Partial hybridization in wide crosses between cultivated sunflower and the perennial Helianthus species H. mollis and H. orgyalis.

To obtain introgressed sunflower lines with improved disease resistance, interspecific crosses were performed with foreign perennial species. We report on several unusual features displayed by these hybrid plants. The methods used to produce the kernels affected yield and genotypes of progeny. Phenotypic traits and DNA markers were investigated in 97 plants derived from cross-pollination between annual diploid cultivated sunflower (Helianthus annuus) and the perennial diploid species H. mollis or H. orgyalis, and the reverse reciprocal crosses. The level of hybridization in progeny was determined using RAPD and RFLP markers. Hybridization was performed by leaving embryos to develop normally on the head (classical crossing) or using embryo rescue. All observed plants derived from H. mollis were diploid (2n = 34). Phenotypes were predominantly similar to the female when cultivated sunflower was the female parent. Progeny from crosses using a wild species as the female parent resembled that parent. Thus, reciprocal crosses led to different progeny. F1 sister progeny shared different sets of molecular markers representing a few of those of the wild species used as the pollen donor. Our results indicate mechanisms leading to the unusual event of partial hybridization. Possible mechanisms behind these unusual events and their possible impact on evolution are discussed.

Occurrence of partial hybrids in wide crosses between sunflower ( Helianthus annuus) and perennial species H. mollisand H. orgyalis.

Hybridisation between the annual diploid sunflower ( Helianthus annuus)and the perennial diploid species Helianthus mollis and Helianthus orgyalis was obtained by means of a normal crossing procedure or embryo rescue. Hybridisation success was low. All plants examined cytologically appeared to be diploid. However, the phenotypes of these diploids were not intermediate between the parents and, despite great variation, they resembled the female parent-type predominantly. Thirty five percent of plants issued from sunflower pollinated with perennial Helianthus had a phenotype resembling the female sunflower parent. On average, only 5% of the minimum number of expected RAPD and RFLP bands from male parents were recovered in plants produced from mature seeds after pollination of sunflower by H. mollis. More hybrids were found among plants obtained from embryo rescue, with an average of 25% of the male parent bands recovered per plant. Analysis of individual plants indicated the occurrence of various levels of hybridisation. There was a significant positive correlation between the number of phenotype traits related to hybrid status and the number of bands derived from the male parent. A single hybrid plant might possibly represent the product of a 'normal' hybridisation event. The mechanisms behind these unusual events and the consequences for the breeder are discussed.

Specifying the introgressed regions from H. argophyllus in cultivated sunflower (Helianthus annuus L.) to mark Phomopsis resistance genes.

A method based upon targetting of introgressed markers in a Phomopsis-resistant line (R) of cultivated sunflower, issuing from a H. argophyllus cross was used to mark the Phomopsis resistance regions. Our study was based upon 203 F(3) families derived from a cross between an inbred line susceptible to Phomopsis (S1) and the introgressed resistant line (R). Families were checked for Phomopsis resistance level in a design with replicated plots and natural infection was re-inforced by pieces of contaminated stems. Thirty four primers were employed for RAPD analysis. Out of 102 polymorphic fragments between (S1) and H. argophyllus, seven were still present in (R) suggesting that they marked introgressions of H. argophyllus into (R). The F(2) plants were scored for the presence or absence of 19 fragments obtained from five primers, and the relationships between the presence/absence of fragments in F(2) plants and Phomopsis resistance/susceptiblity in the F(3) progenies was determined by using an analysis of variance. We found that at least two introgressed regions, as well as favourable factors from sunflower, contributed to the level of Phomopsis resistance in cultivated sunflower.

Mapping genetic factors controlling pollen viability in an interspecific cross in Helianthus sect. Helianthus.

Segregation of 48 genetic markers, including one CMS restorer gene, one morphological character gene, six isozymes and 40 RAPD loci, was scored in a backcross progeny of an interspecific hybrid H. argophyllusxH. annuus cv RHA274. A linkage map was generated taking into account segregation distortions for 11 of the 48 loci in the frame of two different models considering locus-pair segregation in the context of either independent selection pressures or non-equilibrated parental classes. The map consists of nine linkage groups and nine isolated markers covering 390 cM. Approximately half of the plants of the BC1 were male fertile as expected for the segregation of one dominant male-fertility restorer gene; however, these displayed a large range of variation for pollen viability. About 80% of this variation was explained by three genomic regions located on linkage groups 1, 2 and 3. The observation of meiotic chromosomes revealed a significant rate of mispairing (rod bivalents and tetravalents) in tight correlation with pollen viability, indicating that chromosome rearrangements (translocations) are the preponderant factors reducing pollen viability in this progeny. Cytogenetic and mapping data suggest that the three genomic regions involved in pollen-viability variation are located close to translocation points which differentiate the parental-species karyotypes. Segregation distortion was observed for loci correlated with pollen-viability variation. These were most likely the result of two possible suggested mechanisms.

Mitochondrial DNA RFLP and genetical studies of cytoplasmic male sterility in the sunflower (Helianthus annuus).

Fifteen sunflower (Helianthus annuus L.) cytoplasmic male-sterile, and a single male-fertile, cytotypes were studied by both mtDNA (mitochondrial DNA) restriction fragment length polymorphism (RFLP) and genetical analysis of male-fertility restoration patterns. It was found by multivariate analysis that the two methods of identification of cytoplasmic male sterility (CMS) should be of use in sunflower breeding programs. The RFLP study distinguished 13 groups based on differences in mtDNA organization. DNA molecular diversity occurs both within and between the Helianthus species from which the steriles originate. The mitochondrial genes analyzed present specific molecular configurations for each type of sterility studied. The analysis of male-fertility restoration separated the cytotypes into 12 groups. The associations of CMS and inbred restorer lines indicated the presence of specific nuclear genes involved in cytoplasmic male-sterility restoration.

Cytoplasmic male sterility in sunflower: comparison of molecular biology and genetic studies.

The genetics of male fertility restoration and the RFLP of mitochondrial DNA were studied for 16 sunflower cytoplasms (15 male-sterile and a male-fertile). Male fertility restoration/male sterility maintenance patterns distinguished 12 cytotypes. Four cytoplasms were completely unrestored so they were not distinguished genetically. The sunflower lines, tested for their restorer/maintenance reaction, showed that there was a continuous range between 0% and 100% of restorer genotypes according to the CMS considered. Restoration/maintenance patterns indicated that at least some restorer genes are specific to certain CMS. RFLP of mitochondrial DNA revealed specific differences between the cytotypes studied. Three restriction enzymes and 12 probes permitted distinction of 13 cytotypes. No relationship exists between CMS cytotypes and the species from which they originated. For genetical and mitochondrial RFLP studies, phenograms were constructed according to the similarity indexes between cytotypes. Most of the CMS defined by restoration patterns correspond with a restriction fragment pattern of mitochondrial DNA.