Quantitative real-time-PCR: fast and efficient technology for knowledge-based breeding
2008
Steinhauer, D., GenXPro GmbH, Frankfurt (Germany) | Khan, F., Jamia Hamdard University, New Delhi (India) | Fatnassi, N., Centre de Biotechnologie de Borj Cedria, Hammam-Lif (Tunisia) | Molina, C., University of Frankfurt, Frankfurt (Germany). Biocentre | Horres, R., GenXPro GmbH, Frankfurt (Germany) | Abdelly, C., Centre de Biotechnologie de Borj Cedria, Hammam-Lif (Tunisia) | Delgado, M.-J., Estacion Experimental del Zaidin c/Prof. Alboreda, Granada (Spain) | Kahl, G., University of Frankfurt, Frankfurt (Germany). Biocentre | Winter, P., GenXPro GmbH, Frankfurt (Germany)
Dehydration stresses such as drought and salinity constrain yield of chickpea throughout the world. Using SuperSAGE whole-genome transcription profiling, we analyzed more than 600.000 root- and nodule transcript SuperTags from salt and drought tolerant chickpea varieties and compared them to non-stressed control plant to identify potential molecular targets for knowledge-based stress-tolerance breeding. Amongst expression profiles differentiating between tolerant and susceptible varieties those involved in signal perception and transduction and also transcripts coding for down-stream effectors such as reactive-oxygen-(ROS)-related proteins provided interesting candidates deserving further evaluation for their value in breeding. However, not only transcripts coding for proteins of potentially known function but also transcripts coding for proteins of unknown function as well as their different splicing variants were differentially expressed in tolerant susceptible varieties. We used commercially available, splicing-variant0specific quantitative Real-Time (qRT)-PCR assays to investigate the temporal expression of the different splicing variants in stressed and non-stressed roots and nodules of salt and drought tolerant and susceptible chickpea varieties. All splicing variants were higher expressed in at last 1 tolerant variety as compared to susceptible varieties already under non-stress conditions. Further whereas in tolerant varieties all splicing were generally up-regulated under stress, susceptible varieties reacted by their down-regulation. The fact that putative Narbonin transcripts were already much stronger expressed in tolerant genotypes than in susceptible ones even before the onset of stress suggests a necessity of priming for stress-tolerance. Therefore, it may be possible to pre-screen germplasm and breeding lines for stress-tolerance using qRT-PCR even without stressing it.
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