Construction and validation of cDNA-based Mt6k-RIT macro- and microarrays to explore root endosymbioses in the model legume Medicago truncatula.
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.
Küster
Helge
H
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Postfach 100131, Bielefeld D-33501, Germany. helge.kuester@genetik-uni-bielefeld.de
Hohnjec
Natalija
N
Krajinski
Franziska
F
El
Yahyaoui Fikri
YF
Manthey
Katja
K
Gouzy
Jéôme
J
Dondrup
Michael
M
Meyer
Folker
F
Kalinowski
Jörn
J
Brechenmacher
Laurent
L
van Tuinen
Diederik
D
Gianinazzi-Pearson
Vivienne
V
Pühler
Alfred
A
Gamas
Pascal
P
Becker
Anke
A
D004867:Equipment Design; D019544:Equipment Failure Analysis; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling / Q000295:instrumentation* / Q000379:methods; D015966:Gene Expression Regulation, Fungal; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D029902:Medicago / Q000235:genetics* / Q000382:microbiology; D038821:Mycorrhizae / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics* / Q000382:microbiology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013559:Symbiosis / Q000235:genetics*
D010940:Plant Proteins