Abstract
Conifer somatic embryogenesis has long been recognized as an advanced vegetative propagation technology. Somatic embryogenesis provides a platform for capturing and long-term preservation of elite genotypes and developing commercial scale-up systems for mass production of plants. Although, significant success has been reported in improving conifer somatic embryogenic protocols, little has been presented to describe the complexity of integrating in vitro (laboratory) and ex-vitro (greenhouse) programs for developing a commercial production system capable of delivering tens of millions of conifer somatic seedlings. This integration requires both programs to run in concert and produce propagules capable of surviving and growing under greenhouse conditions at very early stages. It also requires the integration of seedling development events to ensure the production of quality seedlings that meet needs of the forest restoration program.
 This paper describes the importance of protocol optimization for scaling-up the in vitro and ex vitro programs. The ‘biology of scaling’ is discussed in view of plant cell, embryo, germinant and somatic seedling requirements throughout the program. Logistical issues related to protocol optimization and scale-up are addressed. Specific control points for monitoring and controlling the commercial process are presented. The importance of developing standard operating procedures, media batch records, and quality control systems are discussed. Ultimately, a fully integrated system capable of producing tens of millions of conifer somatic seedlings is presented.
Highlights
Somatic embryogenesis is the preferred in vitro process for the large-scale commercial production of conifer seedlings
Major challenges in commercializing this technology include: a) Low and inconsistent initiation and multiplication rates delivering variable quantities and quality of somatic embryos, b) Losses throughout the program associated with cryopreservation and variable quality of the material removed from cryopreservation, c) Low number and poor quality of mature somatic embryos due to insufficient protocol optimization, d) Losses due to contamination and poor laboratory practices, e) Low conversion rates in nursery due to poor germinant quality attributable to the absence of in vitro germinant selection criteria, f)
This paper examines issues that need to be considered during the scale-up of a somatic embryogenesis tissue culture program
Summary
Somatic embryogenesis is the preferred in vitro process for the large-scale commercial production of conifer seedlings. Protocol optimization must precede scale-up activities to maximize the in vitro and initial ex vitro performance for each genotype This paper addresses both - process optimization and scale-up requirements that have to be met in order to create a commercial system for conifer somatic seedling production. Whether one is addressing the plant material in the in vitro or ex vitro program phase, the creation of optimum environmental conditions is critical for plant cell, embryo, germinant and seedling development during the scale-up of any tissue culture program. This paper presents some of the critical program steps that need to be considered if one is going to create a successful and cost efficient process for the large-scale commercial production of somatic seedlings from a somatic embryogenesis tissue culture program
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