Mycobiology.  2010 Jun;38(2):89-96.

Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

• Alam N
• Kim JH
• Shim MJ
• Lee UY
• Lee TS

Abstract

This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at 25degrees C and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Keyword

Agrocybe cylindracea; ITS; Mycelial growth; RAPD; rDNA

MeSH Terms

Agar
Agrocybe
Alanine
Arginine
Carbon
China
DNA
DNA, Ribosomal
Fructose
Genetic Variation
Glucose
Glycine
Histidine
Hydrogen-Ion Concentration
Korea
Maltose
Methionine
Nitrogen
Polymerase Chain Reaction
Solanum tuberosum
Taiwan
Urea
Agar
Alanine
Arginine
Carbon
DNA
DNA, Ribosomal
Fructose
Glucose
Glycine
Histidine
Maltose
Methionine
Nitrogen
Urea