Effect of passage through the plant on virulence and endophytic behavioural adaptation in the entomopathogenic fungus Beauveria bassiana

Systemic crop protection using endophytic isolates of entomopathogenic fungi is at the forefront of IPM. Nonetheless, any potential trade-offs between virulence and endophytic behaviour must be elucidated if they are to be effectively used in pest management strategies. Here we investigated endophytic adaptation in an isolate of Beauveria bassiana following successive passage through melon, tomato and cotton tissues. Plants were sprayed with a suspension of B. bassiana endophytic isolate EABb 04/01-Tip to initiate endophytic colonization. Once colonization was established, the fungus was re-isolated from the plant, applied to another plant and re-isolated again; this was repeated to achieve three passages. After each passage, a conidial suspension of each isolate was used in bioassays to evaluate both virulence against 4th instar larvae of the model insect Galleria mellonella and to quantify the extent of endophytic activity in each respective host plant species. When sprayed leaves were inspected for fungal colonization, differences in percentage tissue colonization amongst the plant species were detected after the first re-isolation. Endophytic colonization rates in melon and tomato, which varied from 70 to 100%, were higher than those observed in cotton, which ranged from 40 to 50%; endophytic colonization in cotton increased to 75-100% after the third passage. This improvement in endophytic behaviour in cotton, an apparently suboptimal plant for fungal colonization, suggests an evolutionary adaptation to localized or transient endophytic colonization, while further assays are needed. Meanwhile, when endophytic colonization of nonsprayed leaves distant from the sprayed ones was investigated, endophytic activity was evident in all three crop species suggesting that movement within plants after successive passage increased the extent of endophytic colonization from transient to systemic. The present research highlights the potential for adaptation to