Articles
The effect of recombinant S-RNases on in vitro pollen germination and pollen tube growth in Prunus avium
Article number
1408_9
Pages
63 – 70
Language
English
Abstract
Sweet cherry (Prunus avium) exhibits S-RNase-based gametophytic self-incompatibility (GSI), which uses S-RNase as the pistil determinant.
S-RNase is a T2/S-type ribonuclease protein secreted in the stigma and the style transmitting tract.
It functions as a cytotoxin against incompatible pollen tubes, arresting them at the middle part of the styles.
The development of an in vitro assay for incompatible pollen inhibition will help to investigate the mechanism of the incompatible reaction in sweet cherry.
We previously demonstrated that the S-RNase fraction of the cation-exchange chromatograph of the ‘Satonishiki’ (S3S6) style extracts specifically inhibited the growth of cognate pollen tubes in vitro.
In this study, we tested the inhibitory effect of the mixture of recombinant PavS3– and PavS6-RNases, produced in a silkworm-baculovirus system, against ‘Satonishiki’ pollen.
Semi-quantification using protein blot indicated that the ‘Satonishiki’ S-RNase fraction contained the PavS3– and PavS6-RNases at 17.4 and 8.7%, respectively.
The mixture of recombinant PavS3– and PavS6-RNases at the equivalent concentration to 0.5 µg µL-1 ‘Satonishiki’ S-RNase fraction inhibited the growth of both ‘Satonishiki’ and ‘Rainier’ (S1S4, compatible) pollen tubes, while specifically inhibiting the germination of ‘Satonishiki’ pollen.
A lower concentration of the recombinant S-RNase mixture explicitly inhibited the growth of ‘Satonishiki’ pollen tubes with no effect on pollen germination.
These results indicated that recombinant S-RNase was successfully applied to an in vitro assay for incompatible reaction of sweet cherry.
S-RNase is a T2/S-type ribonuclease protein secreted in the stigma and the style transmitting tract.
It functions as a cytotoxin against incompatible pollen tubes, arresting them at the middle part of the styles.
The development of an in vitro assay for incompatible pollen inhibition will help to investigate the mechanism of the incompatible reaction in sweet cherry.
We previously demonstrated that the S-RNase fraction of the cation-exchange chromatograph of the ‘Satonishiki’ (S3S6) style extracts specifically inhibited the growth of cognate pollen tubes in vitro.
In this study, we tested the inhibitory effect of the mixture of recombinant PavS3– and PavS6-RNases, produced in a silkworm-baculovirus system, against ‘Satonishiki’ pollen.
Semi-quantification using protein blot indicated that the ‘Satonishiki’ S-RNase fraction contained the PavS3– and PavS6-RNases at 17.4 and 8.7%, respectively.
The mixture of recombinant PavS3– and PavS6-RNases at the equivalent concentration to 0.5 µg µL-1 ‘Satonishiki’ S-RNase fraction inhibited the growth of both ‘Satonishiki’ and ‘Rainier’ (S1S4, compatible) pollen tubes, while specifically inhibiting the germination of ‘Satonishiki’ pollen.
A lower concentration of the recombinant S-RNase mixture explicitly inhibited the growth of ‘Satonishiki’ pollen tubes with no effect on pollen germination.
These results indicated that recombinant S-RNase was successfully applied to an in vitro assay for incompatible reaction of sweet cherry.
Publication
Authors
D. Matsumoto, S. Suzuki, T. Takeda, R. Tao
Keywords
in vitro assay, gametophytic self-incompatibility, pollen germination medium, silkworm-baculovirus system, sweet cherry
Groups involved
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