Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions

Yannic Müller; Payal Patwari; Tyll Stöcker; Viktoria Zeisler-Diehl; Ulrike Steiner; Chiara Campoli; Lea Grewe; Magdalena Kuczkowska; Maya Marita Dierig; Sarah Jose; Alistair M. Hetherington; Ivan F. Acosta; Heiko Schoof; Lukas Schreiber; Peter Dörmann

doi:10.1111/nph.19063

Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions

Yannic Müller, Payal Patwari, Tyll Stöcker, Viktoria Zeisler-Diehl, Ulrike Steiner, Chiara Campoli, Lea Grewe, Magdalena Kuczkowska, Maya Marita Dierig, Sarah Jose, Alistair M. Hetherington, Ivan F. Acosta, Heiko Schoof, Lukas Schreiber, Peter Dörmann^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

3 Citations (Scopus)

Abstract

The cuticle is a protective layer covering aerial plant organs. We studied the function of waxes for the establishment of the cuticular barrier in barley (Hordeum vulgare). The barley eceriferum mutants cer-za.227 and cer-ye.267 display reduced wax loads, but the genes affected, and the consequences of the wax changes for the barrier function remained unknown. Cuticular waxes and permeabilities were measured in cer-za.227 and cer-ye.267. The mutant loci were isolated by bulked segregant RNA sequencing. New cer-za alleles were generated by genome editing. The CER-ZA protein was characterized after expression in yeast and Arabidopsis cer4-3. Cer-za.227 carries a mutation in HORVU5Hr1G089230 encoding acyl-CoA reductase (FAR1). The cer-ye.267 mutation is located to HORVU4Hr1G063420 encoding β-ketoacyl-CoA synthase (KAS1) and is allelic to cer-zh.54. The amounts of intracuticular waxes were strongly decreased in cer-ye.267. The cuticular water loss and permeability of cer-za.227 were similar to wild-type (WT), but were increased in cer-ye.267. Removal of epicuticular waxes revealed that intracuticular, but not epicuticular waxes are required to regulate cuticular transpiration. The differential decrease in intracuticular waxes between cer-za.227 and cer-ye.267, and the removal of epicuticular waxes indicate that the cuticular barrier function mostly depends on the presence of intracuticular waxes.

Original language	English
Pages (from-to)	1903-1918
Number of pages	16
Journal	New Phytologist
Volume	239
Issue number	5
Early online date	22 Jun 2023
DOIs	https://doi.org/10.1111/nph.19063
Publication status	Published - 1 Aug 2023

Bibliographical note

Funding Information:
Funding for this project was provided by the Deutsche Forschungsgemeinschaft (DFG Research Training Group, GRK2064, ‘Water use efficiency and drought stress responses in Arabidopsis and /barley’; and Germany's Excellence Strategy, EXC 2070, PhenoRob grant no. 390732324), DFG program Major Research Instrumentation (Scanning Electron Microscope with EDX, grant no. 471591895), and by University of Bonn. CC acknowledges funding from the Biotechnological and Biological Sciences Research Council (BB/M018466/1) to Robbie Waugh. We would like to thank Marlies Becker and Christiane Buchholz for barley transformation and Penny von Wettstein‐Knowles (University of Copenhagen) for her insightful guidance on the work with the barley mutants. Open Access funding enabled and organized by Projekt DEAL.

Publisher Copyright:
© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.

Keywords

barley
cuticle
cutin
eceriferum
permeability
wax

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Müller, Y., Patwari, P., Stöcker, T., Zeisler-Diehl, V., Steiner, U., Campoli, C., Grewe, L., Kuczkowska, M., Dierig, M. M., Jose, S., Hetherington, A. M., Acosta, I. F., Schoof, H., Schreiber, L., & Dörmann, P. (2023). Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions. New Phytologist, 239(5), 1903-1918. https://doi.org/10.1111/nph.19063

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title = "Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions",

abstract = "The cuticle is a protective layer covering aerial plant organs. We studied the function of waxes for the establishment of the cuticular barrier in barley (Hordeum vulgare). The barley eceriferum mutants cer-za.227 and cer-ye.267 display reduced wax loads, but the genes affected, and the consequences of the wax changes for the barrier function remained unknown. Cuticular waxes and permeabilities were measured in cer-za.227 and cer-ye.267. The mutant loci were isolated by bulked segregant RNA sequencing. New cer-za alleles were generated by genome editing. The CER-ZA protein was characterized after expression in yeast and Arabidopsis cer4-3. Cer-za.227 carries a mutation in HORVU5Hr1G089230 encoding acyl-CoA reductase (FAR1). The cer-ye.267 mutation is located to HORVU4Hr1G063420 encoding β-ketoacyl-CoA synthase (KAS1) and is allelic to cer-zh.54. The amounts of intracuticular waxes were strongly decreased in cer-ye.267. The cuticular water loss and permeability of cer-za.227 were similar to wild-type (WT), but were increased in cer-ye.267. Removal of epicuticular waxes revealed that intracuticular, but not epicuticular waxes are required to regulate cuticular transpiration. The differential decrease in intracuticular waxes between cer-za.227 and cer-ye.267, and the removal of epicuticular waxes indicate that the cuticular barrier function mostly depends on the presence of intracuticular waxes.",

keywords = "barley, cuticle, cutin, eceriferum, permeability, wax",

author = "Yannic M{\"u}ller and Payal Patwari and Tyll St{\"o}cker and Viktoria Zeisler-Diehl and Ulrike Steiner and Chiara Campoli and Lea Grewe and Magdalena Kuczkowska and Dierig, {Maya Marita} and Sarah Jose and Hetherington, {Alistair M.} and Acosta, {Ivan F.} and Heiko Schoof and Lukas Schreiber and Peter D{\"o}rmann",

note = "Funding Information: Funding for this project was provided by the Deutsche Forschungsgemeinschaft (DFG Research Training Group, GRK2064, {\textquoteleft}Water use efficiency and drought stress responses in Arabidopsis and /barley{\textquoteright}; and Germany's Excellence Strategy, EXC 2070, PhenoRob grant no. 390732324), DFG program Major Research Instrumentation (Scanning Electron Microscope with EDX, grant no. 471591895), and by University of Bonn. CC acknowledges funding from the Biotechnological and Biological Sciences Research Council (BB/M018466/1) to Robbie Waugh. We would like to thank Marlies Becker and Christiane Buchholz for barley transformation and Penny von Wettstein‐Knowles (University of Copenhagen) for her insightful guidance on the work with the barley mutants. Open Access funding enabled and organized by Projekt DEAL. Publisher Copyright: {\textcopyright} 2023 The Authors. New Phytologist {\textcopyright} 2023 New Phytologist Foundation.",

year = "2023",

month = aug,

day = "1",

doi = "10.1111/nph.19063",

language = "English",

volume = "239",

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Müller, Y, Patwari, P, Stöcker, T, Zeisler-Diehl, V, Steiner, U, Campoli, C, Grewe, L, Kuczkowska, M, Dierig, MM, Jose, S, Hetherington, AM, Acosta, IF, Schoof, H, Schreiber, L & Dörmann, P 2023, 'Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions', New Phytologist, vol. 239, no. 5, pp. 1903-1918. https://doi.org/10.1111/nph.19063

Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions. / Müller, Yannic; Patwari, Payal; Stöcker, Tyll et al.
In: New Phytologist, Vol. 239, No. 5, 01.08.2023, p. 1903-1918.

Research output: Contribution to journal › Article (Academic Journal) › peer-review

TY - JOUR

T1 - Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions

AU - Müller, Yannic

AU - Patwari, Payal

AU - Stöcker, Tyll

AU - Zeisler-Diehl, Viktoria

AU - Steiner, Ulrike

AU - Campoli, Chiara

AU - Grewe, Lea

AU - Kuczkowska, Magdalena

AU - Dierig, Maya Marita

AU - Jose, Sarah

AU - Hetherington, Alistair M.

AU - Acosta, Ivan F.

AU - Schoof, Heiko

AU - Schreiber, Lukas

AU - Dörmann, Peter

N1 - Funding Information: Funding for this project was provided by the Deutsche Forschungsgemeinschaft (DFG Research Training Group, GRK2064, ‘Water use efficiency and drought stress responses in Arabidopsis and /barley’; and Germany's Excellence Strategy, EXC 2070, PhenoRob grant no. 390732324), DFG program Major Research Instrumentation (Scanning Electron Microscope with EDX, grant no. 471591895), and by University of Bonn. CC acknowledges funding from the Biotechnological and Biological Sciences Research Council (BB/M018466/1) to Robbie Waugh. We would like to thank Marlies Becker and Christiane Buchholz for barley transformation and Penny von Wettstein‐Knowles (University of Copenhagen) for her insightful guidance on the work with the barley mutants. Open Access funding enabled and organized by Projekt DEAL. Publisher Copyright: © 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.

PY - 2023/8/1

Y1 - 2023/8/1

N2 - The cuticle is a protective layer covering aerial plant organs. We studied the function of waxes for the establishment of the cuticular barrier in barley (Hordeum vulgare). The barley eceriferum mutants cer-za.227 and cer-ye.267 display reduced wax loads, but the genes affected, and the consequences of the wax changes for the barrier function remained unknown. Cuticular waxes and permeabilities were measured in cer-za.227 and cer-ye.267. The mutant loci were isolated by bulked segregant RNA sequencing. New cer-za alleles were generated by genome editing. The CER-ZA protein was characterized after expression in yeast and Arabidopsis cer4-3. Cer-za.227 carries a mutation in HORVU5Hr1G089230 encoding acyl-CoA reductase (FAR1). The cer-ye.267 mutation is located to HORVU4Hr1G063420 encoding β-ketoacyl-CoA synthase (KAS1) and is allelic to cer-zh.54. The amounts of intracuticular waxes were strongly decreased in cer-ye.267. The cuticular water loss and permeability of cer-za.227 were similar to wild-type (WT), but were increased in cer-ye.267. Removal of epicuticular waxes revealed that intracuticular, but not epicuticular waxes are required to regulate cuticular transpiration. The differential decrease in intracuticular waxes between cer-za.227 and cer-ye.267, and the removal of epicuticular waxes indicate that the cuticular barrier function mostly depends on the presence of intracuticular waxes.

AB - The cuticle is a protective layer covering aerial plant organs. We studied the function of waxes for the establishment of the cuticular barrier in barley (Hordeum vulgare). The barley eceriferum mutants cer-za.227 and cer-ye.267 display reduced wax loads, but the genes affected, and the consequences of the wax changes for the barrier function remained unknown. Cuticular waxes and permeabilities were measured in cer-za.227 and cer-ye.267. The mutant loci were isolated by bulked segregant RNA sequencing. New cer-za alleles were generated by genome editing. The CER-ZA protein was characterized after expression in yeast and Arabidopsis cer4-3. Cer-za.227 carries a mutation in HORVU5Hr1G089230 encoding acyl-CoA reductase (FAR1). The cer-ye.267 mutation is located to HORVU4Hr1G063420 encoding β-ketoacyl-CoA synthase (KAS1) and is allelic to cer-zh.54. The amounts of intracuticular waxes were strongly decreased in cer-ye.267. The cuticular water loss and permeability of cer-za.227 were similar to wild-type (WT), but were increased in cer-ye.267. Removal of epicuticular waxes revealed that intracuticular, but not epicuticular waxes are required to regulate cuticular transpiration. The differential decrease in intracuticular waxes between cer-za.227 and cer-ye.267, and the removal of epicuticular waxes indicate that the cuticular barrier function mostly depends on the presence of intracuticular waxes.

KW - barley

KW - cuticle

KW - cutin

KW - eceriferum

KW - permeability

KW - wax

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DO - 10.1111/nph.19063

M3 - Article (Academic Journal)

C2 - 37349864

AN - SCOPUS:85162874283

SN - 0028-646X

VL - 239

SP - 1903

EP - 1918

JO - New Phytologist

JF - New Phytologist

IS - 5

ER -

Isolation and characterization of the gene HvFAR1 encoding acyl-CoA reductase from the cer-za.227 mutant of barley (Hordeum vulgare) and analysis of the cuticular barrier functions

Abstract

Bibliographical note

Keywords

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