Detection of Flavescence doree phytoplasma in grapevine by reverse-transcription PCR

被引:15
作者
Margaria, P.
Rosa, C.
Marzachi, C.
Turina, M.
Palmano, S. [1 ]
机构
[1] CNR, Ist Virol Vegetale, I-10135 Turin, Italy
[2] Univ Calif Davis, Dept Plant Pathol, Davis, CA 95616 USA
[3] CNR, Ist Virol Vegetale, Turin, Italy
关键词
D O I
10.1094/PDIS-91-11-1496
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Flavescence doree (FD) is the most serious phytoplasma disease of grapevine. This report describes a novel method of detecting FD phytoplasma based on reverse-transcription polymerase chain reaction (RT-PCR) on 16S ribosomal RNA (16SrRNA) which will greatly improve mass screening of infected grapevines. A rapid protocol for extracting sap from whole leaves or mid-veins and successive one-tube amplification by RT-PCR was applied to grapevine samples with or without symptoms collected from different areas of Piedmont (northwestern Italy). Results were compared with those obtained using one of the current diagnostic methods that utilizes nested PCR on phytoplasma DNA-enriched preparations. A Cohen's kappa index of 0.76 indicated a substantial agreement between the two sets of results, The RT-PCR method has the advantage of being a rapid, reliable, and sensitive assay for large-scale screening of grapevines.
引用
收藏
页码:1496 / 1501
页数:6
相关论文
共 41 条
[1]   DETECTION OF DNA OF PLANT PATHOGENIC MYCOPLASMALIKE ORGANISMS BY A POLYMERASE CHAIN-REACTION THAT AMPLIFIES A SEQUENCE OF THE 16S RNA GENE [J].
AHRENS, U ;
SEEMULLER, E .
PHYTOPATHOLOGY, 1992, 82 (08) :828-832
[2]   BASIC LOCAL ALIGNMENT SEARCH TOOL [J].
ALTSCHUL, SF ;
GISH, W ;
MILLER, W ;
MYERS, EW ;
LIPMAN, DJ .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410
[3]  
Angelini E, 2001, VITIS, V40, P79
[4]  
[Anonymous], 1996, Phytopathologia Mediterranea
[5]  
Barba M., 1998, NOTIZIARIO PROTEZION, V9, P263
[6]   Occurrence, distribution and epidemiology of Grapevine Yellows in Spain [J].
Batlle, A ;
Martínez, MA ;
Laviña, A .
EUROPEAN JOURNAL OF PLANT PATHOLOGY, 2000, 106 (09) :811-816
[7]  
Batlle A, 1997, VITIS, V36, P211
[8]   Range of phytoplasma concentrations in various plant hosts as determined by competitive polymerase chain reaction [J].
Berges, R ;
Rott, M ;
Seemüller, E .
PHYTOPATHOLOGY, 2000, 90 (10) :1145-1152
[9]  
BIANCO PA, 2003, P 14 INT C GRAP LOC, P55
[10]  
Boudon-Padieu E, 2003, VITIS, V42, P141