Reverse genetics analysis of the 55-kDa B regulatory subunit of 2A serine/threonine protein phosphatase (PP2A) related to self-incompatibility in Chinese cabbage
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The type 2A serine/threonine protein phosphatases (PP2As) are key components in regulating signal transduction and controlling cell metabolism. In particular, PP2A has a role in pollen-pistil interaction during pollination in Brassica. In this study, reverse genetics screening was used to obtain a T-DNA-inserted Brassica rapa mutant line with the self-compatible phenotype. The type 2A PP2A 55-kDa B regulatory subunit gene (PR55/B) in this mutant was knocked out by T-DNA insertion, and its expression level was decreased, which led to self-compatible phenotypes, such as seed setting by flower self-pollination. For functional analysis of this self-compatible mutant line, the full-length PR55/B gene related to the self-incompatibility mechanism was isolated and the down-regulation vector (pPPi) was constructed for introduction into Chinese cabbage. Chinese cabbage lines transformed with pPPi showed significantly decreased PR55/B mRNA accumulation, which yielded self-compatible phenotypes. These transgenic lines were found to set pods and seeds by the self-pollination in both buds and flowers. Down-regulation of PP2A, which is related to self-incompatibility signal transduction, led to higher levels of the phosphorylated S-locus receptor kinase (SRK) substrate, which phosphorylated SRK blocks activation of armadillo-repeat-containing 1 (ARC1). Thus, the self-incompatibility response by ARC1 was not activated, and this led to the self-compatible response. Finally, PP2A is proposed to be a significant factor in self-incompatibility of Chinese cabbage.
KeywordsBrassica rapa Flower-pollination Insertional mutant line Reverse genetics Self-incompatibility
This work was carried out with the support of the “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ01365201)”, Rural Development Administration, Republic of Korea.
G-HL performed the majority of the experiment and data analysis. N-RS contributed to fertility experiment and data analysis. Y-DP designed the experiment and analyzed data. G-HL and Y-DP wrote manuscript. All authors contributed to and corrected the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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