First report of post-harvest tuber rot of Apios americana Medikus caused by Penicillium expansum in South Korea

Plant Dis. 2022 Nov 2. doi: 10.1094/PDIS-06-22-1290-PDN. Online ahead of print.

Abstract

Apios americana Medikus, a perennial vine legume native to North America, is known as 'Indian potato' or 'Apios' for their underground tubers that are used for human consumption in Korea (Choi et al., 2017). These tubers are known to be rich in isoflavones as well as other secondary products to have known several medicinal properties (Chu et al., 2019). The harvested tubers in 2020 were observed to rot during storage of tubers for 4 months at 4°C in the genetic resource storage room at the Bioenergy Crop Research Institute, National Institute of Crop Science, Muan, Jeollanam-do, South Korea. The incidence of rot symptoms with blue mold was less than 1% per 20 kg box. Ten infected tubers were collected from several boxes and the lesions were cut into small pieces, and then surface sterilized in 1.5% NaClO for 2 minutes, followed by rinsing 3 times with sterilized water. To investigate their morphological characteristics, ten isolates were cultured in Malt Extract Agar (MEA) medium at 25° C for 5 days (Pitt and Hocking, 1988). The surface morphology of the mycelium had white or light green fluffy, and completely blue spores were formed after about 5 days. The conidia were one-stage branched with an elliptical shape, about 3.5 to 4.3 × 2.9 to 3.6 (mean 3.8 ± 0.3 × 3.2 ± 0.2) ㎛ in diameter (n=30). Genomic DNAs of the isolates were extracted using Solgent DNA Extraction Kit (Solgent, Daejeon, Korea), and then PCR products of the internal transcribed spacer (ITS1/ITS4) region and the beta-tubulin gene (Bt2a/Bt2b) were sequenced and analyzed (Glass and Donaldson, 1995). The BLASTn showed that the representative isolate had 99% homology with reference Penicillium expansum strain ICMP 2708 (ITS region and TUB2 gene) in NCBI GenBank. The sequences of the isolate were deposited in GenBank as accession numbers MZ636667 and MZ702813 for ITS and TUB2 genes, respectively. Based on the morphological characteristics and molecular analysis, the isolate was identified as P. expansum. Pathogenicity assays of the isolate were also performed using three tubers in three replicates inoculated with spore suspension (concentration, 1×106 conidia/mL) and compared with a control group inoculated with sterilized water. The inoculated Apios tubers were placed in a plastic box maintained in conditions of high humidity at 25°C. Five days after inoculation, the typical symptoms were observed on inoculated tubers, and no symptoms were observed in the control one. P. expansum was again isolated from artificially inoculated tubers to complete Koch's assumption. This is the first report of P. expansum causing tuber rot in A. americana in South Korea. As the cultivated area of Apios is increasing in Korea, it will be necessary to develop effective storage methods and management strategies for the control of storage diseases such as blue mold.

Keywords: Apios americana; Penicillium expansum; South Korea.