First Report of Stunt Nematode, Tylenchorhynchus zeae, on Corn in Republic of Korea

Abstract

Corn (Zea mays), one of the major food sources it contains rich in fiber and many vitamins, is one of the most widely consumed cereal grains in Republic of Korea. A survey of plant-parasitic nematodes (PPNs) was carried out in corn fields in Goesan, Republic of Korea from August in 2021. PPNs were extracted from the corn roots and soil using modified Baermann funnel methods and were identified using morphological and molecular analyses. Among the roots and soil samples of 21 fields, 5 fields (23.8%) were infected with stunt nematodes. Tylenchorhynchus zeae was originally described in India from soil around corn and is reported to dwarf plants, yellow leaves (Sethi and Swarup, 1968). Morphologically, characteristics of females were very similar to T. zeae with cylindrical body and slightly ventrally arcuated after fixation. Lip region slightly offset from body with four annuli. Stylet with anteriorly flattened knobs, the vulva was located in the center of the body, didelphic-amphidelphic reproductive system and tail conoid, tail terminus with obtuse smooth, with four incisures areolated throughout body. Bodies of males were similar to females but with shaper tails, with relatively strong bursa and spicules (Fig. S1). The morphology of Korean populations was in agreement with the described populations of India and China (Alvani et al., 2017; Xu et al., 2020). Measurements and micrographs with the light-microscope (DM5000; Leica[Germany]) and camera (DFC450; Leica[Germany]) were taken from females (n=10) for mean, standard deviation and range of body length: 553.2 ± 41.2 (492.7-643.6) µm, maximum body width: 19.4 ± 1.0 (17.6-21.0) µm, stylet length: 18.1 ± 0.4 (17.5-18.7) µm, percent of distance from anterior end to vulva / body length: 58.5 ± 1.3 (56.1-60.9), tail length: 31.7 ± 1.2 (30.3-34.0) µm, and distance of anterior to excretory pore: 96.5 ± 1.8 (94.1-99.4) µm. In addition, PCR was performed for the 28S rDNA D2-D3 segments using the primers D2A and D3B, and ITS region with the primers TW81 and AB28. The newly obtained sequences were submitted to GenBank database under accession numbers ON909086, ON909087 and ON909088 of 28S rDNA D2-D3 segments, and ON909123, ON909124 and ON909125 of ITS region. The resulting 28S rDNA D2-D3 segment sequences were 100% identical to KJ461565 and the BLASTn search of the ITS region sequences was most similar to T. zeae (KJ461599), which is the species isolated from corn in Spain. The identities of ITS region sequences on these populations were 99.89% (893/894), with no insertions/deletions. The phylogenetic relationships of the population strongly support T. zeae (Fig. S2). Phylogenetic relation analysis based on the two genes was constructed using PAUP version 4.0 and MrBayes 3.1.2 programs. To confirm pathogenicity, a modified version of Koch’s postulates was conducted in the greenhouse by inoculating 100 females and males onto each of five pots of seedling corn (cv. Daehakchal) filled with the sterilized sandy soil and maintained for 60 days at 25℃ under the conditions. Tylenchorhynchus zeae reproduction factor was 2.21 ± 0.37 was observed at the end of the trial in soil on pots. The stunted and swollen roots and dwarfed and yellowing leaf shoots symptoms in the greenhouse pots trial were confirmed the same as those typical damage symptoms. To the best of our knowledge, this is the first report of T. zeae in Republic of Korea. The host range of T. zeae includes some economic crops such as cabbage, cauliflower, grapevine, and olive (Chen at al., 2007; Handoo et al., 2014). It is necessary to investigate the damage to economic crops in the Republic of Korea to this nematode.