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- Title
Pt-CaM 基因克隆及对菌核侧耳耐热性影响.
- Authors
罗 璐; 张书会; 孙雪言; 盛 丽; 马爱民
- Abstract
The calmodulin (CaM) gene of Pleurotus tuber-regium Pt-CaM was cloned through conservative sequence alignment, rapid amplification of 3′cDNA ends (3′ RACE), and fusion primer and nested integrated PCR (FPNI-PCR). The full length of Pt-CaM was found to be 856 bp and its cDNA was 477 bp, encoding 158 amino acids. The overexpression and interference vectors of Pt-CaM were constructed and successfully transformed into P. tuber-regium mycelium by Agrobacterium tumefaciens mediated transformation, respectively. Compared with the wild-type strain, the Pt-CaM overexpression strain showed denser mycelia under 38 ℃, and its heat shock protein 70 gene HSP70 expression, superoxide dismutase (SOD) activity, and catalase (CAT) activity increased by 2. 89 times, 1. 49 times and 83%, respectively. In contrast, the Pt-CaM interference strain exhibited a decrease in mycelial growth, aggravated heat stress damage, a decrease in HSP70 expression by 23%, and a decrease in peroxidase (POD) activity by 30 % under 38 ℃. The activities of SOD and CAT remained the same in the interference strain as in the wild-type strain. The results suggested that Pt-CaM may participate in heat tolerance of P. tuber-regium mycelium via regulating HSP70 expression and activities of SOD, CAT and POD.
- Subjects
HSP70 heat-shock proteins; CATALASE; HEAT shock proteins; SUPEROXIDE dismutase; GENE expression; SEQUENCE alignment
- Publication
Acta Edulis Fungi, 2022, Vol 29, Issue 5, p1
- ISSN
1005-9873
- Publication type
Article
- DOI
10.16488/j.cnki.1005-9873.2022.05.001