|
| Molecular Identification and Analysis of New Cultivar WuzhiNo. 2 Based on Ribosomal DNA ITS and ITS2 Sequence |
| |
| DOI：10.16768/j.issn.1004-874X.2022.02.002 |
|
| Hits: 1330 |
| Download times: 891 |
| Abstract: |
| 【Objective】The study was carried out to explore the molecular identification of Wuzhi No. 2 (original
name: Zizhi S2), a new cultivar of Ganoderma sp., by rDNA-ITS sequence analysis and ITS2 secondary structure model.
【Method】The sequences of Wuzhi No. 2 were acquired by the first and third generation sequencing techniques
respectively, and the phylogenetic tree was constructed based on ITS sequences of related species of genus Ganoderma for
molecular classification and identification, the rDNA-ITS and related primer sequences were aligned by BLAST with the
whole genome, and the secondary structure of ITS2 was predicted online.【Result】Sanger sequencing showed that the ITS sequence of Ganoderma sp. strain Wuzhi No. 2 (GenBank: MG282563.1, 649 bp) displayed high similarity (99.53% -100%)
with five known Ganoderma sinense strains in the GenBank, while Wuzhi No. 2 and four known strains or voucher specimens
of G. sinense (strain) were clustered into a same branch on the Ganoderma spp. phylogenetic tree. Two known short sequences
of specific primers (GS_22bp and GS2_22bp) appeared in the forward position of 119-140 bp and the reverse position of 588-
567 bp of ITS sequence of Wuzhi No. 2, respectively. Four tandem repeat ITS regions were found from the rDNA on Scaffold
No. 49 of Wuzhi No. 2 genome, which was highly consistent with the single ITS sequence by Sanger method. In terms of the
secondary structure of ITS2 sequence, three helix regions in Wuzhi No. 2 and Ganoderma sinense were the same, but only the
included angle in Helix IV region was different.【Conclusion】Based on ITS/ITS2 molecular identification, it was confirmed
that the new cultivar Wuzhi No. 2 and the known strains of Ganoderma sinense belonged to the same population, there were
angle differences in the Helix IV region of secondary structure of ITS2, and the effectiveness of two known specific amplification
primers were also verified. |
|
View Full Text
View/Add Comment Download reader |
|
|
|
