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The susceptibility of Lasioderma serricorne to phosphine (PH3), ethyl formate (EF) and their combination (PH3 + EF) was evaluated in this study. Eggs, larvae, pupae and adults were subjected to treatment with fumigants to determine the 90% lethal concentration time (LCt90) values. Treatment with PH3 for 20 h resulted in LCt90 values of 1.15, 1.39, 14.97 and 1.78 mg h/L while treatment with EF resulted in values of 157.96, 187.75, 126.06 and 83.10 mg h/L, respectively. By contrast, the combination of PH3 + EF resulted in LCt90 values of 36.05, 44.41, 187.17 and 35.12 mg h/L after 4 h. These results show that, through treatment with PH3 + EF, control can be achieved at lower concentrations than for treatment with EF alone and at lower exposure times than for treatment with PH3 alone. The sorption rates of the fumigants on cured tobacco leaves were determined for filling ratios of 2.5%, 5.0% and 10.0% (w/v). Cured tobacco leaves were treated with either 2 mg/L PH3, 114 mg/L EF or 0.5 mg/L PH3 + 109 mg/L EF. Treatment with PH3 showed sorption rates of 0.0%, 7.1% and 14.3%. EF, however, showed higher sorption rates of 64.9%, 68.5% and 75.5%, respectively, for the indicated filling ratios. When PH3 and EF were combined, the sorption rate of PH3 was 0.0%, while the sorption rates of EF were lower (9.1%, 12.0% and 23.2%) than treatment with only EF. EF required a ventilation time of longer than 22 h to desorb from cured tobacco leaves. Therefore, PH3 + EF can effectively control L. serricorne in cured tobacco leaves, with sufficient ventilation time required after treatment for the safety of workers.

Alcea rosea, in the family Malvaceae, is a biennial plant native to China and is grown typically for gardening in Korea (Lee 2003). Seven microcyclic Puccinia species have been reported on A. rosea: P. heterogenea, P. heterospora, P. lobata, P. malvacearum, P. platyspora, P. sherardiana, and P. modiolae (Demers et al. 2015; Aime and Abbasi 2018). In early May 2022, characteristic symptoms of rust were observed on four of ten seedlings of A. rosea purchased at a wholesale nursery (36°50′19.8″N, 128°55′28.7″E) in Bonghwa, Korea. Rust spots were present on almost 90% of the 1,000 seedlings of A. rosea in that nursery during our survey in late May. Through a distribution survey from June to July 2022, similar symptomatic leaves were additionally collected from the leaves of A. rosea grown in gardens at five sites in Gimcheon (two sites), Gumi (one), Seongju (one), and Busan (one). Spots were yellow-orange the center surrounded by chlorotic haloes on the adaxial leaf surface, and reddish-brown or dark brown pustules on the abaxial leaf surface. Over time, the spots enlarged and coalesced, causing the decay of large sections of the leaves, and heavily infected leaves fell early. Spermogonia, produced at the center of the chlorotic spot on the adaxial leaf surface, were subepidermal, obovoid, and 113.2–164.5 × 97.6–153.3 μm in size. Telia were reddish-brown to dark brown, round, mostly grouped, 0.28–0.61 mm in diameter, and mainly formed on the abaxial leaf surface but sometimes on the adaxial leaf surface also. Teliospores were two-celled, but rarely one- or three-celled, and were fusoid and 37. 6–110 × 12.4–21.5 μm in size; the wall was yellowish or almost colorless, smooth, 1.2–2.6 μm thick at the sides, and up to 7.4 μm thick at the apex. The morphological characteristics were similar to those of P. modiolae, although the teliospores in our study were longer than those observed by Aime and Abbasi (2018). For phylogenetic analysis, genomic DNA was extracted from the teliospores of each regional specimen. Partial 18S, internal transcribed spacer (ITS), and partial 28S sequences were amplified using primers NS1, ITS4, ITS5, and LR11. The PCR products were sequenced (Celemics, Seoul, Korea) and deposited in GenBank. The ITS-partial large subunit (LSU) sequence and 28S sequences had 100% homology with other P. modiolae sequences deposited in GenBank (accession numbers are shown in Fig. 2). In the phylogenetic trees of the ITS and LSU sequences, the isolates collected in this study were grouped with the reference sequences of P. modiolae, including the Korean isolate (ON631218) recently reported on Malva verticillata by Lee et al. (2022). For the pathogenicity test, the teliospores with germinating basidiospores were suspended in sterile distilled water and smeared on the upper surface of asymptomatic A. rosea leaves in August. The inoculated plants were sprayed with distilled water and kept in the dark with saturated moisture for 24 h in an isolated glass house of the Animal & Plant Quarantine Agent. After 2 weeks, typical rust spots and telia of P. modiolae were observed on the leaves of the inoculated plants, but not in the control plants, which were only sprayed with distilled, sterilized water and otherwise treated similarly to the inoculated plants. The results of this study show that the casual fungus is P. modiolae, which has been commonly found in A. rosea in Korea. To the best of our knowledge, this is the first report of P. modiolae in A. rosea in Korea.

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Malva sylvestris (Malvaceae), known as common mallow, is native to Europe, western Asia, and northern Africa. It was intentionally introduced to Korea as an ornamental plant in the early 20th century, and has become partly naturalized in several areas including the woods (Jung et al. 2017). Among nine microcylic Puccinia species attacking the Malvaceae plants, three species of P. heterospora, P. malvacearum, and P. modiolae have been reported on M. sylvestris (Classen et al. 2000, Colenso 1885, McKenzie 1998 and Melo et al. 2012). Only P. modiolae has been found on Alcea rosea and M. verticillata, and not M. sylvestris in Korea (Lee et al. 2022; Ryu et al. 2022). In August 2022, rust disease symptoms of a Puccinia fungus were observed on some overgrown seedlings of M. sylvestris, which were neglected in containers after sales at a wholesale nursery (36°50′19.8″N, 128°55′28.7″E) in Bonghwa, Korea. Typical rust spots were observed around 60% (on 111 seedlings of the 186 seedlings of M. sylvestris). The brown spots were produced on round chlorotic haloes on the adaxial leaf surface, and brown to dark brown pustules on the abaxial. Subepidermal spermogonia on the adaxial, were obovoid, and 112.1–160.0 × 88.7–149.3 μm in size. Telia were golden-brown to dark brown, round, mostly grouped, and 0.30–0.72 mm in diameter, and mainly hypophyllus. Fusoid teliospores were two-celled, rarely one- or three-celled, 36.2–92.3 × 10.6–19.3 μm in size, with many anomalies appearing notched at apex; wall was yellowish or almost colorless, smooth, 1.0–2.6 μm thick at the sides, and up to 6.8 μm thick at the apex; pedicel was hyaline, thick wall, persistent, and (39.3–)60.4–154.6(–189.9) μm long. Based on these morphological features together with the results of the phylogenetic analyses (e-Xtra 2) using internal transcribed spacer (ITS) and partial large subunit (LSU) sequences according to the method described by Ryu et al. (2022), the fungus was identified as an autoecious P. modiolae, recently reported on M. verticillate and A. rosea in Korea (Lee et al. 2022; Ryu et al. 2022). A representative sample was deposited in the Animal and Plant Quarantine Agency Herbarium (PQK220818). Pathogenicity tests were done using three host plants: M. sylvestris, M. verticillate and A. rosea. Three to four leaf discs with basidiospore-bearing telia were placed on the upper surfaces of healthy young leaves of the seedlings. Three replicates of each host plant set including an untreated control were tested. The plants were kept in an isolated glass house. At ten to twelve days after inoculation, typical telial spots of P. modiolae were recovered, but not in the control plants, showing all three tested species were highly susceptible (e-Xtra 1). The ITS and LSU sequences obtained from the genomic DNAs of each newly recovered rust spot were consistent with that of the inoculum (accession no. OQ542745). The previous A. rosea isolate (OP369290 by Ryu et al. 2022) also showed the pathogenesis on M. sylvestris and M. verticillata by the same tests, mentioned above (e-Xtra 1). To date, only one collection of P. modiolae on M. sylvestris has been reported in Louisiana, the United States (Aime and Abbasi 2018). The results of this study show that P. modiolae is firstly confirmed as the causal rust fungus of M. sylvestris and the same causal agent of M. verticillate and A. rosea rust disease, recently reported in Korea.