Maria Corinna Sanguineti

Quantitative Trait Loci for Grain Yield and Adaptation of Durum Wheat (Triticum durum Desf.) Across a Wide Range of Water Availability

Grain yield is a major goal for the improvement of durum wheat, particularly in drought-prone areas. In this study, the genetic basis of grain yield (GY), heading date (HD), and plant height (PH) was investigated in a durum wheat population of 249 recombinant inbred lines evaluated in 16 environments (10 rainfed and 6 irrigated) characterized by a broad range of water availability and GY (from 5.6 to 58.8 q ha−1). Among the 16 quantitative trait loci (QTL) that affected GY, two major QTL on chromosomes 2BL and 3BS showed significant effects in 8 and 7 environments, with R2 values of 21.5 and 13.8% (mean data of all 16 environments), respectively. In both cases, extensive overlap was observed between the LOD profiles of GY and PH, but not with those for HD. QTL specific for PH were identified on chromosomes 1BS, 3AL, and 7AS. Additionally, three major QTL for HD on chromosomes 2AS, 2BL, and 7BS showed limited or no effects on GY. For both PH and GY, notable epistasis between the chromosome 2BL and 3BS QTL was detected across several environments.

Association mapping in durum wheat grown across a broad range of water regimes

Association mapping was used to dissect the genetic basis of drought-adaptive traits and grain yield (GY) in a collection of 189 elite durum wheat accessions evaluated in 15 environments highly different for water availability during the crop cycle (from 146 to 711 mm) and GY (from 9.9 to 67.3 q ha(-1)). For highly heritable traits (e.g. heading date, kernel weight, etc.) several significant experiment-wise marker-trait associations were detected across five or more (up to 13 for kernel weight) environments, with R(2) values ranging from ca. 5 to 10%. As to GY, significant associations (R(2) from 2.5 to 4.2%) were mostly detected in one environment only (56 markers), while decreasing rapidly from two to five environments (from 20 to three markers, respectively) and with only one marker (Xbarc197 on chr. 5A) found significant in six environments (ranging from low- to high-yielding). These results are probably due to the complex genetic basis of GY and its interaction with environmental conditions. The number of markers significantly affecting GY decreased considerably under drought conditions, suggesting a limited effectiveness of association mapping to identify loci for GY under low-moisture conditions, most likely because different genotypes can attain similar phenotypes via different morpho-physiological traits and corresponding gene networks. Our study confirmed the role of major loci for phenology previously described in biparental mapping populations, highlighted a novel set of loci for drought-adaptive traits, and provided information on the agronomic value of the alleles at such loci across a broad range of soil moisture conditions.

RFLP mapping of quantitative trait loci controlling abscisic acid concentration in leaves of drought-stressed maize (Zea mays L.)

Abstract Abscisic acid (ABA) concentration in leaves of drought-stressed plants is a quantitatively inherited trait. In order to identify quantitative trait loci (QTLs) controlling leaf ABA concentration (L-ABA) in maize, leaf samples were collected from 80 F3:4 families of the cross Os420 (high L-ABA)×IABO78 (low L-ABA) tested under drought conditions in field trials conducted over 2 years. In each year, leaf samples were collected at stem elongation and near anthesis. The genetic map obtained with 106 restriction fragment length polymorphism (RFLP) loci covered 1370 cM, which represented approximately 85% of the UMC maize map. Sixteen different QTLs with a LOD>2.0 were revealed in at least one sampling. Across samplings, only four QTLs significantly influenced L-ABA, accounting for 66% of the phenotypic variation and 76% of the genetic variation among families. At these QTLs, the alleles which increased L-ABA were contributed by Os420. The two most important QTLs were mapped on chromosome 2 near csu133 and csu109a. The effects associated with the QTL near csu133 were more pronounced near anthesis. The support intervals of the four primary QTLs for L-ABA did not overlap the presumed map position of mutants impaired in ABA biosynthesis.

Searching for quantitative trait loci controlling root traits in maize: a critical appraisal

The identification of quantitative trait loci (QTLs) for root traits can provide useful indications on their genetic basis and the associated effects on grain yield under different water regimes. Furthermore, the availability of molecular markers linked to QTLs controlling variation for root traits and grain yield will allow for the implementation of marker-assisted selection to improve productivity. In maize (Zea mays L.), four mapping populations have been investigated to locate QTLs for root traits under controlled conditions and/or in the field. A comparative analysis of the QTL results was carried out based on the availability of molecular markers common to the investigated populations and the UMC maize reference map. Several chromosome regions affected root traits in two or even three populations. A number of these regions also affected grain yield under well-watered and/or drought-stressed conditions. The most important QTL effects were detected on chromosome bins 1.03, 1.06, 1.08, 2.03, 2.04, 7.02, 8.06 and 10.04. Exploiting the syntenic information available for maize and rice, a number of QTLs for root traits described in rice were found to map in regions syntenic to a number of the listed maize chromosome bins (e.g., bin 2.04). The development of near isogenic lines (NILs) for the most important QTLs will allow to investigate whether the concomitant effects of the QTLs on root traits and grain yield are due to linkage and/or pleiotropy and, ultimately, may offer the opportunity to clone the genes underlying such QTLs. Although QTL analysis remains a resource-demanding undertaking, its integration with genomics and post-genomics approaches will play an increasingly important role for the identification of genes affecting root characteristics and grain yield in maize and for harnessing the favourable allelic variation at such loci.

Microsatellite analysis reveals a progressive widening of the genetic basis in the elite durum wheat germplasm

Abstract. It has been argued that the level of genetic diversity in the modern durum wheat (Triticum turgidum L. var. durum) elite germplasm may have declined due to the high selection pressure applied in breeding programs. In this study, 58 accessions covering a wide spectrum of genetic diversity of the cultivated durum wheat gene pool were characterized with 70 microsatellite loci (or simple sequence repeats, SSRs). On average, SSRs detected 5.6 different allelic variants per locus, with a mean diversity index (DI) equal to 0.56, thus revealing a diversity content comparable to those previously observed with SSRs in other small-grain cereal gene pools. The mean genetic similarity value was equal to 0.44. A highly diagnostic SSR set has been identified. A high variation in allele size was detected among SSR loci, suggesting a different suitability of these loci for estimating genetic diversity. The B genome was characterized by an overall polymorphism significantly higher than that of the A genome. Genetic diversity is organised in well-distinct sub-groups identified by the corresponding foundation-genotypes. A large portion (92.7%) of the molecular variation detected within the group of 45 modern cvs was accounted for by SSR alleles tracing back to ten foundation-genotypes; among those, the most recent CIMMYT-derived founders were genetically distant from the old Mediterranean ones. On the other hand, rare alleles were abundant, suggesting that a large number of genetic introgressions contributed to the foundation of the well-diversified germplasm herein considered. The profiles of recently released varieties indicate that the level of genetic diversity present in the modern durum wheat germplasm has actually increased over time.

Characterization of root-yield-1.06, a major constitutive QTL for root and agronomic traits in maize across water regimes

A previous study on maize F(2:3) families derived from Lo964xLo1016 highlighted one QTL in bin 1.06 (hereafter named root-yield-1.06) affecting root and agronomic traits of plants grown in well-watered (WW) and water-stressed (WS) conditions. Starting from different F(4) families, two pairs of near isogenic lines (NILs) were developed at root-yield-1.06. The objective of this study was to evaluate root-yield-1.06 effects across different water regimes, genetic backgrounds, and inbreeding levels. The NILs per se and their crosses with Lo1016 and Lo964 were tested in 2008 and 2009 near to Bologna, with the well-watered (WW) and water-stressed (WS) treatments providing, on average, 70 mm and 35 mm of water, respectively. For NILs per se, the interactions QTL x water regime and QTL x family were negligible in most cases; the QTL additive effects across families were significant for several traits, especially root clump weight. For NILs crosses, analogously to NILs per se, the interactions were generally negligible and the additive effects across water regimes and families were significant for most traits, especially grain yield. A meta-analysis carried out considering the QTLs described in this and previous studies inferred one single locus as responsible for the effects on roots and agronomic traits. Our results show that root-yield-1.06 has a major constitutive effect on root traits, plant vigour and productivity across water regimes, genetic backgrounds, and inbreeding levels. These features suggest that root-yield-1.06 is a valuable candidate for cloning the sequence underlying its effects and for marker-assisted selection to improve yield stability in maize.

A panel of elite accessions of durum wheat (Triticum durum Desf.) suitable for association mapping studies

The effectiveness of association mapping (AM) based on linkage disequilibrium (LD) is currently beingtestedinanumberofcrops.AnimportantprerequisitefortheapplicationofAMistheavailabilityofcollectionsofaccessionswithasuitablelevelofgeneticvariationfortargettraitsandwith limited spurious LD due to the presence of population structure. Herein, the results of a genomewide molecular characterization of a collection of elite durum wheat accessions well-adapted to Mediterranean environments are presented. Ninety-seven highly polymorphic simple sequence repeats and 166 amplified fragment length polymorphism markers were used to characterize 189 durum accessions, mainly cultivars and advanced breeding lines. Genome-wide significant and sizeable LD indices at a centimorgan scale were observed, while LD mainly decayed within 10cM. On the other hand, effects due to spurious LD were notably lower than those previously observed in a durum wheat collection sampling durum gene pools of more diverse origin.

Sequence-based SNP genotyping in durum wheat.

Marker development for marker-assisted selection in plant breeding is increasingly based on next-generation sequencing (NGS). However, marker development in crops with highly repetitive, complex genomes is still challenging. Here we applied sequence-based genotyping (SBG), which couples AFLP®-based complexity reduction to NGS, for de novo single nucleotide polymorphisms (SNP) marker discovery in and genotyping of a biparental durum wheat population. We identified 9983 putative SNPs in 6372 contigs between the two parents and used these SNPs for genotyping 91 recombinant inbred lines (RILs). Excluding redundant information from multiple SNPs per contig, 2606 (41%) markers were used for integration in a pre-existing framework map, resulting in the integration of 2365 markers over 2607 cM. Of the 2606 markers available for mapping, 91% were integrated in the pre-existing map, containing 708 SSRs, DArT markers, and SNPs from CRoPS technology, with a map-size increase of 492 cM (23%). These results demonstrate the high quality of the discovered SNP markers. With this methodology, it was possible to saturate the map at a final marker density of 0.8 cM/marker. Looking at the binned marker distribution (Figure 2), 63 of the 268 10-cM bins contained only SBG markers, showing that these markers are filling in gaps in the framework map. As to the markers that could not be used for mapping, the main reason was the low sequencing coverage used for genotyping. We conclude that SBG is a valuable tool for efficient, high-throughput and high-quality marker discovery and genotyping for complex genomes such as that of durum wheat.

Genetic dissection of maize phenology using an intraspecific introgression library

BackgroundCollections of nearly isogenic lines where each line carries a delimited portion of a donor source genome into a common recipient genetic background are known as introgression libraries and have already shown to be instrumental for the dissection of quantitative traits. By means of marker-assisted backcrossing, we have produced an introgression library using the extremely early-flowering maize (Zea mays L.) variety Gaspé Flint and the elite line B73 as donor and recipient genotypes, respectively, and utilized this collection to investigate the genetic basis of flowering time and related traits of adaptive and agronomic importance in maize.ResultsThe collection includes 75 lines with an average Gaspé Flint introgression length of 43.1 cM. The collection was evaluated for flowering time, internode length, number of ears, number of nodes (phytomeres), number of nodes above the ear, number and proportion of nodes below the ear and plant height. Five QTLs for flowering time were mapped, all corresponding to major QTLs for number of nodes. Three additional QTLs for number of nodes were mapped. Besides flowering time, the QTLs for number of nodes drove phenotypic variation for plant height and number of nodes below and above the top ear, but not for internode length. A number of apparently Mendelian-inherited phenotypes were also observed.ConclusionsWhile the inheritance of flowering time was dominated by the well-known QTL Vgt1, a number of other important flowering time QTLs were identified and, thanks to the type of plant material here utilized, immediately isogenized and made available for fine mapping. At each flowering time QTL, early flowering correlated with fewer vegetative phytomeres, indicating the latter as a key developmental strategy to adapt the maize crop from the original tropical environment to the northern border of the temperate zone (southern Canada), where Gaspé Flint was originally cultivated. Because of the trait differences between the two parental genotypes, this collection will serve as a permanent source of nearly isogenic materials for multiple studies of QTL analysis and cloning.

Utilization of SSR and AFLP markers for the assessment of distinctness in durum wheat

Molecular techniques provide new possibilities to characterize advanced genetic materials for registration purposes and for the protection of breeders’ rights. The objective of this work was to compare the simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) markers with morphological descriptors, currently in use, for the assessment of variety distinctness in durum wheat. Fifty-six F8 or F9 lines or sublines at different levels of relatedness derived from four crosses, two BC1- and 11 BC3-derived lines, were characterized with 27 morphological traits, including 17 official Community Plant Variety Office (CPVO) descriptors, seven AFLP primer combinations, and 98 SSR primer pairs. The similarities based on all three marker classes reflected, on average, the degree of relatedness of the materials. However, molecular markers (MMs) and in particular SSRs produced classifications more closely representing the relationships among lines, allowing us to discriminate even tightly related genotypes that morphological descriptors failed to distinguish. The moderate correlations between similarities based on morphological and molecular data, and the wide range of MM differences corresponding to few or no morphological differences, imply that at present MMs should only be used as a complementary tool to assess distinctness. Based on these results, it seems that a set of 28 SSRs (one per chromosome arm) represents a useful prescreening tool to identify the entry pairs sufficiently different at MM level (≥13 polymorphisms) for which a field evaluation could be avoided, with relevant savings in resources and optimization of the field trial design.