First Report of Stem Canker Caused by Diaporthe tulliensis on jasmine in Taiwan

Plant Dis. 2022 Jul 13. doi: 10.1094/PDIS-09-21-1908-PDN. Online ahead of print.

Abstract

Diaporthe species can infect forest trees, ornamentals, and crops, causing root and fruit rots, stem cankers, leaf spots, etc. (Yang et al. 2018). In February 2021, about 10-20% of jasmine plants showing stem canker, foot rot, and wilting were observed in Changhua (24°01'57.7"N 120°34'54.7"E), Taiwan. The diseased plants initially showed chlorosis, leaf drop, and dieback. Sunken lesions were observed on the infected stem and kept expanding gradually. Eventually, plants wilted and black spots formed on the lesions. The margin of healthy and infected tissues of six samples were cut into 4 pieces, disinfected with 10% NaOCl for 30 seconds, rinsed twice in sterilized distilled water for 1 minute, and cultured on water agar at 28℃ under 12 h light / 12 h dark cycle. Hyphae grown out from isolated tissues were sub-cultured on potato dextrose agar (PDA). All tissues grown out of fungi showed similar colony morphology. Two hyphal tips from different tissues were isolated as representatives and deposited in Bioresource Collection and Research Center, Hsinchu, Taiwan, under BCRC numbers FU31566 and FU31567. The colonies on PDA were white to pale gray and produced black pycnidial conidiomata. The two-week-old conidiomata scattered or aggregated in small groups, exuded cream to pale yellow conidial droplets, 0.3-1.1 mm (n=50). The α-conidia were one-celled, hyaline, ovoid to cylindrical with one or two droplets, 3.8-6.3 × 2.5-3.8 μm (n=50). β-conidia were absent. The internal transcribed spacer (ITS), translational elongation factor subunit 1-α (EF1α), and β-tubulin of the two isolates were amplified using primer pairs ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (Glass and Donaldson, 1995), respectively. The ITS (MZ389113, MZ389114), EF1α (MZ419338, MZ419339), and β-tubulin (MZ408893, MZ408894) sequences of two isolates showed 98.55-98.56% (KR936130), 98.82% (KR936133), and 99.11-99.33% (KR936132) match to those of Diaporthe tulliensis R.G. Shivas, Vawdery & Y.P. Tan ex-type isolate BRIP 62248a (Dissanayake et al. 2017), respectively. Based on the morphological and molecular characters, this fungus is identified as D. tulliensis. To confirm the pathogenicity, the needle-wounded stem bases of eight-month-old cutting jasmine seedlings were inoculated with BCRC FU31566 by two PDA disks with actively grown fungal edges or conidial suspension at the concentration of approximately 2 × 105 conidia/ml. Each method inoculated five seedlings, performed in the greenhouse at 25 ± 2°C. Non-inoculated plants served as control. Two weeks after inoculation, three plants inoculated with PDA disks of the fungal culture showed wilting, and conidiomata formed on the stem base. The same symptoms were observed in one plant inoculated with the conidial suspension 3 weeks after inoculation. By contrast, the controls remained symptomless. Koch's postulates were completed by re-isolating the fungus from the inoculated plant. The re-isolated pathogen showed similar morphology and molecular characteristics to the original. D. tulliensis has been reported to cause cocoa rotted stem in Australia, kiwifruit stem canker in China, and Boston ivy leaf spot in Taiwan (Crous et al. 2015; Bai et al. 2017; Huang et al. 2021; Farr and Rossman 2021). To our knowledge, this is the first report of stem canker on jasmine associated with D. tulliensis in Taiwan. Furthermore, this is the first record of jasmine as a host of D. tulliensis worldwide.

Keywords: Diaporthe tulliensis; Canker; Causal Agent; Crop Type; Fungi; Jasmine; Ornamentals; Pathogen detection; Pathogen diversity; Subject Areas; Taiwan; herbaceous/flowering plants.