A multiplex RT-PCR assay for simultaneous detection of Tomato spotted wilt virus and Tomato yellow ring virus in tomato plants

被引:6
作者
Zarzynska-Nowak, Aleksandra [1 ]
Hasiow-Jaroszewska, Beata [1 ]
Korbecka-Glinka, Grazyna [2 ]
Przybys, Marcin [2 ]
Borodynko-Filas, Natasza [1 ]
机构
[1] Natl Res Inst, Inst Plant Protect, Dept Virol & Bacteriol, Ul Wl Wegorka 20, PL-60318 Poznan, Poland
[2] State Res Inst, Inst Soil Sci & Plant Cultivat, Dept Plant Breeding & Biotechnol, Ul Czartoryskich 8, PL-24100 Pulawy, Poland
关键词
co-infection; detection; multiplex RT-PCR; TSWV; TYRV; TOSPOVIRUS; DIFFERENTIATION; ISOLATE; POTATO; IRAN;
D O I
10.1080/07060661.2018.1503195
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Orthotospoviruses have become one of the major threats to vegetable crops worldwide. Recently, severe outbreaks of Tomato spotted wilt virus (TSWV) and Tomato yellow ring virus (TYRV) have been observed in tomato production regions in Poland. In this study, a multiplex RT-PCR assay was developed and optimized for the simultaneous detection of these two viruses. Primers were designed to amplify 666 bp and 328 bp of the nucleocapsid (N) genes of TYRV and TSWV, respectively. The multiplex RT-PCR detection limit was 1 pgL(-1) of total RNA, and it was successfully applied for the detection of TSWV and TYRV in 21 samples of tomato, tobacco and weed plants collected in Poland. Four samples were found to be co-infected with both viruses, whereas infection solely with TSWV and TYRV was found in 9 and 8 samples, respectively. Overall, this study presents a rapid, specific and sensitive method for orthotospoviruses identification in plants that can be applied to mixed infections.
引用
收藏
页码:580 / 586
页数:7
相关论文
共 35 条
[11]  
Hall T. A., NUCL ACIDS S SER, V41, P95, DOI DOI 10.1021/BK-1999-0734.CH008
[12]   A new tomato-infecting tospovirus from Iran [J].
Hassani-Mehraban, A ;
Saaijer, J ;
Peters, D ;
Goldbach, R ;
Kormelink, R .
PHYTOPATHOLOGY, 2005, 95 (08) :852-858
[13]   Molecular and biological comparison of two Tomato yellow ring virus (TYRV) isolates:: challenging the Tospovirus species concept [J].
Hassani-Mehraban, A. ;
Saaijer, J. ;
Peters, D. ;
Goldbach, R. ;
Kormelink, R. .
ARCHIVES OF VIROLOGY, 2007, 152 (01) :85-96
[14]   Development of a generic method for inspection of tospoviruses [J].
Huang, Kuo-Shiou ;
Li, Siang-ng ;
Sun, Jing-Hua ;
Wang, Yun-Chi ;
Jan, Fuh-Jyh ;
Chen, Tsung-Chi .
EUROPEAN JOURNAL OF PLANT PATHOLOGY, 2018, 150 (02) :457-469
[15]  
Kamiska M, 1997, PROG PLANT PROT, V37, P374
[16]  
Kulshrestha S, 2013, J APPL HORT, V15, P7180
[17]   Improved multiplex reverse transcription-polymerase chain reaction to detect and identify five tospovirus species simultaneously [J].
Kuwabara, Katsuya ;
Yokoi, Naoto ;
Ohki, Takehiro ;
Tsuda, Shinya .
JOURNAL OF GENERAL PLANT PATHOLOGY, 2010, 76 (04) :273-277
[18]  
Laskowska D., 2008, STUD REP I SOIL SCI, V13, P43
[19]   Development of a microarray for simultaneous detection and differentiation of different tospoviruses that are serologically related to Tomato spotted wilt virus [J].
Liu, Lu-Yuan ;
Ye, He-Yi ;
Chen, Tsang-Hai ;
Chen, Tsung-Chi .
VIROLOGY JOURNAL, 2017, 14 :1-10
[20]  
Mortazavi N., 2013, Linzer Biologische Beitraege, V45, P2011