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The main objective in this research was the genetic analysis of heterosis in rapeseed at the QTL level. A linkage map comprising 235 SSR and 144 AFLP markers covering 2045 cM was constructed in a doubled-haploid population from a cross between the cultivar “Express” and the resynthesized line “R53.” In field experiments at four locations in Germany 250 doubled-haploid (DH) lines and their corresponding testcrosses with Express were evaluated for grain yield and three yield components. The heterosis ranged from 30% for grain yield to 0.7% for kernel weight. QTL were mapped using three different data sets, allowing the estimation of additive and dominance effects as well as digenic epistatic interactions. In total, 33 QTL were detected, of which 10 showed significant dominance effects. For grain yield, mainly complete dominance or overdominance was observed, whereas the other traits showed mainly partial dominance. A large number of epistatic interactions were detected. It was concluded that epistasis together with all levels of dominance from partial to overdominance is responsible for the expression of heterosis in rapeseed.

Hybrid breeding relies on the combination of parents from two differing heterotic groups. However, the genetic diversity in adapted oilseed rape breeding material is rather limited. Therefore, the use of resynthesized Brassica napus as a distant gene pool was investigated. Hybrids were derived from crosses between 44 resynthesized lines with a diverse genetic background and two male sterile winter oilseed rape tester lines. The hybrids were evaluated together with their parents and check cultivars in 2 years and five locations in Germany. Yield, plant height, seed oil, and protein content were monitored, and genetic distances were estimated with molecular markers (127 polymorphic RFLP fragments). Resynthesized lines varied in yield between 40.9 dt/ha and 21.5 dt/ha, or between 85.1 and 44.6% of check cultivar yields. Relative to check cultivars, hybrids varied from 91.6 to 116.6% in yield and from 94.5 to 103.3% in seed oil content. Mid-parent heterosis varied from −3.5 to 47.2% for yield. The genetic distance of parental lines was not significantly correlated with heterosis or hybrid yield. Although resynthesized lines do not meet the elite rapeseed standards, they are a valuable source for hybrid breeding due to their large distance from present breeding material and their high heterosis when combined with European winter oilseed rape.

Genetic variation in N uptake and utilization was investigated in a set of genetically diverse oilseed rape (Brassica napus L.) genotypes, including hybrid, modern, and old cultivars and resynthesized lines. A total of 36 genotypes were grown at seven locations with two levels of N supply: no fertilization and optimal fertilization (between 160 and 220 kg ha–1 N, depending on regional recommendations). The hybrid cultivars had the highest yields and the old land races and resynthesized lines had the lowest yields. This ranking of the various groups was consistent and independent of the N level. Genetic variation exists for both the uptake efficiency and the utilization efficiency. In the presence of low N levels, variation in N use efficiency is mainly due to differences in uptake efficiency whereas differences in utilization efficiency were more important in the presence of high N fertilization.

The rape stem weevil, Ceutorhynchus napi Gyll., is a serious pest of winter oilseed rape (Brassica napus L.) crops in Europe causing severe yield loss. In currently used oilseed rape cultivars no resistance to C. napi has been identified. Resynthesized lines of B. napus have potential to broaden the genetic variability and may improve resistance to insect pests. In this study, the susceptibility to C. napi of three cultivars, one breeding line and five resynthesized lines of oilseed rape was compared in a semi-field plot experiment under multi-choice conditions. Plant acceptance for oviposition was estimated by counting the number of C. napi larvae in stems. The larval instar index and the dry body mass were assessed as indicators of larval performance. The extent of larval feeding within stems was determined by the stem injury coefficient. Morphological stem traits and stem contents of glucosinolates were assessed as potential mediators of resistance. The resynthesized line S30 had significantly fewer larvae than the cultivars Express617 and Visby and the resynthesized lines L122 and L16. The low level of larval infestation in S30 was associated with a low larval instar and stem injury index. Low numbers of larvae were not correlated with the length or diameter of stems, and the level of stem glucosinolates. As indicated by the low larval infestation and slow larval development the resistance of S30 to C. napi is based on both antixenotic and antibiotic properties of the genotypes. The resynthesized line S30 should therefore be introduced into B. napus breeding programs to enhance resistance against C. napi.

Circulating tumor DNA (ctDNA) has demonstrated great potential as a noninvasive biomarker to assess minimal residual disease (MRD) and profile tumor genotypes in patients with non‐small‐cell lung cancer (NSCLC). However, little is known about its dynamics during and after tumor resection, or its potential for predicting clinical outcomes. Here, we applied a targeted‐capture high‐throughput sequencing approach to profile ctDNA at various disease milestones and assessed its predictive value in patients with early‐stage and locally advanced NSCLC. We prospectively enrolled 33 consecutive patients with stage IA to IIIB NSCLC undergoing curative‐intent tumor resection (median follow‐up: 26.2 months). From 21 patients, we serially collected 96 plasma samples before surgery, during surgery, 1–2 weeks postsurgery, and during follow‐up. Deep next‐generation sequencing using unique molecular identifiers was performed to identify and quantify tumor‐specific mutations in ctDNA. Twelve patients (57%) had detectable mutations in ctDNA before tumor resection. Both ctDNA detection rates and ctDNA concentrations were significantly higher in plasma obtained during surgery compared with presurgical specimens (57% versus 19% ctDNA detection rate, and 12.47 versus 6.64 ng·mL−1, respectively). Four patients (19%) remained ctDNA‐positive at 1–2 weeks after surgery, with all of them (100%) experiencing disease progression at later time points. In contrast, only 4 out of 12 ctDNA‐negative patients (33%) after surgery experienced relapse during follow‐up. Positive ctDNA in early postoperative plasma samples was associated with shorter progression‐free survival (P = 0.013) and overall survival (P = 0.004). Our findings suggest that, in early‐stage and locally advanced NSCLC, intraoperative plasma sampling results in high ctDNA detection rates and that ctDNA positivity early after resection identifies patients at risk for relapse. ctDNA profiling in plasma revealed higher levels and detection rates of ctDNA during tumor resection as compared to pretreatment time points in patients with early‐stage or locally advanced NSCLC. Moreover, patients testing positive for ctDNA immediately after tumor resection had worse clinical outcomes than patients with undetectable ctDNA. Our research highlights the role of ctDNA assessment for guiding treatment in patients with respectable NSCLC.

In oilseed rape (Brassica napus) semi-dwarf hybrid varieties from crosses between bzh dwarf and normal-type lines are of increasing interest. They have improved nitrogen (N) uptake, N-utilization and N-use efficiency compared to normal types. This study aimed to elucidate whether these N-related effects can be explained by the bzh shoot growth-type alone or also by differences in root traits. Root system size was measured using root electrical capacitance (EC) in field trials with two N levels in two sets of genotypes segregating for the bzh-locus: (1) 108 doubled haploid (DH) test hybrids in two seasons, 2010-2012, and (2) 16 near-isogenic hybrids in the 2016-17 season. Quantitative trait loci (QTL) for root EC were estimated in DH test hybrids. Seedling root architecture parameters were monitored in vitro. In vitro root growth showed a higher root: shoot ratio in bzh semi-dwarf hybrids. Root EC in field trials was higher at high N supply than at zero N fertilization. In most trials semi-dwarf hybrids had higher EC than normal-type hybrids, but they reduced root EC in response to N limitation more than normal types. Root EC was more heritable at the end of flowering (h2 = 0.73) than at the beginning of flowering (h2 = 0.36) in near-isogenic hybrids and had a lower heritability in trials of DH test hybrids (h2 = 0.27). A QTL for root EC in the genomic region of the bzh-locus on linkage group A06 was significant at zero N fertilization. Root EC proved to be a meaningful method in oilseed rape breeding programmes targeting root system size. The greater reduction of semi-dwarf root EC compared to the normal type under low N supply with simultaneous increase in N efficiency implies that in roots it is not a question of 'the more the merrier' and that the bzh root system reacts highly economically when N is scarce.

Resynthesized (Resyn) Brassica napus L. can be used to broaden the genetic diversity and to develop a heterotic genepool for rapeseed hybrid breeding. Domesticated vegetable types are usually employed as B . oleracea parents. We sought to evaluate the potential of wild species as parents for Resyn lines. Fifteen Resyn lines were derived by crossing wild B . oleracea ssp. oleracea and oilseed B. rapa , and 29 Resyn lines were generated from 10 wild Brassica species ( B. bourgaei, B. cretica, B. incana, B. insularis , B. hilarionis , B. macrocarpa , B. montana , B. rupestris , B. taurica , B. villosa ). Genetic distances were analyzed with AFLP markers for 71 Resyn lines from wild and domesticated B. oleracea , and compared with 55 winter, spring, vegetable, and Asian B. napus genotypes. The genetic distances clearly showed that Resyn lines with wild species provide a genetic diversity absent from the breeding material or Resyn lines from domesticated species. Forty-two Resyn lines were crossed with one or two winter oilseed rape testers, resulting in 64 hybrids that were grown in one year and four locations in Germany and France. The correlation between hybrid yield and genetic distance was slightly negative ( r  = −0.29). Most of the hybrids with Resyn lines from wild B. oleracea were lower in yield than hybrids with Resyn lines from domesticated B. oleracea . It is promising that Resyn lines descending from unselected wild B. oleracea accessions produced high-yielding hybrids when crossed with adapted genotypes: these Resyn lines would be suited to develop heterotic pools in hybrid breeding.

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease. Circulating tumor cells (CTC) in the blood are hypothesized as the means of systemic tumor spread. Blood obtained from healthy donors and patients with PDAC was therefore subject to size-based CTC-isolation. We additionally compared Kirsten rat sarcoma viral oncogene homolog ( KRAS ) mutations in pancreatic CTC and corresponding tumors, and evaluated their significance as prognostic markers. Samples from 68 individuals (58 PDAC patients, 10 healthy donors) were analyzed; CTCs were present in patients with UICC stage IA-IV tumors and none of the controls (p < 0.001). Patients with >3 CTC/ml had a trend for worse median overall survival (OS) than patients with 0.3–3 CTC/ml ( P  = 0.12). Surprisingly, CTCs harbored various KRAS mutations in codon 12 and 13. Patients with a KRAS G12V mutation in their CTC (n = 14) had a trend to better median OS (24.5 months) compared to patients with other (10 months), or no detectable KRAS mutations (8 months; P  = 0.04). KRAS mutations in CTC and corresponding tumor were discordant in 11 of 26 “tumor-CTC-pairs” (42%), while 15 (58%) had a matching mutation; survival was similar in both groups ( P  = 0.36). Genetic characterization, including mutations such as KRAS , may prove useful for prognosis and understanding of tumor biology.

Established allopolyploids are known to be genomically stable and fertile. However, in contrast, most newly resynthesized allopolyploids are infertile and meiotically unstable. Identifying the genetic factors responsible for genome stability in newly formed allopolyploid is key to understanding how 2 genomes come together to form a species. One hypothesis is that established allopolyploids may have inherited specific alleles from their diploid progenitors which conferred meiotic stability. Resynthesized Brassica napus lines are often unstable and infertile, unlike B. napus cultivars. We tested this hypothesis by characterizing 41 resynthesized B. napus lines produced by crosses between 8 Brassica rapa and 8 Brassica oleracea lines for copy number variation resulting from nonhomologous recombination events and fertility. We resequenced 8 B. rapa and 5 B. oleracea parent accessions and analyzed 19 resynthesized lines for allelic variation in a list of meiosis gene homologs. SNP genotyping was performed using the Illumina Infinium Brassica 60K array for 3 individuals per line. Self-pollinated seed set and genome stability (number of copy number variants) were significantly affected by the interaction between both B. rapa and B. oleracea parental genotypes. We identified 13 putative meiosis gene candidates which were significantly associated with frequency of copy number variants and which contained putatively harmful mutations in meiosis gene haplotypes for further investigation. Our results support the hypothesis that allelic variants inherited from parental genotypes affect genome stability and fertility in resynthesized rapeseed.

Resynthesized Brassica napus L. could be employed to increase the genetic variation in the narrow oilseed rape genepool and to establish a genetically distant genepool for hybrid breeding. One important criterion for selecting appropriate resynthesized lines is their genetic distance to adapted breeding material. In this study we estimated the genetic distances in a wide collection of 142 resynthesized lines and 57 winter and spring rapeseed cultivars from Europe, North America and Asia using RFLP marker data. The highest in-group genetic distance (0.36) was observed in 142 resynthesized lines compared to 0.21 in winter, 0.23 in summer, and 0.28 in Asian genotypes. The group of adapted breeding material clustered into three groups of winter-, spring-type, and Asian genotypes. Resynthesized lines did not form distinct subgroups in the cluster analysis, but large differences were revealed in the genetic distance of resynthesized lines to the adapted winter oilseed rape genepool. The highest distance to winter oilseed rape was found in resynthesized lines with parental lines from the Asian genepool as B. oleracea convar. botrytis var. alboglabra (Bail.) Sun or B. rapa ssp. chinensis (L.) Hanelt.