First Record of the Cereal Cyst Nematode Heterodera filipjevi in China.

Cereal cyst nematode (CCN) is now recognized as a widespread and often damaging parasite of wheat in China. Only Heterodera avenae has been reported in China (4). However, molecular analysis of four samples from Beijing and one from Shanxi Province indicated genetic differences from H. avenae and other named species (3). Here we report the detection of H. filipjevi at a site in Henan Province that was not included in any previous study or report. The infested crop was rainfed winter wheat (Triticum aestivum) cv. Wenmai 19 in a field near Banpopu Village in Xuchang County (34.0447°N, 113.7415°E) with a long-established maize-wheat semiannual crop rotation. During the winter growing season, the crop was patchy with uneven growth and cyst nematode females were observed on the roots. In June 2009, soil was collected and mature cysts were extracted for morphological and molecular identification. Cysts were also kept at 4°C for 2 months and then incubated in shallow water at 15°C for a month to obtain second-stage juveniles (J2). Measurements (range; mean ± sd) of 10 cysts were body length including neck (569 to 786 µm; 699 ± 56), body width (403 to 600 µm; 523 ± 55), length:width ratio (1.3 to 1.5; 1.3 ± 0.1), neck length (61 to 125 µm; 106 ± 19) and width (49 to 83 µm; 69 ± 13), fenestra length (52 to 59 µm; 57 ± 2.9) and width (24.5 to 34.4; 27.9 ± 3.5), underbridge (64 to 101 µm; 85 ± 10), and vulval slit (7.4 to 10.0 µm; 9.6 ± 1.0). Lemon-shaped cysts were brown with a surface zigzag pattern. The vulval cone was bifenestrate with horseshoe-shaped semifenestra, with heavy underbridge and many bullae. The J2 (n = 22) measurements were body length (496 to 590 µm; 552 ± 24), body width (20.0 to 23.8; 21.5 ± 0.9), stylet (22.8 to 25.3; 24.0 ± 1.0) with anchor-shaped basal knobs, tail (47 to 64; 61.6 ± 4.4), and hyaline tail terminus (32 to 43; 40.2 ± 3.0). The J2 had up to four lateral lines, but the inner two were often the only lines clearly visible, and the shape of the stylet knobs, tail, and tail terminus were consistent with H. filipjevi. All morphological data and characters were consistent with H. filipjevi (1). Specimens have been lodged with the Australian National Insect Collection. DNA from single cysts was extracted to amplify the internal transcribed spacer region of rDNA by PCR with forward primer TW81 (5'-GTTTCCGTAGGTGAACCTGC-3') and reverse primer AB28 (5'-ATATGCTTAAGTTCAGCGGGT-3') (2). The PCR product was sequenced (Genbank Accession No. HM027892) and digested by restriction enzymes (AluI, CfoI, HaeI, HinfI, PstI, RsaI, TaqI, and Tru9I) to obtain restriction fragment length polymorphism profiles (2). Profiles for the Xuchang population consistently matched those published for H. filipjevi and were distinct from those of H. avenae and other species (3). Phylogenic analysis of the sequence further indicated conspecificity with H. filipjevi. These morphological and molecular data confirmed that the specimens from Xuchang were H. filipjevi, which represents the first detection of H. filipjevi in China, and extends the known distribution of the species from Europe, North America, South Asia, and West Asia to East Asia. This finding adds complexity to the management of CCN in China, especially for control by host resistance, which now must consider both species and pathotype diversity. References: (1) Z. A. Handoo. J. Nematol. 34:250, 2002. (2) S. A. Subbotin et al. Nematology 2:153, 2000. (3) S. A. Subbotin et al. Nematology 5:515, 2003. (4) H. X. Yuan et al. Australas. Plant Pathol. 39:107, 2010.