Abstract
A subtracted library was constructed from planthopper-infested wild rice (Oryza minuta) by suppression subtractive hybridization in combination with mirror orientation selection. To screen the differentially expressed transcripts in the library, we applied a cDNA microarray containing 960 random clones in a reverse Northern blot analysis using cDNA probes prepared from the mRNAs of control and planthopper-infested samples. On the basis of the signal intensities and expression ratios obtained from experiments performed in triplicate, we selected 383 clones. The elevated expression levels and overall profiles over time were verified by Northern blot analysis. Although Southern blot analysis showed similar copy numbers of the screened genes in O. minuta and O. sativa, it also revealed that the expression profiles had a different pattern . Functional categorization placed the identified transcripts in the categories of subcellular localization, metabolism, and protein fate. The presence of these expressed sequence tags implies that resistance of O. minuta to insect infestation can be achieved not only by an elevated expression of defense-related genes but also by enhanced metabolic activities.
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Abbreviations
- blast :
-
Basic local alignment search tool
- EST:
-
Expressed sequence tag
- MIPS:
-
Munich Information Center for Protein Sequences
- MOS:
-
Mirror orientation selection
- omii :
-
Oryza minuta insect-infested
- SSH:
-
Suppression subtractive hybridization
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Acknowledgements
This project was funded by a grant (CG1212) from the Crop Functional Genomics Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology (MOST) and Rural Development Administration (RDA), and supported by a grant from the BioGreen21 Program of Rural Development Administration (RDA) of Republic of Korea
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Communicated by I.S. Chung
S.K. Cho and K.W. Jung contributed equally to this work.
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Cho, S.K., Jung, K.W., Jeung, J.U. et al. Analysis of differentially expressed transcripts from planthopper-infested wild rice (Oryza minuta). Plant Cell Rep 24, 59–67 (2005). https://doi.org/10.1007/s00299-004-0905-9
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DOI: https://doi.org/10.1007/s00299-004-0905-9