Short Communication |
Corresponding author: Kyeong-Yeoll Lee ( leeky@knu.ac.kr ) Academic editor: Sergei Subbotin
© 2016 Dongwoo Kim, Jae-Yong Chun, Kyeong-Yeoll Lee.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Kim D, Chun J-Y, Lee K-Y (2016) Morphometric and molecular characterization of populations of Pratylenchus kumamotoensis and P. pseudocoffeae (Nematoda, Pratylenchidae) newly recorded in Korea. ZooKeys 600: 1-5. https://doi.org/10.3897/zookeys.600.8508
|
At least 70 species of root-lesion nematodes, Pratylenchus spp., have been identified worldwide, many of which are serious pests of various agricultural crop plants. In Korea, only 14 species have been recorded in vegetable and fruit fields. Here, we report two new records of root-lesion nematode species in Korea based on morphometric and molecular methods. Soil samples were collected from chrysanthemum cultivars in various regions. Morphometric diagnosis showed that two new records for Korea: Pratylenchus kumamotoensis in Chilgok County and P. pseudocoffeae in Geumsan County. In addition, molecular diagnosis using the two sequences of the internal transcribed spacer (ITS) and the D2–D3 region of ribosomal DNA showed that these two species were most similar with those from Japan, Costa Rica and USA. The similarities of the ITS and D2–D3 sequences were 99.85 and 99.74%, respectively, for P. kumamotoensis and 99.99 with Costa Rica populations and 99.86% with USA populations, respectively, for P. pseudocoffeae. To our knowledge, this is the first report of two species in Korea.
Chrysanthemum cultivars, ITS, Root-lesion nematodes
In 2012–2014, amphimictic root-lesion nematode populations were detected in soil and root samples from chrysanthemum (Chrysanthemum sp.) fields located in Chilgok and Geumsan Counties in Korea. The population from Chilgok Co. was identified as Pratylenchus kumamotoensis and the other from Geumsan Co. as P. pseudocoffeae using morphological and molecular analyses. The phylogenetic relationship of these Korean root-lesion nematodes with other populations reported on chrysanthemum in Japan and distant geographical areas was also analyzed.
Nematodes were extracted from the soil samples using Cobb’s sieving and gravity method and the modified Baermann funnel method. Extracted nematodes in water suspension were killed by gentle heating, dehydrated by using the rapid lactophenol method and transferred in glycerin in permanent slides (
The sequence of the ITS region, including ITS1, 5.8S, and ITS2, was amplified using the forward TW81 primer (5’-GTT TCC GTA GGT GAA CCT GC-3’) and reverse AB28 primer (5'-ATA TGC TTA AGT TCA GCG GGT-3') (
The morphometric characters of the Korea population of P. kumamotoensis females that were measured included (n = 29) : body length = 511.3 ± 30.87 (451.9 – 564.1 μm), a = 26.7 ± 2.10 (23.2 – 31.9 μm), b = 6.9 ± 0.55 (6.0 – 7.9 μm), b’ = 5.0 ± 0.40 (4.3 – 5.8 μm), c = 20.3 ± 1.87 (17.2 – 23.9 μm), c’ = 2.4 ± 0.19 (2.0 – 2.6 μm), V(%) = 75.7 ± 1.85 (71.4 – 81.8), stylet = 14.8 ± 0.55 (13.4 – 15.7 μm).
The morphometric characters obtained for the Korean P. pseudocoffeae females included (n = 36) : body length = 565.9 ± 51.99 (475.3 – 660.4 μm), a = 23.1 ± 1.67 (20.7 – 29.2 μm), b = 6.8 ± 0.46 (6.1 – 7.7 μm), b’ = 3.7 ± 0.24 (3.2 – 4.1 μm), c = 19.4 ± 1.40 (17.3 – 21.8 μm), c’ = 2.0 ± 0.23 (1.6 – 2.4 μm), V(%) = 79.6 ± 0.80 (78.2 – 81.2), stylet = 17.2 ± 0.97 (15.9 – 19.3 μm).
These morphometrics matched those reported by
The ITS regions, including ITS1, 5.8S, and ITS2 of the Korean P. kumamotoensis and P. pseudocoffeae were 664 and 849 bp, respectively (Suppl. material
The results of the phylogenetic analysis using ITS sequences of our sample (Chilgok population) and eight populations of P. kumamotoensis from Kagosima, Kumamoto, and Oita Prefectures of Japan showed that they clustered in two clades, which were not geographically separated (Suppl. material
To our knowledge, this is the first record of occurrence of P. kumamotoensis and P. pseudocoffeae in chrysanthemum fields in Korea.
Analysis of ITS2 and D2-d3 segment sequences of Pratylenchus kumamotoensis and P. pseudocoffeae.
Data type: Nucleotide sequences and cladograms
Explanation note:
Figure S1. Alignment of the ITS region of Pratylenchus kumamotoensis.
Figure S2. Alignment of the ITS region of Pratylenchus pseudocoffeae.
Figure S3. Alignment of the D2-D3 expansion segment of Pratylenchus kumamotoensis.
Figure S4. Alignment of the D2-D3 expansion segment of Pratylenchus pseudocoffeae.
Figure S5. Neighbor-joining tree for Pratylenchus species based on ITS sequences. Numbers adjacent to branches denote the bootstrap values (>50%) of 1000 replicates. Sequences identified in this study were indicated in bold.
Figure S6. Neighbor-joining tree for Pratylenchus species based on D2-D3 sequences. Number adjacent to branches denote the bootstrap values (>50%) of 1000 replicates. Sequences identified in this study are indicated in bold.