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

Abstract

False flax (Camelina sativa L.) is currently under-exploited but highly promising oilseed crop. Combining Camelina’s attractive agronomic traits with its unprecedented ease for genetic engineering makes it an ideal plant chassis for biotechnology applications, in particular synthetic biology strategies. For targeted expression of transgene particularly to seeds requires identification and application of seed specific promoters. In the present study two cultivars of Camelina, namely Zuzana and Smilowska, were used for transformation at early flowering stage using the floral dip method. The plants were inoculated with Agrobacterium bearing a construct for expression of red fluorescent protein (RFP) under the control of the seed specific cruciferin promoter CRUC from Arabidopsis. Transgenic seeds and plants were identified on the basis of red fluorescence (RFP) and kanamycin resistance. Relatively high transformation efficiency of 8 % was achieved particularly for the cultivar Zuzana. However, many of regenerants exerted developmental deformations such as lack of shoot apical meristem, deformed or absent cotyledons, etc. Furthermore, the activity of the CRUC promoter was still active also in true leaves rendering this promoter as inappropriate for seed targeting of the transgene. Nevertheless, genetic transformation remains a tool for direct modulation of pathways for oil synthesis in oilseed crops.

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