Molecular characterizations of somatic hybrids developed between Pleurotus florida and Lentinus squarrosulus through inter-simple sequence repeat markers and sequencing of ribosomal RNA-ITS gene

Pijush Mallick; Shruti Chattaraj; Samir Ranjan Sikdar

doi:10.1007/s13205-017-0931-2

Molecular characterizations of somatic hybrids developed between Pleurotus florida and Lentinus squarrosulus through inter-simple sequence repeat markers and sequencing of ribosomal RNA-ITS gene

3 Biotech. 2017 Oct;7(5):298. doi: 10.1007/s13205-017-0931-2. Epub 2017 Sep 6.

Authors

Pijush Mallick^{1

2}, Shruti Chattaraj¹, Samir Ranjan Sikdar¹

Affiliations

¹ Division of Plant Biology, Bose Institute, P-1/12, C.I.T., Scheme VII M, Kolkata, 700 054 India.
² Present Address: Unidad Académica Sistemas Arrecifales, Instituto de Ciencias del Mar y Limnología, UNAM, Prol. Avenida Niños Heroes S/N, 77580 Puerto Morelos, Quintana Roo Mexico.

Abstract

The 12 pfls somatic hybrids and 2 parents of Pleurotus florida and Lentinus squarrosulus were characterized by ISSR and sequencing of rRNA-ITS genes. Five ISSR primers were used and amplified a total of 54 reproducible fragments with 98.14% polymorphism among all the pfls hybrid populations and parental strains. UPGMA-based cluster exhibited a dendrogram with three major groups between the parents and pfls hybrids. Parent P. florida and L. squarrosulus showed different degrees of genetic distance with all the hybrid lines and they showed closeness to hybrid pfls 1m and pfls 1h, respectively. ITS1(F) and ITS4(R) amplified the rRNA-ITS gene with 611-867 bp sequence length. The nucleotide polymorphisms were found in the ITS1, ITS2 and 5.8S rRNA region with different number of bases. Based on rRNA-ITS sequence, UPGMA cluster exhibited three distinct groups between L. squarrosulus and pfls 1p, pfls 1m and pfls 1s, and pfls 1e and P. florida.

Keywords: ISSR; Mushroom somatic hybrid; Polymorphism; rRNA-ITS gene.