High population densities of the plant-parasitic nematode Paratylenchus shenzhenensis have been observed in association with stunted anthurium in Shenzhen, Guangdong Province, China. P. shenzhenensis is now recognized as a new pathogen that causes severe damage to anthurium. Commercial fields of anthurium infected by P. shenzhenensis present a patchy distribution of plants that are usually stunted and in decline and have yellowing leaves. Roots that are heavily infected by this nematode present brown, dark-brown, or black rot, and the entire root mass is often destroyed. In this study, a method of culturing P. shenzhenensis on carrot disks was established, and the effects of temperature and time on the reproduction rate (Rr = final number of nematodes/initial number of nematodes) of this nematode were assessed. The optimum temperature for culturing P. shenzhenensis on carrot disks is 25°C and, after inoculation with 20 females and 10 males at 25°C for 10 weeks, Rr reached 3,834. At the optimum temperature, a suitable extension culture time or an increase in initial inoculum density will improve the reproductive efficiency of P. shenzhenensis. Koch’s postulates were fulfilled by inoculating P. shenzhenensis on the roots of anthurium and demonstrating the pathogenicity of the nematode. Histological analyses showed that P. shenzhenensis feed endoparasitically on the roots of anthurium; whole nematode bodies were observed in the outermost epidermal cells and root hairs, and the cell walls and middle lamellae were partially dissolved because of nematode migration and feeding.

Soybean (Glycine max L.) is a very important commercial crop in China (Li et al. 2019). Pratylenchus coffeae (Zimmermann, 1898) Filipjev & Schuurmans Stekhoven, 1941, is one of the most important root-lesion nematodes that invade the roots of many crops. In August 2018, five root and soil samples were collected in a soybean field, near Xipan village in Linshu county of Linyi City, Shandong Province, China (Fig. S1), to investigate the occurrence of root-lesion nematodes. The collected plants (cv. Lindou No.10) were growing poorly and the roots showed distinct brown lesions (Fig. S2). Pratylenchus spp. were extracted using the modified Baermann funnel method for 2 days (Hooper et al. 2005). On average, 395 root-lesion nematodes per kg of soil and 36 root-lesion nematodes per gram of fresh roots were extracted. The extracted root-lesion nematodes were disinfected with 0.3% streptomycin sulfate and cultured on carrot disks for propagation at 25°C. The species identification was based on morphological and molecular criteria. Key morphological features were determined for females and males. Measurements of females (n = 16) included body length = 561.0 μm ± 37.6 (standard deviation) (520.5 to 654.0 μm), tail length = 30.0 μm ± 1.9 (27.0 to 33.5 μm), stylet = 16.0 μm ± 0.6 (15.0 to 17.5 μm), a = 28.2 ± 2.3 (23.7 to 31.5), b = 6.4 ± 0.5 (5.7 to 7.3), c = 18.7 ± 1.8 (15.7 to 23.8), and V = 80.8% ± 2.1 (76.5 to 83.8%). Measurements of males (n = 16): body length = 511.0 μm ± 28.1 (range= 475.5 to 566.0 μm), tail length = 26.0 μm ± 1.3 (23.5 to 28.5 μm), stylet = 15.0 μm ± 0.5 (14.5 to 16.0 μm), spicule length = 17.0 μm ± 0.9 (16.0 to 18.5 μm), a = 30.8 ± 1.5 (28.0 to 33.2), b = 6.1 ±0.4 (5.6 to 6.9), and c = 19.8 ± 1.3 (18.1 to 22.2) (Fig. S3). All the morphological features of this population matched the description of P. coffeae (Castillo and Vovlas, 2007). DNA was extracted from an individual female as described previously (Wang et al. 2011). The rDNA-internal transcribed spacer (ITS) region and the D2/D3 region of the 28S rRNA gene were amplified by primers 18S/26S (Vrain et al. 1992) and D2A/D3B (De Ley et al. 1999), respectively. The PCR products were purified and sequenced. The obtained sequences of the ITS region (1,253 bp) and the D2/D3 region of 28S rRNA (781 bp) were deposited in GenBank. The ITS sequences of the root-lesion nematode obtained in this study (GenBank Accession no. MT879294) exhibited 99% identity with several P. coffeae sequences available in the GenBank (e.g., KR106219, MT586756, KY424205, and MN749379), and the obtained D2/D3 region sequence (MT879295) exhibited 100% identity with several P. coffeae sequences (e.g., MT586754, MN750755, MK829009, and MH730447). Both morphological and molecular data confirmed the presence of P. coffeae. To confirm reproduction on soybean, the obtained root-lesion nematode population was used in a greenhouse (25°C) assay to fulfill modified Koch's postulates. About 20 days after sowing, eight pots, each with one soybean plant (Lindou No.10) were inoculated with 1000 P. coffeae. The inoculated plants were kept in 1.5 L pots containing 1.2 L sterilized soil. Eight pots of uninoculated soybeans were used as the control. Ten weeks later, the inoculated roots were washed and brown lesions were observed. The number of nematodes/pot was approximately 7360 in soil and 796 in roots, and the reproduction factor was 8.16. Root-lesion nematodes and symptoms were not observed in control groups. P. coffeae has only been reported on soybean in Zhejiang (Wei et al. 2013) and Henan Province (Li et al. 2019) of China. To our knowledge, this is the first report of P. coffeae infecting soybean in Shandong Province, China. Since the root-lesion nematode can cause considerable damage to soybean, care should be taken to prevent the spread of P. coffeae to other regions in China.

Ten populations of Radopholus similis from different ornamental hosts were tested for their parasitism and pathogenicity to water spinach (Ipomoea aquatic), malabar spinach (Basella rubra), and squash (Cucurbita moschata) in pots. The results showed all three plants were new hosts of R. similis. Growth parameters of plants inoculated with nematodes were significantly lower than those of healthy control plants. All R. similis populations were pathogenic to the three plants, but pathogenicity differed among populations from different hosts. The same R. similis populations also showed different pathogenic effects in the three different plants. RadN5 population from Anthurium andraeanum had the highest pathogenicity to the three studied plants. RadN1 from A. andraeanum had the lowest pathogenicity to squash and RadN7 from Chrysalidocarpus lutesens had the lowest pathogenicity to water spinach and malabar spinach. R. similis is usually associated with root tissues, but here we report that it could be found to move and feed in the stem bases of all three studied plants. Sequence and phylogenetic analyses of DNA markers of the 18S rRNA, 28S rRNA, ITS rRNA, and mitochondrial DNA gene sequences of ten R. similis populations revealed significant genetic diversity. RadN5 and RadN6 populations from anthurium showed a close genetic relationship and could be distinguished from other populations by PCR-RFLP. At the same time, RadN5 and RadN6 populations were the most pathogenic to three studied plants. These results confirm the existence of large biological variability and molecular diversity among R. similis populations from the same or different hosts, and these characteristics are related to pathogenic variability.

The cereal cyst nematode (CCN), Heterodera filipjevi, is a widespread nematode species that causes serious damage to wheat and other cereals (Smiley and Nicol 2009). Since its first report in China in a winter wheat field in 2010 (Li et al. 2010; Peng et al. 2010), H. filipjevi has been a major limiting factor in wheat production. Here we report the first occurrence of H. filipjevi on winter barley in an irrigated field in Zhoukou City, Henan Province, China. In May 2017, cysts of a previously unidentified species of Heterodera were observed on the knotted and shallow roots of winter barley. Mature cysts were extracted from soil by flotation. The cysts were lemon shaped with a prominent neck and bifenestrate vulva cone. Strongly developed bullae and a heavy underbridge were also observed. The key morphometrics of cysts (n=10) included a mean cyst length of 741.5 ± 45.5 (688.2 to 838.1) μm excluding the neck, cyst width 545.2 ± 38.1 (468.5 to 623.0) μm, fenestral length 50.2 ± 3.6 (46.6 to 55.5) μm, fenest...