Details
| Original language | English |
|---|---|
| Pages (from-to) | 95-113 |
| Number of pages | 19 |
| Journal | Journal of biotechnology |
| Volume | 108 |
| Issue number | 2 |
| Publication status | Published - 4 Mar 2004 |
| Externally published | Yes |
Abstract
To construct macro- and microarray tools suitable for expression profiling in root endosymbioses of the model legume Medicago truncatula, we PCR-amplified a total of 6048 cDNA probes representing genes expressed in uninfected roots, mycorrhizal roots and young root nodules [Nucleic Acids Res. 30 (2002) 5579]. Including additional probes for either tissue-specific or constitutively expressed control genes, 5651 successfully amplified gene-specific probes were used to grid macro- and to spot microarrays designated Mt6k-RIT (M. truncatula 6k root interaction transcriptome). Subsequent to a technical validation of microarray printing, we performed two pilot expression profiling experiments using Cy-labeled targets from Sinorhizobium meliloti-induced root nodules and Glomus intraradices-colonized arbuscular mycorrhizal roots. These targets detected marker genes for nodule and arbuscular mycorrhiza development, amongst them different nodule-specific leghemoglobin and nodulin genes as well as a mycorrhiza-specific phosphate transporter gene. In addition, we identified several dozens of genes that have so far not been reported to be differentially expressed in nodules or arbuscular mycorrhiza thus demonstrating that Mt6k-RIT arrays serve as useful tools for an identification of genes relevant for legume root endosymbioses. A comprehensive profiling of such candidate genes will be very helpful to the development of breeding strategies and for the improvement of cultivation management targeted at increasing legume use in sustainable agricultural systems.
Keywords
- Arbuscular mycorrhiza, Expression profiling, Nodulin genes, PCR-based cDNA array, Phosphate transporter, Root nodule symbiosis
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Journal of biotechnology, Vol. 108, No. 2, 04.03.2004, p. 95-113.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Construction and validation of cDNA-based Mt6k-RIT macro- and microarrays to explore root endosymbioses in the model legume Medicago truncatula
AU - Küster, Helge
AU - Hohnjec, Natalija
AU - Krajinski, Franziska
AU - El Yahyaoui, Fikri
AU - Manthey, Katja
AU - Gouzy, Jérôme
AU - Dondrup, Michael
AU - Meyer, Folker
AU - Kalinowski, Jörn
AU - Brechenmacher, Laurent
AU - Van Tuinen, Diederik
AU - Gianinazzi-Pearson, Vivienne
AU - Pühler, Alfred
AU - Gamas, Pascal
AU - Becker, Anke
N1 - Funding information: This work was supported by the European Union project MEDICAGO (QLG2-CT-2000-00676), the Deutsche Forschungsgemeinschaft SPP 1084 “Mykorrhiza”, and the Deutsche Forschungsgemeinschaft grant BIZ 7. M. Dondrup, N. Hohnjec and H. Küster acknowledge financial support by the “International NRW Graduate School in Bioinformatics and Genome Research”. L. Brechenmacher was supported by an INRA/Conseil de Bourgogne grant. We are grateful to Gabriella Endre and Kate vandenBosch (University of Minnesota, St. Paul, MN, USA), Julie Cullimore (INRA-CNRS Toulouse, France), and Jeff Landgraf (Plant Research Laboratory, Michigan State University, USA) for sharing cDNA clones for array construction. Eva Schulte-Berndt and Silvia Rüberg (Institute of Genome Research and Chair of Genetics, Bielefeld University) are acknowledged for their expert support in the initial phase of array construction.
PY - 2004/3/4
Y1 - 2004/3/4
N2 - To construct macro- and microarray tools suitable for expression profiling in root endosymbioses of the model legume Medicago truncatula, we PCR-amplified a total of 6048 cDNA probes representing genes expressed in uninfected roots, mycorrhizal roots and young root nodules [Nucleic Acids Res. 30 (2002) 5579]. Including additional probes for either tissue-specific or constitutively expressed control genes, 5651 successfully amplified gene-specific probes were used to grid macro- and to spot microarrays designated Mt6k-RIT (M. truncatula 6k root interaction transcriptome). Subsequent to a technical validation of microarray printing, we performed two pilot expression profiling experiments using Cy-labeled targets from Sinorhizobium meliloti-induced root nodules and Glomus intraradices-colonized arbuscular mycorrhizal roots. These targets detected marker genes for nodule and arbuscular mycorrhiza development, amongst them different nodule-specific leghemoglobin and nodulin genes as well as a mycorrhiza-specific phosphate transporter gene. In addition, we identified several dozens of genes that have so far not been reported to be differentially expressed in nodules or arbuscular mycorrhiza thus demonstrating that Mt6k-RIT arrays serve as useful tools for an identification of genes relevant for legume root endosymbioses. A comprehensive profiling of such candidate genes will be very helpful to the development of breeding strategies and for the improvement of cultivation management targeted at increasing legume use in sustainable agricultural systems.
AB - To construct macro- and microarray tools suitable for expression profiling in root endosymbioses of the model legume Medicago truncatula, we PCR-amplified a total of 6048 cDNA probes representing genes expressed in uninfected roots, mycorrhizal roots and young root nodules [Nucleic Acids Res. 30 (2002) 5579]. Including additional probes for either tissue-specific or constitutively expressed control genes, 5651 successfully amplified gene-specific probes were used to grid macro- and to spot microarrays designated Mt6k-RIT (M. truncatula 6k root interaction transcriptome). Subsequent to a technical validation of microarray printing, we performed two pilot expression profiling experiments using Cy-labeled targets from Sinorhizobium meliloti-induced root nodules and Glomus intraradices-colonized arbuscular mycorrhizal roots. These targets detected marker genes for nodule and arbuscular mycorrhiza development, amongst them different nodule-specific leghemoglobin and nodulin genes as well as a mycorrhiza-specific phosphate transporter gene. In addition, we identified several dozens of genes that have so far not been reported to be differentially expressed in nodules or arbuscular mycorrhiza thus demonstrating that Mt6k-RIT arrays serve as useful tools for an identification of genes relevant for legume root endosymbioses. A comprehensive profiling of such candidate genes will be very helpful to the development of breeding strategies and for the improvement of cultivation management targeted at increasing legume use in sustainable agricultural systems.
KW - Arbuscular mycorrhiza
KW - Expression profiling
KW - Nodulin genes
KW - PCR-based cDNA array
KW - Phosphate transporter
KW - Root nodule symbiosis
UR - http://www.scopus.com/inward/record.url?scp=10744226442&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2003.11.011
DO - 10.1016/j.jbiotec.2003.11.011
M3 - Article
C2 - 15129719
AN - SCOPUS:10744226442
VL - 108
SP - 95
EP - 113
JO - Journal of biotechnology
JF - Journal of biotechnology
SN - 0168-1656
IS - 2
ER -