NDLI: Rapid expression and validation of seed-specific constructs in transgenic LEC2 induced somatic embryos of Brassica napus

Content Provider	Springer Nature Link
Author	Belide, Srinivas Zhou, Xue Rong Kennedy, Yoko Lester, Geraldine Shrestha, Pushkar Petrie, James R. Singh, Surinder P.
Copyright Year	2013
Abstract	Seed-specific modification of crop plants is a common goal in plant metabolic engineering. Ability for early assessment and validation of seed-specific constructs for metabolic engineering in crop plants is desirable. We describe a method in which the Arabidopsis thaliana LEAFY COTYLEDON2 (LEC2) transcription factor is constitutively expressed to generate somatic embryos that can generate a seed-like fatty acid profile from cotyledonary petiole of Brassica napus. We also demonstrate that the super-transformation of 35S-LEC2 construct in B. napus lines that are already stably-transformed with seed-specific constructs producing arachidonic acid (ARA) or ricinoleic acid (RA) in mature seed results in the generation of somatic embryos that actively expressed these seed-specific transgenes and produced ARA and RA similar to transgenic T$_{2}$ seeds. Stable transformation of seed-specific construct and analysis of T$_{1}$ seed in B. napus requires around 7–8 months. We have used this method to functionally assess single (RA) or multi-gene (ARA) seed specific construct in somatic embryos within 6–7 weeks of transformation and have found the results to be reasonably indicative of mature (T$_{2}$) seed fatty acid profiles. A similar strategy can also be extended to induce somatic embryogenesis in oil seed crops for the testing and validation seed specific traits quickly before stable transformation.
Starting Page	543
Ending Page	553
Page Count	11
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	113
Issue Number	3
e-ISSN	15735044
Language	English
Publisher	Springer Netherlands
Publisher Date	2013-02-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Arachidonic acid Ricinoleic acid Unusual fatty acid Co-transformation Super-transformation Somatic embryogenesis Fatty acid analysis Plant Sciences Plant Physiology Plant Genetics & Genomics Plant Pathology
Content Type	Text
Resource Type	Article
Subject	Horticulture

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