Transmission of 'Candidatus Liberibacter solanacearum' in carrot seeds
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AutorBertolini,E.; Teresani,G. R.; Loiseau,M.; Tanaka,F. A. O.; Barbe,S.; Martinez,C.; Gentit,P.; Lopez,M. M.; Cambra,M.
A protocol for the specific detection and quantification of Candidatus Liberibacter solanacearum' in carrot seeds using real-time PCR was developed. The bacterium was detected in 23 out of 54 carrot seed lots from 2010 to 2014, including seeds collected from diseased mother plants. The average total number of Ca. L. solanacearum' cells in individual seeds ranged from 48 +/- 33 to 210 +/- 67cells per seed from three seed lots, but using propidium monoazide to target live cells, 95% of the cells in one seed lot were found to be dead. Liberibacter-like cells were observed in the phloem sieve tubes of the seed coat and in the phloem of carrot leaf midrib from seedlings. The bacterium was detected as early as 30days post-germination, but more consistently after 90days, in seedlings grown from PCR positive seed lots in an insect-proof P2 level containment greenhouse. Between 12% and 42% of the seedlings from positive seed lots tested positive for Ca. L. solanacearum'. After 150days, symptoms of proliferation were observed in 12% of seedlings of cv. Maestro. Candidatus Liberibacter solanacearum' haplotype E was identified in the seeds and seedlings of cv. Maestro. No phytoplasmas were detected in seedlings with symptoms using a real-time assay for universal detection of phytoplasmas. The results show that to prevent the entry and establishment of the bacterium in new areas and its potential spread to other crops, control of Ca. L. solanacearum' in seed lots is required.