Stem canker caused by the fungus Leptosphaeria maculans is a major disease of Brassica napus. Quantitative resistance factors appear to be important components for effective and durable control of this pathogen. Quantitative trait loci (QTL) for stem canker resistance have previously been identified in the Darmor variety. However, before these QTL can be used in marker-assisted selection (MAS) to breed resistant varieties, they must be validated in a wide range of genetic backgrounds. We used an association mapping approach to confirm the markers located within the QTL previously identified in Darmor and establish their usefulness in MAS. For this, we characterized the molecular diversity of an oilseed rape collection of 128 lines showing a large spectrum of responses to infection by L. maculans, using 72 pairs of primers for simple sequence repeat and other markers. We used different association mapping models which either do or do not take into account the population structure and/or family relatedness. In all, 61 marker alleles were found to be associated with resistance to stem canker. Some of these markers were associated with previously identified QTL, which confirms their usefulness in MAS. Markers located in regions not harbouring previously identified QTL were also associated with resistance, suggesting that new QTL or allelic variants are present in the collection. All of these markers associated with stem canker resistance will help identify accessions carrying desirable alleles and facilitate QTL introgression
The most common and effective way to control phoma stem canker (blackleg) caused by Leptosphaeria maculans in oilseed rape (Brassica napus) is by breeding resistant cultivars. Specific resistance genes have been identified in B. napus and related species but in some B. napus cultivars resistance is polygenic [mediated by quantitative trait loci (QTL)], postulated to be race non-specific and durable. The genetic basis of quantitative resistance in the French winter oilseed rape 'Darmor', which was derived from 'Jet Neuf', was previously examined in two genetic backgrounds. Stable QTL involved in blackleg resistance across year and genetic backgrounds were identified. In this study, near isogenic lines (NILs) were produced in the susceptible background 'Yudal' for four of these QTL using marker-assisted selection. Various strategies were used to develop new molecular markers, which were mapped in these QTL regions. These were used to characterize the length and homozygosity of the 'Darmor-bzh' introgressed segment in the NILs. Individuals from each NIL were evaluated in blackleg disease field trials and assessed for their level of stem canker in comparison to the recurrent line 'Yudal'. The effect of QTL LmA2 was clearly validated and to a lesser extent, QTL LmA9 also showed an effect on the disease level. This work provides valuable material that can be used to study the mode of action of genetic factors involved in L. maculans quantitative resistance.
Improvement of effectiveness and durability of disease resistance in crops most often relies on the use of quantitative resistance, with the hypothesis that a wide range of quantitative resistance factors (QTL) makes the overcoming of the resistance by the pathogen more difficult. For an optimum use of these QTL in effective and durable strategies of resistance deployment, there is a need to precisely know their localization but also their stability/specificity and their allelic effects in various genetic backgrounds. Stem canker caused by the fungus Leptosphaeria maculans is one of the most important diseases in oilseed rape. In this Brassica napus-L. maculans pathosystem, QTL were previously identified by linkage analysis using populations derived from biparental crosses that were analyzed separately. In this study, we explored new quantitative resistance factors using a multi-cross connected design derived from four resistant lines crossed with a single susceptible line. Independent and connected mapping analyses revealed to be complementary to get an overview of QTL organization. We validated different QTL across different years and genetic backgrounds and identified novel QTL which had not yet been mapped. Population-common and population-specific QTL were identified. Knowledge of QTL organization and effects should help in the rational choice of relevant factors in breeding resistant genotypes to be integrated with other control means such as cultural practices and rotations for durable management of the disease.
<span style="font-size:10pt;font-family:'Tahoma','sans-serif';">The accurate mapping of quantitative trait loci (QTL) depends notably on the number of recombination events occurring in the segregating population. The cost of phenotyping often limits the sample size used in QTL mapping. To get round this problem, we assessed a selective phenotyping method, called <i>qtlRec</i> sampling. In order to improve the accuracy of QTL mapping, a subset of individuals was selected to maximize the number of recombination events at putative QTL positions; the usefulness of this subset was compared to a selected sample built to maximize the recombination rate over the whole genome. We assessed this method on the quantitative oil content trait in <i>Brassica napus</i>. We showed that the <i>qtlRec</i> strategy could allow increasing accuracy (both support interval and position) of QTL location while it maintained a similar power of detection. We then applied this approach to the <i>B. napus</i>—<i>Leptosphaeria maculans</i> pathosystem for which resistance QTL with minor effect were previously identified. This allowed the validation of the QTL in six genomic regions. The <i>qtlRec</i> method is an attractive strategy for validating QTL in multiple year and/or location trials for a trait which requires costly and time-consuming phenotyping.</span><span style="font-size:9pt;font-family:'Tahoma','sans-serif';"> </span>
Blackleg, caused by Leptosphaeria maculans, is a major disease of oilseed rape (Brassica napus), worldwide, including Australia and France. The aims of these studies were first, to determine if higher levels of resistance to L. maculans could be generated in double haploid (DH) lines derived from spring-type B. napus cv. Grouse, which has a good level of field resistance to blackleg; and second, to determine whether the resistance to blackleg disease of individual DH lines responds differentially to different L. maculans field populations within and between the two countries. DH lines were extracted from cv. Grouse and tested in field experiments carried out in both France and Australia against natural L. maculans populations. Extracting and screening DH lines were an effective means to select individual lines with greatly improved expression of resistance to blackleg crown canker disease in comparison with the original parental population. However, relative disease resistance rankings for DH lines were not always consistent between sites. The higher level of resistance in France was shown to be because of a high expression level of quantitative resistance in the French growing conditions. Big differences were observed for some DH lines between the 2004 and the 2005 field sites in Australia where the L. maculans populations differed by their virulence on single dominant gene-based resistant lines derived from Brassica rapa ssp. sylvestris. This differential behaviour could not be clearly explained by the specific resistance genes until now identified in these DH lines. This investigation highlights the potential to derive DH lines with superior levels of resistance to L. maculans compared with parental populations. However, in locations with particularly high pathogen diversity, such as in southern Australia, multiyear and multisite evaluations should be performed to screen for the most efficient material in different situations.
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