Cloning and expression analysis of PpSAMS gene of Pleurotus pulmonarius

doi:10.3969/j.issn.1004-1524.2021.01.08

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (1): 62-68.DOI: 10.3969/j.issn.1004-1524.2021.01.08

• Horticultural Science • Previous Articles Next Articles

Cloning and expression analysis of PpSAMS gene of Pleurotus pulmonarius

WANG Weike(), LU Na, YAN Jing, SONG Jiling, YUAN Weidong, ZHOU Zufa

Hangzhou Academy of Agricultural Sciences, Hangzhou 310024,China

Received:2020-04-17 Online:2021-01-25 Published:2021-01-25

Abstract

Abstract:

Based on the high-throughput transcriptome sequencing of different growth stages of Pleurotus pulmonarius samples, the full-length gene of S-adenosylmethionine synthetase was obtained by analysis and screening. In order to explore the function and expression difference of PpSAMS in the formation of primordium of Pleurotus pulmonarius after low temperature stimulation, the gene of PpSAMS was cloned and analyzed systematically in order to explore the function and expression difference of PpSAMS in the primordium formation of Pleurotus pulmonarius after low temperature stimulation. The expression of the gene in different growth stages was analyzed by real time fluorescence quantitative PCR. Bioinformatics analysis showed that PpSAMS gene was composed of 1 152 nucleotides and encoded 384 amino acids. It had the function of binding ATP and metal ions, and had the activity of S-adenosylmethionine synthetase. The phylogenetic tree analysis of amino acid sequence showed that the protein encoded by PpSAMS gene was closely related to Pleurotus ostreatus. Real time fluorescence quantitative PCR indicated that the expression of PpSAMS was the highest when the mycelium recovered to room temperature after low temperature induction (prophase of primordium formation). It might play an important role in the process of DNA methylation and primordium formation, and participate in the development of fruiting body.

Key words: Pleurotus pulmonarius, clone, real time fluorescence quantitative PCR, PpSAMS, expression analysis

CLC Number:

S646

WANG Weike, LU Na, YAN Jing, SONG Jiling, YUAN Weidong, ZHOU Zufa. Cloning and expression analysis of PpSAMS gene of Pleurotus pulmonarius[J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 62-68.

Figures/Tables 5

Table 1 Primers for real time fluorescence quantitative PCR

引物名称 Primer name	引物序列 Primer sequence(5'-3')
PpSAMS-F	GCAACAATCTCCTGATATTGC
PpSAMS-R	GACGAAGCGGCCAGAGGGTTG
Actin-F	CGTTCATTGCCGACGGTGATG
Actin-R	CTCTGAACCCCAAGGCAAACC

Fig.1 ORF and transcription of PpSAMS

Fig.2 Phylogenetic tree of amino acid sequence of PpSAMS between Pleurotus pulmonarius and other fungi

Fig.3 KEGG pathway analysis of PpSAMS

Fig.4 Expression profile of PpSAMS in Pleurotus pulmonarius at different growth stages

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Cloning and expression analysis of PpSAMS gene of Pleurotus pulmonarius

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