The Use of Toluidine Blue Staining Combined with Paraffin Sectioning and the Optimization of Freeze-thaw Counting Methods for Analysing Root-Knot Nematodes in Tomato
2017
Xiaoman Li, Beijing University of Agriculture, Beijing, China | Wenchao Zhao, Beijing University of Agriculture, Beijing, China | Xiaoxuan Zhou, Beijing University of Agriculture, Beijing, China | Jiaping Feng, Beijing University of Agriculture, Beijing, China | Yingjian Gao, Beijing University of Agriculture, Beijing, China | Xuehui Yao, Beijing University of Agriculture, Beijing, China | Yang Liu, Beijing University of Agriculture, Beijing, China | Jie Liu, Beijing University of Agriculture, Beijing, China | Rui Yang, Beijing University of Agriculture, Beijing, China | Fukuan Zhao, Beijing University of Agriculture, Beijing, China | Shaohui Wang, Beijing University of Agriculture, Beijing, China
Root-knot nematodes (RKNs) (Meloidogyne spp.) are one of the most damaging crop pests and cause severe yield losses worldwide. Easier and more efficient methods are needed for assessing nematode infestations and host plant resistance. In the present study, we combined toluidine blue staining and paraffin sectioning to observe the establishment of giant cells (GCs; redifferentiated cells induced by nematode feeding) and the developmental status of RKNs in tomato (Solanum lycopersicum). The number of RKNs was counted using a modified freeze-thaw counting method. Finally, three tomato lines with different levels of RKN resistance, but otherwise identical genetic backgrounds, were tested to verify the applicability of the combined method. The results suggested that using toluidine blue staining combined with paraffin sectioning and freeze-thaw counting accurately reflects the level of RKN resistance of tomato plants.
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