基于rDNA - ITS序列建立LAMP检测水稻潜根线虫的方法
A loop-mediated isothermal amplification method for detection of Hirschmanniella oryzae based on rDNA-ITS sequences
查看参考文献27篇
|
文摘
|
水稻潜根线虫(Hirschmanniella oryzae)在水稻产区普遍存在,是水稻(Oryza spp.)的主要植物寄生线虫,在世界范围内造成产量损失。本研究通过比较GenBank数据库中潜根属线虫相关序列,以水稻潜根线虫ITS-rDNA序列为靶标,设计了水稻潜根线虫LAMP的一组特异性引物,包括一对外引物(F3:5'-ATCTTGTCCTTTGGCACG-3',B3:5'-CGGTTGAACAAACAACGT- 3')和一对内引物(FIP:5'-CAGCATAGCAACAGAATGAATTCACGGTCGTAAACCTAATACGCG-3',BIP:5'-TTGTACTACAATGGATTGTTTTCGCCTGATCCATCCACCCATG-3');用琼脂凝胶电泳法和SYBR Green I染色法筛选合适的引物和扩增条件,建立了优化的水稻潜根线虫LAMP检测条件:扩增温度为57℃,反应时间为45 min,反应体系中dNTPs和MgSO_4的浓度分别为1.4 mmol·L~(-1)和7 mmol·L~(-1)。本研究建立的检测方法可以检测鉴定水稻潜根线虫不同发育时期虫态(雌虫、雄虫、幼虫)个体,还可以从近缘种和其他植物寄生线虫混合的样品及水稻根组织样品中直接检测出水稻潜根线虫,且检测灵敏度达到1/1 000条成虫DNA。 |
|
其他语种文摘
|
Hirschmanniella oryzae is widely distributed in rice producing areas,and it has been affecting the growth of rice and causing some yield losses worldwide.The specific primers for the loop-mediated isothermal amplification (LAMP) were designed based on rDNA ITS sequences of H.oryzae by comparing with related sequences of Hirschmanniella spp.deposited in GenBank.Two external Primers (F3:5'-ATCTTGTCCTTTGGCACG- 3',B3:5'-CGGTTGAACAAACAACGT-3') and two inner primers (FIP:5'-CAGCATAGCAACAGAATGAATTCACGGTCGTAAACCTAATACGCG- 3',BIP:5'-TTGTACTACAATGGATTGTTTTCGCCTGATCCATCCACCCATG- 3') were designed.The optimal reaction conditions were screened based on electrophoresis detection and visual observation with SYBR Green I staining method,and the optimal conditions are:LAMP amplification under 57℃ for 45 min,the concentration of dNTPs and MgSO_4 used in the reaction volume is 1.4 mmol·L~(-1) and 7 mmol·L~(-1),respectively.The results showed that the LAMP method could detect and identify single juvenile,female or male of H.oryzae,and also could directly detect H.oryzae from mixed nematode species and rice root samples,the sensitivity of LAMP assay was 1/1 000 of single nematode DNA. |
|
来源
|
植物病理学报
,2021,51(4):626-635 【核心库】
|
|
DOI
|
10.13926/j.cnki.apps.000538
|
|
关键词
|
水稻潜根线虫
;
LAMP
;
rDNA-ITS
;
快速检测
|
|
地址
|
华南农业大学农学院/华南农业大学植物检疫线虫检测与防疫研究中心, 广州, 510642
|
|
语种
|
中文 |
|
文献类型
|
研究性论文 |
|
ISSN
|
0412-0914 |
|
学科
|
植物保护 |
|
基金
|
国家自然科学基金
;
科技部基础资源调查专项
|
|
文献收藏号
|
CSCD:7033786
|
参考文献 共
27
共2页
|
|
1.
Webster J M.
Economic nematology,1972:125-143
|
CSCD被引
1
次
|
|
|
|
|
2.
Sher A S. Revision of thegenus Hirschmanniella Luc & Goodey, 1963 (Nematoda: Tylenchoidea).
Nematologica,1968,14(2):243-275
|
CSCD被引
5
次
|
|
|
|
|
3.
Smart G C.
Tropical nematology,1968:68-80
|
CSCD被引
1
次
|
|
|
|
|
4.
Luc M.
Plant parasitic nematodes in subtropical and tropical Agriculture (2nd Edition),1990:87-130
|
CSCD被引
1
次
|
|
|
|
|
5.
Hu X Q. Preliminary study on species of Hirshchamanniella in Yunnan China (in Chinese).
西南农业大学学报,2004,26(1):40-43
|
CSCD被引
1
次
|
|
|
|
|
6.
Zhang S S. Identification of rice-root nematodes (Hirschmanniella spp.) in Fujian,China (in Chinese).
福建农林大学学报(自然科学版),1987,16(2):155-159
|
CSCD被引
1
次
|
|
|
|
|
7.
Chen D Y. Distribution of rice root nematode Hirschmanniella oryzae and a new recorded H. mucronata (Nematoda: Pratylenchidae) in Taiwan.
Plant Pathology Bulletin,2006,15:197-210
|
CSCD被引
2
次
|
|
|
|
|
8.
Tandingan De Ley. Nematodes from vernal pools inthe Santa Rosa Plateau Ecological Reserve,California I. Hirschmanniella santarosae sp. n. (Nematoda: Pratylenchidae), a cryptic sibling species of H. pomponiensis AbdelRahman & Maggenti,1987.
Nematology,2007,9:405-429
|
CSCD被引
1
次
|
|
|
|
|
9.
Berg E V D. Hirschmanniella kwazuna sp. n. from South Africa with notes on a new record of H. spinicaudata (Schuurmans Stekhoven,1944) Luc & Goodey,1964 (Nematoda: Pratylenchidae) and on the molecular phylogeny of Hirschmanniella Luc & Goodey,1964.
Nematology,2009,11(4):523-540
|
CSCD被引
2
次
|
|
|
|
|
10.
Feng H. Morphological and molecular characterization of the rice root nematode, Hirschmanniella mucronata,from Jiangsu,China (in Chinese).
植物病理学报,2016,46(4):474-484
|
CSCD被引
1
次
|
|
|
|
|
11.
Katsuta A. Development of real-time PCR primers for the quantification of Meloidogyne graminicola,Hirschmanniella oryzae and Heterodera cajani,pests of the major crops in Myanmar.
Nematology,2016,18(3):257-263
|
CSCD被引
1
次
|
|
|
|
|
12.
Notomi T. Loopmediated isothermal amplification of DNA.
Nucleic Acids Research,2000,28(12):63
|
CSCD被引
991
次
|
|
|
|
|
13.
Mori Y. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation.
Biochemical and Biophysical Research Communications,2001,289(1):150-154
|
CSCD被引
163
次
|
|
|
|
|
14.
Kikuchi T. A Rapid and precise diagnostic method for detecting the pinewood nematode Bursaphelenchus xylophilus by loop-mediated isothermal amplification.
Nematology,2009,99(12):1365-1369
|
CSCD被引
20
次
|
|
|
|
|
15.
Meng F. A loop-mediated isothermal amplification-based method for detecting Bursaphelenchus xylophilus from Monochamus alternatus.
Forest Pathology,2018,48(2):1
|
CSCD被引
1
次
|
|
|
|
|
16.
Niu J H. Rapid detection of Meloidogyne spp. by LAMP assay in soil and roots.
Crop Protection,2011,30(8):1063-1069
|
CSCD被引
13
次
|
|
|
|
|
17.
He X F. Loop-mediated isothermal amplification assay for rapid diagnosis of Meloidogyne enterolobii directly from infected plants (in Chinese).
中国农业科学,2013,46(3):534-544
|
CSCD被引
2
次
|
|
|
|
|
18.
Wei H Y. Loop-mediated isothermal amplification assay for rapid diagnosis of Meloidogyne mali (in Chinese).
植物保护学报,2016,43(2):260-266
|
CSCD被引
2
次
|
|
|
|
|
19.
Cai Y. Rapid and sensitive detection of Meloidogyne camelliae by LAMP-LFD (in Chinese).
农业生物技术学报,2016,24(5):770-780
|
CSCD被引
1
次
|
|
|
|
|
20.
Zhou Q J. Rapid and sensitive detection of Meloidogyne mali by loop-mediated isothermal amplification combined with a lateral flow dipstick.
European Journal of Plant Pathology,2017,148(4):755-769
|
CSCD被引
2
次
|
|
|
|
|
了解气象卫星更多信息，请访问