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| Establishment and Application of Molecular Detection Technology System for Two Kinds Pathogens of Sugarcane (Saccharum officinarum) Pokkah boeng |
| LI Jie1, LI Yin-Hu1, PU Jia-Rong2, ZHANG Rong-Yue1, SHAN Hong-Li1, LI Wen-Feng1, WANG Xiao-Yan1, PU Jin-An2, HUANG Ying-Kun1,* |
1 Sugarcane Research Institute, Yunnan Province Academy of Agricultural Science/Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan 661699, China;
2 Economic Crop Workstation in Xinping County, Xinping 653400, China |
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Abstract Sugarcane (Saccharum officinarum) is the most important sugar crop in China. Sugarcane Pokkah boeng disease poses a serious threat to the high-quality development of the sucrose industry in China. Therefore, specific primers were designed based on the nuclear ribosomal DNA internal transcribed spacer (rDNA-ITS) sequences of 2 main pathogens of sugarcane Pokkah boeng Fusarium verticillioides and Fusarium proliferatum, respectively. The results showed that primers Fv-F3/Fv-R3 could amplify a specific amplification fragment of 400 bp from F. verticillioides and its infected sugarcane, but not other pathogens of sugarcane diseases. The detection sensitivity for genomic DNA of F. verticillioides was 30 pg/µL. Primers Fp-F4/Fp-R4 could amplify a specific amplification fragment of 362 bp from F. proliferarum and its infected sugarcane, but not other pathogens of sugarcane diseases. The detection sensitivity for genomic DNA of F. proliferarum was 30 pg/µL. A total of 117 samples with typical Pokkah boeng symptoms were collected from different main cultivated sugarcane varieties in different sugarcane areas of Yunnan Province were detected by PCR with these two pairs of specific primers. And the occurrence of the two main pathogens of sugarcane Pokkah boeng in this area was clarified. The results showed that 112 (95.7%) of the 117 samples were infected by F. verticillioides, 103 (88%) were infected by F. proliferatum. 103 (88%) were complex infected by F. verticillioides+F. proliferatum. The two pathogens were not detected in other 5 samples, which may be caused by other species. The PCR products of 23 F. verticillioides and 19 F. proliferatum from different main sugarcane varieties in different sugarcane regions were selected for sequencing and analysis. The results showed that the F. verticillioides and F. proliferatum amplification product sequences had 99.45%~100% similarity with F. verticillioides (KU508286) and 99.26%~100% similarity with F. proliferatum (MK252904), respectively. Some sequences were selected to construct phylogenetic tree, they were divided into F. verticillioides group and F. proliferatum group. The results of this study showed that the established detection system could accurately and quickly detect F. verticillioides and F. proliferatum. The detection rates of F. verticillioides and F. proliferatum were high from different main sugarcane varieties in different sugarcane areas of Yunnan Province. They were important pathogens of sugarcane Pokkah boeng in Yunnan province, and the compound infection was common. Among them, F. verticillioides was the dominant species in sugarcane areas such as Puer, Lincang, Honghe, and Yuxi sugarcane areas. However, the detection rate of F. proliferatum was also high in these sugarcane areas, and maybe there were other species of sugarcane Pokkah boeng pathogens might be present. This study provides technical support and practical basis for early diagnosis and timely prevention and control of these diseases.
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Received: 23 July 2021
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Corresponding Authors:
* huangyk64@163.com
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