High-Efficient and Transient Transformation of Moso Bamboo ( Phyllostachys edulis ) and Ma Bamboo ( Dendrocalamus latiflorus Munro)

Moso bamboo (Phyllostachys edulis) and Ma bamboo (Dendrocalamus latiflorus Munro) are two major species, which have significant economic and research application value. At present, methods for interrogation of gene function in bamboo are limited and time consuming. Therefore, establishing an efficient transient expression system for bamboo is very useful for dissecting the function of candidate genes. In this study, we developed protocols for the transient transformation of protoplasts from different tissues in Moso bamboo and Ma bamboo and established the optimized protocol and materials for improving the efficiency of transient transformation. The results indicated that the transformation efficiency of protoplasts from 15-day-old etiolated seedlings was higher than other tissues, which were 44.7% in Moso bamboo and 35.2% in Ma bamboo. In addition, we also established an experimental approach for optimizing overexpression of external proteins using Agrobacterium-mediated transient transformation of bamboo leaves and whole seedlings. Finally, we utilized the RUBY reporter for monitoring successful transformation samples with red color for downstream experiments in Moso bamboo. In summary, this study provides optimized protocols for the over-expression of candidate genes in protoplasts or bamboo leaves and whole seedlings.