Use of intersimple sequence repeats markers to develop strain-specific SCAR markers for Lentinula edodes.
Although Lentinula edodes is the second most important cultivated mushroom worldwide, most industrially cultivated strains have been identified only through traditional phenotypic analysis. Here, we report for the first time the use of sequence characterized amplified region (SCAR) markers for strain differentiation. SCAR markers were created by first generating and sequencing single intersimple sequence repeats fragments, and then designing primers based on these sequences to amplify strain-specific fragments of a certain size. One SCAR primer pair, ISL450F/R7 (amplifying a band of c. 450 bp), was designed to identify one strain of L. edodes (strain No. 7). The SCAR primer pair was then used to correctly amplify the single unique fragment from DNA samples taken from a total of 85 strains representing three separate species. Our data provide the foundation for a precise and rapid PCR-based strain-diagnostic system for L. edodes.
College of Life Sciences, Nanjing Agricultural University, Nanjing, China.
Lian-Hua
LH
Qin
Qi
Q
Tan
Ming-Jie
MJ
Chen
Ying-Jie
YJ
Pan
D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis / Q000737:chemistry; D008969:Molecular Sequence Data; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D017422:Sequence Analysis, DNA; D020942:Shiitake Mushrooms / Q000145:classification* / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D004271:DNA, Fungal