Identification of Stylar RNases Associated with Gametophytic Self-Incompatibility in Almond (Prunus dulcis)

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Stylar proteins of 13 almond (Prunus dulcis) cultivars with known S-genotypes were surveyed by IEF and 2D-PAGE combined with immunoblot and N-terminal amino acid sequence analyses to identify S-RNases associated with gametophytic self-incompatibility (SI) in this plant species. RNase activities corresponding to S_a and S_b, two of the four S-alleles tested, were identified by IEF and RNase activity staining. The S_a-RNase band reacted with the anti-S_4-serum prepared from Japanese pear (Pyrus serotina); no reaction with the antiserum was observed with the S_b-RNase band. When the S_a-RNase band was excised from an IEF gel stained for RNase activity, subjected to SDS-PAGE, and detected by immunoblotting, it appeared that this band consisted of a single protein that reacted with the anti-S_4-serum with M_r of about 28 kDa. With 2D-PAGE and silver staining of the stylar extracts, all four S-proteins could be successfully distinguished from each other in the highly basic zone of the gel. Although S_b-, S_c-, and S_d-Proteins had roughly the same M_r of about 30 kDa, the S_c-protein seemed to be slightly smaller than the S_b-Protein and slightly larger than the S_d-protein. In 2D-PAGE profiles as well, the S_a-protein had M_r of about 28 kDa, apparently smaller than the other three proteins. A bud sport, in which one of the two S-alleles of the original cultivar is impaired, was visualized as a loss of S_c-protein, which is consistent with the previous pollination study. All four S-proteins reacted with the anti-S_4-serum, probably because of the differing conformations of these S-proteins in the IEF and 2D-PAGE gels. The S_a-protein in 2D-PAGE appeared to be identical to S_a-RNase in IEF; both had the same M_r and were reactive with the anti-S_4-serum. N-terminal amino acid sequence analysis of the four S-proteins revealed that they were highly homologous to each other and similar to the S-RNases of Malus. Pyrus, Scrophulariaceae, and Solanaceae. Taken together, RNases in the style are strongly suggested to be associated with the gametophytic SI of almond. This is the first report identifying and characterizing S-RNase in almond.
日本植物生理学会の論文

Identification of Stylar RNases Associated with Gametophytic Self-Incompatibility in Almond (Prunus dulcis)

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