Detection of tomato yellow leaf curl virus by loop-mediated isothermal amplification reaction.
The genomic DNA molecule of tomato yellow leaf curl virus (TYLCV), a whitefly-transmitted geminivirus, was amplified from total DNA extracts of TYLCV-infected tomato (Lycopersicon esculentum) by the use of loop-mediated isothermal amplification (LAMP). The procedure was also used to amplify TYLCV DNA from total DNA extracts of individual whiteflies (Bemisia tabaci) that had fed on TYLCV-infected plants. One of the characteristics of the LAMP method is its ability to synthesize an extremely large amount of DNA. Accordingly, a large amount of by-product, pyrophosphate ion, is produced yielding a white precipitate of magnesium pyrophosphate in the reaction mixture. The presence or absence of this white precipitate allows easy detection of amplification of TYLCV genomic DNA without gel electrophoresis.
Fukuta
Shiro
S
Aichi-ken Agricultural Research Center, 1-1 Sagamine, Yazako, Nagakute, Aichi 480-1193, Japan. sfukuta@agri-rc.pref.aichi.jp
Kato
Shinro
S
Yoshida
Keiko
K
Mizukami
Yuko
Y
Ishida
Akira
A
Ueda
Junnichi
J
Kanbe
Michio
M
Ishimoto
Yoshiyuki
Y
D000595:Amino Acid Sequence; D000818:Animals; D011232:Chemical Precipitation; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D017791:Geminiviridae / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D005784:Gene Amplification; D006430:Hemiptera / Q000502:physiology / Q000821:virology*; D018551:Solanum lycopersicum / Q000821:virology*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010935:Plant Diseases / Q000821:virology; D016133:Polymerase Chain Reaction
D004279:DNA, Viral