Evaluation of 'Hamlin' sweet orange plus 'Montenegrina' mandarin somatic hybrid for tolerance to Xanthomonas axonopodis pv. citri and Xylella fastidiosa

Asiatic citrus canker (ACC), caused by Xanthomonas axonopodis Starr & Garces pv. citri (Hasse) Vauterin et al., and citrus variegated chlorosis (CVC), caused by Xylella fastidiosa Wells et al., are considered the main diseases affecting sweet orange scion varieties in Brazil. Among commercial varieties, mandarins and tangerines are recognized as tolerant to these pathogens. We report herein the production of 'Hamlin' sweet orange (Citrus sinensis L. Osbeck) + 'Montenegrina' mandarin (Citrus deliciosa Ten.) allotetraploid somatic hybrid plants by protoplast fusion with improved disease tolerance that could be used as a donor of resistance genes in interploid hybridisation. Somatic hybridisation was confirmed by leaf morphology, flow cytometry and RAPD analyses. The somatic hybrid was propagated by grafting and cultivated in a screenhouse for tolerance assays. For X. axonopodis pv. citri assays, buds were collected from both 'Hamlin' sweet orange and the somatic hybrid and grafted onto 'Cleopatra' mandarin (Citrus reshni hort. ex Tanaka). As a negative control, buds from 'Mexerica Tardia' mandarin (C. deliciosa) were collected and grafted onto 'Cleopatra' mandarin. Two-month old plants with at least one young vegetative flush were individually spray-inoculated with a 106 CFU mL(-1) X. axonopodis pv. citri suspension and incubated in a growth chamber, at 27 degrees C, under 16-h photoperiod. The somatic hybrid showed a statistically significant reduction in susceptibility to ACC 30 days after inoculation. Compared to 'Hamlin' sweet orange, disease severity was reduced by 70%, with similar tolerance to that of the mandarin negative control. For X. fastidiosa assays, buds were collected from the somatic hybrid and its parental plants and grafted onto 'Rangpur' lime (Citrus limonia Osbeck). The developed plants were needle-inoculated with a X. fastidiosa suspension (8.7 x 10(10) CFU mL-1)