Abstract

ZAMORA-RODRIGUEZ, Victoria et al. Use of shoot-tip grafting in vitro to eliminate ´Candidatus liberibacter asiaticus´ in citrus cultivars in Cuba. Rev. Protección Veg. [online]. 2015, vol.30, n.2, pp. 123-132. ISSN 2224-4697.

The aim of this research was to optimize the in vitro shoot-tip grafting (STG) technique to eliminate Candidatus Liberibacter asiaticus (CLas) from Citrus infected cultivars. Five citrus cultivars (Satsuma Tropical and Parson's Special mandarins, Sunburst IVIA 200-C-1 and Nova IVIA 74-7 hybrids of mandarins and Persian SRA-58 lime) showing symptoms of huanglongbing (HLB) were used as bud source:. Three different apex sizes, composed of the apical meristem with two, three and four leaf primordia (lp) (0.1 - 0.4 mm), were tested to standardize the apex size required to eliminate CLas. The presence of the pathogen was verified by the conventional PCR in the symptomatic plants and the nested PCR in the asymptomatic ones. The final success percentage of the shoot-tip grafting was 71.79%; 46.15% for the apex with 2 lp and over 80% for apexes with 3 and 4 lp. Sixty per cent of all the shoot-tip grafting grew normally, and this parameter was over 40% when apexes with 2 and 3 lpwere used, whereas it reached 80.61% when the apex with 4 lp was used. The 100% of the shoot-tip grafted plants were diagnosed as CLas free and this status was kept for four years regardless the shoot-tip size used. The results confirmed the reliability of the STG for removing CLas from citrus infected cultivars.

Keywords : shoot-tip grafting; sanitation; huanglongbing; PCR; nPCR.

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