Improving resistance to the European corn borer: a comprehensive study in elite maize using QTL mapping and genome-wide prediction.

KEY MESSAGE: The efficiency of marker-assisted selection for native resistance to European corn borer stalk damage can be increased when progressing from a QTL-based towards a genome-wide approach. Marker-assisted selection (MAS) has been shown to be effective in improving resistance to the European corn borer (ECB) in maize. In this study, we investigated the performance of whole-genome-based selection, relative to selection based on individual quantitative trait loci (QTL), for resistance to ECB stalk damage in European elite maize. Three connected biparental populations, comprising 590 doubled haploid (DH) lines, were genotyped with high-density single nucleotide polymorphism markers and phenotyped under artificial and natural infestation in 2011. A subset of 195 DH lines was evaluated in the following year as lines per se and as testcrosses. Resistance was evaluated based on stalk damage ratings, the number of feeding tunnels in the stalk and tunnel length. We performed individual- and joint-population QTL analyses and compared the cross-validated predictive abilities of the QTL models with genomic best linear unbiased prediction (GBLUP). For all traits, the GBLUP model consistently outperformed the QTL model despite the detection of QTL with sizeable effects. For stalk damage rating, GBLUP's predictive ability exceeded at times 0.70. Model training based on DH line per se performance was efficient in predicting stalk breakage in testcrosses. We conclude that the efficiency of MAS for ECB stalk damage resistance can be increased considerably when progressing from a QTL-based towards a genome-wide approach. With the availability of native ECB resistance in elite European maize germplasm, our results open up avenues for the implementation of an integrated genome-based selection approach for the simultaneous improvement of yield, maturity and ECB resistance.

Polymorphisms in O-methyltransferase genes are associated with stover cell wall digestibility in European maize (Zea mays L.).

BACKGROUND: OMT (O-methyltransferase) genes are involved in lignin biosynthesis, which relates to stover cell wall digestibility. Reduced lignin content is an important determinant of both forage quality and ethanol conversion efficiency of maize stover. RESULTS: Variation in genomic sequences coding for COMT, CCoAOMT1, and CCoAOMT2 was analyzed in relation to stover cell wall digestibility for a panel of 40 European forage maize inbred lines, and re-analyzed for a panel of 34 lines from a published French study. Different methodologies for association analysis were performed and compared. Across association methodologies, a total number of 25, 12, 1, 6 COMT polymorphic sites were significantly associated with DNDF, OMD, NDF, and WSC, respectively. Association analysis for CCoAOMT1 and CCoAOMT2 identified substantially fewer polymorphic sites (3 and 2, respectively) associated with the investigated traits. Our re-analysis on the 34 lines from a published French dataset identified 14 polymorphic sites significantly associated with cell wall digestibility, two of them were consistent with our study. Promising polymorphisms putatively causally associated with variability of cell wall digestibility were inferred from the total number of significantly associated SNPs/Indels. CONCLUSIONS: Several polymorphic sites for three O-methyltransferase loci were associated with stover cell wall digestibility. All three tested genes seem to be involved in controlling DNDF, in particular COMT. Thus, considerable variation among Bm3 wildtype alleles can be exploited for improving cell-wall digestibility. Target sites for functional markers were identified enabling development of efficient marker-based selection strategies.

Characterization of phenylpropanoid pathway genes within European maize (Zea mays L.) inbreds.

BACKGROUND: Forage quality of maize is influenced by both the content and structure of lignins in the cell wall. Biosynthesis of monolignols, constituting the complex structure of lignins, is catalyzed by enzymes in the phenylpropanoid pathway. RESULTS: In the present study we have amplified partial genomic fragments of six putative phenylpropanoid pathway genes in a panel of elite European inbred lines of maize (Zea mays L.) contrasting in forage quality traits. Six loci, encoding C4H, 4CL1, 4CL2, C3H, F5H, and CAD, displayed different levels of nucleotide diversity and linkage disequilibrium (LD) possibly reflecting different levels of selection. Associations with forage quality traits were identified for several individual polymorphisms within the 4CL1, C3H, and F5H genomic fragments when controlling for both overall population structure and relative kinship. A 1-bp indel in 4CL1 was associated with in vitro digestibility of organic matter (IVDOM), a non-synonymous SNP in C3H was associated with IVDOM, and an intron SNP in F5H was associated with neutral detergent fiber. However, the C3H and F5H associations did not remain significant when controlling for multiple testing. CONCLUSION: While the number of lines included in this study limit the power of the association analysis, our results imply that genetic variation for forage quality traits can be mined in phenylpropanoid pathway genes of elite breeding lines of maize.

High levels of linkage disequilibrium and associations with forage quality at a phenylalanine ammonia-lyase locus in European maize (Zea mays L.) inbreds.

Forage quality of maize is influenced by both the content and structure of lignin in the cell wall. Phenylalanine Ammonia-Lyase (PAL) catalyzes the first step in lignin biosynthesis in plants; the deamination of L-phenylalanine to cinnamic acid. Successive enzymatic steps lead to the formation of three monolignols, constituting the complex structure of lignin. We have cloned and sequenced a PAL genomic sequence from 32 maize inbred lines currently employed in forage maize breeding programs in Europe. Low nucleotide diversity and excessive linkage disequilibrium (LD) was identified at this PAL locus, possibly reflecting selective constrains resulting from PAL being the first enzyme in the monolignol, and other, pathways. While the association analysis was affected by extended LD and population structure, several individual polymorphisms were associated with neutral detergent fiber (not considering population structure) and a single polymorphism was associated with in vitro digestibility of organic matter (considering population structure).