Protoplast Production, Regeneration and Transformation of Neurospora crassa

doi:10.7506/spkx1002-6630-201517032

Abstract

Abstract:

This study aimed to establish protoplast-mediated genetic transformation system of Neurospora crassa. Several
major factors influencing the production and regeneration of N. crassa protoplast, such as mycelium incubation time,
macerozyme and digestion time, were investigated. The results showed that mycelium incubation in liquid PDA medium at
30 ℃ for 10 h, and enzymatic hydrolysis at 30 ℃ for 10 h with a macerozyme solution containing 0.5 g/100 mL snailase,
0.5 g/100 mL lyticase and 1.0 g/100 mL cellulase were optimal for the production and regeneration of N. crassa protoplast. Under
these conditions, the number of production and regeneration in 38# regeneration medium of Neurospora crassa protoplast were
4.53 × 106 and 1.41 × 106, respectively, and the regeneration rate was 31.71%. Transformation efficiency with PAKHB
plasmid by PEG-mediated was more than 8 transformants per μg of plasmid DNA.

Key words: Neurospora crassa, protoplasts, production, regeneration, transformation

CLC Number:

TS201

YANG Jianyuan, LI Jing, FAN Yawei, ZHANG Binghuo, DENG Zeyuan. Protoplast Production, Regeneration and Transformation of Neurospora crassa[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201517032.

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Protoplast Production, Regeneration and Transformation of Neurospora crassa

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