Quantitative trait analysis of seed yield and other complex traits in hybrid spring rapeseed (Brassica napus L.): 2. Identification of alleles from unadapted germplasm

Unadapted germplasm may contain alleles that could improve hybrid cultivars of spring oilseed Brassica napus. Quantitative trait loci (QTL) mapping was used to identify potentially useful alleles from two unadapted germplasm sources, a Chinese winter cultivar and a re-synthesized B. napus, that increase seed yield when introgressed into a B. napus spring hybrid combination. Two populations of 160 doubled haploid (DH) lines were created from crosses between the unadapted germplasm source and a genetically engineered male-fertility restorer line (P1804). A genetically engineered male-sterile tester line was used to create hybrids with each DH line (testcrosses). The two DH line populations were evaluated in two environments and the two testcross populations were evaluated in three or four environments for seed yield and other agronomic traits. Several genomic regions were found in the two testcross populations which contained QTL for seed yield. The map positions of QTL for days to flowering and resistance to a bacterial leaf blight disease coincided with QTL for seed yield and other agronomic traits, suggesting the occurrence of pleiotropic or linked effects. For two hybrid seed yield QTL, the favorable alleles increasing seed yield originated from the unadapted parents, and one of these QTL was detected in multiple environments and in both populations. In this QTL region, a chromosome rearrangement was identified in P1804, which may have affected seed yield.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

Confirmation of QTL controlling seed yield in spring canola (Brassica napus L.) hybrids

Allelic effects observed in QTL discovery experiments must be confirmed to be useful in subsequent breeding efforts. Two QTL affecting seed yield of spring hybrid canola (Brassica napus L.) were previously identified in two populations of inbred backcross lines (IBLs) containing germplasm introgressed from a winter cultivar. The effects of favorable alleles at these QTL were retested by crossing two selected IBLs (M5 and M31) to three spring canola lines having different genetic backgrounds. Doubled haploid (DH) lines derived from each F₁ were genotyped with RFLP markers flanking the QTL and grouped into the four possible QTL genotypes. For the first field experiment, DH lines derived by crossing the M5 line to one spring line were crossed to two female testers and evaluated as individual testcross progenies in one environment. QTL genotypes had large variances and were not significantly different. A second field experiment was conducted using the DH lines from the first experiment and two other sets of DH lines derived from the M31 line crossed to two different spring canola lines. Individual lines within each QTL genotype of each set were bulked and crossed to the same testers used in Experiment 1. Bulked hybrid seeds of each QTL genotype were planted in a split-split plot randomized block design and 12 replicates. QTL genotypes had smaller variances in this experiment, and the effects of one QTL were confirmed in some genetic backgrounds. These results suggest that bulking of QTL genotypes and use of an appropriate experimental design with many replicates are needed to detect small differences between QTL genotypes.     

Evaluation of QTL alleles from exotic sources for hybrid seed yield in the original and different genetic backgrounds of spring-type <Emphasis Type="Italic">Brassica napus</Emphasis> L.

Previously identified alleles at quantitative trait loci (QTL) for hybrid seed yield were re-evaluated in the same genetic background (in hybrid combination with the same tester) as the original QTL mapping study and also evaluated in a different genetic background (in hybrid combination with two different testers). The QTL were identified from wide crosses of exotic germplasm sources with spring-type Brassica napus L., in which alleles from the exotic germplasm sources increased hybrid seed yield. Results from the re-evaluation of six QTL, in the same genetic background and hybrid combination, indicate that several of the exotic donor QTL alleles did increase hybrid seed yield and could be successfully used for improving the original single-cross hybrid. However, results from the evaluation of seven QTL (including the same six previous QTL) in a new genetic background, in combination with two new testers, indicate that the exotic QTL alleles were often no different or produced significantly lower hybrid seed yield than the spring QTL alleles. In all studies, the QTL were also very sensitive to environmental interactions. Thus, our results indicate that although these exotic sources contain favorable QTL alleles when introgressed into one spring hybrid background, the effects are not predictive of other genetic backgrounds or hybrid combinations. Although QTL affecting hybrid seed yield have been identified, comparisons of multiple QTL alleles are needed to determine the most favorable allele at each locus. Characterization of QTL complementation across testers will be required to predict their effects in multiple hybrid combinations.     

Mapping of days to flower and seed yield in spring oilseed <Emphasis Type="Italic">Brassica napus</Emphasis> carrying genome content introgressed from <Emphasis Type="Italic">Brassica oleracea</Emphasis>

Earliness of flowering and maturity and high seed yield are important objectives of breeding spring Brassica napus canola. Previously, we have introgressed earliness of flowering from Brassica oleracea into spring B. napus canola through interspecific crossing between these two species. In this paper, we report quantitative trait locus (QTL) mapping of days to flower and seed yield by use of publicly available markers and markers designed based on flowering time genes and a doubled haploid population, derived from crossing of the spring canola parent and an early flowering line developed from a B. napus × B. oleracea cross, tested in nine field trials for over 5 years. Five genomic regions associated with days to flower were identified on C1, C2, C3, and C6 of which the single QTL of C1 was detected in all trials; in all cases, the allele introgressed from B. oleracea reduced the number of days to flower. BLASTn search in the Brassica genomes located the physical position of the QTL markers and identified putative flowering time genes in these regions. In the case of seed yield, ten QTL from eight linkage groups were detected; however, none could be consistently detected in all trials. The QTL region of C1 associated with days to flower did not show significant association with seed yield in more than 80% of the field trials; however, in a single trial, the allele introgressed from B. oleracea exerted a negative effect on seed yield. Thus, the genomic regions and molecular markers identified in this research could potentially be used in breeding for the development of early flowering B. napus canola cultivars without affecting seed yield in a majority of the environments.     

Population-based resequencing analysis of improved wheat germplasm at wheat leaf rust resistance locus Lr21

Little is known about the genetic control of heterosis in the complex polyploid crop species oilseed rape (Brassica napus L.). In this study, two large doubled-haploid (DH) mapping populations and two corresponding sets of backcrossed test hybrids (THs) were analysed in controlled greenhouse experiments and extensive field trials for seedling biomass and yield performance traits, respectively. Genetic maps from the two populations, aligned with the help of common simple sequence repeat markers, were used to localise and compare quantitative trait loci (QTL) related to the expression of heterosis for seedling developmental traits, plant height at flowering, thousand seed mass, seeds per silique, siliques per unit area and seed yield. QTL were mapped using data from the respective DH populations, their corresponding TH populations and from mid-parent heterosis (MPH) data, allowing additive and dominance effects along with digenic epistatic interactions to be estimated. A number of genome regions containing numerous heterosis-related QTL involved in different traits and at different developmental stages were identified at corresponding map positions in the two populations. The co-localisation of per se QTL from the DH population datasets with heterosis-related QTL from the MPH data could indicate regulatory loci that may also contribute to fixed heterosis in the highly duplicated B. napus genome. Given the key role of epistatic interactions in the expression of heterosis in oilseed rape, these QTL hotspots might harbour genes involved in regulation of heterosis (including fixed heterosis) for different traits throughout the plant life cycle, including a significant overall influence on heterosis for seed yield.     

Quantitative trait loci for resistance to Sclerotinia sclerotiorum and its association with a homeologous non-reciprocal transposition in Brassica napus L.

Sclerotinia stem rot, caused by fungus Sclerotinia sclerotiorum, is one of the most devastating diseases in rapeseed (Brassica napus L.). We report the identification of Quantitative trait loci (QTL) involved in the resistance to S. sclerotiorum in two segregating populations of DH lines: the HUA population, derived from a cross between a partially resistant Chinese winter line (Hua dbl2) and a susceptible European spring line (P1804); and the MS population, derived from a partially resistant French winter cultivar (Major) and a susceptible Canadian spring cultivar (Stellar). A petiole inoculation technique and two scoring methods, days to wilt (DW) and stem lesion length (SLL), were used for the resistance assessment. A total of eight genomic regions affecting resistance were detected in the HUA population, with four of these regions affecting both measures of resistance. Only one region, which affected both measurements, was detected in the MS population. Individual QTL explained 6–22% of the variance. At five of the QTL from both populations, alleles from the resistant parent contributed to the resistance. QTL on N2 from the HUA population had the highest LOD score and R ² value and was detected for SLL in the first evaluation. The N12 resistance allele in Hua dbl2 was detected in a region containing a homeologous non-reciprocal transposition (HNRT) from the resistance-containing portion of N2. This result suggests that QTL in the N12.N2 HNRT enhanced the resistance of Hua dbl2 by increasing the dosage of resistance genes. The relationship of QTL from different genetic backgrounds and their associations with other agronomic traits are discussed.     

Mapping of QTL for tiller number at different stages of growth in wheat using double haploid and immortalized F<Subscript>2</Subscript> populations

Effective tiller number is one of the most important traits for wheat (Triticum aestivum L.) yield, but the inheritance of tillering is poorly understood. A set of 168 doubled haploid (DH) lines derivatives of a cross between two winter wheat cultivars (Huapei 3 and Yumai 57), and an immortalized F₂ (IF₂) population generated by randomly permutated intermating of these DHs were investigated, and QTLs of tillering related to the maximum tillering of pre-winter (MTW), maximum tillering in spring (MTS), and effective tillering in harvest (ETH) were mapped. Phenotypic data were collected for the two populations from two different environments. Using inclusive composite interval mapping (ICIM), a total of 9 and 18 significant QTL were detected across environments for tillering in the DH and IF₂ populations, respectively. Four QTLs were common between two populations. A major QTL located on the 5D chromosome with the allele originating from Yumai 57 was detected and increased 1.92 and 3.55 tillers in MTW and MTS, respectively. QTLs (QMts6D, QEth6D) having a neighbouring marker interval at Xswes679.1 and Xcfa2129 on chromosome 6D was detected in MTS and ETH. These results provide a better understanding of the genetic factors for selectively expressing the control of tiller number in different growth stages and facilitate marker-assisted selection strategy in breeding.     

Detection of QTL for six yield-related traits in oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>) using DH and immortalized F<Subscript>2</Subscript> populations

The inheritance of yield-related traits in rapeseed (Brassica napus) is poorly understood, and the investigations on mapping of quantitative trait loci (QTL) for such traits are only few. QTL related to six traits were mapped which include plant height (PH), height of lowest primary effective branch (HPB), length of main inflorescence (LMI), silique length (SL), number of primary branches (FB) and silique density (SD). A set of 258 doubled haploid (DH) lines derivatives of a cross between a canola variety Quantum and a resynthesized B. napus line No.2127-17, and a fixed immortalized F₂ (designated as IF₂) population generated by randomly permutated intermating of these DHs were investigated. A genetic linkage map was constructed using 208 SSR and 189 SRAP markers for the DH population. Phenotypic data were collected from three environments for the two populations. Using composite interval mapping analyses, 30 and 22 significant QTL were repeatedly detected across environments for the six traits in the DH and IF₂ populations, respectively. Twenty-nine QTL were common between the two populations. The directions of parental contribution for all common QTL were the same, showing a great potential for marker-assisted selection in improving these traits. Some chromosomal regions harbor QTL for multiple traits, which were consistent with significant phenotypic correlations observed among traits. The results provided a better understanding of the genetic factors controlling yield-related traits in rapeseed. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Genetic analysis and mapping of genes controlling freezing tolerance in oilseedBrassica

Freezing tolerance is the ability of plants to survive subfreezing temperatures and is a major component of winter survival. In order to study the genetic regulation of freezing tolerance, an F2 population ofBrassica rapa and a doubled haploid population ofBrassica napus were assayedin vitro for relative freezing tolerance of acclimated and nonacclimated plants. Linkage maps developed previously were used to identify putative quantitative trait loci (QTL). Genomic regions with significant effects on freezing tolerance were not found for theB. napus population, but forB. rapa four regions were associated with acclimated freezing tolerance (FTA) and acclimation ability (FTB), and two unliked regions were associated with nonacclimated freezing tolerance (FTN). Acclimation ability was regulated by genes with very small additive effects and both positive and negative dominance effects. The allele from the winter parent at the FTN QTL had positive additive effects, but negative dominance effects. RFLP loci detected by a cold-induced and a stress-related cDNA fromArabidopsis thaliana mapped near two QTL for FTA/FTB. Further tests are needed to determine if alleles at these loci are responsible for the QTL effects we detected.     

Broadening the avenue of intersubgenomic heterosis in oilseed Brassica

Accumulated evidence has shown that each of the three basic Brassica genomes (A, B and C) has undergone profound changes in different species, and has led to the concept of the “subgenome”. Significant intersubgenomic heterosis was observed in hybrids between traditional Brassica napus and first generation lines of new type B. napus. The latter were produced by the partial introgression of subgenomic components from different species into B. napus. To increase the proportion of exotic subgenomic components and thus achieve stronger heterosis, lines of first generation new type B. napus were intercrossed with each other, and subjected to intensive marker-assisted selection to develop the second generation of new type B. napus. The second generation showed better agronomic traits and a higher proportion of introgression of subgenomic components than did the first generation. Compared with the commercial hybrid and the hybrids produced with the first generation new type B. napus, the novel hybrids showed stronger heterosis for seed yield during the 2 years of field trials. The extent of heterosis showed a significant positive correlation with the introgressed subgenomic components in the parental new type B. napus. To increase the content of the exotic subgenomic components further and to allow sustainable breeding of novel lines of new type B. napus, we initiated the development of a gene pool for new type B. napus that contained a substantial amount of genetic variation in the A^r and C^c genome. We discuss new approaches to broaden the avenue of intersubgenomic heterosis in oilseed Brassica.     

Hybridisation and introgression between Brassica napus and B. rapa in the Netherlands

We used flow cytometry, chromosome counting and AFLP markers to investigate gene flow from the crop plant oilseed rape, Brassica napus (AACC) to wild B. rapa (AA) in the Netherlands. From 89 B. napus source populations investigated, all near cropping fields or at transhipment sites, only 19 contained a B. rapa population within a 2.5‐km radius. During our survey we found only three populations with F1 hybrids (AAC), as recognized by their nine extra chromosomes and by flow cytometry. These hybrids were all collected in mixed populations where the two species grew in close proximity. Populations with F1 hybrids were not close to crops, but instead were located on road verges with highly disturbed soils, in which both species were probably recruited from the soil seed bank. Many plants in the F2, BC1 or higher backcrosses are expected to carry one to eight C chromosomes. However, these plants were not observed among the hybrids. We further investigated introgression with molecular markers (AFLP) and compared sympatric B. rapa populations (near populations of B. napus) with control populations of B. rapa (no B. napus within at least 7 km). We found no difference between sympatric and control populations in the number of C markers in B. rapa, nor did we find that these sympatric populations closely resembled B. napus. Our data show that hybrids occur but also suggest no recent introgression of alleles from the crop plant B. napus into wild B. rapa in the Dutch populations studied.     

Genetic analysis of phenylpropanoid metabolites associated with resistance against Verticillium longisporum in Brassica napus

Verticillium longisporum is a major threat to production of oilseed rape (Brassica napus) in Europe. The aim of the study was to develop new markers and obtain insights into putative mechanisms and pathways involved in the resistance reaction. A genetic approach was used to identify quantitative trait loci (QTL) for V. longisporum resistance and metabolic traits potentially influencing resistance in a B. napus mapping population. Resistance to V. longisporum was mapped in a doubled haploid (DH) population from a cross between the partially resistant winter oilseed rape variety Express 617 and a resistant resynthesized B. napus line, R53. One major resistance QTL contributed by R53 was identified on chromosome C5, while a further, minor QTL contributed by Express 617 was detected on chromosome C1. Markers flanking the QTL also significantly correlated with V. longisporum resistance in four further DH populations derived from crosses between elite oilseed rape cultivars and other resynthesized B. napus lines originating from genetically and geographically diverse brassica A and C genome donors. The tightly-linked markers developed enable the combination of favorable alleles for novel resistance loci from resynthesized B. napus materials with existing resistance loci from commercial breeding lines. HPLC analysis of hypocotyls from infected DH lines revealed that concentrations of a number of phenylpropanoids were correlated with V. longisporum resistance. QTL for some of these phenylpropanoids were also found to co-localize with the QTL for V. longisporum resistance. Genes from the phenylpropanoid pathway are suggested as candidates for V. longisporum resistance.     

Identification of molecular markers associated with linoleic acid desaturation in Brassica napus

 Linolenic acid is a component of canola oil that is readily oxidized, which results in a reduced frying stability and shelf life of the oil. The reduction of linolenic acid in canola seed has therefore been an important breeding objective for many years. The inheritance of linolenic acid concentrations in seed oil is polygenic and is also strongly influenced by the environment. For these reasons, molecular markers are sought to assist in early and reliable selection of desired low linolenic acid genotypes in breeding programmes. Molecular markers associated with low linolenic acid loci were identified in a doubled-haploid population derived from a cross between the Brassica napus lines, ‘Apollo’ (low linolenic)×YN90-1016 (high linolenic) using RAPDs and bulked segregant analysis. A total of 16 markers were distributed over three linkage groups, which individually accounted for 32%, 14% and 5% of the phenotypic variation in linolenic acid content. The rapeseed fad3 gene was mapped near the locus controlling 14% of the variation. The mode of inheritance appeared to be additive, and a QTL analysis showed that collectively the three loci explained 51% of the phenotypic variation within this population. PCR fragments for low linolenic acid ‘Apollo’ alleles (3% linolenic acid) were identified at all three loci. Simultaneous selection for low linolenic acid ‘Apollo’ alleles at each locus resulted in a group of DH lines with 4.0% linolenic acid. The use of these makers in the breeding programme will enhance the breeding of low linolenic acid B. napus cultivars for production in Canada. Received: 23 September 1997 / Accepted: 21 October 1997     

Genetic basis of soybean adaptation to North American vs. Asian mega-environments in two independent populations from Canadian × Chinese crosses

One of the goals of plant breeding is to increase yield with improved quality characters. Plant introductions (PI) are a rich source of favorable alleles that could improve different characters in modern soybean [Glycine max (L.) Merril] including yield. The objectives of this study were to identify yield QTL underlying the genetic basis for differential adaptation of soybeans to the Canadian, United States or Chinese mega-environments (ME) and to evaluate the relationship and colocalization between yield and agronomic traits QTL. Two crosses between high-yielding Canadian cultivars and elite Chinese cultivars, OAC Millennium × Heinong 38 and Pioneer 9071 × #8902, were used to develop two recombinant inbred line (RIL) populations. Both populations were evaluated at different locations in Ontario, Canada; Minnesota, United States (US), Heilongjiang and Jilin, China, in 2009 and 2010. Significant variation for yield was observed among the RILs of both populations across the three hypothetical ME. Two yield QTL (linked to the interval Satt364–Satt591 and Satt277) and one yield QTL (linked to marker Sat_341) were identified by single-factor ANOVA and interval mapping across all ME in populations 1 and 2, respectively. The most frequent top ten high-yielding lines across all ME carried most of the high-yielding alleles of the QTL that were identified in two and three ME. Both parents contributed favorable alleles, which suggests that not only the adapted parent but also the PI parents are potential sources of beneficial alleles in reciprocal environments. Other QTL were detected also at two and one ME. Most of the yield QTL were co-localized with a QTL associated with an agronomic trait in one, two, or three ME in just one or in both populations. Results suggested that most of the variation observed in seed yield can be explained by the variation of different agronomic traits such a maturity, lodging and height. Novel alleles coming from PI can favorably contribute, directly or indirectly, to seed yield and the utilization of QTL detected across one, two or three ME would facilitate the new allele introgression into breeding populations in both North America and China.     

Molecular dissection of interspecific variation between Gossypium hirsutum and Gossypium barbadense (cotton) by a backcross-self approach: I. Fiber elongation

The current study is the first installment of an effort to explore the secondary gene pool for the enhancement of Upland cotton (Gossypium hirsutum L.) germplasm. We developed advanced-generation backcross populations by first crossing G. hirsutum cv. Tamcot 2111 and G. barbadense cv. Pima S6, then independently backcrossing F₁ plants to the G. hirsutum parent for three cycles. Genome-wide mapping revealed introgressed alleles at an average of 7.3% of loci in each BC₃F₁ plant, collectively representing G. barbadense introgression over about 70% of the genome. Twenty-four BC₃F₁ plants were selfed to generate 24 BC₃F₂ families of 22–172 plants per family (totaling 2,976 plants), which were field-tested for fiber elongation and genetically mapped. One-way analysis of variance detected 22 non-overlapping quantitative trail loci (QTLs) distributed over 15 different chromosomes. The percentage of variance explained by individual loci ranged from 8% to 28%. Although the G. barbadense parent has lower fiber elongation than the G. hirsutum parent, the G. barbadense allele contributed to increased fiber elongation at 64% of the QTLs. Two-way analysis of variance detected significant (P<0.001) among-family genotype effects and genotype×family interactions in two and eight regions, respectively, suggesting that the phenotypic effects of some introgressed chromosomal segments are dependent upon the presence/absence of other chromosomal segments.Electronic Supplementary Material Supplementary material is available for this article at     

Alien introgression in rice

Rice (Oryza sativa L.) productivity is affected by several biotic and abiotic stresses. The genetic variability for some of these stresses is limited in the cultivated rice germplasm. Moreover, changes in insect biotypes and disease races are a continuing threat to increased rice production. There is thus an urgent need to broaden the rice gene pool by introgressing genes for such traits from diverse sources. The wild species of Oryza representing AA, BB, CC, BBCC, CCDD, EE, FF, GG and HHJJ genomes are an important reservoir of useful genes. However, low crossability and limited recombination between chromosomes of cultivated and wild species limit the transfer of such genes. At IRRI, a series of hybrids and monosomic alien addition lines have been produced through embryo rescue following hybridization between rice and several distantly related species. Cytoplasmic male sterility and genes for resistance to grassy stunt virus and bacterial blight have been transferred from A genome wild species into rice. Similarly, genes for resistance to brown planthopper, bacterial blight and blast have also been introgressed across crossability barriers from distanly related species into rice. Some of the introgressed genes have been mapped via linkage to molecular markers. One of the genes Xa-21 introgressed from O. longistaminata has been cloned and physically mapped on chromosome 11 of rice using BAC library and flourescence in-situ hybridization. RFLP analysis revealed introgression from 11 of the 12 chromosomes of C genome species into rice. Introgression has also been obtained from other distant genomes (EE, FF, GG) into rice and in majority of the cases one or two RFLP markers were introgressed. Reciprocal replacement of RFLP alleles of wild species with the alleles of O. sativa indicates alien gene transfer through crossing over. The rapid recovery of recurrent phenotypes in BC₂ and BC₃ generations from wide crosses is an indication of limited recombination. Further cytogenetic and molecular investigations are required to determine precisely the mechanism of introgression of small chromosome segments from distant genomes in the face of limited homoeologous chromosome pairing. Future research should focus on enhancing recombination between homoeologous chromosomes. Introgression of QTL from wild species should be attempted to increase the yield potential of rice.     

Identification of positive yield QTL alleles from exotic soybean germplasm in two backcross populations

Increasing seed yield is an important breeding goal of soybean [Glycine max (L.) Merr.] improvement efforts. Due to the small number of ancestors and subsequent breeding and selection, the genetic base of current soybean cultivars in North America is narrow. The objective of this study was to map quantitative trait loci (QTL) in two backcross populations developed using soybean plant introductions as donor parents. The first population included 116 BC(2)F(3)-derived lines developed using "Elgin" as the recurrent parent and PI 436684 as the donor parent (E population). The second population included 93 BC(3)F(3)-derived lines developed with "Williams 82" as the recurrent parent and PI 90566-1 as the donor parent (W population). The two populations were evaluated with 1,536 SNP markers and during 2?years for seed yield and other agronomic traits. Genotypic and phenotypic data were analyzed using the programs MapQTL and QTLNetwork to identify major QTL and epistatic QTL. In the E population, two yield QTL were identified by both MapQTL and QTLNetwork, and the PI 436684 alleles were associated with yield increases. In the W population, a QTL allele from PI 90566-1 accounted for 30?% of the yield variation; however, the PI region was also associated with later maturity and shorter plant height. No epistasis for seed yield was identified in either population. No yield QTL was previously reported at the regions where these QTL map indicating that exotic germplasm can be a source of new alleles that can improve soybean yield.     

QTL analysis reveals context-dependent loci for seed glucosinolate trait in the oilseed <Emphasis Type="Italic">Brassica juncea</Emphasis>: importance of recurrent selection backcross scheme for the identification of ‘true’ QTL

Seed glucosinolate content in Brassica juncea is a complex quantitative trait. A recurrent selection backcross (RSB) method with a doubled haploid (DH) generation interspersing backcross generations was used for the introgression of low glucosinolate alleles from an east European gene pool B. juncea line, Heera into an Indian gene pool variety, Varuna. Phenotypic comparisons among the DH populations derived from early to advanced backcrosses revealed a shift in the mean values for various glucosinolates with the advancement of backcrossing, indicating a change in the selective values of the alleles with change in the genetic background due to the existence of epistasis and context dependencies. QTL mapping for various seed glucosinolates from early (F₁DH) and advanced generation (BC₄DH) populations confirmed the presence of epistasis and context dependency. The common QTL detected in both F₁DH and BC₄DH changed their R ² values from the former to the later generation. Some of the QTL detected in the F₁DH became irrelevant in the BC₄DH population. Further, new QTL were detected in the BC₄DH population for various glucosinolates. A validation study on a population of low glucosinolate DH lines derived from all the backcross generations of the RSB breeding programme revealed that the QTL detected in BC₄DH were the ‘true’ QTL. Using glucosinolate as an example, the study provides strong evidence for the importance of the RSB method for the identification of the ‘true’ QTL which would be significant for marker assisted introgression of a complex quantitative trait whose expression is influenced by epistatic interactions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Authors N. Ramchiary, N. C. Bisht, V. Gupta, A. Mukhopadhyay and N. Arumugam have contributed equally to this work.     

Identification of putative quantitative trait loci associated with a flavonoid related to resistance to cabbage seedpod weevil (Ceutorhynchus obstrictus) in canola derived from an intergeneric cross,Sinapis alba × Brassica napus

Key message

Kaempferol 3- O -sinapoyl-sophoroside 7- O -glucoside was putatively identified as the major component of a characteristic HPLC peak previously correlated with the reduction of cabbage seedpod weevil larval infestation in a novel canola genotype.

Abstract

The cabbage seedpod weevil (Ceutorhynchus obstrictus [Marsham]) (CSPW) is a serious pest of brassicaceous oilseed crops such as canola in both Europe and more recently in North America. At present, the only control strategy against CSPW is the application of insecticides. As an alternative more environmentally-friendly control strategy, we developed novel canola germplasm resistant to weevil attack through introgression of Sinapis alba DNA into Brassica napus by making the wide cross followed by embryo rescue and backcrossing to the B. napus parent. We have previously characterized resistant canola lines by metabolic profiling and were able to correlate reduction of larval infestation to the presence of a characteristic HPLC peak. In this study, we have putatively identified the major component in the peak using mass spectrometry as kaempferol 3-O-sinapoyl-sophoroside 7-O-glucoside (KSSG). We have also identified quantitative trait loci (QTL) associated with this HPLC peak in a mapping population consisting of more than 200 individual doubled haploid (DH) lines derived from a cross between CSW428 (the resistant parent) and SC030686 (the susceptible parent). This QTL accounted for approximately 9.5 % of the phenotypic variation in KSSG content. The observation that only one QTL was identified as surpassing the LOD threshold of 3.0 suggests that both parents may possess the positive alleles for other QTL that have not been detected in our study. This finding also indicates a complex regulatory mechanism for KSSG levels and provides an appropriate explanation for the large transgressive segregation observed in the DH lines of the QTL mapping population.