[1]李华伟,林志坚,张 鸿,等.甘薯薯瘟病菌RPA检测方法的建立及应用[J].福建农林大学学报(自然科学版),2020,49(05):583-588.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2020.05.002]
 LI Huawei,LIN Zhijian,ZHANG Hong,et al.Establishment and application of RPA detection for bacterial wilt of sweet potato caused by Ralstonia solanacearum[J].,2020,49(05):583-588.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2020.05.002]
点击复制

甘薯薯瘟病菌RPA检测方法的建立及应用()
分享到:

福建农林大学学报(自然科学版)[ISSN:1671-5470/CN:35-1255/S]

卷:
49卷
期数:
2020年05期
页码:
583-588
栏目:
植物保护
出版日期:
2020-09-18

文章信息/Info

Title:
Establishment and application of RPA detection for bacterial wilt of sweet potato caused by Ralstonia solanacearum
文章编号:
1671-5470(2020)05-0583-06
作者:
李华伟 林志坚 张 鸿 刘中华 李国良 汤 浩 邱思鑫
福建省农业科学院作物研究所/农业部南方薯类科学观测实验站/福建省特色旱作物品种选育工程技术研究中心,福建 福州 350013
Author(s):
LI Huawei LIN Zhijian ZHANG Hong LIU Zhonghua LI Guoliang TANG Hao QIU Sixin
Crop Research Institute, Fujian Academy of Agricultural Sciences/Scientific Observing and Experimental Station of Tuber and Root Crops in South China, Ministry of Agriculture/Technical Research Center of Specialty Dry Crop Variety Breeding of Fujian, Fuzhou, Fujian 350013, China
关键词:
甘薯 薯瘟病 青枯菌 重组酶聚合酶等温扩增方法
Keywords:
sweet potato bacterial wilt Ralstonia solanacearum RPA
分类号:
S531; S411
DOI:
10.13323/j.cnki.j.fafu(nat.sci.).2020.05.002
文献标志码:
A
摘要:
根据甘薯薯瘟病菌特异基因(orf428)序列设计重组酶聚合酶等温扩增(RPA)引物,通过特异性和灵敏性验证,建立了能够快速检测甘薯薯瘟病菌的RPA方法.该方法检测结果与PCR检测结果一致,且灵敏度高、特异性强,在39 ℃的恒温条件下反应20 min即可完成核酸扩增,大大提高了检测效率.此外,该方法可用于检测甘薯发病组织、带菌土壤和水体中的薯瘟病菌.
Abstract:
In order to develop a quick detection method for sweet potato bacterial wilt arising from Ralstonia solanacearum, specific primers for recombinase polymerase amplification(RPA)were designed from the specific gene(orf428)of sweet potato bacterial wilt pathogen. The results of RPA were consistent with those of PCR, with high sensitivity and specificity. It only took 20 minutes to complete nucleic acid amplification at 39 ℃, which greatly improved the detection efficiency. Moreover, the RPA method can be applied to detect infected sweet potato tissue, soil and water body.

参考文献/References:

[1] CHEN Y J, LIN Y S, CHUNG W H. Bacterial wilt of sweet potato caused by Ralstonia solanacearum in Taiwan[J]. Journal of General Plant Pathology, 2012,78(1):80-84.
[2] JIANG G F, WEI Z, XU J, et al. Bacterial wilt in China: history, current status, and future perspectives[J]. Frontiers in Plant Science, 2017,8:1-10.
[3] CLARK C A, WILDER-AYERSJ A, DUARTE V. Resistance of sweet potato to bacterial root and stem rot caused by Erwinia chrysanthemi[J]. Plant Disease, 1989,73(12):984-987.
[4] WANG W, RUAN M H, QIU Y X, et al. Phenylaprapanoid metabolism of sweet potato against Pseudomonas solanacearum[J]. Chinese Journal of Eco-Agriculture, 2009,17(5):944-948.
[5] SCHAADN W, BRENNER D. A bacterial wilt and root rot of sweet potato caused by Erwinia chrysanthemi[J]. Phytopathology, 1977,67(3):302-308.
[6] GRAHAM J, LIOYD A B. Survival of potato strain(race 3)of Pseudomonas solanacearum in the deeper soil layers[J]. Australian Journal of Agricultural Research, 1979,30(3):489-496.
[7] JANSEJ D. A detection method for Pseudomonas solanacearum in symptomless potato tubers and some data on its sensitivity and specificity[J]. EPPO Bulletin, 1988,18(3):343-351.
[8] 王俊,王金正,苏佳威,等.一种制备桑青枯菌抗体的方法及其在DAS-ELISA检测方法上的应用:CN110386979A[P].2019-10-29.
[9] NOTOMI T, OKAYAMA H, MASUBUCHI H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Research, 2000,28(12):e63.
[10] 王帅,徐进,许景升,等.PMA-qPCR定量检测青枯菌活菌方法的建立[J].植物保护,2018,44(6):122-128,135.
[11] 黄雯,徐进,张昊,等.植物青枯菌LAMP检测方法的建立[J].中国农业科学,2016,49(11):2093-2102.
[12] 张海燕,张小芳,魏兰芳,等.土壤中茄科雷尔氏菌实时荧光定量PCR快速检测体系的建立与应用[J].江苏农业科学,2017,45(14):17-20.
[13] 许文建,丁良,肖逍,等.一种基于噬菌体的青枯菌检测试剂盒:CN209442998U[P].2019-09-27.
[14] 周大祥,熊书,谢桂香.利用MTT建立一种快速检测青枯菌活菌的方法[J].生物学杂志,2019,36(1):103-106.
[15] PIEPENBURG O, WILLIAMS C H, STEMPLE D L, et al. DNA detection using recombination proteins[J]. PLoS Biology, 2006,4(7):1115-1121.
[16] CUI J, ZHAO Y N, SUN Y, et al. Detection of Babesia gibsoni in dogs by combining recombinase polymerase amplification(RPA)with lateral flow(LF)dipstick[J]. Parasitology Research, 2018,117(12):3945-3951.
[17] KERSTING S, RAUSCH V, BIERF F, et al. Multiplex isothermal solid-phase recombinase polymerase amplification for the specific and fast DNA-based detection of three bacterial pathogens[J]. Microchimica Acta, 2014,181(13/14):1715-1723.
[18] FENG X Y, SHEN L B, WANG W Z, et al. Development of a reverse transcription-recombinase polymerase amplification assay for detection of Sugarcane yellow leaf virus[J]. Sugar Tech, 2018,20(6):700-707.
[19] OPINA N, TAVNER F, HOLLWAY G, et al. A novel method for development of species and strain-specific DNA probes and PCR primers for identifying Burkholderia solanacearum(formerly Pseudomonas solanacearum)[J]. Asia-Pacific Journal of Molecular Biology and Biotechnology, 1997,5(1):19-30.
[20] WANG T M, YANG J T. Visual DNA diagnosis of Tomato yellow leaf curl virus with integrated recombinase polymerase amplification and a gold-nanoparticle probe[J]. Scientific Reports, 2019,9(1):e15146.
[21] 魏梅生,田茜,赵文军,等.番茄细菌性叶斑病菌RPA检测技术[J].植物保护,2016,42(1):150-153.
[22] GLAIS L, JACQUOT E. Detection and characterization of viral species/subspecies using isothermal recombinase polymerase amplification(RPA)assays[J]. Methods Mol Biol, 2015,1302:207-225.
[23] JU Y, LIN Y, YANG G, et al. Development of recombinase polymerase amplification assay for rapid detection of Meloidogyne incognita, M. javanica, M. arenaria, and M. enterolobii[J]. European Journal of Plant Pathology, 2019,155(4):1155-1163.

相似文献/References:

[1]陈选阳,张招娟.高淀粉能源甘薯新品种“金山208”的选育及特性鉴定[J].福建农林大学学报(自然科学版),2012,41(06):570.
 CHEN Xuan-yang,ZHANG Zhao-juan.Breeding of new sweet potato cultivar “Jinshan 208” with high starch content and identification of its characteristics[J].,2012,41(05):570.

备注/Memo

备注/Memo:
收稿日期:2019-12-09 修回日期:2020-01-29
基金项目:福建省公益类科研专项(2017R1026-4); 现代农业产业技术体系(CARS-10-B14); 福建省农业科学院创新团队项目(STIT2017-2-3).
作者简介:李华伟(1982-),男,助理研究员.研究方向:植物病虫害防控.Email:hwlpoto@126.com.通信作者邱思鑫(1974-),男,研究员,博士.研究方向:植物病理学.Email:qiusixin@faas.cn.
更新日期/Last Update: 2020-09-20