The most common and effective way to control phoma stem canker (blackleg) caused by Leptosphaeria maculans in oilseed rape (Brassica napus) is through the breeding of resistant cultivars. Race specific major genes that mediate resistance from the seedling stage have been identified in B. napus or have been introgressed from related species. Many race specific major genes have been described and some of them are probably identical in B. napus (allotetraploid AACC) and the parental species B. rapa (diploid AA). More work is needed using a set of well-characterised isolates to determine the number of different major resistance genes available. In some B. napus cultivars, there is resistance which is polygenic (mediated by Quantitative Trait Loci) and postulated to be race non-specific. Many of these major genes and Quantitative Trait Loci for resistance to L. maculans have been located on B. napus genetic maps. Genes involved in race specific and polygenic resistance are generally distinct.
Blackleg disease, caused by the fungus Leptosphaeria maculans, is the major disease of canola (Brassica napus) worldwide. A set of 12 Australian L. maculans isolates was developed and used to characterise seedling resistance in 127 Australian cultivars and advanced breeding lines. Plant mortality data used to assess the effectiveness of seedling resistance in canola growing regions of Australia showed that Rlm3 and Rlm4 resistance genes were less effective than other seedling resistance genes. This finding was consistent with regional surveys of the pathogen, which showed the frequency of Rlm4-attacking isolates was >70% in fungal populations over a 10-year period. Differences in adult plant resistance were identified in a subset of Australian cultivars, indicating that some adult gene resistance is isolate-specific.
Data from advanced breeding experiments between 1985 and 1994 were used to determine the effects of region, year and environment on the quality of canola grown across Victoria. Estimates from these unbalanced data were made using residual maximum likelihood. Environmental effects were large relative to cultivar effects for oil and protein content, while the reverse occurred for glucosinolate content. High oil contents (and low seed protein contents) were correlated with cooler spring temperatures and higher spring rainfall. Oil contents were lowest, on average, in canola grown in dry years, or from the hotter regions, such as the Mallee, and were highest in canola from the cooler, wetter regions, such as south-western and north-eastern Victoria. Fatty acid composition varied with year and region. Means for saturated fatty acid content averaged 6.4 0.1%. The oleic acid content averaged 60.3 0.4% and was higher in canola grown in central Victoria and the Wimmera, and in most years, in north-eastern Victoria compared with other regions. Low temperatures and low rainfall reduced oleic acid content. Linoleic acid content averaged 19.7 0.3% and linolenic acid averaged 10.4 0.3%, with the content of these fatty acids negatively correlated with the content of oleic acid. Erucic acid levels were below 0.6% in all regions.
BackgroundField pea (Pisum sativum L.) is a self-pollinating, diploid, cool-season food legume. Crop production is constrained by multiple biotic and abiotic stress factors, including salinity, that cause reduced growth and yield. Recent advances in genomics have permitted the development of low-cost high-throughput genotyping systems, allowing the construction of saturated genetic linkage maps for identification of quantitative trait loci (QTLs) associated with traits of interest. Genetic markers in close linkage with the relevant genomic regions may then be implemented in varietal improvement programs.ResultsIn this study, single nucleotide polymorphism (SNP) markers associated with expressed sequence tags (ESTs) were developed and used to generate comprehensive linkage maps for field pea. From a set of 36,188 variant nucleotide positions detected through in silico analysis, 768 were selected for genotyping of a recombinant inbred line (RIL) population. A total of 705 SNPs (91.7%) successfully detected segregating polymorphisms. In addition to SNPs, genomic and EST-derived simple sequence repeats (SSRs) were assigned to the genetic map in order to obtain an evenly distributed genome-wide coverage. Sequences associated with the mapped molecular markers were used for comparative genomic analysis with other legume species. Higher levels of conserved synteny were observed with the genomes of Medicago truncatula Gaertn. and chickpea (Cicer arietinum L.) than with soybean (Glycine max [L.] Merr.), Lotus japonicus L. and pigeon pea (Cajanus cajan [L.] Millsp.). Parents and RIL progeny were screened at the seedling growth stage for responses to salinity stress, imposed by addition of NaCl in the watering solution at a concentration of 18 dS m-1. Salinity-induced symptoms showed normal distribution, and the severity of the symptoms increased over time. QTLs for salinity tolerance were identified on linkage groups Ps III and VII, with flanking SNP markers suitable for selection of resistant cultivars. Comparison of sequences underpinning these SNP markers to the M. truncatula genome defined genomic regions containing candidate genes associated with saline stress tolerance.ConclusionThe SNP assays and associated genetic linkage maps developed in this study permitted identification of salinity tolerance QTLs and candidate genes. This constitutes an important set of tools for marker-assisted selection (MAS) programs aimed at performance enhancement of field pea cultivars.
Key message “We identified both quantitative and quantitative resistance loci to Leptosphaeria maculans, a fungal pathogen, causing blackleg disease in canola. Several genome-wide significant associations were detected at known and new loci for blackleg resistance. We further validated statistically significant associations in four genetic mapping populations, demonstrating that GWAS marker loci are indeed associated with resistance to L. maculans. One of the novel loci identified for the first time, Rlm12, conveys adult plant resistance in canola.” Blackleg, caused by Leptosphaeria maculans, is a significant disease which affects the sustainable production of canola (Brassica napus). This study reports a genome-wide association study based on 18,804 polymorphic SNPs to identify loci associated with qualitative and quantitative resistance to L. maculans. Genomic regions delimited with 694 significant SNP markers, that are associated with resistance evaluated using 12 single spore isolates and pathotypes from four canola stubble were identified. Several significant associations were detected at known disease resistance loci including in the vicinity of recently cloned Rlm2/LepR3 genes, and at new loci on chromosomes A01/C01, A02/C02, A03/C03, A05/C05, A06, A08, and A09. In addition, we validated statistically significant associations on A01, A07, and A10 in four genetic mapping populations, demonstrating that GWAS marker loci are indeed associated with resistance to L. maculans. One of the novel loci identified for the first time, Rlm12, conveys adult plant resistance and mapped within 13.2 kb from Arabidopsis R gene of TIR-NBS class. We showed that resistance loci are located in the vicinity of R genes of Arabidopsis thaliana and Brassica napus on the sequenced genome of B. napus cv. Darmor-bzh. Significantly associated SNP markers provide a valuable tool to enrich germplasm for favorable alleles in order to improve the level of resistance to L. maculans in canola.
Due to the large increase of canola production in Australia, current blackleg cultural control recommendations (extended rotation length and isolation distance from canola stubble) are not adhered to by farmers in many canola-producing regions. Canola crops are increasingly being sown in short rotation and, in many instances, adjacent to paddocks containing canola stubble. In this study, the level of disease in commercial canola crops was determined for different rotations and distances from canola stubble. There was a strong relationship between the presence of canola stubble from the previous year (6-month-old stubble) and distance to current canola crops, but no relationship between the presence of older (18–42 month old) stubble and distance to current canola crops. Blackleg severity was highest where canola crops had been sown adjacent to 6-month-old canola stubble, with the level of blackleg severity decreasing markedly in the first 100 m. Disease severity then generally declined up to 500 m. Plants 500–1000 m from 6-month-old stubble had similar levels of blackleg infection. Blackleg severity was similar between canola crops sown into 18-month-old canola stubble (short rotation) and crops sown into paddocks that had no history of canola for at least the previous 3 years (long rotation). Based on these findings, we recommend that canola crops should be sown at distances greater than 100 m and preferably 500 m from last season's canola stubble, rather than extending rotation length between crops.
BackgroundResistance to the blackleg disease of Brassica napus (canola/oilseed rape), caused by the hemibiotrophic fungal pathogen Leptosphaeria maculans, is determined by both race-specific resistance (R) genes and quantitative resistance loci (QTL), or adult-plant resistance (APR). While the introgression of R genes into breeding material is relatively simple, QTL are often detected sporadically, making them harder to capture in breeding programs. For the effective deployment of APR in crop varieties, resistance QTL need to have a reliable influence on phenotype in multiple environments and be well defined genetically to enable marker-assisted selection (MAS).ResultsDoubled-haploid populations produced from the susceptible B. napus variety Topas and APR varieties AG-Castle and AV-Sapphire were analysed for resistance to blackleg in two locations over 3 and 4 years, respectively. Three stable QTL were detected in each population, with two loci appearing to be common to both APR varieties. Physical delineation of three QTL regions was sufficient to identify candidate defense-related genes, including a cluster of cysteine-rich receptor-like kinases contained within a 49 gene QTL interval on chromosome A01. Individual L. maculans isolates were used to define the physical intervals for the race-specific R genes Rlm3 and Rlm4 and to identify QTL common to both field studies and the cotyledon resistance response.ConclusionThrough multi-environment QTL analysis we have identified and delineated four significant and stable QTL suitable for MAS of quantitative blackleg resistance in B. napus, and identified candidate genes which potentially play a role in quantitative defense responses to L. maculans.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0877-2) contains supplementary material, which is available to authorized users.
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