Dragon fruit (Hylocereus spp.) is a tropical perennial plant (Family Cactaceae) that is popular for consumption in India. Originated from Central and South America and currently became popular in India. Hylocereus undatus and H. polyrhizus are the most commonly cultivated species in India. During August to December, 2021, stem cankers were observed in commercial orchards within the Satara, Pune and Solapur districts of Maharashtra. The disease incidence across four orchards was approx. 40% and severity ranged between 30 and 80%. Initial symptoms on infected cladodes were minute, circular, depressed chlorotic spots often with a brick red flecks followed by elevation of the centre of the lesion. Later the lesions turned necrotic and contained black, erumpent pycnidia, followed by chlorosis and stem rot. Twelve diseased cladodes from different orchards were collected and isolations were conducted. Edges of the lesions (5 to10 mm2) were excised and surface sterilized by exposure to 1% sodium hypochlorite (2 min) followed by triple rinsing with sterilized distilled water. Excess moisture was removed with sterilized blotter paper and pieces were plated on potato dextrose agar (PDA) amended with streptomycin sulphate (30 mg/L) for 3 days at 27 ± 2°C with a 12 h photoperiod. All twelve isolation attempts yielded uniform fungal colonies and out of these, purified colonies from each location viz., SLNeo, LNeo, MGNeo and KNeo were selected for etiology study. On PDA, initially white mycelial colonies turned to olive green to grayish with dark gray to black pigmentation. Colony growth was rapid (30 mm/day). Brown, branched septate hyphae fragmented to produce abundant arthroconidia in chains that were hyaline to dark brown, thick walled, 0 to 1 septate, ellipsoid to ovoid (10.1 ± 1.4 × 4.88 ± 1.1 μm), rod (9.0 ± 0.8 μm × 5.3 ± 0.8 μm), round (7.9 ± 1.6 μm) and capsule (14.2 ± 3.1 × 6.1 ± 0.8 μm) shape. Conidia from pycnidia developed on host tissues were aseptate, hyaline and ellipsoid-cylindrical (8.2 ± 0.8 × 2.2 ± 0.3 μm) in shape. Based on morphological characteristics, the isolates were identified as Neoscytalidium dimidiatum (Penz.) Crous & Slippers (Crous et al. 2006). Molecular characterization was done by amplifying the partial internal transcribed spacer (ITS) region, and translation elongation factor 1-α and β-tubulin genes of four isolates using ITS1/ITS4, EF1-728F/EF1-986R and BT2A/BT2B primers (White et al. 1990, Glass and Donaldson 1995, Carbone and Kohn 1999). Sequences were deposited in GenBank accesions (ITS: OM884028, OM884029, OM884030, OM899800; TUB2: OM927962, OM927963, ON099066; TEF1-α: OM927965, OM927966, OM927964, OM984744) showed 99 to 100% identity with the epitype CBS 499.66 accession numbers KF531820 for ITS and KF531798 for TEF1-α and KF531800 for TUB2. N. dimidiatum isolate designated as SLNeo has been deposited at Microbial Type Culture Collection (MTCC), Chandigarh, India with MTCC 13250. Phylogenetic analysis using maximum likelihood (ML) revealed these isolates were clustered with N. dimidiatum clade. To evaluate pathogenicity, stems of one year old dragon fruit plants were wounded with a sterilized needle and inoculated with 7 mm fresh mycelial discs. For each isolate three replications were kept. Non-inoculated controls were plants receiving only non-colonized PDA discs. Inoculated and control plants were kept separately at ambient temperature (35 ± 2°C during day and 22 ± 2°C at night). Symptom started to develop at 2 days after inoculation (DAI) and canker lesions with chlorotic halos and rotting were observed at 15 DAI. No symptoms were observed on the negative control. Pathogenicity testing was repeated twice and the pathogen was re-isolated from symptomatic tissues with 100% re-isolation frequency and found identical with N. dimidiatum. N. dimidiatum has been reported on H. undatus and H. polyrhizus in subtropical and tropical countries worldwide (Chuang et al. 2012; Ezra et al. 2013; Lan et al. 2012; Sanahuja et al. 2016; Serrato-Diaz and Goenaga 2021). To our knowledge, this is the first report of dragon fruit stem canker caused by N. dimidiatum from India. Since the disease poses a major threat to dragon fruit plantations, additional epidemiological studies may assist in developing management strategies.
Keywords: Causal Agent; Crop Type; Fruit; Fungi; Pathogen detection; Subject Areas.