Unlocking the Genetic Diversity of Maize Landraces with Doubled Haploids Opens New Avenues for Breeding

Landraces are valuable genetic resources for broadening the genetic base of elite germplasm in maize. Extensive exploitation of landraces has been hampered by their genetic heterogeneity and heavy genetic load. These limitations may be overcome by the in-vivo doubled haploid (DH) technique. A set of 132 DH lines derived from three European landraces and 106 elite flint (EF) lines were genotyped for 56,110 single nucleotide polymorphism (SNP) markers and evaluated in field trials at five locations in Germany in 2010 for several agronomic traits. In addition, the landraces were compared with synthetic populations produced by intermating DH lines derived from the respective landrace. Our objectives were to (1) evaluate the phenotypic and molecular diversity captured within DH lines derived from European landraces, (2) assess the breeding potential (usefulness) of DH lines derived from landraces to broaden the genetic base of the EF germplasm, and (3) compare the performance of each landrace with the synthetic population produced from the respective DH lines. Large genotypic variances among DH lines derived from landraces allowed the identification of DH lines with grain yields comparable to those of EF lines. Selected DH lines may thus be introgressed into elite germplasm without impairing its yield level. Large genetic distances of the DH lines to the EF lines demonstrated the potential of DH lines derived from landraces to broaden the genetic base of the EF germplasm. The comparison of landraces with their respective synthetic population showed no yield improvement and no reduction of phenotypic diversity. Owing to the low population structure and rapid decrease of linkage disequilibrium within populations of DH lines derived from landraces, these would be an ideal tool for association mapping. Altogether, the DH technology opens new opportunities for characterizing and utilizing the genetic diversity present in gene bank accessions of maize.     

Variation of DNA fingerprints among accessions within maize inbred lines and implications for identification of essentially derived varieties.

Genetic distances (GDs) based on molecular markers are important parameters for identifying essentially derived varieties (EDVs). In this context information about the variability of molecular markers within maize inbred lines is essential. Our objectives were to (1) determine the variation in the size of simple sequence repeat (SSR) fragments among different accessions of maize inbreds and doubled haploid (DH) lines, (2) attribute the observed variation to genetic and marker system-specific sources, and (3) investigate the effect of SSR fragment size differences within maize lines on the GD between maize lines and their consequences for the identification of essentially derived varieties. Two to five accessions from nine inbred lines and five DH lines were taken from different sources or drawn as independent samples from the same seed lot. Each accession was genotyped with 100 SSR markers that evenly covered the whole maize genome. In total, 437 SSR fragments were identified, with a mean of 4.4 alleles per locus. The average polymorphic information content (PIC) was 0.58. GD estimates between two accessions of the same genotype ranged from 0.00 to 0.12 with an average of 0.029 for inbred lines and 0.001 for DH lines. An average of 11.1 SSRs was polymorphic between accessions of the same inbred line due to non-amplification (8.1 SSRs), heterogeneity (4.0 SSRs) or unknown alleles (2.6 SSRs). In contrast to lab errors, heterogeneity contributed considerably to the observed variation for GD. In order to decrease the probability to be suited for infringing an EDV threshold by chance, we recommend to increase the level of homogeneity of inbred lines before applying for plant variety protection.     

Re-evaluation of the prospects of marker-assisted selection for improving insect resistance against Diatraea spp. in tropical maize by cross validation and independent validation

Cross validation (CV) and validation with an independent sample (IV) are new biometric approaches in QTL analysis to obtain unbiased estimates of QTL effects and the proportion of the genetic variance explained by the detected marker-QTL association (p). Our objective with these methods was to obtain a realistic picture on the prospects of marker-assisted selection (MAS) for improving the resistance of maize against the tropical stem borer species Diatraea grandiosella (SWCB) and Diatraea saccharalis (SCB). Published QTL mapping studies on leaf-damage ratings (LDR) with populations of F_2:3 lines and recombinant inbred lines (RIL) from crosses CML131×CML67 and Ki3× CML139 of tropical maize inbreds were re-analyzed with CV and IV. With CV, the reduction in p for LDR compared to p obtained with the whole data set varied between 41.0 and 79.6% in the populations of F_2:3 lines and between 30.1 and 65.2% in the two populations of RIL. Estimates of p for SCB LDR were similar for CV and IV. For SWCB LDR, p estimates obtained with IV were larger than those obtained with CV in CML131× CML67. The reverse was observed for Ki3×CML139. Under the assumption of identical selection intensities, and based on the re-estimates of p, MAS using only molecular marker information is less-efficient than conventional phenotypic selection (CPS). MAS combining marker and phenotypic data increases the relative efficiency by only 4% in comparison to CPS. In conclusion, MAS for improving SWCB and SCB LDR seems not-promising unless additional QTLs with proven large effects are available or the costs of marker assays are considerably reduced. Received: 7 December 2000 / Accepted: 5 February 2001     

Chilling tolerance of Central European maize lines and their factorial crosses

BACKGROUND AND AIMS: Chilling-stress tolerance is a prerequisite for maize production under cool climatic conditions. The main goal of this study was to evaluate the Central European dent and flint pools for chilling tolerance during heterotrophic and early autotrophic growth in field trials and growth chamber experiments. METHODS: Five European flint and five dent inbreds and their 25 factorial crosses were evaluated in six natural environments, where chilling occurred, for chlorophyll concentration and plant height at the three-leaf stage, and plant height and fresh weight at the seven-leaf stage. In growth chambers, leaf 3 growth was analysed under cold and control conditions. KEY RESULTS: Comparing the field and growth chamber data, the strongest association was found between leaf elongation rate during cold nights and plant height at the three-leaf stage, with a weaker association with the seven-leaf stage. In the field, moderate correlations were observed between plant height at the three-leaf stage, and plant height and fresh weight at the seven-leaf stage, respectively. Furthermore, mid-parent and hybrid performance were only moderately correlated. CONCLUSIONS: The results suggest that heterotrophic and early autotrophic growth stages are controlled by different genetic factors or that maternal effects play a role. In addition, the findings showed that mid-parent performance is a poor predictor of hybrid performance. Consequently, test cross performance should be the target in quantitiative trait locus (QTL) mapping studies with the final goal of establishing marker-assisted breeding programmes for chilling-tolerant hybrids.     

Temporal changes in allele frequencies in two European F2 flint maize populations under modified recurrent full-sib selection

Selection and random genetic drift are the two main forces affecting the selection response of recurrent selection (RS) programs by changes in allele frequencies. Therefore, detailed knowledge on allele frequency changes attributable to these forces is of fundamental importance for assessing RS programs. The objectives of our study were to (1) estimate the number, position, and genetic effect of quantitative trait loci (QTL) for selection index and its components in the base populations, (2) determine changes in allele frequencies of QTL regions due to the effects of random genetic drift and selection, and (3) predict allele frequency changes by using QTL results and compare these predictions with observed values. We performed QTL analyses, based on restriction fragment length polymorphisms (RFLPs) and simple sequence repeats (SSRs), in 274 F_2:3 lines of cross KW1265 × D146 (A × B) and 133 F_3:4 lines of cross D145 × KW1292 (C × D) originating from two European flint maize populations. Four (A × B) and seven (C × D) cycles of RS were analyzed with SSRs for significant allele frequency changes due to selection. Several QTL regions for selection index were detected with simple and composite interval mapping. In some of them, flanking markers showed a significant allele frequency change after the first and the final selection cycles. The correlation between observed and predicted allele frequencies was significant only in A × B. We attribute these observations mainly to (1) the high dependence of the power of QTL detection on the population size and (2) the occurrence of undetectable QTL in repulsion phase. Assessment of allele frequency changes in RS programs can be used to detect marker alleles linked to QTL regions under selection pressure. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Potential causes of linkage disequilibrium in a European maize breeding program investigated with computer simulations

Knowledge about the forces generating and conserving linkage disequilibrium (LD) is important for drawing conclusions about the prospects and limitations of association mapping. The objectives of our research were to examine the importance of (1) selection, (2) mutation, and (3) genetic drift for generating LD in a typical maize breeding program. We conducted computer simulations based on genotypic data of Central European maize open-pollinated varieties which have played an important role as founders of the European flint heterotic group. The breeding scheme and the dimensioning underlying our simulations reflect essentially the maize breeding program of the University of Hohenheim. Results suggested that in a plant breeding program of the examined dimension and breeding scheme, genetic drift and selection are major forces generating LD. The currently used population-based association mapping tests do not explicitly correct for LD caused by these two forces. Therefore, increased type I error rates are expected if these tests are applied to plant breeding populations. As a consequence, we recommend to use family-based association tests for association mapping approaches in plant breeding populations.     

Impact of interpopulation divergence on additive and dominance variance in hybrid populations

We present a theoretical proof that the ratio of the dominance vs. the additive variance decreases with increasing genetic divergence between two populations. While the dominance variance is the major component of the variance due to specific combining ability (sigma(SCA)(2)), the additive variance is the major component of the variance due to general combining ability (sigma(GCA)(2)). Therefore, we conclude that interpopulation improvement becomes more efficient with divergent than with genetically similar heterotic groups, because performance of superior hybrids can be predicted on the basis of general combining ability effects.     

Precision of recombination frequency estimates after random intermating with finite population sizes

Random intermating of F2 populations has been suggested for obtaining precise estimates of recombination frequencies between tightly linked loci. In a simulation study, sampling effects due to small population sizes in the intermating generations were found to abolish the advantages of random intermating that were reported in previous theoretical studies considering an infinite population size. We propose a mating scheme for intermating with planned crosses that yields more precise estimates than those under random intermating.