Construction and validation of cDNA-based Mt6k-RIT macro- and microarrays to explore root endosymbioses in the model legume Medicago truncatula

被引:77
作者
Küster, H
Hohnjec, N
Krajinski, F
El Yahyaoui, F
Manthey, K
Gouzy, J
Dondrup, M
Meyer, F
Kalinowski, J
Brechenmacher, L
van Tuinen, D
Gianinazzi-Pearson, V
Pühler, A
Gamas, P
Becker, A
机构
[1] Univ Bielefeld, Fak Biol, Lehrstuhl Genet, D-33501 Bielefeld, Germany
[2] Univ Bielefeld, Inst Genome Res, CeBiTec, D-33594 Bielefeld, Germany
[3] Univ Bielefeld, Bioinformat Resources Facil, CeBiTec, D-33594 Bielefeld, Germany
[4] Univ Bielefeld, Int NRW Grad Sch Bioinformat & Genome Res, CeBiTec, D-33594 Bielefeld, Germany
[5] Univ Bielefeld GmbH, Inst Innovationstransfer, D-33501 Bielefeld, Germany
[6] Leibniz Univ Hannover, Lehrgebiet Molekulargenet, D-30419 Hannover, Germany
[7] INRA, UMR, CNRS, Interact Plantes Microorganismes, F-31326 Castanet Tolosan, France
[8] Univ Bourgogne, UMR 1088, INRA, F-21065 Dijon, France
[9] INRA, CNRS Plante Microbe Environm, FRE 2625, CMSE, F-21065 Dijon, France
关键词
arbuscular mycorrhiza; expression profiling; nodulin genes; PCR-based cDNA array; phosphate transporter; root nodule symbiosis;
D O I
10.1016/j.jbiotec.2003.11.011
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:95 / 113
页数:19
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