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Lasiodiplodia theobromae as a Causal Agent of Kumquat Dieback in Taiwan

Kumquat (Fortunella margarita (Lour.) Swingle) is an important citrus fruit crop of Iland County in northeastern Taiwan. Fruit produced in this area are mainly for making preserves, which is a well-known product in Taiwan. Decline of kumquat was first noticed in 1990. Since 1995, it has become an important problem affecting ≈80% of kumquat orchards. The problem was especially serious after the passage of a typhoon. Some orchards were abandoned due to death of many declining trees. Initial symptoms were yellowing and browning of leaves on some branches of affected trees. Abscission of leaves and fruits occurred, subsequently resulting in the appearance of dieback of affected branches. Disease symptoms could progress to other branches of the same tree, gummosis on the trunk, and eventually death of the tree. Previously, Phytophthora citrophthora (Smith & Smith) Leonian was reported to be a causal organism of kumquat decline (1). Recently, isolations from declining kumquat trees in several orchards failed to recover P. citrophthora and instead we isolated Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (syn. Botryodiplodia theobromae Pat.) from tissue taken from the margin of discolored bark and wood on symptomatic branches. The fungus produced grayish black colonies on V8 agar and black ostiolate pycnidia (125 to 650 μm in diameter) with ovoid to elongate conidia (20 to 32 × 12 to 16 μm) on autoclaved whole wheat grains that were placed on V8 agar. Young conidia were hyaline and nonseptate, whereas mature conidia were brown, one septate, and striate. Pathogenicity tests were carried out on healthy kumquat trees located at the Taiwan Agricultural Research Institute experimental farm. Three isolates of L. theobromae were cultured on wheat-oat medium. Colonized grains (≈5 g) were held against a wound made by lightly scrapping a branch (5 to 7 mm in diameter) 200 to 250 cm above the ground by wrapping a sheet of clear plastic around the branch (2). Eighteen branches were used for each isolate, and the same number of branches similarly inoculated with sterile grains as a control. Healthy branches inoculated with L. theobromae showed disease symptoms similar to those observed on naturally infected plants. Discoloration of leaves on inoculated branches occurred within 1 week. Subsequently, all the leaves fell and infected branches died. The numbers of inoculated branches killed by the three isolates of L. theobromae tested were 18, 9, and 14 after 1 month. All control branches remained disease free. L. theobromae was reisolated from symptomatic tissues, completing Koch's postulates. The same isolates were used to inoculate trunks of kumquat trees with the method described above. Six trunks were inoculated with each isolate, and the same number of trunks similarly inoculated with sterile grains was used as the control. Gummosis on inoculated trunks occurred in 1 week. The numbers of inoculated trunks showing gummosis induced by the three isolates of L. theobromae tested were 6, 5, and 6 after 1 month. L. theobromae was reisolated from symptomatic tissue. All control trunks remained free of gummosis. Our results show that in addition to P. citrophthora, L. theobromae can also cause a dieback on kumquat. To our knowledge, this is the first report of disease caused by L. theobromae on kumquat or on any species in the Rutaceae family in Taiwan.

References: (1) P. J. Ann et al. Plant Pathol. Bull. (Taiwan) 6:198, 1997. (2) W. H. Ko et al. Plant Pathol. 35:254, 1986.