Xylella fastidiosa can be graft transmitted to healthy plants or

Xylella fastidiosa can be graft transmitted to healthy plants or vectored by several species Selleck CX 5461 of xylem-feeding leafhoppers (Redak et al., 2004). Antimicrobial peptides (AMPs) are defensive substances widespread in nature, being produced from bacteria to mammals (Sang & Blecha, 2008). The majority display common features such as a low molecular

mass, a positive net charge at physiological pH and an amphipathic structure (Bulet et al., 2004). Generally, AMPs disrupt the plasmatic membrane, causing the rapid death of microorganisms. Nevertheless, some AMPs can cross the microbial membrane acting on internal cellular targets (Brogden, 2005). The elucidation of the exact mode of action of AMPs is essential to allow the use of these substances to develop a new generation of antibiotics to control infections of both plants and animals. Gomesin is a short β-hairpin AMP (2270.4 Da) isolated from the hemocytes of the tarantula spider Acanthoscurria gomesiana (Silva et al.,

2000). Similar to LEE011 molecular weight most AMPs, gomesin is cationic and possesses an amphipatic structure (Mandard et al., 2002). Bacteria and fungi synthesize and secrete AMPs to inhibit the growth of neighboring microorganisms that compete for both space and nutrients (Sang & Blecha, 2008). Several bacteria and fungi species have been reported to compose an endophytic community that cohabits the xylem vessels of citrus plants along with X. fastidiosa (Araujo et al., 2002). Moreover, many tissues of insects, including salivary glands and the lining epithelial cells, produce AMPs to prevent infection (Bulet et al., 2004). Therefore, it is likely that X. fastidiosa may be exposed to AMPs during colonization of both the host plant and the insect vector. To verify whether and how X. fastidiosa would respond to AMPs, we evaluated the effect of gomesin on its global gene expression profile and on Dichloromethane dehalogenase X. fastidiosa colonization success of

tobacco plants. Ampicillin, streptomycin and tetracycline were from Sigma-Aldrich (St. Louis, MO). Gomesin (Silva et al., 2000) was synthesized as described previously (Fazio et al., 2006). Lyophilized gomesin and conventional antibiotic were reconstituted in sterile deionized water immediately before use. Virulent 9a5c and avirulent J1a12 X. fastidiosa strains (Li et al., 1999; Koide et al., 2004) were grown in periwinkle wilt (PW) broth medium (Davis et al., 1981) supplemented with 20% glucose as described previously (Zaini et al., 2008). Mid-log suspensions (OD600 nm=0.400) of either 9a5c or J1a12 strains were incubated with different concentrations of gomesin (0.14–9 μg mL−1), ampicillin (0.31 × 103–5 × 103 μg mL−1), streptomycin (0.19–100 μg mL−1), tetracycline (0.97–500 μg mL−1) or water as control. After 18 h at 28 °C, cells were harvested by centrifugation at 4000 g for 5 min at 25 °C, suspended in fresh PW medium and plated on 2% agar PW.

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