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Quantification of the arbuscular mycorrhizal fungus Glomus intraradices in host tissue using real‐time polymerase chain reaction
Author(s) -
Alkan Noam,
Gadkar Vijay,
Coburn Joel,
Yarden Oded,
Kapulnik Yoram
Publication year - 2004
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1046/j.1469-8137.2004.00975.x
Subject(s) - biology , fungus , medicago truncatula , polymerase chain reaction , glomus , real time polymerase chain reaction , primer (cosmetics) , colonization , mycorrhiza , botany , host (biology) , spore , hypha , microbiology and biotechnology , gene , symbiosis , bacteria , biochemistry , genetics , chemistry , organic chemistry
Summary• A rapid method to quantify the colonization of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices in planta using quantitative real‐time polymerase chain reaction (qRT‐PCR) technique. • Specific PCR primers for the fungus (28S rDNA sequence) and host root tissue (chitinase and chalcone synthase gene) were developed and their respective specificity determined. • The plant specific primers for Lycopersicon esculentum , Medicago truncatula amplified linearly over a concentration range of: 6.4 pg to 20 ng. The G. intraradices ‐specific primer amplified as low as 1 pg of its target DNA, which allowed us to detect a single spore of the fungus. High degrees of correlation were obtained when threshold cycle (Ct) was plotted against vesicular, hyphal and total colonization using microscopically quantified host roots. • This is the first report of the application of the qRT‐PCR technique for quantification of AMF colonization in planta . The success of its application should open up the possibility of its wider application in AM research.