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Li Yong-Hong, Qi Mei-Yan, Fang Yuan, Fu An, Wang Xiao-Qin, He Yi-Kun. Functional analysis of PpAux/IAA2 in Physcomitrella patens (Hedw.) Mitt. during protoplast regeneration[J]. Plant Science Journal, 2021, 39(5): 515-525. DOI: 10.11913/PSJ.2095-0837.2021.50515
Citation: Li Yong-Hong, Qi Mei-Yan, Fang Yuan, Fu An, Wang Xiao-Qin, He Yi-Kun. Functional analysis of PpAux/IAA2 in Physcomitrella patens (Hedw.) Mitt. during protoplast regeneration[J]. Plant Science Journal, 2021, 39(5): 515-525. DOI: 10.11913/PSJ.2095-0837.2021.50515
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Functional analysis of PpAux/IAA2 in Physcomitrella patens (Hedw.) Mitt. during protoplast regeneration

  • 1. Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing University of Agriculture, Beijing 102206, China;

  • 2. Beijing Bei Nong Enterprise Management Co., Ltd., Beijing 102206, China;

  • 3. College of Life Sciences, Capital Normal University, Beijing 100048, China

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This work was supported by grants from the National Natural Science Foundation of China (31871235, 31371243)。

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  • Received Date: May 16, 2021
  • Revised Date: June 29, 2021
  • Available Online: October 31, 2022
  • Published Date: October 27, 2021
    Graphical Abstract
    • Abstract
  • We used wild-type (WT) and auxin-resistant mutant Ppiaa2-87 strains of Physcomitrella patens to explore the regulation mechanism of the early auxin-response gene PpAux/IAA2 in protoplast regeneration. Gene expression, survival rate, cell cycle progression, chromosome remodeling, and DNA methylation during protoplast regeneration were analyzed separately using quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), fluorescein diacetate (FDA) staining, flow cytometry of DNA ploidy, 4',6-diamidino-2-phenylindole (DAPI) staining, and methylation-sensitive amplified polymorphism analysis. Results showed that the expression levels of three PpAux/IAA homologous genes in 0-h protoplasts were significantly higher than that in protonemata, 48-h, and 96-h protoplasts; survival rate of Ppiaa2-87 protoplasts decreased significantly with culture time; entry into the S phase of Ppiaa2-87 protoplasts was inhibited; chromosomes of some Ppiaa2-87 0-h protoplasts were not remodeled; expression of four genes in the SWI/SNF protein family in the chromatin remodeling complex decreased in 0-h protoplasts compared with that in protonemata; and methylation of Ppiaa2-87 0-h protoplasts was higher than that of WT protoplasts. This study indicates that the PpAux/IAA2-related auxin signaling pathway plays an important role in protoplast regeneration. Mutation of the PpAux/IAA2 gene affects protoplast DNA methylation and chromatin remodeling, which is involved in cell development reprogramming, leading to protoplast death due to lack of pluripotency.
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    • References (39)
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