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

다국어 초록 (Multilingual Abstract)

Background: The patatin-related phospholipase AIII family (pPLAIIIs) genes alter cell elongation and cellwall composition in Arabidopsis and rice plant, suggesting diverse commercial purposes of theeconomically important medicinal ginseng plant. Herein, we show the functional characterization of aginseng pPLAIII gene for the first time and discuss its potential applications.
Methods: pPLAIIIs were identified from ginseng expressed sequence tag clones and further confirmed bysearch against ginseng database and polymerase chain reaction. A clone showing the highest homologywith pPLAIIIb was shown to be overexpressed in Arabidopsis using Agrobacterium. Quantitative polymerasechain reaction was performed to analyze ginseng pPLAIIIb expression. Phenotypes were observedusing a low-vacuum scanning electron microscope. Lignin was stained using phloroglucinol and quantifiedusing acetyl bromide.
Results: The PgpPLAIIIb transcripts were observed in all organs of 2-year-old ginseng. Overexpression ofginseng pPLAIIIb (PgpPLAIIIb-OE) in Arabidopsis resulted in small and stunted plants. It shortened thetrichomes and decreased trichome number, indicating defects in cell polarity. Furthermore, OE linesexhibited enlarged seeds with less number per silique. The YUCCA9 gene was downregulated in the OElines, which is reported to be associated with lignification. Accordingly, lignin was stained less in the OElines, and the expression of two transcription factors related to lignin biosynthesis was also decreasedsignificantly.
Conclusion: Overexpression of pPLAIIIb 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 auxinresponsivegenes, and its stunted phenotype showed decreased lignin content.

참고문헌 (Reference)

1 Zhong R, "Transcriptional activation of secondary wall biosynthesis by rice maize NAC and MYB transcription factors" 52 : 1856-1871, 2011

2 Hentrich M, "The jasmonic acid signaling pathway is linked to auxin homeostasis through the modulation of YUCCA8 and YUCCA9 gene expression" 74 : 626-637, 2013

3 Yan L, "The heterologous expression in Arabidopsis thaliana of sorghum transcription factor SbbHLH1 downregulates lignin synthesis" 64 : 3021-3032, 2012

4 Hobbie L, "The axr4 auxin-resistant mutants of Arabidopsis thaliana define a gene important for root gravitropism and lateral root initiation" 7 : 211-200, 1995

5 Rietz S, "Roles of Arabidopsis patatin-related phospholipases A in root development are related to auxin responses and phosphate deficiency" 3 : 524-538, 2010

6 Kumar M, "Protocol : a medium-throughput method for determination of cellulose content from single stem pieces of Arabidopsis thaliana" 11 : 46-, 2015

7 Wilkins MR, "Protein identification and analysis tools in the ExPASy server" 122 : 531-552, 1999

8 Murthy HN, "Production of ginsenosides from adventitious root cultures of panax ginseng" 625-651, 2014

9 Wang X, "Plant phospholipases" 52 : 211-231, 2001

10 Ryu SB, "Phospholipid-derived signaling mediated by phospholipase A in plants" 9 : 229-235, 2004

11 Meijer HJ, "Phospholipid-based signaling in plants" 54 : 265-306, 2003

12 Li M, "Patatin-related phospholipase pPLAIIIδ increases seed oil content with long-chain fatty acids in Arabidopsis" 162 : 39-51, 2013

13 Li M, "Patatin-related phospholipase pPLAIIIβ-induced changes in lipid metabolism alter cellulose content and cell elongation in Arabidopsis" 23 : 1107-1123, 2011

14 Liu G, "Patatin-related phospholipase A, pPLAIIIα, modulates the longitudinal growth of vegetative tissues and seeds in rice" 66 : 6945-6955, 2015

15 Scherer GFE, "Patatin-related phospholipase A : nomenclature, subfamilies and functions in plants" 15 : 693-700, 2010

16 Li M, "Overexpression of patatinrelated phospholipase AIIIδ altered plant growth and increased seed oil content in camelina" 13 : 766-778, 2013

17 Ngoc Quy Nguyen, "Overexpression of ginseng UGT72AL1 causes organ fusion in the axillary leaf branch of Arabidopsis" 고려인삼학회 41 (41): 419-427, 2017

18 Pomar F, "O-4-linked coniferyl and sinapyl aldehydes in lignifying cell walls are the main targets of the Wiesner(phloroglucinol-HCl)reaction" 220 : 17-28, 2002

19 Holk A, "Molecular identification of cytosolic, patatin-related phospholipases A from Arabidopsis with potential functions in plant signal transduction" 130 : 90-101, 2002

20 Zhou J, "MYB58 and MYB63 are transcriptional activators of the lignin biosynthetic pathway during secondary cell wall formation in Arabidopsis" 21 : 248-266, 2009

21 Matos AR, "Lipid deacylating enzymes in plants : old activities, new genes" 47 : 491-503, 2009

22 Moore KJ, "Lignin and fiber digestion" 54 : 420-430, 2001

23 Bechtold N, "In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration" 82 : 259-266, 1998

24 Qiao Y, "Fine mapping and candidate gene analysis of dense and erect panicle 3, DEP3, which confers high grain yield in rice(Oryza sativa L.)" 122 : 1439-1449, 2011

25 Rietz S, "Expression of patatin-related phospholipase A gene AtPLA IIA in Arabidopsis thaliana is up-regulated by salicylic acid, wounding, ethylene, and iron and phosphate deficiency" 219 : 743-753, 2004

26 Lin CC, "Expression of an Oncidium gene encoding a patatin-like protein delays flowering in Arabidopsis by reducing gibberellin synthesis" 52 : 421-435, 2011

27 Kim MK, "Comparative analysis of expressed sequence tags(ESTs)of ginseng leaf" 25 : 599-606, 2006

28 Huang S, "Cloning of an Arabidopsis patatinlike gene, STURDY, by activation T-DNA tagging" 125 : 573-584, 2001

29 Andrews DL, "Characterization of the lipid acyl hydrolase activity of the major potato(Solanum tuberosum)tuber protein, patatin, by cloning and abundant expression in a baculovirus vector" 252 : 199-206, 1988

30 Yang W, "AtPLAIIs an acyl hydrolase involved in basal jasmonic acid production and Arabidopsis resistance to Botrytis cinerea" 282 : 18116-18128, 2007

31 Higginson T, "AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana" 35 : 177-192, 2003

32 Zhao Y, "A role for flavin monooxygenase-like enzyme in auxin biosynthesis" 291 : 306-309, 2001

33 La Camera S, "A pathogen-inducible patatin-like lipid acyl hydrolase facilitates fungal and bacterial host colonization in Arabidopsis" 44 : 810-825, 2005