ZmGrp3: identification of a novel marker for root initiation in maize and development of a robust assay to quantify allele-specific contribution to gene expression in hybrids.
This study comprises a comprehensive gene expression analysis of the root tip specific maize gene ZmGrp3. In the first part of this paper expression of ZmGrp3 was studied in maize inbred lines. First, RNA in situ hybridization experiments confined the expression of ZmGrp3 to the columella and the epidermis of all embryonic and postembryonic root types. Second, Northern-blot analyses of the maize root initiation mutants rtcs and lrt1 revealed that the ZmGrp3 gene is not expressed prior to root initiation, thus providing a novel marker for this developmental process. Finally, a comprehensive expression profiling in 42 tissues via the Lynx MPSS system revealed almost exclusive expression of ZmGrp3 in maize roots. In the second part of this survey, ZmGrp3 expression was assayed in maize hybrids. In this context, a novel approach to quantify allele-specific contribution to gene expression in maize hybrids was developed. This assay combines RT-PCR amplification of polymorphisms between two alleles and subsequent quantification of allele-specific gene expression via a combination of didesoxyterminator assays and capillary electrophoresis. Allelic expression of the ZmGrp3 gene in six reciprocal hybrids generated from three ZmGrp3 alleles was analyzed via a new statistical mixed model approach.
Woll
Katrin
K
Center for Plant Molecular Biology (ZMBP), Department of General Genetics, Eberhard-Karls-University Tuebingen, Auf der Morgenstelle 28, Tuebingen 72076, Germany.
Dressel
Angela
A
Sakai
Hajime
H
Piepho
Hans-Peter
HP
Hochholdinger
Frank
F
D001483:Base Sequence; D015152:Blotting, Northern; D017931:DNA Primers; D030541:Databases, Genetic; D019075:Electrophoresis, Capillary; D020869:Gene Expression Profiling; D005819:Genetic Markers / Q000235:genetics*; D006824:Hybridization, Genetic; D017403:In Situ Hybridization; D015233:Models, Statistical; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D018517:Plant Roots / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D003313:Zea mays / Q000235:genetics*
D017931:DNA Primers; D005819:Genetic Markers; D010940:Plant Proteins