Development of a film-assisted honeybee egg collection system (FECS)
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Original article
Development of a film-assisted honeybee egg collection system (FECS) Jaeho LEE1 , Si Hyeock LEE1,2 1 Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea
2
Received 1 February 2019 – Revised 21 June 2019 – Accepted 1 September 2019
Abstract – Despite the huge potential of genome editing in honeybee research and breeding programs, only a few successful studies have been reported, implying the presence of several obstacles. One such obstacle is the difficulty in obtaining a large quantity of young eggs. To facilitate the overall procedures for egg collection and microinjection, we developed the film-assisted honeybee egg collection system (FECS), in which a removable transparent film is used as the platform for consecutive collection and injection of embryos. The FECS provides a significantly higher (~ 2-fold) collection efficiency and usability compared with the conventional plug-based queen rearing system while maintaining a high hatching rate. The system can be readily utilized for various studies that require a large number of honeybee eggs, including honeybee genome editing. honeybee / Apis mellifera / embryo / egg collection / genome editing
1. INTRODUCTION The western honeybee, Apis mellifera , plays a pivotal role in the pollination of many crop plants, thereby being one of the key elements of modern agriculture, and produces honey and other useful hive products (Klein et al. 2007). Despite economic importance in agriculture and value as a unique model with unusual biological characteristics, research on honeybee biology has been limited by lack of several classical genetic tools (Robinson et al. 2000). Although transgenesis and genome editing, in particular, have huge potential to be utilized in both honeybee studies and breedElectronic supplementary material The online version of this article (https://doi.org/10.1007/s13592-019-00687-8) contains supplementary material, which is available to authorized users.
Corresponding author: S. Lee, [email protected]. Lee, [email protected] Manuscript editor: Peter Rosenkranz
ing programs, they were not available to honeybee researchers until the first report of a transgenic honeybee by Schulte et al. in 2014 (Schulte et al. 2014). Despite various attempts for germline transformation in honeybees (Robinson et al. 2000; Pen 2018), microinjection into young embryos is the only successful method for delivery of manipulated genetic materials, leaving egg collection as the first prerequisite (Schulte et al. 2014; Kohno et al. 2016; Kohno and Kubo 2018). To date, various methods have been proposed for honeybee egg collection either in small (Taber 1961; Milne Jr et al. 1988; Collins 2002) or large (Omholt et al. 1995; Evans et al. 2010) quantities. Among these, plug-based queen rearing kits have been commonly employed for successful transgenesis and genome editing studies (Schulte et al. 2014; Kohno
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