First Report of Black Stem Caused by Botryosporium longibrachiatum on Statice in Korea.

Abstract

Perennial statice is widely cultivated worldwide. In Korea, hybrid statice (Limonium latifolium × bellidifolium) is grown as a commercial cut flower crop in polyethylene-film-covered greenhouses. In April 2013, hundreds of hybrid statice plants of the cvs. Yellow Cream and Pinky Cream were found symptomatic of a previously unknown disease, with 10 to 15% incidence in 10 greenhouses of 1,600 m2 surveyed in Gochang County, Korea. Affected stems turned dark brown and were usually covered with a fungus resembling the hoar-frost fungus, Botryosporium longibrachiatum (3), especially in a cool and humid environment. Symptoms consisted of stem blackening, as is typical for burley tobacco (1) and sweet basil (2). According to the farmer of the hybrid statice, stems blackened in the winter and spring of January to April when the tunnels were mostly closed, thus reaching 100% relative humidity (RH) every night due to poor ventilation. The fungus had an elongate main axis with lateral fertile branches in acropetal succession. Conidiophores were simple, erect, macronematous, 32 to 79 μm in length, with a terminal cluster of three to five ampullae. Conidiogneous cells were polyblastic. Conidia were ellipsoidal, elliptical-fusiform, hyaline, 7.6 to 9.5 × 3.0 to 4.2 μm. Colonies on potato dextrose agar (PDA) were floccose, non-pigmented, and chalk-white in color. Morphological and cultural characteristics of the fungus were consistent with previous reports of B. longibrachiatum (Oudem.) Maire (2,3). A voucher specimen was deposited in the Korea University Herbarium (KUS). Isolate KUS-F27305 was submitted to the Korean Agricultural Culture Collection (Accession No. KACC47263). Fungal DNA was extracted from isolate KACC47263 with DNeasy Plant Mini Kits. The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 604 bp was deposited in GenBank (Accession No. KF372591). A BLAST search in GenBank exhibited ≥99% nucleotide identity with the ITS sequence of B. longibrachiatum (JX666334) from sweet basil in Korea. To confirm pathogenicity, colonized mycelial agar blocks from isolate KACC47263 were transferred individually onto stem apices and leaves of five statice plants of the cv. Yellow Cream. Five control plants were treated similarly with non-colonized agar blocks. The plants were incubated in a humid chamber at 22 ± 2°C with a 12-h photoperiod for 48 h, and then maintained in 100% RH. After 3 to 4 days, necrotic lesions identical to those observed in the original greenhouses, started to develop on the stem and leaves of inoculated plants, leading to blackened stems covered with the hoar-frost fungus after 14 days. B. longibrachiatum was re-isolated from the lesions of inoculated plants, fulfilling Koch's postulates. No symptoms were observed on control plants. The pathogenicity test was repeated with the cv. Pinky Cream with identical results. To our knowledge, this is the first report of B. longibrachiatum infecting perennial statice globally as well as in Korea. We propose the name black stem of statice for this disease, analogous to the disease on basil (2).