Genetic trends in CIMMYT's tropical maize breeding pipelines.

Fostering a culture of continuous improvement through regular monitoring of genetic trends in breeding pipelines is essential to improve efficiency and increase accountability. This is the first global study to estimate genetic trends across the International Maize and Wheat Improvement Center (CIMMYT) tropical maize breeding pipelines in eastern and southern Africa (ESA), South Asia, and Latin America over the past decade. Data from a total of 4152 advanced breeding trials and 34,813 entries, conducted at 1331 locations in 28 countries globally, were used for this study. Genetic trends for grain yield reached up to 138 kg ha-1 yr-1 in ESA, 118 kg ha-1 yr-1 South Asia and 143 kg ha-1 yr-1 in Latin America. Genetic trend was, in part, related to the extent of deployment of new breeding tools in each pipeline, strength of an extensive phenotyping network, and funding stability. Over the past decade, CIMMYT's breeding pipelines have significantly evolved, incorporating new tools/technologies to increase selection accuracy and intensity, while reducing cycle time. The first pipeline, Eastern Africa Product Profile 1a (EA-PP1a), to implement marker-assisted forward-breeding for resistance to key diseases, coupled with rapid-cycle genomic selection for drought, recorded a genetic trend of 2.46% per year highlighting the potential for deploying new tools/technologies to increase genetic gain.

Maize varietal replacement in Eastern and Southern Africa: Bottlenecks, drivers and strategies for improvement.

Seed security is vital for food security. Rapid-cycle, climate-adaptive breeding programs and seed systems that deliver new, elite varieties to farmers to replace obsolete ones can greatly improve the productivity of maize-based cropping systems in sub-Saharan Africa (SSA). Despite the importance and benefits of accelerated varietal turnover to climate change adaptation and food security, the rate of maize varietal replacement in SSA is slow. This review outlines the major bottlenecks, drivers, risks, and benefits of active replacement of maize varieties in eastern and southern Africa (ESA) and highlights strategies that are critical to varietal turnover. Although there is an upsurge of new seed companies in ESA and introduction of new varieties with better genetics in the market, some established seed companies continue to sell old (over 15-year-old) varieties. Several recently developed maize hybrids in ESA have shown significant genetic gains under farmers' conditions. Empirical evidence also shows that timely replacement of old products results in better business success as it helps seed companies maintain or improve market share and brand relevance. Therefore, proactive management of product life cycles by seed companies benefits both the farmers and businesses alike, contributing to improved food security and adaptation to the changing climate.

Multinutrient Biofortification of Maize (Zea mays L.) in Africa: Current Status, Opportunities and Limitations.

Macro and micronutrient deficiencies pose serious health challenges globally, with the largest impact in developing regions such as subSaharan Africa (SSA), Latin America and South Asia. Maize is a good source of calories but contains low concentrations of essential nutrients. Major limiting nutrients in maize-based diets are essential amino acids such as lysine and tryptophan, and micronutrients such as vitamin A, zinc (Zn) and iron (Fe). Responding to these challenges, separate maize biofortification programs have been designed worldwide, resulting in several cultivars with high levels of provitamin A, lysine, tryptophan, Zn and Fe being commercialized. This strategy of developing single-nutrient biofortified cultivars does not address the nutrient deficiency challenges in SSA in an integrated manner. Hence, development of maize with multinutritional attributes can be a sustainable and cost-effective strategy for addressing the problem of nutrient deficiencies in SSA. This review provides a synopsis of the health challenges associated with Zn, provitamin A and tryptophan deficiencies and link these to vulnerable societies; a synthesis of past and present intervention measures for addressing nutrient deficiencies in SSA; and a discussion on the possibility of developing maize with multinutritional quality attributes, but also with adaptation to stress conditions in SSA.

Beat the stress: breeding for climate resilience in maize for the tropical rainfed environments.

KEY MESSAGE: Intensive public sector breeding efforts and public-private partnerships have led to the increase in genetic gains, and deployment of elite climate-resilient maize cultivars for the stress-prone environments in the tropics. Maize (Zea mays L.) plays a critical role in ensuring food and nutritional security, and livelihoods of millions of resource-constrained smallholders. However, maize yields in the tropical rainfed environments are now increasingly vulnerable to various climate-induced stresses, especially drought, heat, waterlogging, salinity, cold, diseases, and insect pests, which often come in combinations to severely impact maize crops. The International Maize and Wheat Improvement Center (CIMMYT), in partnership with several public and private sector institutions, has been intensively engaged over the last four decades in breeding elite tropical maize germplasm with tolerance to key abiotic and biotic stresses, using an extensive managed stress screening network and on-farm testing system. This has led to the successful development and deployment of an array of elite stress-tolerant maize cultivars across sub-Saharan Africa, Asia, and Latin America. Further increasing genetic gains in the tropical maize breeding programs demands judicious integration of doubled haploidy, high-throughput and precise phenotyping, genomics-assisted breeding, breeding data management, and more effective decision support tools. Multi-institutional efforts, especially public-private alliances, are key to ensure that the improved maize varieties effectively reach the climate-vulnerable farming communities in the tropics, including accelerated replacement of old/obsolete varieties.

Fall-armyworm invasion, control practices and resistance breeding in Sub-Saharan Africa.

Fall armyworm [Spodoptera frugiperda (J.E. Smith); FAW] invasion has exacerbated maize (Zea mays L.) crop yield losses in sub-Saharan Africa (SSA), already threatened by other stresses, especially those that are climate-change induced. The FAW is difficult to control, manage, or eradicate, because it is polyphagous and trans-boundary, multiplies fast, has a short life cycle and migrates easily, and lacks the diapause growth phase. In this study, FAW and its impact in Africa was reviewed, as well as past and present control strategies for this pest. Pesticides, cultural practices, natural enemies, host-plant resistance, integrated pest management (IPM), and plant breeding approaches were examined as possible control strategies. It was concluded that an IPM control strategy, guided by cultural approaches already being used by farmers, and what can be adopted from the Americas, coupled with an insect-resistance management strategy, is the best option to manage this pest in Africa. These strategies will be strengthened by breeding for multi-trait host-plant resistance through stacking of genes for different modes of control of the pest.

Line × tester analysis of maize grain yield under acid and non-acid soil conditions.

Soil acidity has received less attention than other biophysical stresses such as drought and low N, despite accounting for a considerable reduction in maize (Zea mays L.) productivity in many parts of southern Africa. The line × tester mating design was used to determine the general combining ability (GCA) for grain yield of 14 maize inbred lines and the specific combining ability (SCA) of their corresponding crosses. Thirty-three single-cross hybrids were evaluated under acid and optimum soils across 11 environments over three seasons. Across environments, mean grain yield reduction ranged from 11 to 37% due to low pH. Additive gene action was more important than nonadditive gene action for grain yield under both soil conditions. Tester GCA effects were larger for grain yield than GCA effects of lines and SCA effects of crosses for both soil conditions. Tester GCA effects were less sensitive to environmental fluctuations than line GCA effects and SCA effects of crosses. Cross combinations with desirable SCA effects for grain yield were associated with high per se grain yield, which suggests that SCA was a good predictor of grain yield in this study. These crosses consisted of good × good and good × poor general combiners, which indicates that GCA was a good predictor of grain yield. Therefore, priority should be given for yield selection in progenies and hybridization of specific crosses with desirable SCA when breeding acid-soil-tolerant maize.

Impact of adoption of drought-tolerant maize varieties on total maize production in south Eastern Zimbabwe.

Drought is a huge limiting factor in maize production, mainly in the rain-fed agriculture of sub-Saharan Africa. In response to this threat, drought-tolerant (DT) maize varieties have been developed with an aim to ensure maize production under mild drought conditions. We conducted a study to assess the impact of smallholder farmers' adoption of DT maize varieties on total maize production. Data for the study came from a survey of 200 randomly sampled households in two districts of Chiredzi and Chipinge in southeastern Zimbabwe. The study found that 93% of the households were growing improved maize varieties and that 30% of the sampled households were growing DT maize varieties. Total maize yield was 436.5 kg/ha for a household that did not grow DT maize varieties and 680.5 kg/ha for households that grew DT maize varieties. We control for the endogeneity of the DT adoption variable, by using the control function approach to estimate total maize production in a Cobb-Douglas model. The results show that households that grew DT maize varieties had 617 kg/ha more maize than households that did not grow the DT maize varieties. Given that almost all farmers buy their seeds in the market, a change in varieties to DT maize seeds gives an extra income of US$240/ha or more than nine months of food at no additional cost. This has huge implications in curbing food insecurity and simultaneously saving huge amounts of resources at the household and national levels, which are used to buy extra food during the lean season.

Genetic Analysis of Tropical Midaltitude- Adapted Maize Populations under Stress and Nonstress Conditions.

Maize (Zea mays L.) yield in sub-Saharan Africa (SSA) is low because of both abiotic and biotic constraints, and limited availability or use of improved seed in some areas. This study was conducted (i) to estimate combining ability and heterosis among seven stress-tolerant populations, and (ii) to assess diversity among the populations and the relationship between diversity and heterosis. Twenty-one hybrids developed from diallel crosses of seven populations, parents, and two checks were evaluated in 10 optimal and 11 stressed environments (drought, low N, and random stress) in Kenya, Ethiopia, Uganda, and Zimbabwe for 2 yr. Analysis II of Gardner and Eberhart showed that variety and heterosis were significant for grain yield (GY) under optimal and managed stress, and across environments. Heterosis accounted for most of the variation for GY among populations under optimal conditions (67%) and drought stress (53%), which suggested the importance of dominance in inheritance of GY under these conditions. Genetic distance (GD) among populations ranged from 0.328 to 0.477 (mean = 0.404). The correlation between GD and heterosis was low (r = 0.14-0.40) in all environments. The simple sequence repeat (SSR) marker-based and GY-based clustering of parental populations showed similar patterns, with three populations distinct from the rest, suggesting significant differentiation of allelic variation in these three populations. The SSR-based diversity and phenotypic analysis results should be useful in defining breeding strategies and maintaining heterotic patterns among these populations.

High-Throughput Phenotyping of Canopy Cover and Senescence in Maize Field Trials Using Aerial Digital Canopy Imaging.

In the crop breeding process, the use of data collection methods that allow reliable assessment of crop adaptation traits, faster and cheaper than those currently in use, can significantly improve resource use efficiency by reducing selection cost and can contribute to increased genetic gain through improved selection efficiency. Current methods to estimate crop growth (ground canopy cover) and leaf senescence are essentially manual and/or by visual scoring, and are therefore often subjective, time consuming, and expensive. Aerial sensing technologies offer radically new perspectives for assessing these traits at low cost, faster, and in a more objective manner. We report the use of an unmanned aerial vehicle (UAV) equipped with an RGB camera for crop cover and canopy senescence assessment in maize field trials. Aerial-imaging-derived data showed a moderately high heritability for both traits with a significant genetic correlation with grain yield. In addition, in some cases, the correlation between the visual assessment (prone to subjectivity) of crop senescence and the senescence index, calculated from aerial imaging data, was significant. We concluded that the UAV-based aerial sensing platforms have great potential for monitoring the dynamics of crop canopy characteristics like crop vigor through ground canopy cover and canopy senescence in breeding trial plots. This is anticipated to assist in improving selection efficiency through higher accuracy and precision, as well as reduced time and cost of data collection.

Genome-Wide Association Mapping and Genomic Prediction Analyses Reveal the Genetic Architecture of Grain Yield and Flowering Time Under Drought and Heat Stress Conditions in Maize.

Drought stress (DS) is a major constraint to maize yield production. Heat stress (HS) alone and in combination with DS are likely to become the increasing constraints. Association mapping and genomic prediction (GP) analyses were conducted in a collection of 300 tropical and subtropical maize inbred lines to reveal the genetic architecture of grain yield and flowering time under well-watered (WW), DS, HS, and combined DS and HS conditions. Out of the 381,165 genotyping-by-sequencing SNPs, 1549 SNPs were significantly associated with all the 12 trait-environment combinations, the average PVE (phenotypic variation explained) by these SNPs was 4.33%, and 541 of them had a PVE value greater than 5%. These significant associations were clustered into 446 genomic regions with a window size of 20 Mb per region, and 673 candidate genes containing the significantly associated SNPs were identified. In addition, 33 hotspots were identified for 12 trait-environment combinations and most were located on chromosomes 1 and 8. Compared with single SNP-based association mapping, the haplotype-based associated mapping detected fewer number of significant associations and candidate genes with higher PVE values. All the 688 candidate genes were enriched into 15 gene ontology terms, and 46 candidate genes showed significant differential expression under the WW and DS conditions. Association mapping results identified few overlapped significant markers and candidate genes for the same traits evaluated under different managements, indicating the genetic divergence between the individual stress tolerance and the combined drought and HS tolerance. The GP accuracies obtained from the marker-trait associated SNPs were relatively higher than those obtained from the genome-wide SNPs for most of the target traits. The genetic architecture information of the grain yield and flowering time revealed in this study, and the genomic regions identified for the different trait-environment combinations are useful in accelerating the efforts on rapid development of the stress-tolerant maize germplasm through marker-assisted selection and/or genomic selection.

Genome-wide association mapping reveals novel sources of resistance to northern corn leaf blight in maize.

BACKGROUND: Northern corn leaf blight (NCLB) caused by Exserohilum turcicum is a destructive disease in maize. Using host resistance to minimize the detrimental effects of NCLB on maize productivity is the most cost-effective and appealing disease management strategy. However, this requires the identification and use of stable resistance genes that are effective across different environments. RESULTS: We evaluated a diverse maize population comprised of 999 inbred lines across different environments for resistance to NCLB. To identify genomic regions associated with NCLB resistance in maize, a genome-wide association analysis was conducted using 56,110 single-nucleotide polymorphism markers. Single-marker and haplotype-based associations, as well as Anderson-Darling tests, identified alleles significantly associated with NCLB resistance. The single-marker and haplotype-based association mappings identified twelve and ten loci (genes), respectively, that were significantly associated with resistance to NCLB. Additionally, by dividing the population into three subgroups and performing Anderson-Darling tests, eighty one genes were detected, and twelve of them were related to plant defense. Identical defense genes were identified using the three analyses. CONCLUSION: An association panel including 999 diverse lines was evaluated for resistance to NCLB in multiple environments, and a large number of resistant lines were identified and can be used as reliable resistance resource in maize breeding program. Genome-wide association study reveals that NCLB resistance is a complex trait which is under the control of many minor genes with relatively low effects. Pyramiding these genes in the same background is likely to result in stable resistance to NCLB.

Fine mapping of Msv1, a major QTL for resistance to Maize Streak Virus leads to development of production markers for breeding pipelines.

Msv1 , the major QTL for MSV resistance was delimited to an interval of 0.87 cM on chromosome 1 at 87 Mb and production markers with high prediction accuracy were developed. Maize streak virus (MSV) disease is a devastating disease in the Sub-Saharan Africa (SSA), which causes significant yield loss in maize. Resistance to MSV has previously been mapped to a major QTL (Msv1) on chromosome 1 that is germplasm and environment independent and to several minor loci elsewhere in the genome. In this study, Msv1 was fine-mapped through QTL isogenic recombinant strategy using a large F 2 population of CML206 × CML312 to an interval of 0.87 cM on chromosome 1. Genome-wide association study was conducted in the DTMA (Drought Tolerant Maize for Africa)-Association mapping panel with 278 tropical/sub-tropical breeding lines from CIMMYT using the high-density genotyping-by-sequencing (GBS) markers. This study identified 19 SNPs in the region between 82 and 93 Mb on chromosome 1(B73 RefGen_V2) at a P < 1.00E-04, which coincided with the fine-mapped region of Msv1. Haplotype trend regression identified a haplotype block significantly associated with response to MSV. Three SNPs in this haplotype block at 87 Mb on chromosome 1 had an accuracy of 0.94 in predicting the disease reaction in a collection of breeding lines with known responses to MSV infection. In two biparental populations, selection for resistant Msv1 haplotype demonstrated a reduction of 1.03-1.39 units on a rating scale of 1-5, compared to the susceptible haplotype. High-throughput KASP assays have been developed for these three SNPs to enable routine marker screening in the breeding pipeline for MSV resistance.

QTL mapping in three tropical maize populations reveals a set of constitutive and adaptive genomic regions for drought tolerance.

Despite numerous published reports of quantitative trait loci (QTL) for drought-related traits, practical applications of such QTL in maize improvement are scarce. Identifying QTL of sizeable effects that express more or less uniformly in diverse genetic backgrounds across contrasting water regimes could significantly complement conventional breeding efforts to improve drought tolerance. We evaluated three tropical bi-parental populations under water-stress (WS) and well-watered (WW) regimes in Mexico, Kenya and Zimbabwe to identify genomic regions responsible for grain yield (GY) and anthesis-silking interval (ASI) across multiple environments and diverse genetic backgrounds. Across the three populations, on average, drought stress reduced GY by more than 50 % and increased ASI by 3.2 days. We identified a total of 83 and 62 QTL through individual environment analyses for GY and ASI, respectively. In each population, most QTL consistently showed up in each water regime. Across the three populations, the phenotypic variance explained by various individual QTL ranged from 2.6 to 17.8 % for GY and 1.7 to 17.8 % for ASI under WS environments and from 5 to 19.5 % for GY under WW environments. Meta-QTL (mQTL) analysis across the three populations and multiple environments identified seven genomic regions for GY and one for ASI, of which six mQTL on chr.1, 4, 5 and 10 for GY were constitutively expressed across WS and WW environments. One mQTL on chr.7 for GY and one on chr.3 for ASI were found to be 'adaptive' to WS conditions. High throughput assays were developed for SNPs that delimit the physical intervals of these mQTL. At most of the QTL, almost equal number of favorable alleles was donated by either of the parents within each cross, thereby demonstrating the potential of drought tolerant × drought tolerant crosses to identify QTL under contrasting water regimes.

Quality control genotyping for assessment of genetic identity and purity in diverse tropical maize inbred lines.

Quality control (QC) genotyping is an important component in breeding, but to our knowledge there are not well established protocols for its implementation in practical breeding programs. The objectives of our study were to (a) ascertain genetic identity among 2-4 seed sources of the same inbred line, (b) evaluate the extent of genetic homogeneity within inbred lines, and (c) identify a subset of highly informative single-nucleotide polymorphism (SNP) markers for routine and low-cost QC genotyping and suggest guidelines for data interpretation. We used a total of 28 maize inbred lines to study genetic identity among different seed sources by genotyping them with 532 and 1,065 SNPs using the KASPar and GoldenGate platforms, respectively. An additional set of 544 inbred lines was used for studying genetic homogeneity. The proportion of alleles that differed between seed sources of the same inbred line varied from 0.1 to 42.3 %. Seed sources exhibiting high levels of genetic distance are mis-labeled, while those with lower levels of difference are contaminated or still segregating. Genetic homogeneity varied from 68.7 to 100 % with 71.3 % of the inbred lines considered to be homogenous. Based on the data sets obtained for a wide range of sample sizes and diverse genetic backgrounds, we recommended a subset of 50-100 SNPs for routine and low-cost QC genotyping, verified them in a different set of double haploid and inbred lines, and outlined a protocol that could be used to minimize errors in genetic analyses and breeding.

Molecular characterization of diverse CIMMYT maize inbred lines from eastern and southern Africa using single nucleotide polymorphic markers.

BACKGROUND: Knowledge of germplasm diversity and relationships among elite breeding materials is fundamentally important in crop improvement. We genotyped 450 maize inbred lines developed and/or widely used by CIMMYT breeding programs in both Kenya and Zimbabwe using 1065 SNP markers to (i) investigate population structure and patterns of relationship of the germplasm for better exploitation in breeding programs; (ii) assess the usefulness of SNPs for identifying heterotic groups commonly used by CIMMYT breeding programs; and (iii) identify a subset of highly informative SNP markers for routine and low cost genotyping of CIMMYT germplasm in the region using uniplex assays. RESULTS: Genetic distance for about 94% of the pairs of lines fell between 0.300 and 0.400. Eighty four percent of the pairs of lines also showed relative kinship values ≤ 0.500. Model-based population structure analysis, principal component analysis, neighbor-joining cluster analysis and discriminant analysis revealed the presence of 3 major groups and generally agree with pedigree information. The SNP markers did not show clear separation of heterotic groups A and B that were established based on combining ability tests through diallel and line x tester analyses. Our results demonstrated large differences among the SNP markers in terms of reproducibility, ease of scoring, polymorphism, minor allele frequency and polymorphic information content. About 40% of the SNPs in the multiplexed chip-based GoldenGate assays were found to be uninformative in this study and we recommend 644 of the 1065 for low to medium density genotyping in tropical maize germplasm using uniplex assays. CONCLUSIONS: There were high genetic distance and low kinship coefficients among most pairs of lines, clearly indicating the uniqueness of the majority of the inbred lines in these maize breeding programs. The results from this study will be useful to breeders in selecting best parental combinations for new breeding crosses, mapping population development and marker assisted breeding.

Molecular characterization of global maize breeding germplasm based on genome-wide single nucleotide polymorphisms.

Characterization of genetic diversity is of great value to assist breeders in parental line selection and breeding system design. We screened 770 maize inbred lines with 1,034 single nucleotide polymorphism (SNP) markers and identified 449 high-quality markers with no germplasm-specific biasing effects. Pairwise comparisons across three distinct sets of germplasm, CIMMYT (394), China (282), and Brazil (94), showed that the elite lines from these diverse breeding pools have been developed with only limited utilization of genetic diversity existing in the center of origin. Temperate and tropical/subtropical germplasm clearly clustered into two separate groups. The temperate germplasm could be further divided into six groups consistent with known heterotic patterns. The greatest genetic divergence was observed between temperate and tropical/subtropical lines, followed by the divergence between yellow and white kernel lines, whereas the least divergence was observed between dent and flint lines. Long-term selection for hybrid performance has contributed to significant allele differentiation between heterotic groups at 20% of the SNP loci. There appeared to be substantial levels of genetic variation between different breeding pools as revealed by missing and unique alleles. Two SNPs developed from the same candidate gene were associated with the divergence between two opposite Chinese heterotic groups. Associated allele frequency change at two SNPs and their allele missing in Brazilian germplasm indicated a linkage disequilibrium block of 142 kb. These results confirm the power of SNP markers for diversity analysis and provide a feasible approach to unique allele discovery and use in maize breeding programs.