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. 2014 May 7:5:167.
doi: 10.3389/fpls.2014.00167. eCollection 2014.

OsRAD51C is essential for double-strand break repair in rice meiosis

Ding Tang  1 , Chunbo Miao  1 , Yafei Li  1 , Hongjun Wang  1 , Xiaofei Liu  1 , Hengxiu Yu  2 , Zhukuan Cheng  1
Affiliations

OsRAD51C is essential for double-strand break repair in rice meiosis

Ding Tang et al. Front Plant Sci. .

Abstract

RAD51C is one of the RAD51 paralogs that plays an important role in DNA double-strand break repair by homologous recombination. Here, we identified and characterized OsRAD51C, the rice homolog of human RAD51C. The Osrad51c mutant plant is normal in vegetative growth but exhibits complete male and female sterility. Cytological investigation revealed that homologous pairing and synapsis were severely disrupted. Massive chromosome fragmentation occurred during metaphase I in Osrad51c meiocytes, and was fully suppressed by the CRC1 mutation. Immunofluorescence analysis showed that OsRAD51C localized onto the chromosomes from leptotene to early pachytene during prophase I, and that normal loading of OsRAD51C was dependent on OsREC8, PAIR2, and PAIR3. Additionally, ZEP1 did not localize properly in Osrad51c, indicating that OsRAD51C is required for synaptonemal complex assembly. Our study also provided evidence in support of a functional divergence in RAD51C among organisms.

Keywords: OsRAD51C; chromosome fragmentation; double-strand break repair; meiosis; rice.

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Figures

Figure 1
Figure 1
Male meiosis in wild type. (A) Pachytene; (B) Diakinesis; (C) Metaphase I; (D) Anaphase I; (E) Dyad; (F) Tetrad. Bars, 5 μm.
Figure 2
Figure 2
Male meiosis in the Osrad51c mutant. (A) Pachytene; (B) Diakinesis; (C) Early metaphase I; (D) Middle metaphase I; (E) Late metaphase I; (F) Anaphase I; (G) Telophase I; (H) Dyad; (I) Tetrad. Bars, 5 μ m.
Figure 3
Figure 3
Male meiosis in crc1 (A,B) and crc1 Osrad51c (C,D). (A,C) Diakinesis; (B,D) Metaphase I. Bars, 5 μm.
Figure 4
Figure 4
Dual immunolocalization of OsREC8 (red) and OsRAD51C (green) in wild type PMCs. (A) Leptotene; (B) Zygotene; (C) Mid prophase I; (D) Late pachytene. Bars, 5 μm.
Figure 5
Figure 5
Dual immunolocalization of several meiotic elements in Osrad51c PMCs. (A) OsREC8 (red) and ZEP1 (green) signals at early pachytene; (B) PAIR2 (red) and PAIR3 (green) signals at early pachytene. Bars, 5 μ m.
Figure 6
Figure 6
The localization of OsRAD51C at leptotene in different meiotic mutants of rice. (A) Osrec8; (B) pair2; (C) pair3; (D) zep1. The CENH3 signals (red) indicating the PMC in Osrec8, and the OsREC8 signals (red) indicating the PMCs in pair2, pair3, and zep1. OsRAD51C signals in green. Chromosomes stained with DAPI (blue). Bars, 5μ m.
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