Detection of Seed-borne Rice Bakanae Pathogens in Jiangsu Province, China Using Loop-mediated Isothermal Amplification Assays

doi:10.16819/j.1001-7216.2018.7119

Abstract

Abstract:

【Objective】The research aims to reveal the situation of seed-borne rice bakanae pathogens in Jiangsu Province, China. 【Methods】The seeds of 65 major rice cultivars collected from this area were analyzed by using loop-mediated isothermal amplification (LAMP) assays for specific detection of 4 known rice bakanae pathogen species, including Fusarium fujikuroi, F. proliferatum, F. verticilliodes and F. andiyazi. 【Results】There were 54 rice cultivars carrying rice bakanae pathogens, indicating it’s a common phenomenon that rice seeds carrying rice bakanae pathogens in Jiangsu Province. And the most frequent species was F. fujikuroi, representing 83.07% of the total samples tested, followed by F. proliferatum and F. andiyazi, representing 10% and 6% respectively. However, none of the 65 rice cultivars was tested to be carrying F. verticilliodes. 【Conclusions】The method is readily available for detecting seed-borne rice bakanae pathogens.

Key words: rice bakanae, detection of seed-borne pathogens, loop-mediated isothermal amplification

摘要：

【目的】本研究旨在了解江苏省水稻主栽品种种子携带水稻恶苗病原菌的情况。【方法】采用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)对江苏65个水稻主栽品种种子携带藤仓镰孢、层出镰孢、拟轮枝镰孢和Fusarium andiyazi等4种水稻恶苗病菌进行检测。【结果】65个品种中有54个品种的种子样本携带恶苗病菌,表明江苏省稻种携带水稻恶苗病菌是一种普遍现象。其中,藤仓镰孢为优势种,检出率高达83.07%,其次是层出镰孢和Fusarium andiyazi,检出率分别为12.30%、6.15%,但供试的65个品种种子均未检测到携带有拟轮枝镰孢。【结论】该技术可用于水稻种子携带恶苗病菌的快速检测。

关键词: 水稻恶苗病, 种子带菌检测, 环介导等温扩增技术

CLC Number:

Yongtian YUAN, Zhenyang RONG, Wenwu YE, Hongsheng ZHANG, Yiqing ZHUANG, Xiaobo ZHENG. Detection of Seed-borne Rice Bakanae Pathogens in Jiangsu Province, China Using Loop-mediated Isothermal Amplification Assays[J]. Chinese Journal OF Rice Science, 2018, 32(5): 493-500.

袁咏天, 戎振洋, 叶文武, 张红生, 庄义庆, 郑小波. 应用环介导等温扩增技术检测江苏水稻种子携带的水稻恶苗病菌[J]. 中国水稻科学, 2018, 32(5): 493-500.

Figures/Tables 5

References 22

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目标病原菌 Target pathogen	靶标基因 Target gene	引物名称 Primer name	序列 Sequence (5＇-3＇)
藤仓镰孢 Fusarium fujikuroi	IGS	FIP	TCAGCCTACCCTAGACCCTCGCCCTTACCTTTGCTTGA
		BIP	AGCTAGACTACTCTAAGGGTAGGTACAACTTCCACCAATTGCA
		F3	AGACACCGTTTTGTTTTGC
		B3	CTTTTCTCCAACTTCACAGC
		LF	ATCCGGCCAAGACCGTC

目标病原菌 Target pathogen	靶标基因 Target gene	引物名称 Primer name	序列 Sequence (5＇-3＇)
藤仓镰孢 Fusarium fujikuroi	IGS	FIP	TCAGCCTACCCTAGACCCTCGCCCTTACCTTTGCTTGA
		BIP	AGCTAGACTACTCTAAGGGTAGGTACAACTTCCACCAATTGCA
		F3	AGACACCGTTTTGTTTTGC
		B3	CTTTTCTCCAACTTCACAGC
		LF	ATCCGGCCAAGACCGTC

检测技术 LAMP assay	目标病原菌 Objective species	扩增条件 LAMP reaction	灵敏度 Sensitivity/(μg·L^-1)	参考文献 Reference
RED1-Fp-LAMP	F. proliferatum	62℃, 70 min	400	[18]
Pgk-Fv-LAMP	F. verticillioides	62℃, 70 min	100	[20]
TAT-Fan-LAMP	F. andiyazi	64℃, 80 min	100	[21]

检测技术 LAMP assay	目标病原菌 Objective species	扩增条件 LAMP reaction	灵敏度 Sensitivity/(μg·L^-1)	参考文献 Reference
RED1-Fp-LAMP	F. proliferatum	62℃, 70 min	400	[18]
Pgk-Fv-LAMP	F. verticillioides	62℃, 70 min	100	[20]
TAT-Fan-LAMP	F. andiyazi	64℃, 80 min	100	[21]

编号 Number	水稻品种 Rice cultivar	来源地 Origin	检测到的病原菌 Pathogens detected	编号 Number	水稻品种 Rice cultivar	来源地 Origin	检测到的病原菌 Pathogens detected
1	宁粳1号 Ningjing 1	南京Nanjing	Ff	34	徐稻9号 Xudao 9	徐州Xuzhou	Ff
2	宁粳2号 Ningjing 2	南京Nanjing	Ff, Fp	35	镇稻14 Zhendao 14	淮安Huaian	Ff
3	宁粳3号 Ningjing 3	南京Nanjing	Ff	36	淮稻5号 Huaidao 5	淮安Huaian	Ff
4	宁粳4号 Ningjing 4	南京Nanjing	Ff	37	淮稻18 Huaidao 18	淮安Huaian	Ff
5	宁粳5号 Ningjing 5	南京Nanjing	Ff	38	淮糯12 Huaidao 12	淮安Huaian	Ff, Fan
6	宁粳6号 Ningjing 6	南京Nanjing	Ff, Fp	39	淮稻20 Huaidao 20	淮安Huaian	Ff, Fp
7	宁粳7号 Ningjing 7	南京Nanjing	Ff	40	润农4号 Runnong 4	淮安Huaian	—
8	宁粳8号 Ningjing 8	南京Nanjing	Ff	41	隆粳968 Longjing 968	淮安Huaian	Ff
9	武运粳23 Wuyunjing 23	常州Changzhou	Ff	42	淮香粳15号 Huaixiangjing 15	淮安Huaian	Ff
10	武运粳27 Wuyunjing 27	常州Changzhou	—	43	两优0367 Liangyou 0367	盐城Yancheng	—
11	武运粳30 Wuyunjing 30	常州Changzhou	—	44	盐两优1618 Yanliangyou 1618	盐城Yancheng	Ff
12	扬育粳2 Yangyujing 2	镇江Zhenjiang	Ff	45	盐两优888 Yanliangyou 888	盐城Yancheng	—
13	常农粳8 Changnongjing 8	镇江Zhenjiang	—	46	盐两优2208 Yanliangyou 2208	盐城Yancheng	—
14	武运粳23 Wuyunjing 23	镇江Zhenjiang	Ff	47	盐两优2218 Yanliangyou 2218	盐城Yancheng	Ff
15	镇稻11 Zhendao 11	镇江Zhenjiang	Ff	48	盐稻8号 Yandao 8	盐城Yancheng	Ff
16	镇稻14 Zhendao 11	镇江Zhenjiang	Ff	49	盐稻9号 Yandao 9	盐城Yancheng	—
17	镇稻16 Zhendao 16	镇江Zhenjiang	Ff, Fan	50	盐稻10号 Yandao 10	盐城Yancheng	Ff
18	镇稻18 Zhendao 18	镇江Zhenjiang	Ff, Fp	51	盐稻11 Yandao 11	盐城Yancheng	Ff, Fan
19	镇糯19 Zhendao 19	镇江Zhenjiang	Ff	52	盐稻12 Yandao 12	盐城Yancheng	Ff
20	镇糯20 Zhendao 20	镇江Zhenjiang	Ff	53	荃优1393 Quanyou 1393	盐城Yancheng	—
21	镇稻21 Zhendao 21	镇江Zhenjiang	Ff	54	连粳1号 Lianjing 1	连云港Lianyungang	Ff
22	苏秀867 Suxiu 867	镇江Zhenjiang	Ff	55	连粳5号 Lianjing 5	连云港Lianyungang	Ff, Fp
23	皖垦糯1号 Wankennuo 1	镇江Zhenjiang	Ff	56	连粳6号 Lianjing 6	连云港Lianyungang	Ff
24	扬两优013 Yangliangyou 013	扬州Yangzhou	Ff	57	连粳7号 Lianjing 7	连云港Lianyungang	Ff
25	丰优香5号 Fengyouxiang 5	扬州Yangzhou	—	58	连粳9号 Lianjing 9	连云港Lianyungang	Ff
26	扬稻6号 Yangdao 6	扬州Yangzhou	Ff, Fp	59	连粳10号 Lianjing 10	连云港Lianyungang	Ff
27	武运粳23 Wuyunjing 23	扬州Yangzhou	Ff, Fan	60	连粳11 Lianjing 11	连云港Lianyungang	Ff
28	武运粳27 Wuyunjing 27	扬州Yangzhou	Ff	61	连粳12 Lianjing 12	连云港Lianyungang	Ff
29	南粳9108 Nanjing 9108	扬州Yangzhou	Ff	62	连粳13 Lianjing 13	连云港Lianyungang	Ff, Fp
30	扬粳糯1号 Yangjingnuo 1	扬州Yangzhou	Ff, Fp	63	连粳15 Lianjing 15	连云港Lianyungang	Ff
31	南粳52 Nanjing 52	扬州Yangzhou	Ff	64	连糯1号 Liannuo 1	连云港Lianyungang	Ff
32	徐稻3号 Xudao 3	徐州Xuzhou	Ff	65	中稻1号 Zhongdao 1	连云港Lianyungang	Ff
33	徐稻8号 Xudao 8	徐州Xuzhou	—