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Abstract Background Sunflower broomrape (Orobanche cumana Wallr.) is a holoparasitic plant that jeopardizes sunflower production in most areas of Europe and Asia. Recently, populations with increased virulence, classified as race GGV, have been identified in Southern Spain’s Guadalquivir Valley gene pool. These populations overcome resistance genes in hybrids resistant to the predominant race FGV. This study aimed to (i) determine the inheritance and map the avirulence trait segregating in a cross between O. cumana individuals from populations EK23 (FGV) and IN201 (GGV), and (ii) characterize the host effect on the IN201 parental population allelic diversity. Results A segregating population consisting of 144 F2:3 families was evaluated for virulence using a differential sunflower genotype (Hybrid 1, resistant to race FGV and susceptible to race GGV) and genotyped with SNP markers. The ratio of avirulent to virulent F2:3 families was not significantly different to 1:3 (χ2 = 0.93; P = 0.34), indicating monogenic control of the avirulence/virulence trait. The Avr G−GV locus was mapped on the upper end of O. cumana chromosome 2, 9.2 cM distal from the SNP markers OS04791 and OS02805. Secretome analysis in the Avr G−GV region revealed a cysteine-rich CAP superfamily- and a glucan 1,3-beta-glucosidase family 3-encoding genes as possible candidates for Avr G−GV . SNP allelic analysis on the IN201 population parasitizing a highly susceptible genotype or the differential genotype Hybrid 1 showed that (i) IN201 structure was shaped towards virulent alleles at SNP loci linked to Avr G−GV (ii) there were significant allelic frequency differences associated with the host genotype at Avr G−GV –linked loci. Conclusions This study mapped for the first time an avirulence gene in parasitic plants using a classical genetic approach, confirmed a gene-for-gene model in the O.cumana –sunflower system, and showed the implication of this single avirulence gene in determining the structure of broomrape populations subjected to selection pressure posed by a resistant genotype. The results will contribute to a better understanding of the interaction between crops and weedy parasitic plants, and to effectively manage evolution of virulence by sustainable control strategies based on host genetic resistance.

Sunflower broomrape (Orobanche cumana Wallr.) was detected in 2010 for the first time in sunflower fields of Tunisia, in the Béja region. No information is available on the race composition and genetic diversity of the broomrape populations parasitizing sunflower in that area. Plant tissue and seeds were collected from nine populations in Béja Sud (ORD, ORE), Amdoun (ORF, ORG), and Béja Nord (ORH to ORL) areas of the Béja region. Virulence studies on populations ORD, ORH and ORK revealed that the ORD population was essentially race E, whereas race G individuals were present in ORH and ORK poulations, with greater frequency in the ORK population. Cluster analysis of inter-population relatedness indicated that Tunisian populations were more related to Eastern European than to Spanish populations, with the Béja Nord populations clustering separately from the rest of Tunisian populations. Analysis at the intra-population level of the ORD, ORG, ORH and ORK populations revealed the existence of two gene pools (GP1 and GP2), that were present in all populations at different frequencies. GP2 was at a lower frequency (1/14) in ORD and ORG but at higher frequencies in ORH (5/15) and ORK (8/14). This indicates that GP1 was probably an initial introduction of a race E population, while GP2 is most likely a later introduction of a race G population, introduced initially in Béja Nord area but now spreading to other areas in the region. The risk of a generalized expansion of race G of broomrape in this sunflower cultivation area is discussed.

IntroductionThe sunflower broomrape (Orobanche cumana Wallr.) gene pools of the Guadalquivir Valley and Cuenca province in Spain had predominantly race-F virulence. A new race G was observed recently in the Guadalquivir Valley potentially due to the genetic recombination of the avirulence genes of both gene pools.MethodsIn this research, we have studied populations with atypical virulence from Cuenca. These populations parasitize on DEB2 sunflower line, resistant to all race-G populations evaluated. Ten populations collected in Cuenca province were evaluated with sunflower differential lines and genotyped with 67 SNP markers.ResultsAlthough genetic recombination with individuals of the Guadalquivir Valley gene pool has been observed in most populations, recombination of avirulence genes was discarded as the cause of the new virulence because the population with the highest degree of attack on DEB2 showed no introgression from an external gene pool. Accordingly, a point mutation is proposed as the putative cause of the new virulence.DiscussionThe present study provided a detailed characterization of each population, including the accurate classification of the individuals belonging to each of the classical Spanish gene pools, F1 hybrids, and those that evolved from hybridization between both gene pools. This information is essential to understand how sunflower broomrape populations are evolving in Spain, which in turn may be helpful to understand the dynamics of sunflower broomrape populations in other areas of the world and use this information to develop durable strategies for resistance breeding.

IntroductionSunflower breeding for resistance to the parasitic plant sunflower broomrape (Orobanche cumana Wallr.) requires the identification of novel resistance genes. In this research, we conducted a genome-wide association study (GWAS) to identify QTLs associated with broomrape resistance.MethodsThe marker-trait associations were examined across a germplasm set composed of 104 sunflower accessions. They were genotyped with a 600k AXIOM® genome-wide array and evaluated for resistance to three populations of the parasite with varying levels of virulence (races EFR, FGV, and GTK) in two environments.Results and DiscussionThe analysis of the genetic structure of the germplasm set revealed the presence of two main groups. The application of optimized treatments based on the general linear model (GLM) and the mixed linear model (MLM) allowed the detection of 14 SNP markers significantly associated with broomrape resistance. The highest number of marker-trait associations were identified on chromosome 3, clustered in two different genomic regions of this chromosome. Other associations were identified on chromosomes 5, 10, 13, and 16. Candidate genes for the main genomic regions associated with broomrape resistance were studied and discussed. Particularly, two significant SNPs on chromosome 3 associated with races EFR and FGV were found at two tightly linked SWEET sugar transporter genes. The results of this study have confirmed the role of some QTL on resistance to sunflower broomrape and have revealed new ones that may play an important role in the development of durable resistance to this parasitic weed in sunflower.

Orobanche cumana Wall., sunflower broomrape, is one of the major pests for the sunflower crop. Breeding for resistant varieties in sunflower has been the most efficient method to control this parasitic weed. However, more virulent broomrape populations continuously emerge by overcoming genetic resistance. It is thus essential to identify new broomrape resistances acting at various stages of the interaction and combine them to improve resistance durability. In this study, 71 wild sunflowers and wild relatives accessions from 16 Helianthus species were screened in pots for their resistance to broomrape at the late emergence stage. From this initial screen, 18 accessions from 9 species showing resistance, were phenotyped at early stages of the interaction: the induction of broomrape seed germination by sunflower root exudates, the attachment to the host root and the development of tubercles in rhizotron assays. We showed that wild Helianthus accessions are an important source of resistance to the most virulent broomrape races, affecting various stages of the interaction: the inability to induce broomrape seed germination, the development of incompatible attachments or necrotic tubercles, and the arrest of emerged structure growth. Cytological studies of incompatible attachments showed that several cellular mechanisms were shared among resistant Helianthus species.

Sunflower broomrape (Orobanche cumana Wallr.) is a holoparasitic plant that causes major yield losses to sunflower crops in the Old World. Efforts to understand how this parasitic weed recognizes and interacts with sunflowers are important for developing long-term genetic resistance strategies. However, such studies are hampered by the lack of genetic tools for O. cumana. The objectives of this research were to construct a genetic linkage map of this species using SSR and SNP markers, and mapping the Pg locus that is involved in plant pigmentation. The genetic map was developed from the progenies of a cross between the O. cumana inbred lines EK-12 and EK-A1, which originated from populations belonging to two distant and geographically separated gene pools identified in Spain. The inbred lines also differed in plant pigmentation, with EK-A1 lacking anthocyanin pigmentation (pgpg genotype). A genetic map comprising 26 SSR and 701 SNP markers was constructed, which displayed 19 linkage groups (LGs), corresponding to the 19 chromosome pairs of O. cumana. The total length of the map was 1795.7 cM, with an average distance between two adjacent positions of 2.5 cM and a maximum map distance of 41.9 cM. The Pg locus mapped to LG19 between the SNP markers OS02468 and OS01653 at 7.5 and 3.4 cM, respectively. This study constitutes the first linkage map and trait mapping study in Orobanche spp., laying a key foundation for further genome characterization and providing a basis for mapping additional traits such as those having a key role in parasitism.

In Spain, sunflower broomrape (Orobanche cumana Wallr.) has been restricted to Cuenca province in Central Spain, and the Guadalquivir Valley in Southern Spain, that represent different gene pools of the species. This pathogenic plant has now spread to other areas such as Castilla y León region in Northern Spain. The racial status and genetic diversity were investigated in six populations of sunflower broomrape collected in several provinces of Castilla y León. Evaluation of virulence to a set of differential host genotypes classified three of the populations as race F, while the other three populations were classified as a race below F, probably race E. Genetic diversity analysis using a set of 20 SSR markers showed that the broomrape populations from new areas of Northern Spain are mainly derived from the Guadalquivir Valley gene pool. Introgression from the Cuenca gene pool was observed in one of the populations, in which the percentage of polymorphic loci was 31%, Shanon´s diversity index was 0.17, and the average number of pairwise differences was 1.69, compared to zero for the three parameters in the other five populations. The absence of race F individuals in the populations classified as race below F indicated that seed migration took place, probably before the generalized expansion of race F in the Guadalquivir Valley area, at the beginning of the 1990s.

Ethiopian mustard (Brassica carinata A. Braun) line BCT-6, with reduced γ-tocopherol content in the seeds, has been previously developed. The objective of this research was to conduct a genetic analysis of seed tocopherols in this line. BCT-6 was crossed with the conventional line C-101 and the F1, F2, and BC plant generations were analyzed. Generation mean analysis using individual scaling tests indicated that reduced γ-tocopherol content fitted an additive-dominant genetic model with predominance of additive effects and absence of epistatic interactions. This was confirmed through a joint scaling test and additional testing of the goodness of fit of the model. Conversely, epistatic interactions were identified for total tocopherol content. Estimation of the minimum number of genes suggested that both γ- and total tocopherol content may be controlled by two genes. A positive correlation between total tocopherol content and the proportion of γ-tocopherol was identified in the F2 generation. Additional research on the feasibility of developing germplasm with high tocopherol content and reduced concentration of γ-tocopherol is required.

Broomrape, caused by Orobanche cumana, has affected sunflowers since the early 20th century in Eastern Europe. Currently, it limits sunflower oil production in Southern and Eastern Europe and in some areas of Asia, causing around 50% seed losses when susceptible hybrids are grown. Covered in this review are aspects such as: biological processes that are common to Orobanche spp. and/or particular to O. cumana in sunflower, genetic resistance and its mechanisms, races of the parasite identified in different countries throughout the time and their increasing virulence, and breeding for resistance to some herbicides as a novel control option. The main purpose is to present an updated and, as far as possible, complete picture of the way both the parasitic weed and its host crop have evolved in time, and how they co-exist in the current agriculture. Additionally, we propose a system for determining the races of the parasite that can be internationally adopted from now. In the context of minimal harmful effects on the environment, changing patterns of land use in farming systems, and global environment changes, the final goal of this work is to provide all those interested in parasites from field crops and their integrated management compiled information on the sunflower – O. cumana system as a case study.

The objective of this study was to evaluate the dynamics of tocopherols in cotyledons and radicles from sunflower seeds with high and low total tocopherol content, mainly in the α-tocopherol form, and from seeds with increased proportions of β-, γ-, and δ-tocopherol, both under dark and light conditions. Tocopherol content was measured every 24 h from 1 to 12 days after sowing. In all cases, the content of individual tocopherol forms in the cotyledons and radicles was reduced along the sampling period, which was more pronounced under light conditions. The presence of light had a slightly greater effect on α- and γ-tocopherol than on β- and δ-tocopherol. A marked light effect was also observed on total tocopherol content, with light promoting the reduction of tocopherol content in cotyledons and radicles. The study revealed only slight differences in the patterns of tocopherol losses in lines with different tocopherol profiles, both under dark and light conditions, which suggested that the partial replacement of α-tocopherol by other tocopherol forms had no great impact on the protection against oxidative damage in seedlings.

Orobanche cumana is a holoparasitic plant naturally distributed from central Asia to south-eastern Europe, where it parasitizes wild Asteraceae species. It is also an important parasitic weed of sunflower crops. The objective of this research was to investigate genetic diversity, population structure, and virulence on sunflower of O. cumana populations parasitizing wild plants in eastern Bulgaria. Fresh tissue of eight O. cumana populations and mature seeds of four of them were collected in situ on wild hosts. Genetic diversity and population structure were studied with SSR markers and compared to weedy populations. Two main gene pools were identified in Bulgarian populations, with most of the populations having intermediate characteristics. Cross-inoculation experiments revealed that O. cumana populations collected on wild species possessed similar ability to parasitize sunflower to those collected on sunflower. The results were explained on the basis of an effective genetic exchange between populations parasitizing sunflower crops and those parasitizing wild species. The occurrence of bidirectional gene flow may have an impact on wild populations, as new physiological races continuously emerge in weedy populations. Also, genetic variability of wild populations may favour the ability of weedy populations to overcome sunflower resistance mechanisms.