Identification and characterisation of three genes determining
embryogenesis by means of T-DNA mutagenesis in Arabidopsis
thaliana.

D 2004

Identification and characterisation of three genes determining embryogenesis by means of T-DNA mutagenesis in Arabidopsis thaliana.

ŘEPKOVÁ, Jana, Markéta DUDOVÁ and Tomáš KOCÁBEK

Basic information

Original name

Identification and characterisation of three genes determining embryogenesis by means of T-DNA mutagenesis in Arabidopsis thaliana.

Name in Czech

Identifikace a characterizace tří genů determinujících embryogenezi prostřednictvím T-DNA mutací u Arabidopsis thaliana.

Authors

ŘEPKOVÁ, Jana (203 Czech Republic, guarantor), Markéta DUDOVÁ (203 Czech Republic) and Tomáš KOCÁBEK (203 Czech Republic)

Edition

Berlin, Germany, Abstract Book of the 15th International Conference on Arabidopsis Research, July 11-14, Berlin, Germany. p. T01-037, 1 pp. 2004

Publisher

Max Planck Institute of Molecular Plant Physiology

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

Genetics and molecular biology

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/04:00010788

Organization unit

Faculty of Science

Keywords in English

Arabidopsis thaliana; T-DNA

Abstract

ORIG CZ

V originále

One of the possible approaches to a plant gene function study in model plant Arabidopsis thaliana leads across T-DNA mutagenesis. Collection of 2500 T-DNA lines have been obtained by A. thaliana transformation with Agrobacterium tumefaciens containing plasmid pPCVRN4 with HPT selectable gene (Koncz et al. 1994). Screening of individual lines revealed three different mutations with defects in immature seed development (marked 1265, 1293, 1321) which have been characterised and analysed in detail. One T-DNA insert was determined in each line and the mutations were confirmed to be monogenic and recessive-lethal by genetic analysis. Morphological studies of embryo development were performed by clearing in chloral hydrate. Mutant embryos were blocked in certain steps in the process necessary for embryo viability and development. In 1265 mutation defect in globular stage was observed. It was associated with abnormal pattern of cell division connected with shoot apical meristem formation during embryogenesis. In 1293 mutation defect in cotyledon formation was observed and in 1321 mutation defect was connected with late embryogenesis, seed maturation and storage products formation. Cosegregation of T-DNA with the mutation was confirmed only in the 1265 line. The inverse PCR method was applied for isolation of flanking plant DNA. The obtained fragment of DNA was sequenced and subjected to BLAST and The Arabidopsis Information Resource database search, which revealed T-DNA insertion in At1g53330 gene coding pentatricopeptide repeat (PPR) containing protein. The hypothetical function of this gene is connected with transporter activity and probably cell - cell signalling. The other mutations, 1293 and 1321, which did not cosegregated with T-DNA, were subjected to genetic mapping for perspective isolation of the affected genes by map-based cloning strategy. DNA markers, SSR and CAPS, linked to the mutated gene were used to delimit the region containing the gene of interest. 1321 mutation was located on chromosome 2 in tight linkage with marker nga 1126 and 1293 mutation was linked with marker nga 63 on chromosome 1. The next work will be aimed at fine mapping enabling candidate gene choice and the isolation of DNA sequence that underlines the phenotype of interest.

In Czech

Jedním z možných přístupů studia funkcí rostlinných genů u Arabidopsis thaliana je T-DNA mutageneze. Kolekce 2500 T-DNA linií po transformaci Agrobacterium tumefaciens byla využita při vyhledávání mutací s embryonálními defekty. Tři tyto mutace (1265, 1293, 1321) podrobně analyzovány a charakterizovány. Genetická analýza potvrdila monogenní a recesivní charakter těchto mutací.

Links

MSM 143100008, plan (intention)

Name: Genomy a jejich funkce

Investor: Ministry of Education, Youth and Sports of the CR, Genomes and their functions

Citovat

ŘEPKOVÁ, Jana, Markéta DUDOVÁ and Tomáš KOCÁBEK. Identification and characterisation of three genes determining embryogenesis by means of T-DNA mutagenesis in Arabidopsis thaliana. In Abstract Book of the 15th International Conference on Arabidopsis Research, July 11-14, Berlin, Germany. Berlin, Germany: Max Planck Institute of Molecular Plant Physiology, 2004, p. T01-037, 1 pp.

@inproceedings{563651,
   author = {Řepková, Jana and Dudová, Markéta and Kocábek, Tomáš},
   address = {Berlin, Germany},
   booktitle = {Abstract Book of the 15th International Conference on Arabidopsis Research, July 11-14, Berlin, Germany.},
   keywords = {Arabidopsis thaliana; T-DNA},
   language = {eng},
   location = {Berlin, Germany},
   pages = {T01-037},
   publisher = {Max Planck Institute of Molecular Plant Physiology},
   title = {Identification and characterisation of three genes determining embryogenesis by means of T-DNA mutagenesis in Arabidopsis thaliana.},
   year = {2004}
}

TY  - JOUR
ID  - 563651
AU  - Řepková, Jana - Dudová, Markéta - Kocábek, Tomáš
PY  - 2004
TI  - Identification and characterisation of three genes determining embryogenesis by means of T-DNA mutagenesis in Arabidopsis thaliana.
PB  - Max Planck Institute of Molecular Plant Physiology
CY  - Berlin, Germany
KW  - Arabidopsis thaliana
KW  - T-DNA
N2  - One of the possible approaches to a plant gene function study in model plant Arabidopsis thaliana leads across T-DNA mutagenesis. Collection of 2500 T-DNA lines have been obtained by A. thaliana transformation with Agrobacterium tumefaciens containing plasmid pPCVRN4 with HPT selectable gene (Koncz et al. 1994). Screening of individual lines revealed three different mutations with defects in immature seed development (marked 1265, 1293, 1321) which have been characterised and analysed in detail. One T-DNA insert was determined in each line and the mutations were confirmed to be monogenic and recessive-lethal by genetic analysis. Morphological studies of embryo development were performed by clearing in chloral hydrate. Mutant embryos were blocked in certain steps in the process necessary for embryo viability and development. In 1265 mutation defect in globular stage was observed. It was associated with abnormal pattern of cell division connected with shoot apical meristem formation during embryogenesis. In 1293 mutation defect in cotyledon formation was observed and in 1321 mutation defect was connected with late embryogenesis, seed maturation and storage products formation. Cosegregation of T-DNA with the mutation was confirmed only in the 1265 line. The inverse PCR method was applied for isolation of flanking plant DNA. The obtained fragment of DNA was sequenced and subjected to BLAST and The Arabidopsis Information Resource database search, which revealed T-DNA insertion in At1g53330 gene coding pentatricopeptide repeat (PPR) containing protein. The hypothetical function of this gene is connected with transporter activity and probably cell - cell signalling. The other mutations, 1293 and 1321, which did not cosegregated with T-DNA, were subjected to genetic mapping for perspective isolation of the affected genes by map-based cloning strategy. DNA markers, SSR and CAPS, linked to the mutated gene were used to delimit the region containing the gene of interest. 1321 mutation was located on chromosome 2 in tight linkage with marker nga 1126 and 1293 mutation was linked with marker nga 63 on chromosome 1. The next work will be aimed at fine mapping enabling candidate gene choice and the isolation of DNA sequence that underlines the phenotype of interest.
ER  -

ŘEPKOVÁ, Jana, Markéta DUDOVÁ and Tomáš KOCÁBEK. Identification and characterisation of three genes determining embryogenesis by means of T-DNA mutagenesis in Arabidopsis thaliana. In \textit{Abstract Book of the 15th International Conference on Arabidopsis Research, July 11-14, Berlin, Germany.}. Berlin, Germany: Max Planck Institute of Molecular Plant Physiology, 2004, p.~T01-037, 1 pp.

Detailed Information on Publication Record