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Self-compatibility in 'Zaohong' Japanese apricot is associated with the loss of function of pollen S genes.

Wang PP, Gao ZH, Ni ZJ, Zhang Z, Cai BH - Mol. Biol. Rep. (2013)

Bottom Line: Furthermore, expression analysis based on RT-PCR of S-locus genes revealed no differences at the transcript level when compared with 'Xiyeqing', a self-incompatible cultivar with the same S haplotypes.Putative structural regions of PmF-box genes have been described, corresponding to regions in PmSFB alleles, but with some sequence variations.These results suggest that SC in 'Zaohong' occurs in pollen, and that other factors outside the S-locus, including PmF-box genes, might be associated with the loss of function of pollen S genes.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Nanjing Agricultural University, No.1 Weigang, Nanjing City, 210095, Jiangsu Province, People's Republic of China.

ABSTRACT
While most Japanese apricot (Prunus mume Sieb. et Zucc.) cultivars display typical S-RNase-based gametophytic self-incompatibility, some self-compatible (SC) cultivars have also been identified. In this study, we confirmed SC of 'Zaohong' through replicated self-pollination tests. Cross-pollination tests showed that SC of 'Zaohong' was caused by a loss of pollen function, so we determined that the S-genotype of 'Zaohong' was S 2 S 15 . Sequence analysis of the S-haplotypes of 'Zaohong' showed no mutations which were likely to alter gene function. Furthermore, expression analysis based on RT-PCR of S-locus genes revealed no differences at the transcript level when compared with 'Xiyeqing', a self-incompatible cultivar with the same S haplotypes. In addition, except for S-locus genes, a new type of F-box gene encoding a previously uncharacterised protein with high sequence similarity (61.03-64.65 %) to Prunus SFB genes was identified. Putative structural regions of PmF-box genes have been described, corresponding to regions in PmSFB alleles, but with some sequence variations. These results suggest that SC in 'Zaohong' occurs in pollen, and that other factors outside the S-locus, including PmF-box genes, might be associated with the loss of function of pollen S genes.

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RT-PCR analysis of the SFB2, SFB15 and Actin genes in pollen (P), styles (S) and leaves (L) of ‘Zaohong’ (S2S15) and ‘Xiyeqing’ (S2S15)
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Fig2: RT-PCR analysis of the SFB2, SFB15 and Actin genes in pollen (P), styles (S) and leaves (L) of ‘Zaohong’ (S2S15) and ‘Xiyeqing’ (S2S15)

Mentions: RT-PCR of S-RNase, SFB and Actin genes was performed using total RNA from pollen grains, styles and leaves of ‘Zaohong’ with the primers Pru-C2/PCE-R, SFB-C1F/Pm-Vb and ActF1/ActR1, respectively (Fig. 1). RT-PCR analysis of Actin genes amplified fragments of the same size, and which were shorter than the fragments amplified from genomic DNA, due to the absence of introns (Fig. 1d). The results, therefore, confirmed that the RNA preparations were free from genomic DNA contamination. For RT-PCR of S-RNase alleles, amplified fragments were only found in stylar RNA and genomic DNA (Fig. 1a). Furthermore, amplified fragments from the style were shorter those from genomic DNA due to the absence of the second intron. RT-PCR of SFB alleles from pollen grain RNA however, produced a fragment of the same size as that of genomic DNA, but no amplification was seen with leaf RNA or stylar RNA samples (Fig. 1b). Together, these results indicate that the S-RNase and SFB alleles identified in ‘Zaohong’ are specifically expressed in style and pollen, respectively (Fig. 2).Fig. 2


Self-compatibility in 'Zaohong' Japanese apricot is associated with the loss of function of pollen S genes.

Wang PP, Gao ZH, Ni ZJ, Zhang Z, Cai BH - Mol. Biol. Rep. (2013)

RT-PCR analysis of the SFB2, SFB15 and Actin genes in pollen (P), styles (S) and leaves (L) of ‘Zaohong’ (S2S15) and ‘Xiyeqing’ (S2S15)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824209&req=5

Fig2: RT-PCR analysis of the SFB2, SFB15 and Actin genes in pollen (P), styles (S) and leaves (L) of ‘Zaohong’ (S2S15) and ‘Xiyeqing’ (S2S15)
Mentions: RT-PCR of S-RNase, SFB and Actin genes was performed using total RNA from pollen grains, styles and leaves of ‘Zaohong’ with the primers Pru-C2/PCE-R, SFB-C1F/Pm-Vb and ActF1/ActR1, respectively (Fig. 1). RT-PCR analysis of Actin genes amplified fragments of the same size, and which were shorter than the fragments amplified from genomic DNA, due to the absence of introns (Fig. 1d). The results, therefore, confirmed that the RNA preparations were free from genomic DNA contamination. For RT-PCR of S-RNase alleles, amplified fragments were only found in stylar RNA and genomic DNA (Fig. 1a). Furthermore, amplified fragments from the style were shorter those from genomic DNA due to the absence of the second intron. RT-PCR of SFB alleles from pollen grain RNA however, produced a fragment of the same size as that of genomic DNA, but no amplification was seen with leaf RNA or stylar RNA samples (Fig. 1b). Together, these results indicate that the S-RNase and SFB alleles identified in ‘Zaohong’ are specifically expressed in style and pollen, respectively (Fig. 2).Fig. 2

Bottom Line: Furthermore, expression analysis based on RT-PCR of S-locus genes revealed no differences at the transcript level when compared with 'Xiyeqing', a self-incompatible cultivar with the same S haplotypes.Putative structural regions of PmF-box genes have been described, corresponding to regions in PmSFB alleles, but with some sequence variations.These results suggest that SC in 'Zaohong' occurs in pollen, and that other factors outside the S-locus, including PmF-box genes, might be associated with the loss of function of pollen S genes.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Nanjing Agricultural University, No.1 Weigang, Nanjing City, 210095, Jiangsu Province, People's Republic of China.

ABSTRACT
While most Japanese apricot (Prunus mume Sieb. et Zucc.) cultivars display typical S-RNase-based gametophytic self-incompatibility, some self-compatible (SC) cultivars have also been identified. In this study, we confirmed SC of 'Zaohong' through replicated self-pollination tests. Cross-pollination tests showed that SC of 'Zaohong' was caused by a loss of pollen function, so we determined that the S-genotype of 'Zaohong' was S 2 S 15 . Sequence analysis of the S-haplotypes of 'Zaohong' showed no mutations which were likely to alter gene function. Furthermore, expression analysis based on RT-PCR of S-locus genes revealed no differences at the transcript level when compared with 'Xiyeqing', a self-incompatible cultivar with the same S haplotypes. In addition, except for S-locus genes, a new type of F-box gene encoding a previously uncharacterised protein with high sequence similarity (61.03-64.65 %) to Prunus SFB genes was identified. Putative structural regions of PmF-box genes have been described, corresponding to regions in PmSFB alleles, but with some sequence variations. These results suggest that SC in 'Zaohong' occurs in pollen, and that other factors outside the S-locus, including PmF-box genes, might be associated with the loss of function of pollen S genes.

Show MeSH