Abstract
To investigate the protoplast preparation and transformation system of endophytic fungus Falciphora oryzae. F. oryzae strain obtained higher protoplast yield and effective transformation when treated with enzyme digestion solution containing 0.9M KCl solution and 10mgmL-1 glucanase at 30 °C with shaking at 80rpm for 2-3h. When the protoplasts were plated on a regenerations-agar medium containing 1M sucrose, the re-growth rate of protoplasts was the highest. We successfully acquired green fluorescent protein-expressing transformants by transforming the pKD6-GFP vector into protoplasts. Further, the GFP expression in fungal hyphae possessed good stability and intensity during symbiosis in rice roots. This study provided a protoplast transformation system of F. oryzae, creating opportunities for future genetic research in other endophytic fungi.
Highlights
Beneficial relationships between plants and endophytes commonly occur in the ecosystems and play essential roles in improving plant growth and helping plants overcome biotic or abiotic stress
The results showed that F. oryzae strain obtained higher protoplast yield and effective transformation when treated with enzyme digestion solution containing 0.9mol L-1 KCl solution and 10 mg mL−1 glucanase at 30°C with shaking 80 rpm for 2-3 h
In this study, we intend to explore a genetic transformation method of F. oryzae to study the interaction between other endophytic fungus and host plants
Summary
Beneficial relationships between plants and endophytes commonly occur in the ecosystems and play essential roles in improving plant growth and helping plants overcome biotic or abiotic stress. Phylogenetic analyses have shown that F. oryzae has a close relationship to other members in Magnaporthaceae, such as Gaeumannomyces and Magnaporthe, most of which are plant pathogens. The biological role and evolutionary genetics render F. oryzae to be an attractive model for exploring the mutualistic interaction of endophytes and plants. Few endophytic fungi have been sequenced with a whole genome, let alone have stable genetic transformation systems. In this study, we intend to explore a genetic transformation method of F. oryzae to study the interaction between other endophytic fungus and host plants
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