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Somatic embryo cycling: Evaluation of a novel transformation and assay system for seed-specific gene expression in soybean

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Abstract

Somatic embryo cycling, a modification of soybean somatic embryogenic suspension culture, was developed as an efficient and rapid method of producing tissue suitable for stable transformation of soybean germplasm by biolistic particle bombardment. Instead of using immature seed explants, cotyledon-staged somatic embryo hypocotyls were placed on auxin-containing medium, where they initiated new somatic embryos primarily from single epidermal cells. By bombarding hypocotyls prior to initiation of subsequent embryo formation, we have effectively transformed soybean somatic embryos with the reporter genes neomycin phosphotransferase,gb-glucuronidase, and a mammalian stearyl CoA delta-9 desaturase, controlled by a seed-specific promoter. These embryos contain significantly reduced levels of saturated palmitic and stearic fatty acids, and significant amounts of monounsaturated palmitoleic acid, which is not normally abundant in soybean seeds. This study demonstrates the effectiveness of somatic embryo cycling for soybean transformation, and for testing expression of genes for seed-specific proteins. Abnormal flower development in recovered plants is a limitation for application of the technique to produce transgenic seed at present.

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Abbreviations

ER:

endoplasmic reticulum

D9DS:

delta-9 desaturase

GUS:

β-glucuronidase

NPTII:

neomycin phosphotransferase II

CaMV:

cauliflower mosaic virus 35S promoter

CoA:

coenzyme A

SDS:

sodium dodecyl sulfate

PAGE:

polyacrylamide gel electrophoresis

β-ME:

β-mercaptoethanol

2,4-d :

2,4 dichlorophenoxyacetic acid

NAA:

napthaleneacetic acid

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Authors and Affiliations

  1. R.J Reynolds Corp. Bowman Gray Technical Center, Winston-Salem, NC, USA

    Wennuan Liu

  2. Department of Agronomy, N-212 Agric. Sci. Center-North, University of Kentucky, Lexington, KY, USA

    Rebecca S. Torisky, Kay P McAllister, Sergei Avdiushko & David Hildebrand

Authors
  1. Wennuan Liu
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  2. Rebecca S. Torisky
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  3. Kay P McAllister
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  4. Sergei Avdiushko
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  5. David Hildebrand
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  6. Glenn B. Collins
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Liu, W., Torisky, R.S., McAllister, K.P. et al. Somatic embryo cycling: Evaluation of a novel transformation and assay system for seed-specific gene expression in soybean. Plant Cell Tiss Organ Cult 47, 33–42 (1996). https://doi.org/10.1007/BF02318963

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  • DOI: https://doi.org/10.1007/BF02318963

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