Development of Cleaved Amplified Polymorphic Sequence Markers of Lentinula edodes Cultivars Sanbaekhyang and Sulbaekhyang

Suyun Moon1   Chang Pyo Hong2   Hojin Ryu1,*   Hwa-Yong Lee3,*   

1Department of Biology, Chungbuk National University, Cheongju 28644, Korea
2Theragen Bio, Suwon 16229, Korea
3Department of Forest Science, Chungbuk National University, Cheongju 28644, Korea

Abstract

Lentinula edodes (Berk.) Pegler, the most produced mushroom in the world, is an edible mushroom with very high nutritional and pharmacological value. Currently, interest in the protection of genetic resources is increasing worldwide, and securing the distinction between new cultivars is very important. Therefore, the development of efficient molecular markers that can discriminate between L. edodes cultivars is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanbaekhyang and Sulbaekhyang). These markers were developed from whole genome sequencing data from L. edodes monokaryon strain B17 and resequencing data from 40 cultivars. A nucleotide deletion existed in scaffold 19 POS 214449 in Sanbaekhyang (GT→G), and a single nucleotide polymorphism changed in scaffold 7 POS 215801 in Sulbaekhyang (G→A). The restriction enzymes HhaⅠ and HpyCH4Ⅳ distinguished Sanbaekhyang and Sulbaekhyang, respectively, from other cultivars. Thus, we developed two CAPS markers for the identification of the L. edodes cultivars Sanbaekhyang and Sulbaekhyang.

Figures & Tables

Fig. 1. A and B. Partial nucleotide sequences of each strains amplified with RL-LE-133 (A), RL-LE-145 (B) primer set respectively. Highlighted nucleotides were used for developing the cleaved amplified polymorphic sequence (CAPS) marker. The restriction enzyme used for development of CAPS marker and the position of the recognition site in each marker sequence are shown in a schematic diagram. C and D. The cleaved fragment patterns of each CAPS marker among 40 L. edodes cultivars. The name of cultivars corresponding the indicated number was represented in Table 1.