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- Title
基于CRISPR系统的双孢蘑菇基因编辑体系建立及应用.
- Authors
吕阳; 贺晓飞; 王佩; 刘世玲; 韩少鹏; 曾弓剑; 周雁; 沈祥陵
- Abstract
A gene editing system based on CRISPR/Cas9 was established in Agaricus bisporus. First, a guide RNA specific to AbPPO4 was designed and vector pAbGEB1-AbPPO4 carrying the hygromycin resistence gene was constructed, and then the vector was transformed into A. bisporus by Agrobacterium-mediated transformation. Transformants were first screened for hygromycin resistency on PDA medium and then confirmed by detection of the hygromycin resistence gene with PCR. The results showed that transformant No.1 and No.7 contained the hygromycin resistence gene and their AbPPO4 expression levels were (0.28±0.04) and (0.47±0.02), respectively. The AbPPO4 expression level of both transformants were statistically lower compared with that of the original strain. There were 4 bp and 11 bp deletions in the AbPPO4 locus of transformant No.1 and No.7, respectively. Compared with the starting strain, both transformant No.1 and No.7 showed weaker mycelial growth, suggesting that disruption of AbPPO4 inhibited mycelial growth. The success of establishing CRISPR/Cas9 gene editing system in A. bisporus offers a new tool for genetic breeding and understanding molecular mechanisms underlying physiological processes of A. bisporus.
- Subjects
CULTIVATED mushroom; GENOME editing; CRISPRS; GENE expression; POLYPHENOL oxidase; PLANT genetic transformation
- Publication
Acta Edulis Fungi, 2020, Vol 27, Issue 3, p16
- ISSN
1005-9873
- Publication type
Article
- DOI
10.16488/j.cnki.1005-9873.2020.03.003