Effects and side effects of penicillin injection in huanglongbing affected grapefruit trees

Trunk injection with penicillin has been tested to control citrus huanglongbing (HLB), but side effects and environmental safety must be assured before approval of penicillin injection can be considered. We investigated effects of penicillin injection on densities of Candidatus Liberibacter asiaticus (Las) in leaves, as well as culturable bacterial populations in rhizospheres and petioles of grapefruit trees in field and greenhouse experiments. Trees were injected with penicillin G, and leaf and root concentrations were assessed in bioassays with Bacillus subtilis. Las densities were determined by qPCR, and bacteria were isolated on a low carbon medium from roots plus rhizosphere and surface-sterilized petioles at various times after penicillin injection. Selected bacterial isolates were tested for penicillin resistance (20 mg/mL) and glyphosate resistance (7000 mg/mL), because glyphosate is widely used and cross-resistance against antibiotics had been documented. One month after penicillin injection half of the greenhouse trees were inoculated with Phytophthora nicotianae. Cycle threshold (Ct) values of Las in old and young leaves significantly increased 90 days after trunk injection with penicillin. Bacterial populations in petioles and root-rhizospheres initially increased after penicillin injections, probably due to nutrient release, then returned to control levels after one week. Penicillin resistance was common in isolates from penicillininjected and control trees (30e94%). Significantly more glyphosate resistant than sensitive isolates were penicillin resistant (81% versus 52%). Phytophthora root rot was not increased after penicillin injection. Thus, side effects of penicillin injection tested here were minimal, while Las titers were reduced after