Agrobacterium-mediated transformation of Camelina sativa for production of transgenic plants

Abstract

Camelina sativa ( C . sativa ), an oilseed species rich in poly-unsaturated fatty acids, has gained great importance as an industrial oil platform crop in recent years. Despite the potential benefits of C. sativa for bioenergy applications, limited research has been conducted to improve its agronomic qualities. Hence, a simple and efficient technique for production of transgenic C. sativa plants is warranted. In the present study, shoot apical meristems of two C. sativa cultivars (Pl650159 and Pl650161) were transformed with Agrobacterium strain ‘EHA 105’ harboring the enhanced green fluorescent protein (EGFP) and neomycin phosphotransferase II (nptII) genes. After two days of co-cultivation in the dark, explants were transferred to selection medium. Transgenic shoots were identified on the basis of green fluorescence and kanamycin resistance. Shoots were then rooted and transferred to potting mix soil for acclimatization. This protocol describes an efficient method to generate transgenic C. sativa plants in as little as 4 weeks.

Targeting transgene to seed resulted in high rate of morphological abnormalities of Camelina transformants

“…Celine) plants using floral dip technique. Previously, efficient transformation of two C. sativa cultivars (Pl650159 and Pl650161) has been described by Sitther et al (2018), while the authors concluded that shoot tips with apical meristems were the best target tissues for Agrobacteriummediated transformation of C. sativa plants in vitro.…”

“…Camelina can be successfully transformed using both the floral dip and floral vacuum infiltration methods (Lu and Kang 2008;Liu et al 2012). Recently, two C. sativa cultivars were successfully transformed also with Agrobacterium tumefaciens, for which the shoot tips with apical meristems appeared the best target tissue (Sitther et al 2018). Fully developed, intact transgenic Camelina plants can be obtained within 6 -8 weeks (Lu and Kang 2008;Sitther et al 2018).…”

“…Recently, two C. sativa cultivars were successfully transformed also with Agrobacterium tumefaciens, for which the shoot tips with apical meristems appeared the best target tissue (Sitther et al 2018). Fully developed, intact transgenic Camelina plants can be obtained within 6 -8 weeks (Lu and Kang 2008;Sitther et al 2018). Expression of transgenes can result in production of seed oil with attractive physicochemical properties such as reduced viscosity and freezing point, and thus suitable for preparation of biodegradable lubricants, emulsifiers, plasticizers, and second-generation biofuels (Liu et al 2015;Bansal and Durrett 2016;Haslam et al 2016).…”

Production of Transgenic <i>Camelina sativa</i> Plants via <i>Agrobacterium</i>-Mediated Transformation of Shoot Apical Meristems

“…A binary vector containing the enhanced green fluorescent protein (egfp), β glucuronidase and neomycin phosphotransferase II (nptII) genes under the control of a CaMV 35S promoter was used to optimize transformation parameters [18]. The binary plasmid was transferred to Agrobacterium tumefaciens "EHA 105" by freezing in dry ice, thawing at 25˚C and used in transformation studies ( Figure 1).…”

“…A number of tissue culture-based Agrobacterium transformation protocols using C. sativa leaf segments, petioles and hypocotyls as explants have been reported as well [16] [17]. We recently demonstrated the use of C. sativa in vitro shoots as suitable tissues for Agrobacterium-mediated transformation [18]. Such methods have been successfully used in Vitis vinifera for the development of transgenic plants [19] [20].…”