Isolation and characterization of a novel semi-lethal
Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR)
repeat-containing protein

D 2005

Isolation and characterization of a novel semi-lethal Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR) repeat-containing protein

KOCÁBEK, Tomáš, Jana ŘEPKOVÁ, L. VRBA a K. HOYEROVÁ

Základní údaje

Originální název

Isolation and characterization of a novel semi-lethal Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR) repeat-containing protein

Název česky

Izolace a charakterizace nového mutanta v genu pro protein obsahující PPR motiv u Arabidopsis thaliana

Autoři

KOCÁBEK, Tomáš (203 Česká republika), Jana ŘEPKOVÁ (203 Česká republika, garant), L. VRBA (203 Česká republika) a K. HOYEROVÁ (203 Česká republika)

Vydání

České Budějovice, Book of Abstracts, 6th International Symposium in the Series Recent Advances in Plant Biotechnology, 12.-16.9.2005, s. 46-46, 2005

Nakladatel

Institute of Plant Molecular Biology ASCR

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

Genetika a molekulární biologie

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/05:00021384

Organizační jednotka

Přírodovědecká fakulta

ISBN

80-86778-16-9

UT WoS

000241139800034

Klíčová slova anglicky

Arabidopsis thaliana; T-DNA; PPR; gene isolation

Štítky

Arabidopsis thaliana, gene isolation, PPR, T-DNA

Změněno: 2. 10. 2006 13:33, prof. RNDr. Jana Řepková, CSc.

Anotace

ORIG CZ

V originále

A novel Arabidopsis thaliana mutant of one member of the PPR (pentatricopeptide repeat) gene family has been identified during the screening of 2500 Arabidopsis individual insertion T2 lines. The deviations in growth and development of the mutant plants appeared already at the initial growth. The germination was completely inhibited (about 30 to 40% of the T3 seeds in each line) or distinctly delayed in comparison with wt plants. The next development of the mutant seedlings varied further on agar medium. Complex phenotypic expression from post-embryonic lethality associated with abnormal pattern of cell division during globular to heart transition to fertile plants with just subtle phenotypic changes was observed within this individual mutant line. The Southern blot and segregation analysis revealed that the mutant plants contained one insert of T-DNA tightly linked to the mutant locus. To be sure that the ambiguous mutant phenotype was due to a mutation at a single locus, we have mapped simultaneously recessive homozygotes without strong embryonic defects and heterozygotes with embryonic defects caused lethality. In both cases, the location of the mutation was assigned on chromosome 1 in position 91.0 ą 4.9 cM in genetic map derived from recombinant inbred lines. To determine the genomic region flanking T-DNA borders we have provided inverse PCR (iPCR) approach followed by sequencing of the products. The sequencing and BLAST search revealed that the insert was integrated into a gene coding for pentatricopeptide repeat (PPR) containing protein At1g53330 which corresponds to the previously mapped position. This gene is composed of 8 tandem arrays of classical PPR motif characterized by 35 amino acid repeats (Lurin et al., 2004). The PPR protein At1g53330.1 was predicted to be targeted to mitochondria by TargetP program, whereas Predotar did not found any targeting signal peptides. Extensive search for other Arabidiopsis mutants affected in expression of particular PPR proteins as well as a more detailed look at the protein action, the affinity to RNA or other protein molecules are currently in progress to elucidate the exact functions of one of the largest and least understood protein families in plants.

Česky

Skríningem 2500 T-DNA mutantů Arabidopsis thaliana byla zachycena a charakterizována mutace v novém genu pro protein obsahující PPR (pentatricopeptide repeat)motiv. Gen byl lokalizován na chromozomu 1 a byl izolován metodou inverzní PCR.

Návaznosti

MSM0021622415, záměr

Název: Molekulární podstata buněčných a tkáňových regulací

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Molekulární podstata buněčných a tkáňových regulací

Citovat

KOCÁBEK, Tomáš, Jana ŘEPKOVÁ, L. VRBA a K. HOYEROVÁ. Isolation and characterization of a novel semi-lethal Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR) repeat-containing protein. In Book of Abstracts, 6th International Symposium in the Series Recent Advances in Plant Biotechnology, 12.-16.9.2005. České Budějovice: Institute of Plant Molecular Biology ASCR, 2005, s. 46. ISBN 80-86778-16-9.

@inproceedings{583463,
   author = {Kocábek, Tomáš and Řepková, Jana and Vrba, L. and Hoyerová, K.},
   address = {České Budějovice},
   booktitle = {Book of Abstracts, 6th International Symposium in the Series Recent Advances in Plant Biotechnology, 12.-16.9.2005},
   keywords = {Arabidopsis thaliana; T-DNA; PPR; gene isolation},
   language = {eng},
   location = {České Budějovice},
   isbn = {80-86778-16-9},
   pages = {46-46},
   publisher = {Institute of Plant Molecular Biology ASCR},
   title = {Isolation and characterization of a novel semi-lethal Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR) repeat-containing protein},
   year = {2005}
}

TY  - JOUR
ID  - 583463
AU  - Kocábek, Tomáš - Řepková, Jana - Vrba, L. - Hoyerová, K.
PY  - 2005
TI  - Isolation and characterization of a novel semi-lethal Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR) repeat-containing protein
PB  - Institute of Plant Molecular Biology ASCR
CY  - České Budějovice
SN  - 8086778169
KW  - Arabidopsis thaliana
KW  - T-DNA
KW  - PPR
KW  - gene isolation
N2  - A novel Arabidopsis thaliana mutant of one member of the PPR (pentatricopeptide repeat) gene family has been identified during the screening of 2500 Arabidopsis individual insertion T2 lines. The deviations in growth and development of the mutant plants appeared already at the initial growth. The germination was completely inhibited (about 30 to 40% of the T3 seeds in each line) or distinctly delayed in comparison with wt plants. The next development of the mutant seedlings varied further on agar medium. Complex phenotypic expression from post-embryonic lethality associated with abnormal pattern of cell division during globular to heart transition to fertile plants with just subtle phenotypic changes was observed within this individual mutant line. The Southern blot and segregation analysis revealed that the mutant plants contained one insert of T-DNA tightly linked to the mutant locus. To be sure that the ambiguous mutant phenotype was due to a mutation at a single locus, we have mapped simultaneously recessive homozygotes without strong embryonic defects and heterozygotes with embryonic defects caused lethality. In both cases, the location of the mutation was assigned on chromosome 1 in position 91.0 ą 4.9 cM in genetic map derived from recombinant inbred lines. To determine the genomic region flanking T-DNA borders we have provided inverse PCR (iPCR) approach followed by sequencing of the products. The sequencing and BLAST search revealed that the insert was integrated into a gene coding for pentatricopeptide repeat (PPR) containing protein At1g53330 which corresponds to the previously mapped position. This gene is composed of 8 tandem arrays of classical PPR motif characterized by 35 amino acid repeats (Lurin et al., 2004). The PPR protein At1g53330.1 was predicted to be targeted to mitochondria by TargetP program, whereas Predotar did not found any targeting signal peptides. Extensive search for other Arabidiopsis mutants affected in expression of particular PPR proteins as well as a more detailed look at the protein action, the affinity to RNA or other protein molecules are currently in progress to elucidate the exact functions of one of the largest and least understood protein families in plants.
ER  -

KOCÁBEK, Tomáš, Jana ŘEPKOVÁ, L. VRBA a K. HOYEROVÁ. Isolation and characterization of a novel semi-lethal Arabidopsis thaliana mutant of gene for pentatricopeptide (PPR) repeat-containing protein. In \textit{Book of Abstracts, 6th International Symposium in the Series Recent Advances in Plant Biotechnology, 12.-16.9.2005}. České Budějovice: Institute of Plant Molecular Biology ASCR, 2005, s.~46. ISBN~80-86778-16-9.

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