Restriction fragment length polymorphism and sequence analysis of rRNA-ITS region of somatic hybrids produced between Pleurotus florida and Lentinula edodes

Annals of Microbiology - Tập 66 - Trang 389-395 - 2015
Pijush Mallick1, Samir Ranjan Sikdar1
1Division of Plant Biology, Bose Institute, Kolkata, India

Tóm tắt

Nine pfle somatic hybrids and their parental strains Pleurotus florida and Lentinula edodes were characterized by restriction fragment length polymorphism (RFLP) and sequence analysis of ribosomal RNA internal transcribed spacer (rRNA-ITS) regions. The ITS1-5.8S-ITS2 region was amplified by primer pair ITS1 and ITS4. Restriction digestion of the amplified ITS1-5.8S-ITS2 region with AluI, HinfI, HpaII and HaeIII generated a total of 81 fragments in which HaeIII generated the maximum of 26 restriction fragments and HpaII generated the minimum of 14 fragments for all 11 strains studied. All the pfle hybrids and parents were highly polymorphic in fragment sizes as revealed from the RFLP profiles. The pfle hybrids and parental ITS1-5.8S-ITS2 sequences showed length variations (629–742 bp). Maximum sequence similarity was found in the 5.8S rRNA region followed by ITS1, while maximum polymorphism was identified in the ITS2 region. The nucleotide sequences of the highly conserved 5.8S region of all the pfle hybrids were very similar with minor variations while this variation was greater between the parental strains. The dendrogram generated based on rRNA-ITS sequences of the pfle hybrids and parents by UPGMA method of clustering revealed five distinct groups, where parent P. florida did not group directly with any of the hybrid strains. The cluster of pfle 6w and pfle 6z remained as separate group in the dendrogram, having no intra cluster linkage.

Tài liệu tham khảo

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