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Analyses of air samples for ascospores of Leptosphaeria maculans and L.biglobosa by light microscopy and molecular techniques

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Kaczmarek J. , Jedryczka M. , Fitt B. D. L. , Lucas J. A. , Latunde-Dada A. O.

Journal of Applied Genetics

2009

tom 50

nr 4

411-419

Spores of many fungal pathogens are dispersed by wind. Detection of these airborne inocula is important in forecasting both the onset and the risk of epiphytotics. Species-specific primers targeted at the internal transcribed spacer (ITS) region of Leptosphaeria maculans and L. biglobosa ? the causal organisms of phoma stem canker and stem lesions of Brassica spp., including oilseed rape ? were used to detect DNA extracted from particles deposited on tapes obtained from a spore trap operated in Rarwino (northwest Poland) from September to November in 2004 and 2006. The quantities of DNA assessed by traditional end-point PCR and quantitative real-time PCR were compared to microscopic counts of airborne ascospores. Results of this study showed that fluctuations in timing of ascospore release corresponded to the dynamics of combined concentrations of DNA from L. maculans and L. biglobosa, with significant positive correlations between ascospore number and DNA yield. Thus the utilization of PCR-based molecular diagnostic techniques enabled the detection, identification, and accurate quantification of airborne inoculum at the species level. Moreover, real-time PCR was more sensitive than traditional PCR, especially in years with low ascospore numbers.

Transgenic lines of rape resistant to infection of turnip mosaic virus (tum

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Lehmann P. , Kozubek E. , Wojciechowski A.

Biotechnologia

1998

nr 1

129-139

Transgenic Brassica napus var. oleifera cv. Westar plants that express the coat protein gene of the turnip mosaic virus (TuMV) were generated. The transgenic plant lines that contained one, two and at least three transgenes inserts in both homozygous and hemizygous conditions were produced and examined. Twelve different transgenic plant lines (in T3 generation) were analysed for resistance to TuMV. Three different responses were observed when the transgenic plant lines were inoculated with TuMV; 1) some were highly reistant, 2) some were medium resistant and, 3) some were susceptible to TuMV infection. We did not find any correlation between the number of copies of the transgene and virus resistance. The most promissing transgenic line was used to breed a few winter cultivars of rape to induce the pathogen-mediated virus resistance in these cultivars.

The practical use of molecular markers in hybrid breeding of winter oilseed rape (Brassica napus L. var. oleifera)

100%

Poplawska W. , Liersch A. , Bartkowiak-Broda I.

Biotechnologia

2004

nr 2

17-22

Breeding of oilseed rape hybrid varieties in Poland is based on CMS ogura hybrydization system. The marker assisted selection is used in selection of parental lines of F1 hybrids. The markers of alleles of restorer gene Rfo are the most important in breeding programs. Also, the investigations on genetic distance of hybrid parental lines using molecular markers are undertaken aiming at its application for preliminary selection of F1 combinations.