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Overexpression of ginseng patatin-related phospholipase pPLAIII? alters the polarity of cell growth and decreases lignin content in Arabidopsis.

ABSTRACT: Background:The patatin-related phospholipase AIII family (pPLAIIIs) genes alter cell elongation and cell wall composition in Arabidopsis and rice plant, suggesting diverse commercial purposes of the economically important medicinal ginseng plant. Herein, we show the functional characterization of a ginseng pPLAIII gene for the first time and discuss its potential applications. Methods:pPLAIIIs were identified from ginseng expressed sequence tag clones and further confirmed by search against ginseng database and polymerase chain reaction. A clone showing the highest homology with pPLAIII? was shown to be overexpressed in Arabidopsis using Agrobacterium. Quantitative polymerase chain reaction was performed to analyze ginseng pPLAIII? expression. Phenotypes were observed using a low-vacuum scanning electron microscope. Lignin was stained using phloroglucinol and quantified using acetyl bromide. Results:The PgpPLAIII? transcripts were observed in all organs of 2-year-old ginseng. Overexpression of ginseng pPLAIII? (PgpPLAIII?-OE) in Arabidopsis resulted in small and stunted plants. It shortened the trichomes and decreased trichome number, indicating defects in cell polarity. Furthermore, OE lines exhibited enlarged seeds with less number per silique. The YUCCA9 gene was downregulated in the OE lines, which is reported to be associated with lignification. Accordingly, lignin was stained less in the OE lines, and the expression of two transcription factors related to lignin biosynthesis was also decreased significantly. Conclusion:Overexpression of pPLAIII? retarded cell elongation in all the tested organs except seeds, which were longer and thicker than those of the controls. Shorter root length is related to auxin-responsive genes, and its stunted phenotype showed decreased lignin content.

SUBMITTER: Jang JH

PROVIDER: S-EPMC7031755 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Overexpression of ginseng patatin-related phospholipase <i>pPLAIIIβ</i> alters the polarity of cell growth and decreases lignin content in <i>Arabidopsis</i>.

Jang Jin Hoon JH Lee Ok Ran OR

Journal of ginseng research 20190123 2

<h4>Background</h4>The patatin-related phospholipase AIII family (<i>pPLAIIIs</i>) genes alter cell elongation and cell wall composition in <i>Arabidopsis</i> and rice plant, suggesting diverse commercial purposes of the economically important medicinal ginseng plant. Herein, we show the functional characterization of a ginseng <i>pPLAIII</i> gene for the first time and discuss its potential applications.<h4>Methods</h4><i>pPLAIIIs</i> were identified from ginseng expressed sequence tag clones a ...[more]

PMID: 32148415

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Project description:Brassinosteroids (BRs) are signaling molecules that play essential roles in the spatial regulation of plant growth and development. In contrast to other plant hormones BRs act locally, close to the sites of their synthesis, and thus homeostatic mechanisms must operate at the cellular level to equilibrate BR concentrations. Whilst it is recognized that levels of bioactive BRs are likely adjusted by controlling the relative rates of biosynthesis and by catabolism, few factors, which participate in these regulatory events, have as yet been identified. Previously we have shown that the UDP-glycosyltransferase UGT73C5 of Arabidopsis thaliana catalyzes 23-O-glucosylation of BRs and that glucosylation renders BRs inactive. This study identifies the closest homologue of UGT73C5, UGT73C6, as an enzyme that is also able to glucosylate BRs in planta.In a candidate gene approach, in which homologues of UGT73C5 were screened for their potential to induce BR deficiency when over-expressed in plants, UGT73C6 was identified as an enzyme that can glucosylate the BRs CS and BL at their 23-O-positions in planta. GUS reporter analysis indicates that UGT73C6 shows over-lapping, but also distinct expression patterns with UGT73C5 and YFP reporter data suggests that at the cellular level, both UGTs localize to the cytoplasm and to the nucleus. A liquid chromatography high-resolution mass spectrometry method for BR metabolite analysis was developed and applied to determine the kinetics of formation and the catabolic fate of BR-23-O-glucosides in wild type and UGT73C5 and UGT73C6 over-expression lines. This approach identified novel BR catabolites, which are considered to be BR-malonylglucosides, and provided first evidence indicating that glucosylation protects BRs from cellular removal. The physiological significance of BR glucosylation, and the possible role of UGT73C6 as a regulatory factor in this process are discussed in light of the results presented.The present study generates essential knowledge and molecular and biochemical tools, that will allow for the verification of a potential physiological role of UGT73C6 in BR glucosylation and will facilitate the investigation of the functional significance of BR glucoside formation in plants.

Dataset Information

Overexpression of ginseng patatin-related phospholipase pPLAIII? alters the polarity of cell growth and decreases lignin content in Arabidopsis.

Publications

Overexpression of ginseng patatin-related phospholipase <i>pPLAIIIβ</i> alters the polarity of cell growth and decreases lignin content in <i>Arabidopsis</i>.

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