Advertisement

Plant Cell Reports

, Volume 25, Issue 9, pp 920–926 | Cite as

Comparative analysis of 35S and lectin promoters in transgenic soybean tissue using an automated image acquisition system and image analysis

  • Marco Tulio Buenrostro-Nava
  • Peter P. Ling
  • John J. FinerEmail author
Genetic Transformation and Hybridization

Abstract

Expression of the green fluorescent protein (gfp) gene, under regulatory control of either the constitutive 35S promoter or the developmentally-regulated lectin promoter was monitored and quantified using a newly-developed automated tracking system. The automated system consisted of a computer-controlled two-dimensional robotics table and a programmable image acquisition system, which was used to semi-continuously monitor gfp gene expression during development of transgenic soybean [Glycine max (L.) Merrill] somatic embryos. Quantitative analysis of GFP expression showed that, during somatic embryo proliferation and early development, expression of lectin-GFP was not detected. During late embryo development, expression of lectin-GFP gradually increased until the levels were similar to those of 35S-GFP. The use of an automated image collection system and image analysis facilitated the frequent monitoring and quantification of gfp gene expression on a large number of samples over an extended period of time.

Keywords

Green fluorescent protein Image analysis Promoter analysis Robotics 

Notes

Acknowledgments

We would like to acknowledge Jim Haseloff for the gift of the mgfp5-ER gene and Lila Vodkin for kindly providing the pGLe10 plasmid containing the lectin promoter. Salaries and research support were provided by State and Federal funds appropriated to OSU/OARDC, by the United Soybean Board, and from a scholarship from the Mexican government (CONACYT) to MTBN. Mention of trademark or proprietary products does not constitute a guarantee or warranty of the product by OSU/OARDC and also does not imply approval to the exclusion of other products that may also be suitable. H&CS article #HCS 06-02.

Supplementary material

References

  1. Aspuria ET, Ooura C, Chen GQ, Uchimiya H, Oono Y (2002) GFP accumulation controlled by an auxin-responsive promoter as a non-destructive assay to monitor early auxin response. Plant Cell Rep 21:52–57CrossRefGoogle Scholar
  2. Bastar MT, Luthar Z, Škof S, Bohanec B (2004) Quantitative determination of mosaic gfp gene expression in tobacco. Plant Cell Rep 22:939–944CrossRefPubMedGoogle Scholar
  3. Buenrostro-Nava MT, Ling PP, Finer JJ (2005) Development of an automated image acquisition system for monitoring gene expression and tissue growth. Trans ASAE 48:841–847Google Scholar
  4. Cheng Z, Ling PP (1994) Machine vision techniques for somatic coffee embryo morphological feature extraction. Trans ASAE 37:1663–1669Google Scholar
  5. Chiera JM, Finer JJ, Grabau EA (2004) Ectopic expression of a soybean phytase in developing seeds of Glycine max to improve phosphorus availability. Plant Mol Biol 56:895–904CrossRefPubMedGoogle Scholar
  6. Cho MJ, Widholm JM, Vodkin LO (1995) Cassettes for seed-specific expression tested in transformed embryogenic cultures of soybean. Plant Mol Biol Rep 13:255–269CrossRefGoogle Scholar
  7. de la Riva GA, Gonzalez-Cabrera J, Vazquez-Padron R, Ayra-Pardo C (1998) Agrobacterium tumefaciens: a natural tool for plant transformation. Electron J Biotech 1:1–16Google Scholar
  8. Finer JJ, Larkin KM (in press) Genetic transformation of soybean using particle bombardment and SAAT approaches. In: Kirti PB (ed) Handbook of new technologies for genetic improvement of legumes. Haworth Press, Binghampton, New YorkGoogle Scholar
  9. Finer JJ, Vain P, Jones MW, McMullen MD (1992) Development of the particle inflow gun for DNA delivery to plant cells. Plant Cell Rep 11:232–238.CrossRefGoogle Scholar
  10. Finer JJ, Beck SL, Buenrostro-Nava MT, Chi YT, Ling PP (2006) Monitoring gene expression in plant tissues; Using green fluorescent protein with automated image collection and analysis. In: Gupta SD, Ibaraki Y (eds) Plant tissue culture engineering; focus in biotechnology. Springer, Dordrecht, pp 31–46Google Scholar
  11. Goldberg RB, Barker SJ, Perez-Grau L (1989) Regulation of gene expression during plant embryogenesis. Cell 56:149–160CrossRefPubMedGoogle Scholar
  12. Hadi MZ, McMullen MD, Finer JJ (1996) Transformation of 12 different plasmids into soybean via particle bombardment. Plant Cell Rep 15:500–505CrossRefGoogle Scholar
  13. Hämäläinen JJ, Kurtén U, Kauppinen V, Heilala J (1992) Automated classification of somatic plant embryos. Acta Horticulturae 319:601–606Google Scholar
  14. Harrel RC, Hood CF, Moltó E, Munilla R, Bieniek M, Cantliffe DJ (1992) Automatic identification and separation of somatic embryos in vitro. Acta Horticulturae 319:595–600Google Scholar
  15. Haseloff J, Siemering KR (1998) The uses of green fluorescent protein in plants. In: Chalfie M, Kain SR (eds) Green fluorescent protein: properties, application, and protocols. Wiley, Chichester, pp 191–219Google Scholar
  16. Ling PP, Cheng Z, Musacchio DJ (1996) Quantification of somatic coffee embryo growth. Trans ASAE 38:1911–1917Google Scholar
  17. Maximova SN, Dandekar AM, Guiltinan MJ (1998) Investigation of Agrobacterium-mediated transformation of apple using green fluorescent protein: high transient expression and low stable transformation suggest that factors other than T-DNA transfer are rate-limiting. Plant Mol Biol 37:549–559PubMedCrossRefGoogle Scholar
  18. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325PubMedCrossRefGoogle Scholar
  19. Nagatani N, Takuni S, Tomiyama M, Shimada T, Tamiya E (1997) Semi-real time imaging of the expression of a maize polyubiquitin promoter-GFP gene in transgenic rice. Plant Sci 124:49–56CrossRefGoogle Scholar
  20. Noesis SA (1997) Visilog 5.0® Documentation Set. Canada. p 851Google Scholar
  21. Perez-Grau L, Goldberg RB (1989) Soybean seed protein genes are regulated spatially during embryogenesis. The Plant Cell 1:1095–1109CrossRefPubMedGoogle Scholar
  22. Philip R, Darnowski DW, Sundararaman V, Cho M-J, Vodkin LO (1998) Localization of β−glucuronidase in protein bodies of transgenic tobacco seed by fusion to an amino terminal sequence of the soybean lectin gene. Plant Sci 137:191–204CrossRefGoogle Scholar
  23. Ponappa T, Brzozowski AE, Finer JJ (1999) Transient expression and stable transformation of soybean using the jellyfish green fluorescent protein. Plant Cell Rep 19:6–12CrossRefGoogle Scholar
  24. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  25. Santarém ER, Finer JJ (1999) Transformation of soybean (Glycine max (L.) Merrill) using proliferative embryogenic tissue maintained on semi-solid medium. In Vitro Cell Dev Biol 35P:451–455Google Scholar
  26. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517PubMedCrossRefGoogle Scholar
  27. Steel RGD, Torrie JH, Dickey DA (1997) Principles and procedures of statistics a biometrical approach, 3rd edn. McGraw-Hill, New YorkGoogle Scholar
  28. Thompson JM, Parrott WA (1998) pMECA: A size-based, blue/white selection multiple common and rare-cutter general cloning and transcription vector. BioTechniques 24:922–927PubMedGoogle Scholar
  29. Trick HN, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean (Glycine max [L.] Merrill) embryogenic suspension culture tissue. Plant Cell Rep 17:482–488CrossRefGoogle Scholar
  30. Urwin PE, Møller SG, Lilley CJ, McPherson MJ, Atkinson HJ (1997) Continual green fluorescent protein monitoring of cauliflower mosaic virus 35S promoter activity in nematode-induced feeding cells in Arabidopsis thaliana. MPMI 10:394–400PubMedGoogle Scholar
  31. van der Krol AR, Mur LA, Beld M, Mol JNM, Stuitje AR (1990) Flavonoid genes in petunia: addition of a limited number of gene copies may lead to a suppression of gene expression. Plant Cell 2:291–299PubMedCrossRefGoogle Scholar
  32. Vodkin LO, Raikhel NV (1986) Soybean lectin and related proteins in seeds and roots of Le+ and Le- soybean varieties. Plant Physiol 81:558–565PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Marco Tulio Buenrostro-Nava
    • 1
    • 3
  • Peter P. Ling
    • 2
  • John J. Finer
    • 1
    Email author
  1. 1.Department of Horticulture and Crop ScienceThe Ohio State University/OARDCWoosterUSA
  2. 2.Department of Food, Agricultural and Biological EngineeringThe Ohio State University/OARDCWoosterUSA
  3. 3.Department of Plant Pathology and MicrobiologyInstitute for Plant Genomics and Biotechnology, Texas A&M UniversityCollege StationUSA

Personalised recommendations