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Genome stability in the uvh6 mutant of Arabidopsis thaliana

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Abstract

Key message

Plant XPD homolog UVH6 is the protein involved in the repair of strand breaks, and the excision repair and uvh6 mutant is not impaired in transgenerational increase in HRF.

Abstract

While analyzing the transgenerational response to stress in plants, we found that the promoter and gene body of Arabidopsis thaliana (Arabidopsis) XPD homolog UVH6 underwent hypomethylation and showed an increase in the level of transcript. Here, we analyzed the mutant of this gene, uvh6-1, by crossing it to two different reporter lines: one which allows for analysis of homologous recombination frequency (HRF) and another which makes it possible to analyze the frequency of point mutations. We observed that uvh6-1 plants exhibited lower rate of spontaneous homologous recombination but higher frequencies of spontaneous point mutations. The analysis of strand breaks using ROPS and Comet assays showed that the mutant had a much higher level of strand breaks at non-induced conditions. Exposure to stresses such as UVC, heat, cold, flood and drought showed that the mutant was not impaired in an increase in somatic HRF. The analysis of spontaneous HRF in the progeny of control plants compared to that of the progeny of stressed plants demonstrated that uvh6-1 was mildly affected in response to temperature, UV and drought. Our data suggest that UVH6 may be involved in the repair of strand breaks and excision repair, but it is unlikely that UVH6 is required for transgenerational increase in HRF.

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Acknowledgments

We thank Valentina Titova for proofreading the manuscript. IK acknowledges the financial support of National Science and Engineering Research Council of Canada and Alberta Innovates. We thank Jennifer D. Hall for the gift of uvh6-1 seeds.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Lethbridge, University Drive 4401, Lethbridge, AB, T1K 3M4, Canada

    Andriy Bilichak, Youli Yao, Viktor Titov, Andrey Golubov & Igor Kovalchuk

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  1. Andriy Bilichak
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  2. Youli Yao
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  3. Viktor Titov
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  4. Andrey Golubov
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  5. Igor Kovalchuk
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Corresponding author

Correspondence to Igor Kovalchuk.

Additional information

Communicated by K. Chong.

Electronic supplementary material

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299_2014_1580_MOESM1_ESM.ppt

Fig. S1. Analysis of the number of mesophyll cells, leaf thickness and DNA content in wt and uvh6 plants. A. The average number of mesophyll cells per microscopic view. The data are shown as the average (with SD) calculated from 22 views per each line.B. Representative images of mesophyll cell density in wt and uvh6. C. The average leaf thickness (in μM) calculated from 3 measurements per each view (with SD), with over 20 views measurement per each line. D. Representative images of field views used for measuring leaf thickness in wt and uvh6. E. The average genomic DNA content (with SD)—shown in ug per 20 plants used for extraction (PPT 487 kb)

299_2014_1580_MOESM2_ESM.ppt

Fig. S2. Representative images of recombination events in wild-type plants before (A) and after (B) exposure to UVC (PPT 625 kb)

299_2014_1580_MOESM3_ESM.ppt

Fig. S3. The homologous recombination frequency and recombination rate in the progeny of stressed wild-type and uvh6-1 plants. A. The frequency of homologous recombination was calculated in the 3-week-old progeny of plants exposed to UVC (0.6 J/m2; UV0.6), flood (FL), drought (DR), cold (4 °C) and heat (50 °C). The data are shown as the average recombination frequency (of three independent experiments with SD). B. Recombination rate in wt and uvh6-1 plants (PPT 160 kb)

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Bilichak, A., Yao, Y., Titov, V. et al. Genome stability in the uvh6 mutant of Arabidopsis thaliana . Plant Cell Rep 33, 979–991 (2014). https://doi.org/10.1007/s00299-014-1580-0

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  • DOI: https://doi.org/10.1007/s00299-014-1580-0

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