Abstract
Genetic engineering technology has been successfully used in many plant species, but is limited in woody plants, especially in bamboos. Ma bamboo (Dendrocalamus latiflorus Munro) is one of the most important bamboo species in Asia, and its genetic improvement was largely restricted by the lack of an efficient regeneration and transformation method. Here we reported a plantlet regeneration and Agrobacterium-mediated transformation protocol by using Ma bamboo young shoots as explants. Under our optimized conditions, embryogenic calluses were successfully induced from the excised young shoots on callus induction medium and rapidly grew on callus multiplication medium. Shoots and roots were regenerated on shoot induction medium and root induction medium, respectively, with high efficiency. An Agrobacterium-mediated genetic transformation protocol of Ma bamboo was established, verified by PCR and GUS staining. Furthermore, the maize Lc gene under the control of the ubiquitin promoter was successfully introduced into Ma bamboo genome and generated an anthocyanin over-accumulation phenotype. Our methods established here will facilitate the basic research as well as genetic breeding of this important bamboo species.Key achievements: A stable and high efficiency regeneration and Agrobacterium-mediated transformation protocol for Ma bamboo from vegetative organ is established.
Highlights
Bamboo species belong to the grasses family and are well-known for their great economic, social, and cultural value (Buckingham et al, 2014)
After approximately 3 month subculture on callus induction medium (CIM), they were transferred to the callus multiplication medium (CMM) which contains 3/4 MS basal medium supplemented with 30 g/L sucrose, 3 g/L sorbitol, 250 mg/L PVP, 2 mg/L 2,4-D, and 4.2 g/L phytogel for callus multiplication
As the first step for callus induction, the young shoots were sterilized and cut as we described in “Materials and Methods,” and they were placed on CIM
Summary
Bamboo species belong to the grasses family (poaceae) and are well-known for their great economic, social, and cultural value (Buckingham et al, 2014). They cover tropical, subtropical, and temperate zones of the world and are considered as the prime renewable resource for biomass production due to their short rotation period and fast-growing culm (Tanaka et al, 2003). Ma bamboo (Dendrocalamus latiflorus Munro) is one of the most important semitropical clumping bamboo species and forms abundant forests in Asia (Buckingham et al, 2014). The demands for economically important bamboo species like Ma bamboo are on the rise due to its great benefits to human beings (Singh et al, 2013)
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