Impact of irrigation, nitrogen fertilization, and plant density on stay-green and its effects on agronomic traits in maize.

Introduction: The stay-green (SG) or delayed leaf senescence enables crop plants to maintain their green leaves and photosynthetic capacity for a longer time after flowering. It is considered an important trait in maize breeding, which has contributed to gain in grain yield of modern varieties. It has been also used to improve the tolerance to drought and deficiencies in nitrogen fertilization (NF). However, the objective of this study is to evaluate the influence of water irrigation (WI), NF, and plant density (PD) on SG and the effect of SG on agronomic traits in maize. Methods: Four SG lines and four non-stay-green (NSG) lines were evaluated in four contrasting environments under two WI, three NF, and two PD levels. Results and discussion: As expected, the chlorophyll content of leaves at 45 days after flowering (Chlo45) was, on average, higher in the SG group of lines. The difference in Chlo45 between the SG and NSG genotypes was consistent across WI, NF, and PD and the environments. This is indicative that internal or developmental factors were more important than external signals in controlling the senescence. The effect of SG increasing thousand-kernel weight, stover yield at harvest, or moisture was not influenced by WI, NF, or PD but was altered by the background environment. Our results have implications for the application of SG as a secondary trait for enhancing abiotic stress tolerance. Future studies could consider a wider range of environmental conditions to assess the performance of SG traits under different climatic and soil conditions.

Genomic versus phenotypic selection to improve corn borer resistance and grain yield in maize.

Introduction: The study of yield and resistance/tolerance to pest are related traits fundamental for maize breeding programs. Genomic selection (GS), which uses all marker information to calculate genomic breeding values, is presented as an emerging alternative to phenotypic and marker-assisted selections for improving complex traits controlled by many genes with small effects. Therefore, although phenotypic selection (PS) has been effective for increasing resistance and yield under high infestation with maize stem borers, higher genetic gains are expected to be obtained through GS based on the complex architecture of both traits. Our objective was to test whether GS is more effective than PS for improving resistance and/or tolerance to maize stem borers and grain yield. Methods: For this, we compared different selection programs based on phenotype and genotypic value for a single trait, resistance or yield, and for both traits together. Results and discussion: We obtained that GS achieved the highest genetic gain for yield, meanwhile phenotypic selection for yield was the program that achieved the highest reduction of tunnel length, but was ineffective for increasing yield. However, phenotypic or genomic selection for increased resistance may be more effective in improving both traits together; although the gains per cycle would be small for both traits.

Genome-Wide Association Analysis of Senescence-Related Traits in Maize.

Senescence is a programmed process that involves the destruction of the photosynthesis apparatus and the relocation of nutrients to the grain. Identifying senescence-associated genes is essential to adapting varieties for the duration of the cultivation cycle. A genome-wide association study (GWAS) was performed using 400 inbred maize lines with 156,164 SNPs to study the genetic architecture of senescence-related traits and their relationship with agronomic traits. We estimated the timing of senescence to be 45 days after anthesis in the whole plant and specifically in the husks. A list of genes identified in a previous RNAseq experiment as involved in senescence (core senescence genes) was used to propose candidate genes in the vicinity of the significant SNPs. Forty-six QTLs of moderate to high effect were found for senescence traits, including specific QTLs for husk senescence. The allele that delayed senescence primarily increased grain yield and moisture. Seven and one significant SNPs were found in the coding and promoter regions of eight core senescence genes, respectively. These genes could be potential candidates for generating a new variation by genome editing for functional analysis and breeding purposes, particularly Zm00001d014796, which could be responsible for a QTL of senescence found in multiple studies.

Precision Mapping of a Maize MAGIC Population Identified a Candidate Gene for the Senescence-Associated Physiological Traits.

Senescence is an important trait in maize (Zea mais L.), a key crop that provides nutrition values and a renewable source of bioenergy worldwide. Genome-wide association studies (GWAS) can be used to identify causative genetic variants that influence the major physiological measures of senescence, which is used by plants as a defense mechanism against abiotic and biotic stresses affecting its performance. We measured four physiological and two agronomic traits that affect senescence. Six hundred seventy-two recombinant inbred lines (RILs) were evaluated in two consecutive years. Thirty-six candidate genes were identified by genome-wide association study (GWAS), and 11 of them were supported by additional evidence for involvement in senescence-related processes including proteolysis, sugar transport, and sink activity. We identified a candidate gene, Zm00001d043586, significantly associated with chlorophyll, and independently studied its transcription expression in an independent panel. Our results showed that Zm00001d043586 affects chlorophyl rate degradation, a key determinant of senescence, at late plant development stages. These results contribute to better understand the genetic relationship of the important trait senescence with physiology related parameters in maize and provide new putative molecular markers that can be used in marker assisted selection for line development.

Mapping of resistance to corn borers in a MAGIC population of maize.

BACKGROUND: Corn borers constitute an important pest of maize around the world; in particular Sesamia nonagrioides Lefèbvre, named Mediterranean corn borer (MCB), causes important losses in Southern Europe. Methods of selection can be combined with transgenic approaches to increase the efficiency and durability of the resistance to corn borers. Previous studies of the genetic factors involved in resistance to MCB have been carried out using bi-parental populations that have low resolution or using association inbred panels that have a low power to detect rare alleles. We developed a Multi-parent Advanced Generation InterCrosses (MAGIC) population to map with high resolution the genetic determinants of resistance to MCB. RESULTS: We detected multiple single nucleotide polymorphisms (SNPs) of low effect associated with resistance to stalk tunneling by MCB. We dissected a wide region related to stalk tunneling in multiple studies into three smaller regions (at ~ 150, ~ 155, and ~ 165 Mb in chromosome 6) that closely overlap with regions associated with cell wall composition. We also detected regions associated with kernel resistance and agronomic traits, although the co-localization of significant regions between traits was very low. This indicates that it is possible the concurrent improvement of resistance and agronomic traits. CONCLUSIONS: We developed a mapping population which allowed a finer dissection of the genetics of maize resistance to corn borers and a solid nomination of candidate genes based on functional information. The population, given its large variability, was also adequate to map multiple traits and study the relationship between them.

European maize landraces made accessible for plant breeding and genome-based studies.

KEY MESSAGE: Doubled-haploid libraries from landraces capture native genetic diversity for a multitude of quantitative traits and make it accessible for breeding and genome-based studies. Maize landraces comprise large allelic diversity. We created doubled-haploid (DH) libraries from three European flint maize landraces and characterized them with respect to their molecular diversity, population structure, trait means, variances, and trait correlations. In total, 899 DH lines were evaluated using high-quality genotypic and multi-environment phenotypic data from up to 11 environments. The DH lines covered 95% of the molecular variation present in 35 landraces of an earlier study and represent the original three landrace populations in an unbiased manner. A comprehensive analysis of the target trait plant development at early growth stages as well as other important agronomic traits revealed large genetic variation for line per se and testcross performance. The majority of the 378 DH lines evaluated as testcrosses outperformed the commercial hybrids for early development. For total biomass yield, we observed a yield gap of 15% between mean testcross yield of the commercial hybrids and mean testcross yield of the DH lines. The DH lines also exhibited genetic variation for undesirable traits like root lodging and tillering, but correlations with target traits early development and yield were low or nonsignificant. The presented diversity atlas is a valuable, publicly available resource for genome-based studies to identify novel trait variation and evaluate the prospects of genomic prediction in landrace-derived material.

Fine analysis of a genomic region involved in resistance to Mediterranean corn borer.

BACKGROUND: Sesamia nonagrioides Lefebvere (Mediterranean corn borer, MCB) is the main pest of maize in the Mediterranean area. QTL for MCB stalk tunneling and grain yield under high MCB infestation had been located at bin 8.03-8.05 (4-21 cM and 10-30 cM respectively) in a previous analysis of the EP42 x EP39 RILs mapping population. The objective of the present work was to study with higher resolution those QTL, and validating and estimating with higher precision their locations and effects. To achieve this objective, we developed a set of 38 heterogeneous inbred families (HIFs) which were near-homozygous in the genome, except in the region under study. The HIFs were evaluated in multiple environments under artificial infestation with MCB and genotyped with SNPs. RESULTS: The QTL for grain yield under high infestation was confirmed with higher precision and improved reliability at 112.6-116.9 Mb. On the contrary, the location of the QTL for stalk tunneling was not validated probably due to the fixation of some genomic regions during the development of the HIFs. Our study confirmed that the co-localization of the QTL for stalk tunneling and grain yield in the previous study was due to linked genes, not to pleiotropic effects. So, the QTL for grain yield can be used for improving grain yield without undesirable effect on stalk tunneling. CONCLUSIONS: The HIF analysis is useful for validating QTL and for conducting deeper studies in traits related to corn borer resistance.

Chemical Changes during Maize Tissue Aging and Its Relationship with Mediterranean Corn Borer Resistance.

The Mediterranean corn borer (MCB), Sesamia nonagrioides Lef, is an important pest of maize in temperate areas, causing significant stalk lodging and yield losses. The main goals of this study were to determine possible changes in chemical traits (phenols, flavonoids, anthocyanins, sugars, fibers, and lignin) during plant development after the flowering stage and to assess how those traits may differ in diverse genotypes of maize, such as MCB resistant and susceptible. Higher values for some particular traits in more mature tissues seemed to increase their effectiveness against the MCB attack. A decreased amount of borer damage in the field was recorded in the resistant inbred line and in older tissues (7.90 cm vs 31.70 cm as the mean for the stalk tunnel length). In accordance with these results, the resistant inbred line showed a higher degree of hemicellulose cross-linkage (due to ferulic and diferulic acids), higher soluble sugar content, and higher stalk strength. The use of resistant varieties and early sowings is highly recommended as an integrated approach to reduce the yield losses produced by this pest.

Is there an optimum level of diversity in utilization of genetic resources?

KEY MESSAGE: Capitalizing upon the genomic characteristics of long-term random mating populations, sampling from pre-selected landraces is a promising approach for broadening the genetic base of elite germplasm for quantitative traits. Genome-enabled strategies for harnessing untapped allelic variation of landraces are currently evolving. The success of such approaches depends on the choice of source material. Thus, the analysis of different strategies for sampling allelic variation from landraces and their impact on population diversity and linkage disequilibrium (LD) is required to ensure the efficient utilization of diversity. We investigated the impact of different sampling strategies on diversity parameters and LD based on high-density genotypic data of 35 European maize landraces each represented by more than 20 individuals. On average, five landraces already captured ~95% of the molecular diversity of the entire dataset. Within landraces, absence of pronounced population structure, consistency of linkage phases and moderate to low LD levels were found. When combining data of up to 10 landraces, LD decay distances decreased to a few kilobases. Genotyping 24 individuals per landrace with 5k SNPs was sufficient for obtaining representative estimates of diversity and LD levels to allow an informed pre-selection of landraces. Integrating results from European with Central and South American landraces revealed that European landraces represent a unique and diverse spectrum of allelic variation. Sampling strategies for harnessing allelic variation from landraces depend on the study objectives. If the focus lies on the improvement of elite germplasm for quantitative traits, we recommend sampling from pre-selected landraces, as it yields a wide range of diversity, allows optimal marker imputation, control for population structure and avoids the confounding effects of strong adaptive alleles.

QTL Mapping for Yield and Resistance against Mediterranean Corn Borer in Maize.

Introduction: The Mediterranean corn borer (MCB), Sesamia nonagrioides, is a major pest of maize, Zea mays, in Mediterranean countries, inflicting significant kernel yield losses. For that reason, it necessary to know the genetic mechanisms that regulate the agronomic and resistance traits. A quantitative trait loci (QTL) mapping study for yield, resistance against MCB attack, and other relevant agronomic traits was performed using a recombinant inbred line (RIL) population derived from the cross A637 × A509 that is expected to segregate for yield, and ear, and stalk resistance to MCB. 171 RILs were evaluated in 2014 and 2015 at Pontevedra, Spain, along with the two parental inbreds A637 and A509 using a 13 × 14 single lattice design with two replications. A genetic map with 285 SNP markers was used for QTL analysis. Our objectives were to detect QTL for resistance to MCB and tolerance-related agronomic traits, to gain insights on the genetic relationship between resistance to MCB attack and yield, and to establish the best way for simultaneously improving yield and resistance to MCB. Results: Twelve significant QTL were detected for agronomic and resistance traits. QTL at bins 1.10 and 5.04 were especially interesting because the same allelic variant at these QTL simultaneously improved yield and insect resistance. In contrast, in the region 8.04-8.05, QTL showed opposite effects for yield and resistance. Several QTL for indexes which combine yield and resistance traits were found especially in the region 10.02-10.03. Conclusions: Selecting genotypes with the favorable allele of QTL on chromosome 5 (bin 5.01) will decrease tunnel length without affect yield, silking and plant height and QTL on the region 5.04 could be used to improve stalk resistance and yield simultaneously. An allele of QTL on bin 9.07 will increase ear resistance to MCB attack but it could produce later varieties while favorable allele in region 1.10 could improve ear and stalk resistance and yield without secondary negative effects. The region 8.03-8.05 mainly but also the region 10.02-10.03 and 5.04 may play an important role to elucidate the association between yield, other agronomic traits and MCB resistance.

A comprehensive study of the genomic differentiation between temperate Dent and Flint maize.

BACKGROUND: Dent and Flint represent two major germplasm pools exploited in maize breeding. Several traits differentiate the two pools, like cold tolerance, early vigor, and flowering time. A comparative investigation of their genomic architecture relevant for quantitative trait expression has not been reported so far. Understanding the genomic differences between germplasm pools may contribute to a better understanding of the complementarity in heterotic patterns exploited in hybrid breeding and of mechanisms involved in adaptation to different environments. RESULTS: We perform whole-genome screens for signatures of selection specific to temperate Dent and Flint maize by comparing high-density genotyping data of 70 American and European Dent and 66 European Flint inbred lines. We find 2.2 % and 1.4 % of the genes are under selective pressure, respectively, and identify candidate genes associated with agronomic traits known to differ between the two pools. Taking flowering time as an example for the differentiation between Dent and Flint, we investigate candidate genes involved in the flowering network by phenotypic analyses in a Dent-Flint introgression library and find that the Flint haplotypes of the candidates promote earlier flowering. Within the flowering network, the majority of Flint candidates are associated with endogenous pathways in contrast to Dent candidate genes, which are mainly involved in response to environmental factors like light and photoperiod. The diversity patterns of the candidates in a unique panel of more than 900 individuals from 38 European landraces indicate a major contribution of landraces from France, Germany, and Spain to the candidate gene diversity of the Flint elite lines. CONCLUSIONS: In this study, we report the investigation of pool-specific differences between temperate Dent and Flint on a genome-wide scale. The identified candidate genes represent a promising source for the functional investigation of pool-specific haplotypes in different genetic backgrounds and for the evaluation of their potential for future crop improvement like the adaptation to specific environments.

Redefining Agricultural Residues as Bioenergy Feedstocks.

The use of plant biomass is a sustainable alternative to the reduction of CO2 emissions. Agricultural residues are interesting bioenergy feedstocks because they do not compete with food and add extra value to the crop, which might help to manage these residues in many regions. Breeding crops for dual production of food and bioenergy has been reported previously, but the ideal plant features are different when lignocellulosic residues are burnt for heat or electricity, or fermented for biofuel production. Stover moisture is one of the most important traits in the management of agricultural waste for bioenergy production which can be modified by genetic improvement. A delayed leaf senescence or the stay-green characteristic contributes to higher grain and biomass yield in standard, low nutrient, and drought-prone environments. In addition, the stay-green trait could be favorable for the development of dual purpose varieties because this trait could be associated with a reduction in biomass losses and lodging. On the other hand, the stay-green trait could be detrimental for the management of agricultural waste if it is associated with higher stover moisture at harvest, although this hypothesis has been insufficiently tested. In this paper, a review of traits relevant to the development of dual purpose varieties is presented with particular emphasis on stover moisture and stay-green, because less attention has been paid to these important traits in the literature. The possibility of developing new varieties for combined production is discussed from a breeding perspective.

QTL mapping for Mediterranean corn borer resistance in European flint germplasm using recombinant inbred lines.

BACKGROUND: Ostrinia nubilalis (ECB) and Sesamia nonagrioides (MCB) are two maize stem borers which cause important losses in temperate maize production, but QTL analyses for corn borer resistance were mostly restricted to ECB resistance and maize materials genetically related (mapping populations derived from B73). Therefore, the objective of this work was to identify and characterize QTLs for MCB resistance and agronomic traits in a RILs population derived from European flint inbreds. RESULTS: Three QTLs were detected for stalk tunnel length at bins 1.02, 3.05 and 8.05 which explained 7.5% of the RILs genotypic variance. The QTL at bin 3.05 was co-located to a QTL related to plant height and grain humidity and the QTL at bin 8.05 was located near a QTL related to yield. CONCLUSIONS: Our results, when compared with results from other authors, suggest the presence of genes involved in cell wall biosynthesis or fortification with effects on resistance to different corn borer species and digestibility for dairy cattle. Particularly, we proposed five candidate genes related to cell wall characteristics which could explain the QTL for stalk tunnelling in the region 3.05. However, the small proportion of genotypic variance explained by the QTLs suggest that there are also many other genes of small effect regulating MCB resistance and we conclude that MAS seems not promising for this trait. Two QTLs detected for stalk tunnelling overlap with QTLs for agronomic traits, indicating the presence of pleitropism or linkage between genes affecting resistance and agronomic traits.

Mapping of QTL for resistance to the Mediterranean corn borer attack using the intermated B73 x Mo17 (IBM) population of maize.

The Mediterranean corn borer or pink stem borer (MCB, Sesamia nonagrioides Lefebvre) causes important yield losses as a consequence of stalk tunneling and direct kernel damage. B73 and Mo17 are the source of the most commercial valuable maize inbred lines in temperate zones, while the intermated B73 x Mo17 (IBM) population is an invaluable source for QTL identification. However, no or few experiments have been carried out to detect QTL for corn borer resistance in the B73 x Mo17 population. The objective of this work was to locate QTL for resistance to stem tunneling and kernel damage by MCB in the IBM population. We detected a QTL for kernel damage at bin 8.05, although the effect was small and two QTL for stalk tunneling at bins 1.06 and 9.04 in which the additive effects were 4 cm, approximately. The two QTL detected for MCB resistance were close to other QTL consistently found for European corn borer (ECB, Ostrinia nubilalis Hübner) resistance, indicating mechanisms of resistance common to both pests or gene clusters controlling resistance to different plagues. The precise mapping achieved with the IBM population will facilitate the QTL pyramiding and the positional cloning of the detected QTL.

Genetic variation and quantitative trait loci associated with developmental stability and the environmental correlation between traits in maize.

There is limited experimental information about the genetic basis of micro-environmental variance (V(E)) (developmental stability) and environmental correlations. This study, by using a population of maize recombinant inbred lines (RIL) and simple sequence repeat (SSR) polymorphic markers, aims at the following: firstly, to quantify the genetic component of the V(E) or developmental stability for four traits in maize and the environmental correlation between these traits, and secondly, to identify quantitative trait loci (QTLs) that influence these quantities. We found that, when estimating variances and correlations and testing their homogeneity, estimates and tests are needed that are not highly dependent on normality assumptions. There was significant variation among the RILs in V(E) and in the environmental correlation for some of the traits, implying genetic heterogeneity in the V(E) and environmental correlations. The genetic coefficient of variation of the environmental variance (GCV(V(E))) was estimated to be 20%, which is lower than estimates obtained for other species. A few genomic regions involved in the stability of one trait or two traits were detected, and these did not have an important influence on the mean of the trait. One region that could be associated with the environmental correlations between traits was also detected.

Corn borer (Lepidoptera: Noctuidae and Crambidae) resistance of main races of maize from North America.

Resistance to corn borers, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and Sesamia nonagrioides (Lefebvre) (Lepidoptera: Noctuidae) in maize, Zea mays L., populations is partial, and more resistant populations are needed. The objective of this research was to compare resistance to corn borers of the main maize races from North America. Twenty open-pollinated maize populations belonging to the races Southern Dent, Corn Belt Dent, and Northern Flint, and three check populations, were evaluated under artificial infestation with S. nonagrioides and O. nubilalis. None of the populations had complete resistance. Northern Flint had the lowest yield under corn borer infestation, whereas Southern Dent had the highest yield but also the largest damage. Corn Belt Dent had a shorter growing cycle and similar yield of infected plants than Southern Dent. The checks had intermediate yield and resistance and were not significantly different from Corn Belt Dent for any trait. The Southern Dent populations Tennessee Red Cob and White Dent (PI221885 and PI311232) could be used as sources of tolerance to corn borers, although they are not expected to provide great gains compared with the levels of tolerance already present in some Corn Belt Dent and European Flint populations and would require adaptation to short growing cycle. The Corn Belt Dent synthetic BS17 had the highest yield and general agronomic performance under corn borer infestation, along with Rustler and Silver King, and the European Flint composite EPS13.