Kim, T. and Chowdhury, M. K. U. and Wetzstein, H. Y. (1999) A Quantitative and Histological Comparison of GUS Expression With Different Promoter Constructs Used In Microprojectile Bombardment of Peanut Leaf Tissue. Vitro Cellular & Developmental Biology-Plant , 35 (1). pp. 51-56.
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Abstract
~-glucuronidase (GUS) expression driven with different promoter constructs was quantitatively and histologically compared in peanut leaf tissue following microprojectite bombardment. X-Gluc staining patterns varied with the construct used. Tissues bombarded with the pAC2GUS construct had larger foci and a greater percentage of area staining blue. pEmuGN exhibited the greatest numbers of blue spots compared to pAC2GUS and pTRA140. Histological evaluations of blue staining foci showed a diffusion gradient of blue precipitate from a central, prominently-staining cell outward to as many as seven cell layers. The intensity of X-gluc product in centrally-staining cells varied. Gold microprojectile particles were usually located within the three surface cell layers. Depending on the plasmid construct, 72-90% of the centrally-staining ceils had at least one gold particle. However, the presence of GUS expression did not appear to require a microprojectile within the nucleus, which was observed in 37% or fewer of the centrally-staining cells. With the pAC2GUS construct, staining patterns varied with location within leaflets and had an "edge effect," i.e., blue spots were frequently larger at the margin versus central regions. This enhanced activity could be anticipated with an actin promoter in the more mitotically active marginal leaf cells. Total GUS activity as determined by fluorometric analyses was correlated with the percentage of X-gluc stained area. The pAC2GUS construct exhibited the highest total GUS activity among the three constructs.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Peanut, particle gun, transformation, β-glucuronidase, Arachis hypogaea |
| Author Affiliation: | Plant Science Building, Department of Horticulture, University of Georgia, Athens, Georgia 30602-7273 |
| Subjects: | Crop Improvement > Plant Breeding |
| Divisions: | Groundnut |
| Depositing User: | Mr Siva Shankar |
| Date Deposited: | 16 May 2012 08:52 |
| Last Modified: | 16 May 2012 08:52 |
| Official URL: | http://dx.doi.org/10.1007/s11627-999-0009-x |
| URI: | http://eprints.icrisat.ac.in/id/eprint/5375 |
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