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Bacterial wilt caused by Clavibacter michiganensis subsp. insidiosus (Cmi) is an integral part of chronic vascular wilt of alfalfa. The aim of the presented study was to determine the virulence of Cmi strains and later on to evaluate the level of resistance of alfalfa varieties registered in the Czech Republic and of some standard check varieties. Three methods of inoculation were compared at greenhouse conditions. The severity of the disease was determined on the basis of symptoms appearing on the cross section of the roots. The Cmi strain 12/5/98 and its re-isolates showed a high level of virulence. Freeze-dried isolates lost their virulence. The best differentiation of tested varieties according to the level of resistance was obtained by using successive inoculation on cotyledons, stems and roots. Of tested alfalfa varieties, Vernal and Roamer were classified as resistant and Nitranka, Orca and Europe as the most susceptible. Alfalfa varieties (Czech and European) registered in the Czech Republic recently proved lower disease severity in comparison with ones registered formerly (r = –0.538, p (0.05) = 0.553). The above mentioned fact evidences the progress of Cmi resistant alfalfa breeding in Europe
Fungal entomopathogens can naturally regulate populations of various insects. The entomopathogen Beauveria bassiana (Bals.- Criv.) Vuill. is also able to endophytically colonize different plants. Endophytic colonization by entomopathogens may provide a source of indirect interactions between fungi and insects and has been associated with the ability of the fungus to control insect pests. The tomato leaf miner, Tuta absoluta (Meyrick) is considered one of the most devastating pests of tomato (Solanum lycopersicum L.) and its difficult control is due to its miner habit, short life cycle, and high reproductive capacity. The aims of this study were: (i) to use three different techniques, i.e. leaf spraying, seed immersion and root dipping, for the endophytic inoculation of B. bassiana in tomato plants; and (ii) to assess the effect of B. bassiana on tomato leaf consumption and mortality of T. absoluta after inoculation. The percentage of colonization by B. bassiana was assessed 7, 14 and 28 days after inoculation. All inoculation techniques employed allowed the recovery of B. bassiana, although our results showed significant differences between techniques. Leaf spraying was the most effective, with the highest percentage of colonization recorded 7 days after inoculation. We also evaluated (i) the effect on the mortality of T. absoluta by direct contact with conidia of B. bassiana, and (ii) the effect on tomato leaf consumption and mortality by indirect contact through ingestion of inoculated plant tissues with B. bassiana. Mortality bioassays showed that B. bassiana infected T. absoluta, either by direct contact or indirectly, via ingestion of inoculated tomato leaves. Direct contact showed a higher percentage of mortality and a lower median survival time (MST) than indirect contact. Significant differences in the mortality percentages of T. absoluta after exposure with B. bassiana were found among the treatments and the control. Our results suggest that the endophytic inoculation of B. bassiana in tomato crops provides the basis for further investigation, which should focus on the virulence of the endophytic B. bassiana against T. absoluta.
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