Pathogenicity of Lasiodiplodia theobromae isolated from cocoa dieback disease in South Sulawesi, Indonesia

Abstract

Dieback disease is one of the constraints to cocoa production in Indonesia. A newly identified dieback symptom has been observed in cocoa farms in East Luwu since 2015 and later observed in Luwu, Pinrang, Enrekang and Soppeng Regency, South Sulawesi. The potential causal agent identity was determined using morphological characteristics, DNA sequences, and phylogenetic analyses. A total of four fungal isolates out of seven isolates that were recovered from diseased stems morphologically resembled Lasiodiplodia species. ITS and EF1α sequencing confirmed that three of the Lasiodiplodia isolates were Lasiodiplodia theobromae, and one isolate was Lasiodiplodia pseudotheobromae. The aggresiveness of all fungal isolates and control was further determined by a leaf disk assay. Experiments showed that L. theobromae isolate CAS0321 was the most aggressive isolate in a leaf disk assay, while other Lasiodiplodia isolates, Fusarium isolates and Diaporthe isolate, were less aggressive. L. theobromae isolate CAS0321 was further used for a pathogenicity test on cocoa seedlings. Two months after inoculation, L. theobromae isolate CAS0321 caused elongated streaks on seedling stems with dark brown to black streaking of vascular tissues similar to that observed in natural infections. This is the first report of a virulent isolate of L. theobromae associated with cocoa dieback in Sulawesi.