCTURE analysis revealed six subpopulations (fixation index value = 0.58), which depicts a moderate genetic diversity among the materials. Population 2 comprises the highest number of genotypes (102) from both programs. More than 89% of the elite lines had homozygosity exceeding 95%, with the remaining lines requiring further inbreeding through repeated self‐pollination. There was inconsistency in the predetermined heterotic groups' alignment between CIMMYT and IITA elite inbred lines. Analysis of molecular variance revealed that 96% of the total variation was accounted for by differences within groups, with the remaining 4% representing the variation between groups. This suggests that the two programs can benefit from germplasm exchange for the improvement of maize productivity.

Genetic Diversity of Maize (Zea mays L.) Genotypes under Low Level of Nitrogen

Maize (Zea mays L.) is India's second most significant crop, behind rice. As a cross-pollinated crop, hybrid vigour is commonly used to achieve the best production potential. The selection of various genotypes is a primary goal in the maize hybrid breeding programme. Maize is a voracious feeder and requires a lot of fertiliser. The biggest limits on maize production in India are low soil nitrogen and high fertiliser costs. Jharkhand consumes only 82.5 kg of total fertiliser nutrients per acre. In order to discover genetically varied maize genotypes for nitrogen usage efficiency, the current study evaluated 121 maize genotypes in a randomised block design with two replications at low nitrogen levels. Plant height, days to 50% pollen shed, days to 50% silk, anthesis-silking interval (ASI), days to75% maturity, number of leaves per plant, root length, stover yield per plant, grain yield per plant, and harvest index were all recorded. Statistical analysis was performed using the morphological trait data. Analysis of variance revealed substantial (p≤0.05) variations in morphological features between maize genotypes. For statistical analysis, a clustering method was used to locate various genotypes with greater genetic distance in order to develop good heterotic combinations. One hundred twenty one genotypes were divided into six clusters using the dendogram-by-ward minimum variance approach. Cluster III and V had the greatest inter-cluster distance (79.26), followed by cluster III and VI (74.77) and cluster II and III (73.80). The many heterotic combinations developed from these clusters can be employed in subsequent breeding programmes to produce low nitrogen-requiring hybrids with a low yield penalty.

Genomic prediction of yield-related traits and genome-based establishment of heterotic pattern in maize hybrid breeding of Southwest China.

When genomic prediction is implemented in breeding maize (Zea mays L.), it can accelerate the breeding process and reduce cost to a large extent. In this study, 11 yield-related traits of maize were used to evaluate four genomic prediction methods including rrBLUP, HEBLP|A, RF, and LightGBM. In all the 11 traits, rrBLUP had similar predictive accuracy to HEBLP|A, and so did RF to LightGBM, but rrBLUP and HEBLP|A outperformed RF and LightGBM in 8 traits. Furthermore, genomic prediction-based heterotic pattern of yield was established based on 64620 crosses of maize in Southwest China, and the result showed that one of the parent lines of the top 5% crosses came from temp-tropic or tropic germplasm, which is highly consistent with the actual situation in breeding, and that heterotic pattern (Reid+ × Suwan+) will be a major heterotic pattern of Southwest China in the future.

Local business capacity to scale new hybrid maize seeds: Insights from a decade of tracking sales in Mexico

The return on investments from public-sector hybrid maize breeding depends heavily on the capacities of locally owned, small- and medium-sized seed enterprises (maize SMEs) to build market share for new products. In Mexico, between 2011 and 2020, dozens of maize SMEs obtained new genetic material through a hybrid maize breeding program funded by the Mexican government and led by the International Maize and Wheat Improvement Center (CIMMYT). To examine the maize SMEs’ ability to expand sales of new seed products, we tracked sales for 16 maize SMEs between 2014 and 2022. Our analysis captured variation in capacity by grouping the SMEs by seed sales volumes: emerging group (EG), small-sized group (SG), and medium-sized group (MG). The results showed the capacity of the MGs to launch and scale new seed products and remove older products from the market. The EGs and SGs expanded sales of new products, but also maintained production of older products, including open-pollinated varieties. However, during the observation period we detected little growth in total sales volumes across the three groups. The findings suggest that future efforts to promote local maize SMEs will benefit from a balanced approach between technology transfer and seed system development. We recommend that future seed systems investments consider: (1) support for innovation in seed-product marketing, (2) coordinated public-sector purchases that prioritize new products, and (3) innovation in product design for niche market development. These lessons may also serve future efforts to design regional-level programs in support of smallholder rainfed hybrid maize production.

Progressive meristem and single-cell transcriptomes reveal the regulatory mechanisms underlying maize inflorescence development and sex differentiation

Maize develops separate ear and tassel inflorescences with initially similar morphology but ultimately different architecture and sexuality. The detailed regulatory mechanisms underlying these changes still remain largely unclear. In this study, through analyzing the time-course meristem transcriptomes and floret single-cell transcriptomes of ear and tassel, we revealed the regulatory dynamics and pathways underlying inflorescence development and sex differentiation. We identified 16 diverse gene clusters with differential spatiotemporal expression patterns and revealed biased regulation of redox, programmed cell death, and hormone signals during meristem differentiation between ear and tassel. Notably, based on their dynamic expression patterns, we revealed the roles of two RNA-binding proteins in regulating inflorescence meristem activity and axillary meristem formation. Moreover, using the transcriptional profiles of 53 910 single cells, we uncovered the cellular heterogeneity between ear and tassel florets. We found that multiple signals associated with either enhanced cell death or reduced growth are responsible for tassel pistil suppression, while part of the gibberellic acid signal may act non-cell-autonomously to regulate ear stamen arrest during sex differentiation. We further showed that the pistil-protection gene SILKLESS 1 (SK1) functions antagonistically to the known pistil-suppression genes through regulating common molecular pathways, and constructed a regulatory network for pistil-fate determination. Collectively, our study provides a deep understanding of the regulatory mechanisms underlying inflorescence development and sex differentiation in maize, laying the foundation for identifying new regulators and pathways for maize hybrid breeding and improvement.

Participatory breeding in organic systems: Experiences from maize case studies in the United States

Participatory breeding and crop selection can satis­fy the needs of underserved groups of farmers (e.g., organic producers, farmers producing spe­cialty grain for niche markets) neglected by the modern global seed industry. Participatory research methods that value local knowledge and facilitate the active involvement of producers, researchers, and other actors involved in the agri-food system are tactics that can help us achieve sustainable agri­culture. Interest in the use of participatory methods to increase the value of U.S. land-grant universities to society has grown rapidly during the last decade. Interest includes re-engagement in the develop­ment of maize hybrids that perform well in a diverse range of heterogeneous growing environ­ments and that are better suited for sustainability-minded producers, buyers, and consumers. Sys­tems-based breeding aimed at protecting the envi­ronment and providing food, fiber, and energy while considering equity issues, has been proposed as a way to overcome the shortcomings of privat­ized approaches. In this article, we consider recent projects that use collaborative methods for hybrid maize breeding, cultivar testing, and genetic research to develop, identify, and deliver traits associated with crop performance, quality, and sus­tainability. Three case studies consider the efforts focused on developing non-GMO varieties for organic and specialty markets. We find that, unlike many successful efforts focused on the improve­ment of other crops, there are few promising mod­els for participatory breeding of hybrid maize. Even though many projects have sought to involve stakeholders with a variety of methods, all have struggled to meaningfully engage farmers in maize hybrid improvement. Still, our reflection of case studies calls for systems-based breeding and sug­gests a path forward. This route would seek to address the needs, perspectives, and values of a broader range of actors participating in the food system by leveraging technologies and infrastruc­ture in service of the public. Land-grant universities are well positioned to play a crucial role in coordi­nating efforts, facilitating partnerships, and sup­porting breeding programs that satisfy societal wants that include health, equity, and care.

Accelerating haploid induction rate and haploid validation through marker-assisted selection for qhir1 and qhir8 in maize.

Doubled haploid (DH) technology becomes more routinely applied in maize hybrid breeding. However, some issues in haploid induction and identification persist, requiring resolution to optimize DH production. Our objective was to implement simultaneous marker-assisted selection (MAS) for qhir1 (MTL/ZmPLA1/NLD) and qhir8 (ZmDMP) using TaqMan assay in F2 generation of four BHI306-derived tropical × temperate inducer families. We also aimed to assess their haploid induction rate (HIR) in the F3 generation as a phenotypic response to MAS. We highlighted remarkable increases in HIR of each inducer family. Genotypes carrying qhir1 and qhir8 exhibited 1 - 3-fold higher haploid frequency than those carrying only qhir1. Additionally, the qhir1 marker was employed for verifying putative haploid seedlings at 7 days after planting. Flow cytometric analysis served as the gold standard test to assess the accuracy of the R1-nj and the qhir1 marker. The qhir1 marker showed high accuracy and may be integrated in multiple haploid identifications at early seedling stage succeeding pre-haploid sorting via R1-nj marker.

Selection of maize lines for estimating combining ability in topcrossings

Hybrid maize breeding is carried out based on interline hybridization using heterosis in the first generation. Hybridization efficiency can be increased using parental forms with high combining ability in crosses. The purpose of the study was to identify new self-pollinated lines of maize with high general and specific combining ability, to identify the impact on the accuracy of the estimation of related lines included in the study. The current study was carried out at the FSBSI Agricultural Research Center «Donskoy» in 2021 and 2022. The objects of the study were two testers PD 329, KV 399 and 10 self-pollinated lines, including two lines (KV 410 and DS 22/325) related to tester PD 329, twenty topcross maize hybrids. The method for estimating combining ability was complete topcrosses. Estimation of new self-pollinated lines by the topcross method made it possible to identify the lines ‘KV 498’ and ‘DS 498/203-3’ with a consistently high overall combining ability according to the trait ‘grain productivity’. The specific combining ability was characterized by variability depending on the year of study. Only the line ‘KV 401’ had a high SCR in all years of study. The inclusion of lines related to the testers in the set had no effect or only slightly affected the estimates of the TCS of the remaining unrelated lines. The effect on the SCS was not significant, however, the estimation of the SCS would be more accurate if related lines were excluded from the set. There have been identified such new testcross hybrids with large grain productivity as ‘PD 329 × KV 498’ (3.92 t/ha), ‘KV 399 × KV 498’ (4.71 t/ha),‘KV 399 × KV 401’ (4.80 t/ha), ‘KV 399 × DS 498/203-3’ (4.60 t/ha). They were obtained with the participation of new lines characterized by high general or specific combinative ability, which can be recommended to be included in crossbreeding programs to develop highly heterotic maize hybrids.

Comparison of genomic prediction models for general combining ability in early stages of hybrid breeding programs

AbstractThis study evaluates the impact of genomic prediction models on selecting inbred lines as parents in hybrid breeding programs. New parents in a hybrid breeding program are typically selected from early‐stage yield trials based on general combining ability (GCA) from testcrosses. Genomic studies have largely focused on predicting hybrid performance in the late stages of the breeding pipeline and largely ignored the selection of inbred lines as parents of the subsequent breeding cycles. Here, we used stochastic simulations of a maize (Zea mays L.) hybrid breeding program for 20 years to evaluate the performance of genomic prediction models for selecting parents based on their predicted GCA. Five genomic prediction models were evaluated in terms of achieved genetic gain and heterosis under two different single nucleotide polymorphism (SNP) marker densities and the true quantitative trait loci genotypes. The results show that using high‐density SNP markers generated more genetic gain and heterosis than the low‐density SNP markers. The relative performance of genomic prediction models differed across marker scenarios. For genetic gain, we observed more differences between the models at low than high marker density. For heterosis, we observed the opposite, more differences between the models at high than low marker density. Overall, models that fitted the average or additive effects specific to each heterotic pool and dominance effects provide a better fit and hence higher genetic gain in hybrid breeding programs.

Heterotic grouping of provitamin A-enriched maize inbred lines for increased provitamin A content in hybrids

BackgroundThe establishment of heterotic groups of inbred lines is crucial for hybrid maize breeding programs. Currently, there is no information on the heterotic patterns of the Provitamin A (PVA) inbred lines developed in the maize improvement program of the International Institute of Tropical Agriculture (IITA) to form productive PVA enriched hybrids for areas affected by vitamin A deficiency. This study assessed the feasibility of classifying PVA-enriched inbred lines into heterotic groups based on PVA content without compromising grain yield in hybrids. Sixty PVA inbred lines were crossed to two testers representing two existing heterotic groups. The resulting 120 testcrosses hybrids were evaluated for two years at four locations in Nigeria.ResultsThe two testers effectively classified the inbred lines into two heterotic groups. The PVA-based general combining ability and specific combining ability (HSGCA) method assigned 31 and 27 PVA enriched maize inbred lines into HGB and HGA, respectively, leaving two inbred lines not assigned to any group. The yield-based HSGCA method classified 32 inbred lines into HGB and 28 inbred lines into HGA. Both PVA and yield-based heterotic grouping methods assigned more than 40% of the inbred lines into the same heterotic groups. Even though both PVA and yield-based heterotic grouping of the inbred lines differed from the clusters defined by the DArTag SNP markers, more than 40% of the inbred lines assigned to HGA were present in Cluster-1 and 60% of the inbred lines assigned to HGB were present in Cluster-3. Interestingly, the inbred lines assigned to the same heterotic groups based on PVA content and grain yield were distributed across the three Ward’s clusters. The PVA-based HSGCA was identified as the most effective heterotic grouping method for breeding programs working on PVA biofortification.ConclusionsSelecting PVA enriched maize inbred lines with diverse genetic backgrounds from the three marker-based clusters may facilitate the development of productive hybrids with high PVA content and for generating source populations to develop more vigorous maize inbred lines with much higher concentrations of PVA.

Development of Homozygous Lines by In Vivo Doubled Haploid Technique in Hybrid Maize Breeding

The maize breeding programs focuses on the development of homozygous parental lines for hybrid breeding to obtain heterosis. The Improvement of homozygous lines in a effective time is crucial for hybrid maize breeding. Objectives of this research were to obtain homozygous lines of maize in a short time by using of in-vivo maternal haploid (DH) technique and characterizing them morphologically. The experiments were carried out at the Eastern Mediterranean Agricultural Research Institute(EMARI). Inducers, RWS, RWK-76 and their hybrid RWS X RWK-76 were used as male parent. As female parent, 56 F2 segregated material obtained by selfing of 66 F1 hybrids developed in maize breeding program of Eastern Mediterranean Agricultural Research Institute (EMARI) and 2 F2 segregated material selected from the open pollinated plants of 9 commercial maize variety. As a result of the experiment, 29 doubled haploid lines (DH) were developed. In the developed DH lines, days to tasseling, plant height, height of first ear , ear length, ear diameter, number of kernel per ear and thousand kernel weight varied as 57-78 days, 151-248 cm, 43-112 cm, 11.20-24.50 cm, 24-45 mm, 224-537 kernels/ear, and 180-320 g, respectively. In conclusion, in-vivo maternal DH technique is a highly effective method for obtaining homozygous lines

Creation of doubled haploid lines of maize <i>Zea mays</i> L. by resynthesis from a tetraploid population

The search for new, effective methods for broadening the genetic polymorphism of the original breeding material remains one of important problems of hybrid maize breeding. Globally, breeding of commercial maize varieties and hybrids is carried out using diploid genotypes, whereas tetraploid sources of initial material and wild relatives of maize are poorly involved in the breeding process. The direct heteroploid crosses between diploid and tetraploid genotypes lead to the formation of weakly fertile or completely sterile triploid hybrids, which are cytologically unstable in subsequent generations. Tetraploid maize (2n=40), as well as some wild relatives with tetraploid genome, such as Zea perennis Hitchk. (2n=40) and Tripsacum dactiloides (L.) L. (2n=72), are attractive to breeders as sources for improving economically valuable traits. The attractiveness of resynthesized diploid lines is explained by the fact that unequal crossing over between homologous chromosomes forming polyvalent associations of chromosomes, more chromosomal rearrangements occur in tetraploids than in diploid genotypes, the chromosomes of which form bivalents. Synthetic tetraploid populations of maize and its tetraploid wild relatives have great potential of variability for improving diploid maize. The authors proposed a direct method for the resynthesis of doubled haploid lines using haploid induction and an indirect method for obtaining diploid lines by heteroploid crossing and subsequent segregation of a triploid hybrid in the progeny. The method of resynthesizing the tetraploid maize genome to diploid serves as an ideal model for studying the processes of crossing over in meiosis between multivalent associations of homologous chromosomes; it is promising for obtaining diploid lines with an increased frequency of recombination between the homologous chromosomes of different genomes, combined into a common one, and can serve as a source for obtaining aneuploid series.

ZmRPN1 confers quantitative variation in pollen number and boosts hybrid seed production in maize.

The number of pollen grains is a critical determinant of reproductive success in seed plants and varies among species and individuals. However, in contrast with many mutant-screening studies relevant to anther and pollen development, the natural genetic basis for variations in pollen number remains largely unexplored. To address this issue, we carried out a genome-wide association study in maize, ultimately revealing that a large presence/absence variation in the promoter region of ZmRPN1 alters its expression level and thereby contributes to pollen number variation. Molecular analyses showed that ZmRPN1 interacts with ZmMSP1, which is known as a germline cell number regulator, and facilitates ZmMSP1 localization to the plasma membrane. Importantly, ZmRPN1 dysfunction resulted in a substantial increase in pollen number, consequently boosting seed production by increasing female-male planting ratio. Together, our findings uncover a key gene controlling pollen number, and therefore, modulation of ZmRPN1 expression could be efficiently used to develop elite pollinators for modern hybrid maize breeding.

Identification of best testers for heterotic grouping of tropical maize inbred lines using GGE biplot

AbstractAn understanding of the heterotic pattern in the active germplasm is the first and crucial step in any hybrid breeding program. Diallel and line × tester (L × T) mating designs were extensively followed to understand the heterotic pattern based on the combining ability information. The present study adopted GGE biplots, generated using theL × Tmatrix to understand the tropical maize germplasm's heterotic pattern and identify the best tester combinations for heterotic grouping. TheL × Tmatrix and combining ability information was generated by evaluating 985 testcross, generated by crossing 110 diverse tropical inbred lines with 10 inbred testers at two geographically diverse locations. The stability of combining ability effects, the magnitude and direction of combining ability effects, and also the discrimination and representativeness of testers were considered for the identification of best tester combinations for heterotic grouping. The present investigation has shown the usefulness of GGE biplots’ effectiveness in understanding the heterotic pattern and identifying the best tester combinations for heterotic grouping. The results of the present investigation revealed the existing heterotic pattern in the elite tropical maize inbred lines that were extensively used in hybrid maize breeding. The findings of the present study are of immense practical significance, which not only guides different hybrid maize breeding programs but also helps to identify the best testers in their respective hybrid breeding programs. The study showed that the GGE biplot method can be adopted to understand the active germplasm's heterotic pattern and organize the germplasm into different heterotic groups.

Training set designs for prediction of yield and moisture of maize test cross hybrids with unreplicated trials.

Unreplicated field trials and genomic prediction are both used to enhance the efficiency in early selection stages of a hybrid maize breeding program. No results are available on the optimal experimental design when combining both approaches. Our objectives were to investigate the effect of the training set design on the accuracy of genomic prediction in unreplicated maize test crosses. We carried out a cross validation study on basis of an experimental data set consisting of 1436 hybrids evaluated for yield and moisture for which genotyping information of 461 SNP markers were available. Training set designs of different size, implementing within environment prediction, within year prediction, across year prediction, and combinations of data sources across years and environments were compared with respect to their prediction accuracy. Across year prediction did not reach prediction accuracies that are useful for genomic selection. Within year prediction across environments provided useful correlations between observed and predicted breeding values. The prediction accuracies did not improve when adding to the training set data from previous years. We conclude that using all data available from unreplicated tests of the current breeding cycle provides a good accuracy of predicting test crosses, whereas adding data from previous breeding cycles, in which the genotypes are less related to the tested material, has only limited value for increasing the prediction accuracy.

Heterosis and combining ability of iron, zinc and their bioavailability in maize inbred lines under low nitrogen and optimal environments

Iron (Fe) and zinc (Zn) nutrient enrichment of staple crops through biofortification can contribute to alleviating micronutrient deficiency in sub-Saharan Africa. A line × tester mating design was used to determine the general combining ability (GCA), specific combining ability (SCA) and heterosis for grain yield, iron, Zn and phytic concentration of six lines crossed with three testers. Lines and testers were selected for high, intermediate and low mineral content. The F1 hybrids and parental lines were evaluated under low nitrogen (N) and optimum conditions across four environments over two seasons. Under low N conditions, Fe and Zn concentration in grain, and grain yield of genotypes were reduced by 9%, 9%, and 59%, respectively. However, phytic acid concentration in grain was increased by 10% under low N conditions. Both additive and non-additive gene effects were important in controlling Fe, Zn and phytic acid concentration in grain and grain yield of maize under both N conditions. The preponderance of GCA effects indicates the importance of additive gene effects in the inheritance of grain yield. Line GCA effects were more sensitive to N conditions across the environments than the tester GCA. High and significant positive SCA effects for grain yield, Fe and Zn content under low N conditions, would be a good indicator of possible heterosis in these traits. Hybrid CBY101 LM-1600 × CBY358 LM-1857 had high and significant positive SCA for grain yield under low N conditions and is a promising candidate for production in low N environments. CBY358 LM-1857 (tester) and CBY102 LM-1601 (line) are a good general combiners for Fe, Zn and GY can be used as parents in future maize hybrid breeding programs to develop high-yielding maize genotypes with high Fe and Zn content.

Epistasis Activation Contributes Substantially to Heterosis in Temperate by Tropical Maize Hybrids.

Epistasis strongly affects the performance of superior maize hybrids. In this study, a multiple-hybrid population, consisting of three hybrid maize sets with varied interparental divergence, was generated by crossing 28 temperate and 23 tropical inbred lines with diverse genetic backgrounds. We obtained 1,154 tested hybrids. Among these tested hybrids, heterosis increased steadily as the heterotic genetic distance increased. Mid-parent heterosis was significantly higher in the temperate by tropical hybrids than in the temperate by temperate hybrids. Genome-wide prediction and association mapping was performed for grain weight per plant (GWPP) and days to silking (DTS) using 20K high-quality SNPs, showing that epistatic effects played a more prominent role than dominance effects in temperate by tropical maize hybrids. A total of 33 and 420 epistatic QTL were identified for GWPP and DTS, respectively, in the temperate by tropical hybrids. Protein–protein interaction network and gene-set enrichment analyses showed that epistatic genes were involved in protein interactions, which play an important role in photosynthesis, biological transcription pathways, and protein synthesis. We showed that the interaction of many minor-effect genes in the hybrids could activate the transcription activators of epistatic genes, resulting in a cascade of amplified yield heterosis. The multiple-hybrid population design enhanced our understanding of heterosis in maize, providing an insight into the acceleration of hybrid maize breeding by activating epistatic effects.

Cloning of a new allele of ZmAMP1 and evaluation of its breeding value in hybrid maize

Gene resources associated with plant stature and flowering time are invaluable for maize breeding. In this study, using an F2:3 population derived from a natural semi-dwarf mutant grmm and a normal inbred line Si 273, we identified a major pleiotropic QTL on the distal long arm of chromosome 1 (qPH1_dla), and found that qPH1_dla controlled plant height, flowering time, ear and yield traits. qPH1_dla was fine-mapped to a 16 kb interval containing ZmAMP1, which was annotated as a glutamate carboxypeptidase. Allelism tests using two independent allelic mutants confirmed that ZmAMP1 was the causal gene. Real-time quantitative PCR and genomic sequence analysis suggested that a nonsynonymous mutation at the 598th base of ZmAMP1 gene was the causal sequence variant for the dwarfism of grmm. This novel ZmAMP1 allele was named ZmAMP1_grmm. RNA sequencing using two pairs of near isogenic lines (NILs) showed that 84 up-regulated and 68 down-regulated genes in dwarf NILs were enriched in 15 metabolic pathways. Finally, introgression of ZmAMP1_grmm into Zhengdan 958 and Xianyu 335 generated two improved F1 lines. In field tests, they were semi-dwarf, early-flowering, lodging-resistant, and high-yielding under high-density planting conditions, suggesting that ZmAMP1_grmm is a promising Green Revolution gene for maize hybrid breeding.