A LAMP Assay for Rapid Detection of Stem Rot <i>Fusarium oxysporum</i> on Succulent Plant, <i>Echeveria</i> Perle von Nürnberg

YAO Jinai; HUANG Peng; CHEN Hanxin; HOU Xiangyu; YU Deyi

doi:10.19303/j.issn.1008-0384.2020.08.008

Volume 35 Issue 8

Aug. 2020

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Article Navigation > Fujian Journal of Agricultural Sciences > 2020 > 35(8): 863-868

YAO J A, HUANG P, CHEN H X, et al. A LAMP Assay for Rapid Detection of Stem Rot Fusarium oxysporum on Succulent Plant, Echeveria Perle von Nürnberg [J]. Fujian Journal of Agricultural Sciences，2020，35（8）：863−868 doi: 10.19303/j.issn.1008-0384.2020.08.008

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YAO J A, HUANG P, CHEN H X, et al. A LAMP Assay for Rapid Detection of Stem Rot Fusarium oxysporum on Succulent Plant, Echeveria Perle von Nürnberg [J]. Fujian Journal of Agricultural Sciences，2020，35（8）：863−868 doi: 10.19303/j.issn.1008-0384.2020.08.008

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A LAMP Assay for Rapid Detection of Stem Rot Fusarium oxysporum on Succulent Plant, Echeveria Perle von Nürnberg

doi: 10.19303/j.issn.1008-0384.2020.08.008

1.
Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests/Institute of Plant Protection, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China
2.
Zhangzhou Institute of Agricultural Science, Zhangzhou, Fujian　363005, China

Received Date: 2020-01-17
Rev Recd Date: 2020-04-15
Publish Date: 2020-08-19

Abstract

Abstract

Objective A LAMP (loop-mediated isothermal amplification) assay for rapid, visible and sensitive detection of Fusarium oxysporum on the succulent plant, Echeveria Perle von Nürnberg, was developed for early prevention and control of the disease. Method Based on the elongation factor 1α (EF-1α ) sequence of F. oxysporum , LAMP primers were designed. Template DNA from the infected stems were used to optimize the reaction temperature and time. Specificity and sensitivity of the assay were verified on the detection on infected plants from the field. Result The newly developed LAMP assay at 65 °C for 60 min effectively detected F. oxysporum on Echeveria . The assay gave the positive yellowish green result only on F. oxysporum of Echeveria . It had a detection limit of 10 fg·μL⁻¹ and was 100% positive on detecting F. oxysporum from 15 Echeveria samples showing typical stem rot symptoms. Conclusion The established LAMP assay had high specificity and sensitivity with visual and low false positive results. It was considered suitable for rapid detection of F. oxysporum on Echeveria Perle von Nürnberg in the field.
- Echeveria Perle von Nürnberg,
- stem rot,
- Fusarium oxysporum,
- loop-mediated isothermal amplification (LAMP),
- detection

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References(20)

References

[1]	GRIFFITHS H, MALES J. Succulent plants [J]. Current Biology, 2017, 27(17): R890−R896. doi: 10.1016/j.cub.2017.03.021
[2]	吴旭东, 陆辰晨, 沈浩, 等. 大豆茎褐腐病菌环介导等温扩增检测技术的建立 [J]. 南京农业大学学报, 2015, 38(4):568−574. WU X D, LU C C, SHEN H, et al. A rapid detection method for the plant pathogen Phialophora gregata f.sp.sojae based on loop-mediated isothermal amplification (LAMP) [J]. Journal of Nanjing Agricultural University, 2015, 38(4): 568−574.(in Chinese)
[3]	YAO J N, HUANG P, LAN C Z, et al. Stem rot on Cymbidium ensifolium (orchidaceae) caused by Fusarium oxysporum in China [J]. Canadian Journal of Plant Pathology, 2018, 40(1): 105−108. doi: 10.1080/07060661.2017.1411976
[4]	DAI T T, LU C C, LU J, et al. Development of a loop-mediated isothermal amplification assay for detection of Phytophthora sojae [J]. FEMS Microbiology Letters, 2012, 334(1): 27−34. doi: 10.1111/j.1574-6968.2012.02619.x
[5]	HERRERA-VÁSQUEZ J A, PUCHADES A V, ELVIRA-GONZÁLEZ L, et al. Fast detection by loop-mediated isothermal amplification (LAMP) of the three Begomovirus species infecting tomato in Panama [J]. European Journal of Plant Pathology, 2018, 151(1): 243−250.
[6]	TIAN Q, LU C C, WANG S S, et al. Rapid diagnosis of soybean anthracnose caused by Colletotrichum truncatum using a loop-mediated isothermal amplification (LAMP) assay [J]. European Journal of Plant Pathology, 2017, 148(4): 785−793. doi: 10.1007/s10658-016-1132-2
[7]	李华伟, 许泳清, 罗文彬, 等. 马铃薯S病毒PVS~O株系RT-LAMP检测方法的建立及应用 [J]. 园艺学报, 2018, 45(8):1613−1620. LI H W, XU Y Q, LUO W B, et al. Establishment of Loop-mediated isothermal amplification assay for detection of potato virus S ordinary strain [J]. Acta Horticulturae Sinica, 2018, 45(8): 1613−1620.(in Chinese)
[8]	CHANDRA A, KEIZERWEERD A T, QUE Y X, et al. Loop-mediated isothermal amplification (LAMP) based detection of Colletotrichum falcatum causing red rot in sugarcane [J]. Molecular Biology Reports, 2015, 42(8): 1309−1316. doi: 10.1007/s11033-015-3875-9
[9]	LI G R, HUANG G M, ZHU L H, et al. Loop-mediated isothermal amplification (LAMP) detection of Phytophthora hibernalis, P. syringae and P. cambivora [J]. Journal of Plant Pathology, 2019, 101(1): 51−57. doi: 10.1007/s42161-018-0136-5
[10]	SHAN L Y, ABDUL HASEEB H, ZHANG J, et al. A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of toxigenic Fusarium temperatum in maize stalks and kernels [J]. International Journal of Food Microbiology, 2019, 291: 72−78. doi: 10.1016/j.ijfoodmicro.2018.11.021
[11]	宋必. 应用LAMP检测技术快速诊断大豆根茎部真菌病害[D]. 南京: 南京农业大学, 2015. SONG Bi. Application of Loop-mediated isothermal amplification (LAMP) in rapid detection of fungal soybean root and stem disease[D]. Nanjing: Nanjing Agricultural University, 2015. (in Chinese)
[12]	LAN C Z, RUAN H C, YANG X J, et al. Development of a loop-mediated isothermal amplification assay for sensitive and specific detection of Fusarium oxysporum f.sp. cucumerinum Owen [J]. Phytoparasitica, 2018, 46(3): 283−293. doi: 10.1007/s12600-018-0669-3
[13]	ZHANG S Y, DAI D J, WANG H D, et al. One-step loop-mediated isothermal amplification (LAMP) for the rapid and sensitive detection of Fusarium fujikuroi in bakanae disease through NRPS31, an important gene in the gibberellic acid bio-synthesis [J]. Scientific Reports, 2019, 9(1): 3726. doi: 10.1038/s41598-019-39874-z
[14]	GHOSH R, TARAFDAR A, SHARMA M. Rapid detection of Fusarium oxysporum f.sp. ciceris from disease infested chickpea fields by loop-mediated isothermal amplification [J]. Indian Phytopath, 2016, 69(4s): 47−50.
[15]	漆永红, 曹素芳, 李雪萍, 等. 燕麦镰孢菌环介导等温扩增(LAMP)检测方法的建立与应用 [J]. 草地学报, 2018, 26(4):1004−1010. QI Y H, CAO S F, LI X P, et al. Development of a Loop-mediated isothermal amplification assay for detection of Fusarium avenaceum [J]. Acta Agrestia Sinica, 2018, 26(4): 1004−1010.(in Chinese)
[16]	NOTOMI T, OKAYAMA H, MASUBUCHI H, et al. Loop-mediated isothermal amplification of DNA [J]. Nucleic Acids Research, 2000, 28(12): 63. doi: 10.1093/nar/28.12.e63
[17]	LU C, ZHANG H, WANG Y, et al. Rapid diagnosis of Fusarium root rot in soybean caused by Fusarium equiseti or Fusarium graminearum using loop-mediated isothermal amplification (LAMP) assays [J]. Australasian Plant Pathology, 2015, 44(4): 437−443. doi: 10.1007/s13313-015-0361-8
[18]	AYUKAWA Y, KOMATSU K, KASHIWA T, et al. Detection and differentiation of Fusarium oxysporum f.sp.lycopersici race 1 using loop-mediated isothermal amplification with three primer sets [J]. Letters in Applied Microbiology, 2016, 63(3): 202−209. doi: 10.1111/lam.12597
[19]	魏巍, 朱杰华, 张宏磊, 等. 河北和内蒙古马铃薯干腐病菌种类鉴定 [J]. 植物保护学报, 2013, 40(4):296−300. WEI W, ZHU J H, ZHANG H L, et al. Identification of the pathogens causing potato dry rot in Hebei and Inner Mongolia [J]. Acta Phytophylacica Sinica, 2013, 40(4): 296−300.(in Chinese)
[20]	MENG X L, XIE X W, SHI Y X, et al. Evaluation of a loop-mediated isothermal amplification assay based on hrpZ gene for rapid detection and identification of Pseudomonas syringae pv. lachrymans in cucumber leaves [J]. Journal of Applied Microbiology, 2017, 122(2): 441−449. doi: 10.1111/jam.13356