Germplasm Enhancement Of Maize Project Research Articles

Performance and phenotypic stability of maize hybrids containing exotic introgressions in multi‐environment trials

AbstractMaize (Zea mays L.) grown in the US Midwest contains only a small fraction of the genetic diversity present in the species. Maize populations from other parts of the world may contain genetic variation that could be used to improve or increase the diversity of US germplasm. This study was conducted to assess the performance and phenotypic stability of hybrids containing diverse exotic introgressions across North American environments. Doubled haploid (DH) lines were created by the Germplasm Enhancement of Maize project from backcross 1 families that used 27 open‐pollinated populations from Latin America as donor parents and PHZ51, a non‐Stiff Stalk inbred developed in Iowa, as the recurrent parent. DH lines were testcrossed with LH195, a Stiff Stalk inbred developed in Iowa, and hybrid field trials were performed at 24 environments in the United States and Canada. Experimental hybrids had variable flowering time, plant and ear height, and test weight, but none had significantly greater yield than the PHZ51 × LH195 hybrid. The slopes of linear reaction norm models were significantly lower for the experimental hybrids than the US‐adapted reference hybrids for three traits: yield, growing degree units (GDU) to anthesis, and GDU to silking. The results suggest that unfavorable alleles and phenotypic stability should be considered when exotic open‐pollinated populations are used in US maize breeding.

Genomic prediction for the Germplasm Enhancement of Maize project.

The Germplasm Enhancement of Maize (GEM) project was initiated in 1993 as a cooperative effort of public- and private-sector maize (Zea mays L.) breeders to enhance the genetic diversity of the U.S. maize crop. The GEM project selects progeny lines with high topcross yield potential from crosses between elite temperate lines and exotic parents. The GEM project has released hundreds of useful breeding lines based on phenotypic selection within selfing generations and multienvironment yield evaluations of GEM line topcrosses to elite adapted testers. Developing genomic selection (GS) models for the GEM project may contribute to increases in the rate of genetic gain. Here we evaluated the prediction ability of GS models trained on 6 yr of topcross evaluations from the two GEM programs in Raleigh, NC, and Ames, IA, documenting prediction abilities ranging from 0.36 to 0.75 for grain yield and from 0.78 to 0.96 for grain moisture when models were cross-validated within program and heterotic group. Predicted genetic gain from GS ranged from 0.95 to 2.58 times the gain from phenotypic selection. Prediction ability across program and heterotic group was generally poorer than within groups. Based on observed genomic relationships between GEM breeding lines and their tropical ancestors, GS for either yield or moisture would reduce recovery of exotic germplasm only slightly. Using GS models trained within program, the GEM programs should be able to more effectively deliver on its mission to broaden the genetic base of U.S. germplasm.

Association Mapping of Flowering and Height Traits in Germplasm Enhancement of Maize Doubled Haploid (GEM-DH) Lines.

Flowering and height related traits are extensively studied in maize for three main reasons: 1) easily obtained phenotypic measurements, 2) highly heritable, and 3) importance of these traits to adaptation and grain yield. However, variation in flowering and height traits is extensive and findings from previous studies are genotype specific. Herein, a diverse panel of exotic derived doubled haploid lines, in conjunction with genome-wide association analysis, is used to further explore adaptation related trait variation of exotic germplasm for potential use in adapting exotic germplasm to the U.S. Corn-Belt. Phenotypes for the association panel were obtained from six locations across the central-U.S. and genotyping was performed using the genotyping-by-sequencing method. Nineteen flowering time candidate genes were found for three flowering traits. Eighteen candidate genes were found for four height related traits, with the majority of the candidate genes relating to plant hormones auxin and gibberellin. A single gene was discovered for ear height that also had effects on -like flowering gene expression levels. Findings will be used to inform future research efforts of the USDA Germplasm Enhancement of Maize project and eventually aid in the rapid adaptation of exotic germplasm to temperate U.S. environments.

Extensive genetic diversity and low linkage disequilibrium within the COMT locus in maize exotic populations

The caffeic acid 3-O-methytransferase (COMT) gene is a prime candidate for cell wall digestibility improvement based on the characterization of brown midrib-3 mutants. We compared the genetic diversity and linkage disequilibrium at this locus between exotic populations sampled within the Germplasm Enhancement of Maize (GEM) project and 70 inbred lines. In total, we investigated 55 exotic COMT alleles and discovered more than 400 polymorphisms in a 2.2kb region with pairwise nucleotide diversity (π) up to 0.017, much higher than reported π values of various genes in inbred lines. The ratio of non-synonymous to synonymous SNPs was 3:1 in exotic populations, and significantly higher than the 1:1 ratio for inbred lines. Selection tests detected selection signature in this gene in both pools, but with different evolution patterns. The linkage disequilibrium decay in exotic populations was at least four times more rapid than for inbred lines with r2>0.1 persisting only up to 100bp. In conclusion, the alleles sampled in the GEM Project offer a valuable genetic resource to broaden genetic variation for the COMT gene, and likely other genes, in inbred background. Moreover, the low linkage disequilibrium makes this material suitable for high resolution association analyses.

Diallel Analysis of Diverse Maize Germplasm Lines for Resistance to Aflatoxin Accumulation

ABSTRACTIn this study, diverse, novel maize (Zea mays L.) germplasm from the Germplasm Enhancement of Maize (GEM) Project was screened to determine the combining ability and inheritance of ear rot resistance and resistance to aflatoxin accumulation in hybrid combinations. Nine GEM germplasm lines and six known resistant and susceptible inbred lines were selected as parents of a diallel cross that was evaluated over 2 yr at Mississippi State, MS, and 1 yr at College Station, TX. Resistant lines Mp717 and Mp494 each had negative and significant general combining ability effects, and were frequently present as parents in the hybrid combinations with the lowest aflatoxin accumulation. Of the GEM germplasm lines evaluated, GEMN‐0130 was the best source of aflatoxin resistance, with significant and negative GCA values at all three site years. The GEM lines exhibiting significant and negative GCA effects for aflatoxin accumulation in at least one site year include two GEM germplasm lines, GEMS‐0030 (2 yr) and GEMS‐0074 (1 yr), and a nonreleased GEM source derived from the tropical Cuban accession, CUBA117 (1 yr). The GEM germplasm lines significantly contributing to reduced ear rot in at least two out of three environments included CUBA117, GEMS‐0030, GEMS‐0002, and GEMN‐0130. These results demonstrate the GEM Project is a useful and untapped source of diverse germplasm for resistance to Aspergillus flavus ear rot and aflatoxin accumulation.

Genotypic and phenotypic characterization of isogenic doubled haploid exotic introgression lines in maize

We characterized the genotypic and phenotypic variation in cell wall digestibility (CWD) and other agronomic traits of 50 backcross 1 generation doubled haploid (BC1DH) lines developed from the Germplasm Enhancement of Maize project. These lines were generated by introgressing 31 exotic unadapted maize races into PHZ51 and PHB47, temperate inbred lines with expired Plant Variety Protection. The 50 BC1DH lines and five check lines were genotyped with 199 single nucleotide polymorphism markers distributed across the genome. We identified, on average, 11.8% of markers with exotic donor parent alleles. This likely underestimates the proportion of donor introgressions, since we cannot discriminate monomorphic alleles from donor and recurrent parents. The potential roles of natural selection and the doubled haploid process in favouring selection of the recurrent genome are discussed. Although the proportion of donor parent genome was underestimated, donor fragments evaluated across the 50 BC1DH lines covered 92.9% of the recurrent parent genome. The evaluation of BC1DH lines for CWD revealed promising lines with CWD not differing significantly (α = 0.05) from forage quality lines used as checks. The introgression of exotic genome segments, however, was generally associated with higher ears, lodging, and late flowering. Even with limited power, our association analysis revealed quantitative trait polymorphisms associated with CWD, flowering date, and lodging.

Wet‐Milling Characteristics of 10 Lines from Germplasm Enhancement of Maize Project Compared with Five Corn Belt Lines

ABSTRACTThe use of corn (Zea mays L.) hybrids with high grain yield and starch extractability has steadily increased in the processing industry. In light of widespread corn seed industry participation in the Germplasm Enhancement of Maize Project (GEM), which seeks to enhance exotic germplasm, future hybrids may contain more exotic sources in genetic backgrounds. It is necessary to establish and monitor physical, compositional, and milling characteristics of the new exotic breeding materials to determine the processing value. The present study was conducted to determine the wet‐milling characteristics of a set of GEM lines compared with typical Corn Belt lines. Ten GEM lines introgressed with exotic materials from Argentina, Chile, Cuba, Florida, and Uruguay and previously identified as having different starch yields, three commercial inbred lines, and two public inbred lines (B73 and Mo17) were analyzed using both near‐infrared transmittance (NIT) and a 100‐g wet‐milling procedure. There were statistical differences (P < 0.05) in the yield of wet‐milled fractions (starch, fiber, gluten, and germ). The GEM lines AR16035:S19‐227‐1‐B and CUBA117:S1520‐562‐1‐B had similar or better starch yield and starch recovery than B73 and the other adapted inbred lines, indicating that they may be useful in improving the proportion of extractable starch present in kernels of hybrids. Residual protein levels in the starch and gluten fractions were 0.26–0.32% and 38–45%, respectively. The starch yield of GEM lines from wet milling correlated positively with starch content from NIT and was negatively correlated with protein content of the corn kernels. Oil content in the germ varied from 50 to 60%. Our results indicate that incorporating GEM lines in a breeding program can maintain or even improve wet‐milling characteristics of Corn Belt materials if lines with appropriate traits are used.

Evaluation of the grain methionine, lysine and tryptophan contents of maize ( Zea mays L.) germplasm in the Germplasm Enhancement of Maize Project

The Germplasm Enhancement of Maize (GEM) Project is a cooperative effort between the USDA-ARS, private industry and public researchers to broaden and enhance the germplasm base of maize. In this program, selected accessions from the Latin American Maize Project, and seven tropical hybrids donated by DeKalb to the GEM Project, were crossed to elite proprietary inbred lines contributed by commercial plant breeding programs. In most cases, the resulting hybrids were crossed to a second commercial inbred line and the resulting 25% exotic hybrids were used as breeding populations for further development. To identify GEM germplasm with value to protein quality breeding programs, we developed a process for evaluating the content of the essential amino acids methionine, lysine and tryptophan in the grain of GEM germplasm that balances the need for multiple-year evaluations with the constantly changing entry list of this germplasm screening program. This process involves annual field trials with common checks. Weak entries are dropped from the trial each year to make room for new entries, while strong entries are retained. Methionine exhibited the most significant variation, followed by lysine and then tryptophan. A number of GEM lines had methionine or lysine levels that were significantly better than Corn Belt checks and some were competitive with high-amino acid checks.

Registration of 20 GEM Maize Breeding Germplasm Lines Adapted to the Southern USA

Twenty maize breeding germplasm lines were developed cooperatively by the USDA GEM (Germplasm Enhancement of Maize) project (Reg. no. GP-407 to GP-426, PI 639037 to PI 639056). These lines were developed by selfing and selecting variable F1s from variable source × US inbred crosses in North Carolina under standard nursery conditions, followed by a second selfing-selection season in Homestead, Florida, and a third selfing-selection season in a selection nursery in Raleigh (F2S2). The germplasm lines were selected on the basis of resistance to Fusarium ear rot (Gibberella moniliformis and Fusarium proliferatum) and anthracnose (Colletotrichum graminicola), resistance to lodging, early flowering, synchrony of silk and pollen production, and reduced plant and ear height. In trials conducted in 2001 and 2002, the germplasm lines recorded grain yields ranging from 11197 to 13596 kg/ha (compared with 11009 kg/ha for the control) and grain moisture content ranging from 185 to 212 g/kg (compared with 190 g/kg for the control).

Thermal Properties and Paste and Gel Behaviors of Starches Recovered from Accessions Used in the Germplasm Enhancement of Maize Project

ABSTRACTThe objective of this study was to screen starches recovered from the corn accessions used in the Germplasm Enhancement of Maize (GEM) project for unusual thermal properties and paste and gel behaviors, so that they could be exploited in corn breeding programs to enhance traits important to corn utilization. In general, the values for gelatinization temperatures and peak height indices were greater, but heat of gelatinization values were less for the starches recovered from the GEM accessions (particularly BRA 052051 (SE 32)) than for starches from commercial Dent corn hybrids (11.3 vs. 13.6 J/g). Generally, retrogradation properties were similar among the GEM accessions, although there were specific accessions (particularly Lima 13) that possessed modestly lower percentage retrogradation (34 vs. 42%). Generally, peak viscosities, cold paste viscosities, and viscosity breakdowns were greater for the starches of the GEM accessions (particularly ARZM 01150, Antiqua 3, and URZM 01089, respectively) than for the starches of commercial hybrids. Pasting temperatures were about the same for all starches. Both 1‐day and 7‐day gel strengths were considerably greater for the starches recovered from the GEM accessions (particularly BRA 052051 (SE 32), 21.0 g for 1‐day and FS8A(T), 66.2 g for 7‐day). Although the differences in starch properties were statistically different, only the higher gel strengths of the starches recovered from the GEM accessions were of practical significance to the starch industry.

Compositional, Physical, and Wet‐Milling Properties of Accessions Used in Germplasm Enhancement of Maize Project

ABSTRACTForty‐nine accessions used in the Germplasm Enhancement of Maize (GEM) project, two commercial hybrids (Pioneer Brand Hybrids 3394 and 3489), and two Corn Belt inbreds (B73 and Mo17) were evaluated for compositional, physical, and wet‐milling properties. GEM accessions had lower starch contents (65.9–69.1% vs. a mean of 72.2% for the commercial hybrids) and greater protein contents (12.0–14.4% vs. a mean of 8.2% for the commercial hybrids) than did the improved Corn Belt material. Absolute densities were consistently higher for the GEM accessions compared with the commercial hybrids (1.320 vs. 1.265 g/cm3, respectively). The wet‐milling characteristics of the GEM accessions were not nearly as good as for the commercial hybrids. Mean starch yields were only 54.3% for the GEM accessions versus 64.8% for the commercial hybrids. Residual protein levels in the starches recovered from the GEM accessions were much greater (0.45–2.03%) than for commercial corn hybrids (<0.3%).

Identification of Tropical and Temperate Maize Populations Having Favorable Alleles for Disease Resistance

A possible use of nonelite germplasm is as a source of alleles for disease resistance. Our objective was to determine the value of tropical and temperate maize (Zea mays L.) germplasm accessions as sources of alleles to improve disease resistance of the Corn Belt hybrid FR1064 × LH185. A group of tropical populations and hybrids from the Germplasm Enhancement of Maize Project (GEM) crossed to either Mo17 or B73 (temperate inbreds), was studied. In addition, a group of temperate accessions was evaluated. Reaction to southern corn leaf blight (SCLB) [Bipolaris maydis (Nisikado & Miyake) Shoemaker = Helminthosporium maydis Nisikado & Miyake], northern corn leaf spot (NCLS) [Bipolaris zeicola (G. L. Stout) Shoemaker = Helminthosporium carbonum Ullstrup Races 2 and 3], gray leaf spot (GLS) (Cercospora zeae‐maydis Tehon & E.Y. Daniels), northern corn leaf blight (NCLB) [Exserohilum turcicum (Pass.) K. J. Leonard & E. G. Suggs = Helminthosporium turcicum Pass., Races 1, 1, 2, 23N and three unidentified races] and common rust (Puccinia sorghi Schwein) was studied using Dudley's method for identifying populations with favorable alleles. All accessions had favorable dominant alleles for resistance not present in FR1064 × LH185 for common rust, GLS, and SCLB. Four accessions had a number of favorable alleles for resistance not significantly different from the best accession for all five diseases. Crosses of the accessions containing Mo17 to FR1064 and LH185 were less susceptible to SCLB, NCLB, and rust than crosses of accessions containing B73. At least seven of the best 10 populations were tropical × Mo17 crosses for SCLB, NCLB, and rust when populations were ranked for presence of favorable alleles for resistance not present in either LH185 or FR1064. Net value statistics for tropical × B73 and tropical × Mo17 accessions differed in indicating whether backcrossing or selfing from the F1 is more desirable. Therefore, if tropical populations are crossed to Corn Belt inbreds to increase adaptation, comparisons among tropical populations should only be made when all populations being compared have been crossed to the same Corn Belt inbred.

Identification of Tropical and Temperate Maize Populations Having Favorable Alleles for Yield and Other Phenotypic Traits

If tropical germplasm is to be useful in increasing maize (Zea mays L.) grain yield, populations containing favorable alleles not present in elite hybrids need to be identified. Thus, a major objective of this study was to identify tropical and temperate populations containing favorable alleles useful for improving the Corn Belt hybrid FR1064 × LH185 (a commercial hybrid representative of the typical Stiff‐Stalk × Lancaster heterotic pattern). A group of tropical populations and hybrids from the Germplasm Enhancement of Maize Project (GEM) crossed to either Mo17 or B73 (temperate maize inbreds) was studied. In addition, a group of temperate accessions was evaluated. Grain yield, flowering date, plant height, ear height, and penetrometer reading were evaluated using Dudley's method for identifying populations with favorable alleles. Only three accessions of 41 had positive and significant relative numbers of favorable alleles for yield. For most accessions, unexpectedly, the estimate of l plμ′ (an estimator of relative number of favorable alleles in the accessions not present in the hybrid to be improved) for yield was negative. Negative l plμ′ values for yield resulted from low true values of l plμ′ combined with low frequencies of favorable alleles in the accession for loci having favorable alleles in one inbred and not the other. None of the accessions were identified as having favorable alleles for lowering ear height or increasing stalk penetrometer resistance. These results suggest Dudley's method may not be useful when frequencies of favorable alleles in the populations being evaluated are very low.