Efficacy of Event MON 87460 in drought-tolerant maize hybrids under optimal and managed drought-stress in eastern and southern africa.

BACKGROUND: Frequent drought events due to climate change have become a major threat to maize (Zea mays L.) production and food security in Africa. Genetic engineering is one of the ways of improving drought tolerance through gene introgression to reduce the impact of drought stress in maize production. This study aimed to evaluate the efficacy of Event MON 87460 (CspB; DroughtGard®) gene in more than 120 conventional drought-tolerant maize hybrids in Kenya, South Africa, and Uganda for 3-6 years under managed drought-stress and optimal conditions and establish any additional yield contribution or yield penalties of the gene in traited hybrids relative to their non-traited isohybrids. Germplasm used in the study were either MON 87460 traited un-adapted (2008-2010), adapted traited DroughtTEGO® (2011-2013) or a mix of both under confined field trials. RESULTS: Results showed significant yield differences (p < 0.001) among MON 87460 traited and non-traited hybrids across well-watered and managed drought-stress treatments. The gene had positive and significant effect on yield by 36-62% in three hybrids (CML312/CML445; WMA8101/CML445; and CML312/S0125Z) relative to non-traited hybrids under drought, and without significant yield penalty under optimum-moisture conditions in Lutzville, South Africa. Five traited hybrids (WMA2003/WMB4401; CML442/WMB4401; CML489/WMB4401; CML511/CML445; and CML395/WMB4401) had 7-13% significantly higher yield than the non-traited isohybrids out of 34 adapted DroughtTEGO® hybrids with same background genetics in the three countries for ≥ 3 years. The positive effect of MON 87460 was mostly observed under high drought-stress relative to low, moderate, or severe stress levels. CONCLUSION: This study showed that MON 87460 transgenic drought tolerant maize hybrids could effectively tolerate drought and shield farmers against severe yield loss due to drought stress. The study signified that development and adoption of transgenic drought tolerant maize hybrids can cushion against farm yield losses due to drought stress as part of an integrated approach in adaptation to climate change effects.

Performance of Bt maize event MON810 in controlling maize stem borers Chilo partellus and Busseola fusca in Uganda.

Stem borers are major insect pests of maize in Uganda. A study was conducted in 2014-2016 to assess the performance of Bt hybrids expressing Cry1Ab (event MON810) against the two major stem borer species in Uganda - the African stem borer (Busseola fusca) and the spotted stem borer (Chilo partellus) - under artificial infestation. The study comprised 14 non-commercialized hybrids, including seven pairs of Bt and non-Bt hybrids (isolines), three non-Bt commercial hybrids and a conventional stem borer resistant check. All stem borer damage parameters (leaf damage, number of internodes tunneled and tunnel length) were generally significantly lower in Bt hybrids than in their isolines, the conventionally resistant hybrid, and local commercial hybrids. Mean yields were significantly higher by 29.4-80.5% in the Bt hybrids than in the other three categories of non-Bt hybrids. This study demonstrated that Bt maize expressing Cry1Ab protects against leaf damage and can limit entry of stem borers into the stems of maize plants, resulting in higher yield than in the non-transgenic hybrids. Thus, Bt maize has potential to contribute to the overall management package of stem borers in Uganda.

Designing a Pest and Disease Outbreak Warning System for Farmers, Agronomists and Agricultural Input Distributors in East Africa.

Early warnings of the risks of pest and disease outbreaks are becoming more urgent, with substantial increases in threats to agriculture from invasive pests. With geospatial data improvements in quality and timeliness, models and analytical systems can be used to estimate potential areas at high risk of yield impacts. The development of decision support systems requires an understanding of what information is needed, when it is needed, and at what resolution and accuracy. Here, we report on a professional review conducted with 53 professional agronomists, retailers, distributors, and growers in East Africa working with the Syngenta Foundation for Sustainable Agriculture. The results showed that respondents reported fall armyworm, stemborers and aphids as being among the most common pests, and that crop diversification was a key strategy to reduce their impact. Chemical and cultural controls were the most common strategies for fall armyworm (FAW) control, and biological control was the least known and least used method. Of the cultural control methods, monitoring and scouting, early planting, and crop rotation with non-host crops were most used. Although pests reduced production, only 55% of respondents were familiar with early warning tools, showing the need for predictive systems that can improve farmer response.

Development and initial validation of the attitudes toward face mask use scale (ATFMUS).

Despite availability of instruments for measuring attitudes towards mask use, the psychometric properties of many available instruments are not adequately established which limits their research usefulness across contexts. In three studies, we developed the Attitudes Towards Face Mask Use Scale (ATFMUS) in three phases: item generation, scale development, and scale evaluation. Phase one and two were addressed in study 1 while phase three was addressed in studies 2 and 3. In Study 1, a combined online and pen-and-paper sample of 174 (78% university students) completed a questionnaire with 19 items regarding attitudes towards face mask use derived from theory, previous research, and experience. Responses were subjected to item reduction analysis, exploratory factor analysis and reliability analysis. In Study 2, a student sample of 674 (70.5% high school) completed the new scale together with measures of COVID-19 related anxiety and obsession, personality, affect, social media use, and social desirability. Data from the ATFMUS were analyzed using confirmatory factor analysis and pertinent revisions done. The ATFMUS was then validated using correlation analyses, measurement invariance analyses, and known-group comparisons. In study 3, two samples of university students from Ghana (n = 242) and Kenya (n = 199) were involved in testing the cross-country invariance of the ATFMUS. The results reveal that the 5-item ATFMUS is a reliable and valid scale for assessing attitudes towards face mask use. Invariance analysis revealed that the ATFMUS is fair to use across participants of different age, level of education, and countries. The scale is also sensitive to participants' actual use of face masks as well as their beliefs about COVID-19 and efficacy of the facemasks. This study offers a foundation for further psychometric evaluation of the ATFMUS.

Evaluation of early-generation tropical maize testcrosses for grain-yield potential and weevil (Sitophilus zeamais Motschulsky) resistance.

Smallholder maize farmers in Africa experience pre- and post-harvest production stresses either individually or in combination at different stages of the crop cycle. The maize weevil is among the major post-harvest storage pests. A strategy to address this problem is to develop and promote high yielding maize germplasm with resistance to multiple stresses. A study was conducted to: 1) assess yield and agronomic performance of testcross hybrids developed from early generation lines; and 2) assess the response of the testcross hybrids to infestation with Sitophilus zeamais. Fifty-eight drought-tolerant testcross hybrids were evaluated for agronomic performance and weevil resistance at four environments in Uganda in 2016. Hybrid G39 (L2/T2) had the best grain yield performance; it significantly out-performed the best check by 11.4% in all environments. Hybrid grain from field trials was subjected to Sitophilus zeamais infestation in a choice and no choice test under laboratory conditions. Hybrids G56 (L49/T2) and G58 (L51/T2) had the least weevil damage and were rated as resistant to Sitophilus zeamais. The numbers of damaged kernels, number of exit holes and ear aspect were positively correlated with the grain weight loss. The results suggest possibilities for simultaneous selection for high grain yield and storage insect pest resistance among drought-tolerant genotypes. Use of high-yielding and resistant maize hybrids to storage insect pest should be promoted for increased maize production and managing post-harvest losses due to the maize weevil in smallholder farming communities in Africa.

On-farm performance and farmers' participatory assessment of new stress-tolerant maize hybrids in Eastern Africa.

The development and deployment of high-yielding stress tolerant maize hybrids are important components of the efforts to increase maize productivity in eastern Africa. This study was conducted to: i) evaluate selected, stress-tolerant maize hybrids under farmers' conditions; ii) identify farmers' selection criteria in selecting maize hybrids; and iii) have farmers evaluate the new varieties according to those criteria. Two sets of trials, one with 12 early-to-intermediate maturing and the other with 13 intermediate-to-late maturing hybrids, improved for tolerance to multiple stresses common in farmers' fields in eastern Africa (drought, northern corn leaf blight, gray leaf spot, common rust, maize streak virus), were evaluated on-farm under smallholder farmers' conditions in a total of 42 and 40 environments (site-year-management combinations), respectively, across Kenya, Uganda, Tanzania and Rwanda in 2016 and 2017. Farmer-participatory variety evaluation was conducted at 27 sites in Kenya and Rwanda, with a total of 2025 participating farmers. Differential performance of the hybrids was observed under low-yielding (<3 t ha-1) and high-yielding (>3 t ha-1) environments. The new stress-tolerant maize hybrids had a much better grain-yield performance than the best commercial checks under smallholder farmer growing environments but had a comparable grain-yield performance under optimal conditions. These hybrids also showed better grain-yield stability across the testing environments, providing an evidence for the success of the maize-breeding approach. In addition, the new stress- tolerant varieties outperformed the internal genetic checks, indicating genetic gain under farmers' conditions. Farmers gave high importance to grain yield in both farmer-stated preferences (through scores) and farmer-revealed preferences of criteria (revealed by regressing the overall scores on the scores for the individual criteria). The top-yielding hybrids in both maturity groups also received the farmers' highest overall scores. Farmers ranked yield, early maturity, cob size and number of cobs as the most important traits for variety preference. The criteria for the different hybrids did not differ between men and women farmers. Farmers gave priority to many different traits in addition to grain yield, but this may not be applicable across all maize-growing regions. Farmer-stated importance of the different criteria, however, were quite different from farmer- revealed importance. Further, there were significant differences between men and women in the revealed-importance of the criteria. We conclude that incorporating farmers' selection criteria in the stage-gate advancement process of new hybrids by the breeders is useful under the changing maize-growing environments in sub-Saharan Africa, and recommended to increase the turnover of new maize hybrids.

Empirical Comparison of Tropical Maize Hybrids Selected Through Genomic and Phenotypic Selections.

Genomic selection predicts the genomic estimated breeding values (GEBVs) of individuals not previously phenotyped. Several studies have investigated the accuracy of genomic predictions in maize but there is little empirical evidence on the practical performance of lines selected based on phenotype in comparison with those selected solely on GEBVs in advanced testcross yield trials. The main objectives of this study were to (1) empirically compare the performance of tropical maize hybrids selected through phenotypic selection (PS) and genomic selection (GS) under well-watered (WW) and managed drought stress (WS) conditions in Kenya, and (2) compare the cost-benefit analysis of GS and PS. For this study, we used two experimental maize data sets (stage I and stage II yield trials). The stage I data set consisted of 1492 doubled haploid (DH) lines genotyped with rAmpSeq SNPs. A subset of these lines (855) representing various DH populations within the stage I cohort was crossed with an individual single-cross tester chosen to complement each population. These testcross hybrids were evaluated in replicated trials under WW and WS conditions for grain yield and other agronomic traits, while the remaining 637 DH lines were predicted using the 855 lines as a training set. The second data set (stage II) consists of 348 DH lines from the first data set. Among these 348 best DH lines, 172 lines selected were solely based on GEBVs, and 176 lines were selected based on phenotypic performance. Each of the 348 DH lines were crossed with three common testers from complementary heterotic groups, and the resulting 1042 testcross hybrids and six commercial checks were evaluated in four to five WW locations and one WS condition in Kenya. For stage I trials, the cross-validated prediction accuracy for grain yield was 0.67 and 0.65 under WW and WS conditions, respectively. We found similar responses to selection using PS and GS for grain yield other agronomic traits under WW and WS conditions. The top 15% of hybrids advanced through GS and PS gave 21%-23% higher grain yield under WW and 51%-52% more grain yield under WS than the mean of the checks. The GS reduced the cost by 32% over the PS with similar selection gains. We concluded that the use of GS for yield under WW and WS conditions in maize can produce selection candidates with similar performance as those generated from conventional PS, but at a lower cost, and therefore, should be incorporated into maize breeding pipelines to increase breeding program efficiency.

Genetic architecture of maize chlorotic mottle virus and maize lethal necrosis through GWAS, linkage analysis and genomic prediction in tropical maize germplasm.

KEY MESSAGE: Analysis of the genetic architecture of MCMV and MLN resistance in maize doubled-haploid populations revealed QTLs with major effects on chromosomes 3 and 6 that were consistent across genetic backgrounds and environments. Two major-effect QTLs, qMCMV3-108/qMLN3-108 and qMCMV6-17/qMLN6-17, were identified as conferring resistance to both MCMV and MLN. Maize lethal necrosis (MLN) is a serious threat to the food security of maize-growing smallholders in sub-Saharan Africa. The ability of the maize chlorotic mottle virus (MCMV) to interact with other members of the Potyviridae causes severe yield losses in the form of MLN. The objective of the present study was to gain insights and validate the genetic architecture of resistance to MCMV and MLN in maize. We applied linkage mapping to three doubled-haploid populations and a genome-wide association study (GWAS) on 380 diverse maize lines. For all the populations, phenotypic variation for MCMV and MLN was significant, and heritability was moderate to high. Linkage mapping revealed 13 quantitative trait loci (QTLs) for MCMV resistance and 12 QTLs conferring MLN resistance. One major-effect QTL, qMCMV3-108/qMLN3-108, was consistent across populations for both MCMV and MLN resistance. Joint linkage association mapping (JLAM) revealed 18 and 21 main-effect QTLs for MCMV and MLN resistance, respectively. Another major-effect QTL, qMCMV6-17/qMLN6-17, was detected for both MCMV and MLN resistance. The GWAS revealed a total of 54 SNPs (MCMV-13 and MLN-41) significantly associated (P ≤ 5.60 × 10-05) with MCMV and MLN resistance. Most of the GWAS-identified SNPs were within or adjacent to the QTLs detected through linkage mapping. The prediction accuracy for within populations as well as the combined populations is promising; however, the accuracy was low across populations. Overall, MCMV resistance is controlled by a few major and many minor-effect loci and seems more complex than the genetic architecture for MLN resistance.

Genetic Analysis of QTL for Resistance to Maize Lethal Necrosis in Multiple Mapping Populations.

Maize lethal necrosis (MLN) occurs when maize chlorotic mottle virus (MCMV) and sugarcane mosaic virus (SCMV) co-infect maize plant. Yield loss of up to 100% can be experienced under severe infections. Identification and validation of genomic regions and their flanking markers can facilitate marker assisted breeding for resistance to MLN. To understand the status of previously identified quantitative trait loci (QTL)in diverse genetic background, F3 progenies derived from seven bi-parental populations were genotyped using 500 selected kompetitive allele specific PCR (KASP) SNPs. The F3 progenies were evaluated under artificial MLN inoculation for three seasons. Phenotypic analyses revealed significant variability (P ≤ 0.01) among genotypes for responses to MLN infections, with high heritability estimates (0.62 to 0.82) for MLN disease severity and AUDPC values. Linkage mapping and joint linkage association mapping revealed at least seven major QTL (qMLN3_130 and qMLN3_142, qMLN5_190 and qMLN5_202, qMLN6_85 and qMLN6_157qMLN8_10 and qMLN9_142) spread across the 7-biparetal populations, for resistance to MLN infections and were consistent with those reported previously. The seven QTL appeared to be stable across genetic backgrounds and across environments. Therefore, these QTL could be useful for marker assisted breeding for resistance to MLN.

Maize Combined Insect Resistance Genomic Regions and Their Co-localization With Cell Wall Constituents Revealed by Tissue-Specific QTL Meta-Analyses.

Combinatorial insect attacks on maize leaves, stems, and kernels cause significant yield losses and mycotoxin contaminations. Several small effect quantitative trait loci (QTL) control maize resistance to stem borers and storage pests and are correlated with secondary metabolites. However, efficient use of QTL in molecular breeding requires a synthesis of the available resistance information. In this study, separate meta-analyses of QTL of maize response to stem borers and storage pests feeding on leaves, stems, and kernels along with maize cell wall constituents discovered in these tissues generated 24 leaf (LIR), 42 stem (SIR), and 20 kernel (KIR) insect resistance meta-QTL (MQTL) of a diverse genetic and geographical background. Most of these MQTL involved resistance to several insect species, therefore, generating a significant interest for multiple-insect resistance breeding. Some of the LIR MQTL such as LIR4, 17, and 22 involve resistance to European corn borer, sugarcane borer, and southwestern corn borer. Eleven out of the 42 SIR MQTL related to resistance to European corn borer and Mediterranean corn borer. There KIR MQTL, KIR3, 15, and 16 combined resistance to kernel damage by the maize weevil and the Mediterranean corn borer and could be used in breeding to reduce insect-related post-harvest grain yield loss and field to storage mycotoxin contamination. This meta-analysis corroborates the significant role played by cell wall constituents in maize resistance to insect since the majority of the MQTL contain QTL for members of the hydroxycinnamates group such as p-coumaric acid, ferulic acid, and other diferulates and derivates, and fiber components such as acid detergent fiber, neutral detergent fiber, and lignin. Stem insect resistance MQTL display several co-localization between fiber and hydroxycinnamate components corroborating the hypothesis of cross-linking between these components that provide mechanical resistance to insect attacks. Our results highlight the existence of combined-insect resistance genomic regions in maize and set the basis of multiple-pests resistance breeding.

Discovery and validation of genomic regions associated with resistance to maize lethal necrosis in four biparental populations.

In sub-Saharan Africa, maize is the key determinant of food security for smallholder farmers. The sudden outbreak of maize lethal necrosis (MLN) disease is seriously threatening the maize production in the region. Understanding the genetic basis of MLN resistance is crucial. In this study, we used four biparental populations applied linkage mapping and joint linkage mapping approaches to identify and validate the MLN resistance-associated genomic regions. All populations were genotyped with low to high density markers and phenotyped in multiple environments against MLN under artificial inoculation. Phenotypic variation for MLN resistance was significant and heritability was moderate to high in all four populations for both early and late stages of disease infection. Linkage mapping revealed three major quantitative trait loci (QTL) on chromosomes 3, 6, and 9 that were consistently detected in at least two of the four populations. Phenotypic variance explained by a single QTL in each population ranged from 3.9% in population 1 to 43.8% in population 2. Joint linkage association mapping across three populations with three biometric models together revealed 16 and 10 main effect QTL for MLN-early and MLN-late, respectively. The QTL identified on chromosomes 3, 5, 6, and 9 were consistent with the QTL identified by linkage mapping. Ridge regression best linear unbiased prediction with five-fold cross-validation revealed high accuracy for prediction across populations for both MLN-early and MLN-late. Overall, the study discovered and validated the presence of major effect QTL on chromosomes 3, 6, and 9 which can be potential candidates for marker-assisted breeding to improve the MLN resistance.

Grain-yield stability among tropical maize hybrids derived from doubled-haploid inbred lines under random drought stress and optimum moisture conditions.

Drought is a devastating environmental stress in agriculture and hence a common target of plant breeding. A review of breeding progress on drought tolerance shows that, to a certain extent, selection for high yield in stress-free conditions indirectly improves yield in water-limiting conditions. The objectives of this study were to (i) assess the genotype × environment (GE) interaction for grain yield (GY) and other agronomic traits for maize (Zea mays L.) across East African agro-ecologies; and (ii) evaluate agronomic performance and stability in Uganda and Tanzania under optimum and random drought conditions. Data were recorded for major agronomic traits. Genotype main effect plus GE (GGE) biplot analysis was used to assess the stability of varieties within various environments and across environments. Combined analysis of variance across optimum moisture and random drought environments indicated that locations, mean-squares for genotypes and GE were significant for most measured traits. The best hybrids, CKDHH1097 and CKDHH1090, gave GY advantages of 23% and 43%, respectively, over the commercial hybrid varieties under both optimum-moisture and random-drought conditions. Across environments, genotypic variance was less than the GE variance for GY. The hybrids derived from doubled-haploid inbred lines produced higher GY and possessed acceptable agronomic traits compared with the commercial hybrids. Hybrid CKDHH1098 ranked second-best under optimum-moisture and drought-stress environments and was the most stable with broad adaptation to both environments. Use of the best doubled-haploids lines in testcross hybrids make-up, well targeted to the production environments, could boost maize production among farmers in East Africa.

Combining ability and testcross performance of drought-tolerant maize inbred lines under stress and non-stress environments in Kenya.

Drought and poor soil fertility are among the major abiotic stresses affecting maize productivity in sub-Saharan Africa. Maize breeding efforts at the International Maize and Wheat Improvement Center (CIMMYT) have focused on incorporating drought stress tolerance and nitrogen-use efficiency (NUE) into tropical maize germplasm. The objectives of this study were to estimate the general combining ability (GCA) and specific combining ability (SCA) of selected maize inbred lines under drought stress (DS), low-nitrogen (LN) and optimum moisture and nitrogen (optimum) conditions, and to assess the yield potential and stability of experimental hybrids under these management conditions. Forty-nine experimental three-way cross hybrids, generated from a 7 × 7 line by tester crosses, and six commercial checks were evaluated across 11 optimum, DS and LN sites in Kenya in 2014 using an alpha lattice design with two replicates per entry at each site. DS reduced both grain yield (GY) and plant height (PH), while anthesis-silking interval (ASI) increased under both DS and LN. Hybrids 'L4/T2' and 'L4/T1' were found to be superior and stable, while inbreds 'L4' and 'L6' were good combiners for GY and other secondary traits across sites. Additive variance played a greater role for most traits under the three management conditions, suggesting that further progress in the improvement of these traits should be possible. GY under optimum conditions was positively correlated with GY under both DS and LN conditions, but GY under DS and LN was not correlated. Our results suggest the feasibility for simultaneous improvement in grain yield performance of genotypes under optimum, DS and LN conditions.

Genetic analysis of tropical maize inbred lines for resistance to maize lethal necrosis disease.

Maize lethal necrosis (MLN) disease is a recent outbreak in eastern Africa and has emerged as a significant threat to maize production in the region. The disease is caused by the co-infection of Maize chlorotic mottle virus and any member of potyviridae family. A total of 28 maize inbred lines with varying levels of tolerance to MLN were crossed in a half-diallel mating design, and the resulting 340 F1 crosses and four commercial checks were evaluated under MLN artificial inoculation at Naivasha, Kenya in 2015 and 2016 using an alpha lattice design with two replications. The objectives of the study were to (i) investigate the magnitude of general combining ability variance (σGCA 2) and specific combining ability variance (σSCA 2) and their interaction with years; (ii) evaluate the efficiencies of GCA based prediction and hybrid performance by means of a cross-validation procedure; (iii) estimate trait correlations in the hybrids; and (iv) identify the MLN tolerant single cross hybrids to be used as female parents for three-way cross hybrids. Results of the combined analysis of variance revealed that both GCA and SCA effects were significant (P < 0.05) for all traits except for ear rot. For MLN scores at early and late stages, GCA effects were 2.5-3.5 times higher than SCA effects indicating that additive gene action is more important than non-additive gene action. The GCA based prediction efficiency for MLN resistance and grain yield accounted for 67-90% of the variations in the hybrid performance suggesting that GCA-based prediction can be proposed to predict MLN resistance and grain yield prior to field evaluation. Three parents, CKDHL120918, CML550, and CKLTI0227 with significant GCA effects for GY (0.61-1.21; P < 0.05) were the most resistant to MLN. Hybrids "CKLTI0227 × CML550", "CKDHL120918 × CKLTI0138", and "CKDHL120918 × CKLTI0136" ranked among the best performing hybrids with grain yield of 6.0-6.6 t/ha compared with mean yield of commercial check hybrids (0.6 t/ha). The MLN tolerant inbred lines and single cross hybrids identified in this study could be used to improve MLN tolerance in both public and private sector maize breeding programs in eastern Africa.

Resistance of Bt-maize (MON810) against the stem borers Busseola fusca (Fuller) and Chilo partellus (Swinhoe) and its yield performance in Kenya.

A study was conducted to assess the performance of maize hybrids with Bt event MON810 (Bt-hybrids) against the maize stem borer Busseola fusca (Fuller) in a biosafety greenhouse (BGH) and against the spotted stem borer Chilo partellus (Swinhoe) under confined field trials (CFT) in Kenya for three seasons during 2013-2014. The study comprised 14 non-commercialized hybrids (seven pairs of near-isogenic Bt and non-Bt hybrids) and four non-Bt commercial hybrids. Each plant was artificially infested twice with 10 first instar larvae. In CFT, plants were infested with C. partellus 14 and 24 days after planting; in BGH, plants were infested with B. fusca 21 and 31 days after planting. In CFT, the seven Bt hybrids significantly differed from their non-Bt counterparts for leaf damage, number of exit holes, percent tunnel length, and grain yield. When averaged over three seasons, Bt-hybrids gave the highest grain yield (9.7 t ha-1), followed by non-Bt hybrids (6.9 t ha-1) and commercial checks (6 t ha-1). Bt-hybrids had the least number of exit holes and percent tunnel length in all the seasons as compared to the non-Bt hybrids and commercial checks. In BGH trials, Bt-hybrids consistently suffered less leaf damage than their non-Bt near isolines. The study demonstrated that MON810 was effective in controlling B. fusca and C. partellus. Bt-maize, therefore, has great potential to reduce the risk of maize grain losses in Africa due to stem borers, and will enable the smallholder farmers to produce high-quality grain with increased yield, reduced insecticide inputs, and improved food security.

Performance and grain yield stability of maize populations developed using marker-assisted recurrent selection and pedigree selection procedures.

A marker-assisted recurrent selection (MARS) program was undertaken in sub-Saharan Africa to improve grain yield under drought-stress in 10 biparental tropical maize populations. The objectives of the present study were to evaluate the performance of C1S2-derived hybrids obtained after three MARS cycles (one cycle of recombination (C1), followed by two generations of selfing (S2), and to study yield stability under both drought-stress (DS) and well-watered (WW) conditions. For each of the 10 populations, we evaluated hybrids developed by crossing 47-74 C1S2 lines advanced through MARS, the best five S5 lines developed through pedigree selection, and the founder parents with a single-cross tester from a complementary heterotic group. The hybrids and five commercial checks were evaluated in Kenya under 1-3 DS and 3-5 WW conditions with two replications. Combined across DS locations, the top 10 C1S2-derived hybrids from each of the 10 biparental populations produced 0.5-46.3 and 11.1-55.1 % higher mean grain yields than hybrids developed using pedigree selection and the commercial checks, respectively. Across WW locations, the best 10 hybrids derived from C1S2 of each population produced 3.4-13.3 and 7.9-36.5 % higher grain yields than hybrids derived using conventional pedigree breeding and the commercial checks, respectively. Mean days to anthesis of the best 10 C1S2 hybrids were comparable to those of hybrids developed using the pedigree method, the founder parents and the commercial checks, with a maximum difference of 3.5 days among the different groups. However, plant height was significantly (P < 0.01) different in most pairwise comparisons. Our results showed the superiority of MARS over pedigree selection for improving diverse tropical maize populations as sources of improved lines for stress-prone environments and thus MARS can be effectively integrated into mainstream maize breeding programs.

Meta-analyses of QTL for grain yield and anthesis silking interval in 18 maize populations evaluated under water-stressed and well-watered environments.

BACKGROUND: Identification of QTL with large phenotypic effects conserved across genetic backgrounds and environments is one of the prerequisites for crop improvement using marker assisted selection (MAS). The objectives of this study were to identify meta-QTL (mQTL) for grain yield (GY) and anthesis silking interval (ASI) across 18 bi-parental maize populations evaluated in the same conditions across 2-4 managed water stressed and 3-4 well watered environments. RESULTS: The meta-analyses identified 68 mQTL (9 QTL specific to ASI, 15 specific to GY, and 44 for both GY and ASI). Mean phenotypic variance explained by each mQTL varied from 1.2 to 13.1% and the overall average was 6.5%. Few QTL were detected under both environmental treatments and/or multiple (>4 populations) genetic backgrounds. The number and 95% genetic and physical confidence intervals of the mQTL were highly reduced compared to the QTL identified in the original studies. Each physical interval of the mQTL consisted of 5 to 926 candidate genes. CONCLUSIONS: Meta-analyses reduced the number of QTL by 68% and narrowed the confidence intervals up to 12-fold. At least the 4 mQTL (mQTL2.2, mQTL6.1, mQTL7.5 and mQTL9.2) associated with GY under both water-stressed and well-watered environments and detected up to 6 populations may be considered for fine mapping and validation to confirm effects in different genetic backgrounds and pyramid them into new drought resistant breeding lines. This is the first extensive report on meta-analysis of data from over 3100 individuals genotyped using the same SNP platform and evaluated in the same conditions across a wide range of managed water-stressed and well-watered environments.

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.