Abstract

SCR/SP11 encodes the male determinant of recognition specificity of self-incompatibility (SI) in Brassica species and is sporophytically expressed in the anther tapetum. Based on dominance relationships in pollen and nucleotide sequence similarity, the S haplotypes in Brassica have been classified as class I or class II, with class-I S haplotypes being dominant over class-II S haplotypes. Here, we revealed that S-22 in B. rapa belonging to class I is recessive to class-II S-44 and class-I S-36 in pollen, whereas it is dominant over S-60, S-40, and S-29 based on pollination tests. SCR/SP11 of S-22 (SCR-22) was sequenced, revealing that the deduced amino-acid sequence of SCR-22 has the longest C-terminal domain among the SCR/SP11 sequences. The expression of SCR-22 was found to be suppressed in S-22/S-44 and S-22/S-36 heterozygotes. Normal transcription of SCR-44 was considered to be due to the transcription suppression of Smi sRNA of the S-22 haplotype and a very low methylation state of the SCR-44 promoter region in the tapetum of S-22/S-44 heterozygotes. In SCR-22, only the cytosine residue located at the –37 bp position of the promoter region was hypermethylated in the tapetum of S-22/S-44 heterozygotes, and few methylated cytosines were detected in the promoter and coding regions of SCR-22 in S-22/S-36 heterozygotes. SCR-22 was also expressed in microspores in S-22 homozygotes but not in S-22/S-44 and S-22/S-36 heterozygotes. These results suggest that a mechanism different from class-II SCR/SP11 suppression may operate for the suppression of recessive class-I SCR-22 in S heterozygotes.

Highlights

  • Self-incompatibility (SI) is a genetic mechanism exploited by many angiosperm species to prevent inbreeding and to promote outcrossing

  • Pollination tests showed that class-I SCR-22 is recessive to SCR-44 but dominant to SCR-60, SCR-40, and SCR-29 (Table 1)

  • The dominance relationships between SCR-22 and class-II SCR/SP11 alleles were further confirmed by gene expression analysis

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Summary

Introduction

Self-incompatibility (SI) is a genetic mechanism exploited by many angiosperm species to prevent inbreeding and to promote outcrossing. Subsequent studies demonstrated that the expression suppression of recessive class-II SCR/SP11 alleles results from their tissue-specific methylation of promoter sequences in the tapetum[18]. The expression of SCR-22 was suppressed in the S-22/S-44 heterozygote, but the cytosine methylation pattern in the SCR-22 promoter was different from that of recessive class-II SCR alleles.

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