Broomrape (Orobanche Cumana Wallr.) resistance breeding utilizing wild Helianthus species

Wild Helianthus species possess valuable resistance genes for sunflower broomrape (Orobanche cumana Wallr.), especially the 39 largely underutilized perennial species. Resistance to race F has been transferred into a cultivated background via bridging of interspecific amphiploids. More recently, a single dominant gene resistant to race G was identified in annual H. debilis ssp. tardi-florus and transferred into cultivated HA 89. Interspecific crosses between wild annual Helianthus species and cultivated lines are relatively easy compared to those involving wild perennial species, which were made easier only after the development of embryo rescue techniques. Interspecific amphiploids resulting from colchicine treatment of F1 hybrids provide bridging materials for transferring genes without relying on embryo rescue. Among the diploid, tetraploid, and hexaploid perennial species, the speed of gene utilization follows the ploidy level of diploids, tetraploids, and hexaploids due to the time-consuming backcrosses required to eliminate the extra chromosomes in the latter two groups. In the development of pre-breeding materials, the retention rate of genetic material of the wild species is another concern with each additional backcross. For crosses involving tetraploid and hexaploid wild perennials, the use of 2n=51 chromosome F1 or BC1F1 generation, as pollen source, could accelerate chromosome reduction to 2n=34 in BC1F1 or BC2F1, resulting in useful materials with fewer backcrosses for trait selection.This work is partially funded by a National Sclerotinia Grant awarded to C.C. Jan.Peer Reviewe

Phylogenetic relationships and genetic diversity among Orobanche cumana Wallr. and O. cernua L. (Orobanchaceae) populations in the Iberian Peninsula

Orobanche cumana is found in the Iberian Peninsula as an allochthonous species parasitizing exclusively sunflower, in contrast to the closely related species Orobanche cernua, which is an autochthonous species that only parasitizes wild Asteraceae hosts. Ten O. cumana populations were collected in the two traditional areas of sunflower broomrape occurrence, the Guadalquivir Valley, Southern Spain (six populations) and Cuenca province, Central Spain (four populations). Twelve O. cernua populations were collected on wild hosts across its natural distribution area in Southeastern Spain. Genetic relationships within and between both sets of populations were studied using a set of 50 robust and co-dominant SSR markers from O. cumana. The results supported the taxonomic separation of the two species and the existence of two distant genetic groups for O. cumana, one in Guadalquivir Valley and another one in Cuenca province. The inter- and intra-population variability was extremely low for O. cumana, whereas the overall genetic diversity was much higher for O. cernua. The genetic structure of O. cumana populations probably reflects a founder effect, with the two genetically distant groups deriving from separate introduction events. The high degree of genetic differentiation observed in O. cernua is mainly explained on the basis of restricted gene flow due to ecological barriers together with the occurrence of a predominantly self-pollinating mating system. Complementary diversity studies on both species in its current distribution area are required for understanding global genetic variability and evolutionary characteristics of the parasitism.The study was partially funded by Fundación Ramón Areces, Madrid, Spain. R. Pineda-Martos was the recipient of a Ph.D. fellowship from the Spanish National Research Council (CSIC) (JAEPre_08_00370).Peer Reviewe

A novel sunflower broomrape race with unusual virulence potentially caused by a mutation

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

Genetic studies in sunflower broomrape

Much research has been conducted to identify sources of genetic resistance to sunflower broomrape (Orobanche cumana Wallr.) and to study their mode of inheritance. However, studies on the parasite have been scarce. This manuscript reviews three genetic studies in sunflower broomrape. First, the inheritance of the absence of pigmentation in a natural mutant of this species with yellow plant color phenotype was studied. In a first stage, lines from the unpigmented mutant and a normally pigmented population were developed by several generations of self-pollination. Plants of both lines were crossed and the F1, F2, and F3 generations were evaluated. The results indicated that plant pigmentation is controlled by a partially dominant allele at a single locus. Second, the unpigmented mutant was used to evaluate outcrossing potential of the species. Two experiments in which single unpigmented plants were surrounded by normally pigmented plants were conducted under pot and field conditions. The cross-fertilization rate was estimated as the percentage of F1 hybrids in the progenies of unpigmented plants, which averaged 21.5% in the pot and 28.8% in the field experiment. The results indicated that, under the conditions of this study, the species was not strictly self-pollinated. Finally, the inheritance of avirulence was studied in crosses of plants from lines of O. cumana races E and F, developed by several generations of self-pollination. The F1 and F3 generations were evaluated on the differential line P-1380 carrying the race-E resistance gene Or5. The results suggested that race E avirulence and race F virulence on P-1380 are allelic and controlled by a single locus, which confirmed the gene-for-gene theory for the O. cumana-sunflower interaction.The manuscript reviews research partially funded by Fundación Ramón Areces, Madrid. The contribution of Dr. Enrique Quesada Moraga, entomologist from the University of Córdoba, Spain, to taxonomic classification of pollinators is gratefully acknowledged. R. Pineda-Martos was the recipient of a PhD fellowship from the Spanish National Research Council (CSIC) (JAEPre_08_00370)Peer Reviewe

Association mapping for broomrape resistance in sunflower

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

Wild Helianthus species: A reservoir of resistance genes for sustainable pyramidal resistance to broomrape 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.This study was performed in the frame of a 3-year project (ResODiv), funded by “Promosol” (the association of French Sunflower and Rapeseed Breeders for promoting these crops).Peer reviewe

International consortium on sunflower broomrape resistance

15th World Congress on Parasitic Plants, 30th June–5th July, Amsterdam, the Netherland.Peer reviewe

Changes in plastochromanol-8 and tocopherols during germination in Ethiopian mustard lines with contrasting tocopherol levels

Plastochromanol-8 and tocopherols are natural antioxidants with vitamin E activity that protect seeds and plants from oxidative damage. Whereas α-tocopherol is predominant in photosynthetic tissues, γ-tocopherol is more abundant in seeds. Ethiopian mustard (Brassica carinata A. Braun) seeds contain varying levels of both tocopherol forms. The objective of this research was to study changes in plastochromanol-8 and tocopherol contents during seed germination in Ethiopian mustard lines with contrasting seed tocopherol levels, and to evaluate their influence on seed germination and seedling growth. Cotyledons and radicles of seven lines with contrasting seed tocopherol levels and a control line were sampled periodically until 120 h after sowing (HAS) under light and dark conditions, and were analysed for plastochromanol-8 and tocopherol contents. The main factors genotype, tissue, light and HAS were significant for all the traits, excepting the light effect for total tocopherol content. Most of the lines showed a gradual decrease in γ-tocopherol that was nearly completely replaced by α-tocopherol at 120 HAS. However, line BCT-4, with high γ-tocopherol content in seeds, exhibited unusually high γ-tocopherol content in cotyledons and radicles during the whole sampling period. Light treatment and initial seed tocopherol levels had no influence on germination percentage. Despite significant differences for seedling development among genotypes, no clear association between seedling development and seed tocopherols was observed. The results revealed that the unusual presence of high γ-tocopherol levels in cotyledons and radicles of line BCT-4 did not have an effect on seedling growth. © 2014 Cambridge University Press.This work was supported by the Spanish government through Ministerio de Educación y Ciencia and EU FEDER funds, research project AGL2007-62 834/AGR and PhD student fellowship EBES-2008-003080 to E.G.-N.Peer Reviewe