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Polyploidy is widespread in plants and has played a major role in the evolution and diversification of the plant kingdom. Unreduced (2n) gametes are an interesting tool for polyploidisation and the creation of genetic variation in plant breeding. Especially in ornamentals, polyploidisation can broaden attractive features within a species. A Begonia collection was screened on the occurrence of 2n pollen with the aid of four different techniques: pollen size measurement, flow cytometric analysis of nuclei isolated from germinated and non germinated pollen, investigation of the microsporogenesis and analysis of progeny. In ten of the 70 screened genotypes (B. dregei, B. pearcei, B. 'Anna Christina', B. 'Bubbles', B. 'Florence Rita', B. 'Orococo', B. 'Rubaiyat', B. 'Spatflacier', B. 'Tamo' and B276), large pollen were observed with a rather spherical than normal ellipsoidal shape. Flow cytometric data proved that these aberrantly shaped pollen were associated with 2n ploidy levels, although they were not always viable. Meiotic aberrations in these large pollen producers resulted mainly in dyads although also monads, triads and polyads were observed. Successful crosses were obtained with B. dregei, B. 'Orococo', and B276 as pollinators; DNA content had increased in all or a part of the progeny. The results show that the occurrence of 2n pollen is not a rare phenomenon in Begonia

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-A Chlamydophila psittaci species-specific real-time PCR targeting the rDNA ribosomal spacer was developed as well as a genotype-specific real-time PCR targeting the Cp. psittaci outer membrane protein A (ompA) gene. The SYBR Green-based species-specific real-time PCR detected Cp. psittaci genotypes A to F, and the recently discovered E/B genotype. The genotype-specific real-time PCR could easily distinguish genotypes C, D, F by use of TaqMan probes. Genotypes A, B and E could not be distinguished from each other by simply using TaqMan probes. For this purpose, non-fluorescent competitor oligonucleotides, had to be used next to the TaqMan probes. Genotype E/B could only be detected by use of a minor groove binder (MGB) probe. Both real-time PCR assays allowed reproducible, sensitive (10 rDNA or ompA copies/µL DNA extract) and specific detection of Cp. psittaci DNA. The genotype-specific real-time PCR was compared to ompA sequencing and ompA restriction fragment length polymorphism (RFLP) analysis using five Cp. psittaci field isolates (99, 61/8, 7344/2, 8615/1 and 7778B15) each consisting of two different genotypes. The currently developed real-time PCR assays were used in a case study on a veterinary school and a turkey farm. In the veterinary school, Cp. psittaci genotypes D, E/B and F infection were detected in all five groups of turkeys, and one veterinarian who was taking care of all these turkeys. On the turkey farm, the presence of two Cp. psittaci genotype B infection waves was demonstrated in one randomly selected turkey, the first wave at the age of 6 weeks, and the second at the age of 12 weeks.

The powdery mildew fungus Podosphaera pannosa (Wallr.: Fr.) de Bary (syn. Sphaerotheca pannosa) is a major problem on roses worldwide. Twenty-six monoconidial isolates of Podosphaera collected on roses and Prunus spp. in Belgium, Germany, France, Denmark, Israel and The Netherlands were characterized on the basis of differential reactions on in vitro rose genotypes and Prunus avium L. and by DNA sequence analysis of the rDNA ITS (internal transcribed spacer) region. Twenty-four isolates were determined as P. pannosa. Amongst these, different groups could be distinguished. A first group of 18 isolates was highly virulent on rose and avirulent or very weakly virulent on P. avium. A second group of four isolates was highly virulent on both rose and P. avium. Analysis of the ITS sequence could discriminate these two groups of P. pannosa strains by a one base pair difference. Finally, two isolates of powdery mildew collected on Prunus sp. could be classified as P. pannosa based on their ITS sequence, which was identical to the ITS sequence of the isolates only highly virulent on roses. However, these two isolates were not able to infect roses. These results indicate that different strains of P. pannosa exist with varying host specificity. We demonstrated by ITS sequencing and plant reactions that the host range of P. pannosa comprises roses and Prunus spp.www.blackwell-synergy.com

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The genome sizes of a Begonia collection comprising 37 species and 23 hybrids of African, Asiatic, Middle American, and South American origin were screened using flow cytometry. Within the collection, 1C values varied between 0.23 and 1.46 pg DNA. Genome sizes were, in most cases, not positively correlated with chromosome number, but with pollen size. A 12-fold difference in mean chromosome size was found between the genotypes with the largest and smallest chromosomes. In general, chromosomes from South American genotypes were smaller than chromosomes of African, Asian, or Middle American genotypes, except for B. boliviensis and B. pearcei. Cytological chromosome studies in different genotypes showed variable chromosome numbers, length, width, and total chromosome volume, which confirmed the diversity in genome size. Large secondary constrictions were present in several investigated genotypes. These data show that chromosome number and structure exhibit a great deal of variation within the genus Begonia, and likely help to explain the large number of taxa found within the genus.

In this study, treatments of both trifluralin (at 10, 100 and 1000 mu M) and N2O (in the form of gas under pressure) were applied to Begonia flower buds to induce the formation of 2n pollen. Three male fertile species (B. cucullata, B. subvillosa var. leptotricha and B. fischeri) and two male sterile hybrids (B. schmidtiana x B. cucullata and B. subvillosa var. leptotricha x B. cucullata) were treated. Pollen size, which is related to pollen DNA content, increased after both N2O and trifluralin treatments, but the induction of large pollen was genotype dependent. Trifluralin induced large pollen only in the male fertile species, while N2O treatments induced fertile 2n pollen in the male sterile B. schmidtiana x B. cucullata. Cytological studies showed that trifluralin induced multinuclear monads that resulted in 4n gametes in stead of 2n gametes. In general, large pollen obtained after trifluralin treatments showed low germination capability, while large pollen obtained after N2O treatments retained high germination capability. Seedlings with raised ploidy level could only be obtained after crosses were performed with large pollen obtained from N2O treatments. Hence, N2O treatments are preferable to the use of trifluralin to induce 2n gametes in Begonia