OsKu70 is associated with developmental growth and genome stability in rice

Jong Pil Hong, Mi Young Byun, Kyungsook An, Sae Jun Yang, Gynheung An, Woo Taek Kim

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The cellular functions of Ku70 in repair of DNA double-stranded breaks and telomere regulation have been described in a wide range of organisms. In this study, we identified the rice (Oryza sativa) Ku70 homolog (OsKu70) from the rice genome database. OsKu70 transcript was detected constitutively in every tissue and developmental stage examined and also in undifferentiated callus cells in rice. Yeast two-hybrid and in vitro pull-down experiments revealed that OsKu70 physically interacts with OsKu80. We obtained loss-of-function osku70 T-DNA knockout mutant lines and constructed transgenic rice plants that overexpress the OsKu70 gene in the sense (35S:OsKu70) or antisense (35S:anti-OsKu70) orientation. The homozygous G2 osku70 mutant lines were more sensitive than wild-type plants to a DNA-damaging agent (0.01%-0.05% methyl-methane sulfonate), consistent with the notion that OsKu70 participates in the DNA repair mechanism. Terminal restriction fragment analysis revealed that telomeres in homozygous G2 osku70 mutants were markedly longer (10-20 kb) than those in wild-type plants (5-10 kb), whereas telomere length in heterozygous G2 osku70 mutant and T2 OsKu70-overexpressing transgenic (35S: OsKu70) rice resembled that of the wild-type plant. In contrast to what was observed in Arabidopsis (Arabidopsis thaliana) atku70 mutants, homozygous G2 osku70 rice plants displayed severe developmental defects in both vegetative and reproductive organs under normal growth conditions, resulting in sterile flowers. Analysis of meiotic progression in pollen mother cells demonstrated that up to 11.1% (seven of 63) of G2 mutant anaphase cells displayed one or more chromosomal fusions. These results suggest that OsKu70 is required for the maintenance of chromosome stability and normal developmental growth in rice plants.

Original languageEnglish
Pages (from-to)374-387
Number of pages14
JournalPlant physiology
Volume152
Issue number1
DOIs
Publication statusPublished - 2010 Jan 1

Fingerprint

Genomic Instability
Oryza sativa
rice
genome
Growth
telomeres
mutants
DNA repair
Telomere
Zizania aquatica
Oryza
methyl methanesulfonate
genetically modified organisms
aseptic conditions
knockout mutants
Arabidopsis
anaphase
DNA
cells
callus

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Genetics
  • Physiology

Cite this

Hong, Jong Pil ; Byun, Mi Young ; An, Kyungsook ; Yang, Sae Jun ; An, Gynheung ; Kim, Woo Taek. / OsKu70 is associated with developmental growth and genome stability in rice. In: Plant physiology. 2010 ; Vol. 152, No. 1. pp. 374-387.
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abstract = "The cellular functions of Ku70 in repair of DNA double-stranded breaks and telomere regulation have been described in a wide range of organisms. In this study, we identified the rice (Oryza sativa) Ku70 homolog (OsKu70) from the rice genome database. OsKu70 transcript was detected constitutively in every tissue and developmental stage examined and also in undifferentiated callus cells in rice. Yeast two-hybrid and in vitro pull-down experiments revealed that OsKu70 physically interacts with OsKu80. We obtained loss-of-function osku70 T-DNA knockout mutant lines and constructed transgenic rice plants that overexpress the OsKu70 gene in the sense (35S:OsKu70) or antisense (35S:anti-OsKu70) orientation. The homozygous G2 osku70 mutant lines were more sensitive than wild-type plants to a DNA-damaging agent (0.01{\%}-0.05{\%} methyl-methane sulfonate), consistent with the notion that OsKu70 participates in the DNA repair mechanism. Terminal restriction fragment analysis revealed that telomeres in homozygous G2 osku70 mutants were markedly longer (10-20 kb) than those in wild-type plants (5-10 kb), whereas telomere length in heterozygous G2 osku70 mutant and T2 OsKu70-overexpressing transgenic (35S: OsKu70) rice resembled that of the wild-type plant. In contrast to what was observed in Arabidopsis (Arabidopsis thaliana) atku70 mutants, homozygous G2 osku70 rice plants displayed severe developmental defects in both vegetative and reproductive organs under normal growth conditions, resulting in sterile flowers. Analysis of meiotic progression in pollen mother cells demonstrated that up to 11.1{\%} (seven of 63) of G2 mutant anaphase cells displayed one or more chromosomal fusions. These results suggest that OsKu70 is required for the maintenance of chromosome stability and normal developmental growth in rice plants.",
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OsKu70 is associated with developmental growth and genome stability in rice. / Hong, Jong Pil; Byun, Mi Young; An, Kyungsook; Yang, Sae Jun; An, Gynheung; Kim, Woo Taek.

In: Plant physiology, Vol. 152, No. 1, 01.01.2010, p. 374-387.

Research output: Contribution to journalArticle

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