Reverse genetics analysis of the 55-kDa B regulatory subunit of 2A serine/threonine protein phosphatase (PP2A) related to self-incompatibility in Chinese cabbage

  • Gi-Ho Lee
  • Na-Ri Shin
  • Young-Doo ParkEmail author
Research Report


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.


Brassica 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.

Author contributions

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.

Supplementary material

13580_2019_194_MOESM1_ESM.docx (159 kb)
Supplementary material 1 (DOCX 159 kb)


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Copyright information

© Korean Society for Horticultural Science 2019

Authors and Affiliations

  1. 1.Department of Horticultural BiotechnologyKyung Hee UniversityYongin-siRepublic of Korea

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