The Genomic Loci Cer-za and Cer-ye Contribute to Cuticular Wax Biosynthesis in Hordeum vulgare
The Genomic Loci Cer-za and Cer-ye Contribute to Cuticular Wax Biosynthesis in Hordeum vulgare
Dokument öffnen (7.3MB)Art der Hochschulschrift
DissertationPrüfungsdatum
23.06.2022Datum der Veröffentlichung
28.06.2022Erstgutachter
Dörmann, PeterZweitgutachter
Schreiber, LukasMetadaten
Zur Langanzeige
Zitierbare Links 
Inhalt
Changing environmental conditions demand the expansion of the genetic resources to enable the rapid adaption and development of novel crop species. The cuticle as direct interface between plants and their environment is thereby of a special interest. This study aimed to contribute to the knowledge about the genetic background of the cuticular wax biosynthesis in barley (Hordeum vulgare) with the characterisation of the two wax-deficient eceriferum mutants cer-za.227 and cer-ye.267. Both lines showed reduced water-repellence properties due to the reduction of epicuticular wax crystals on the leaf surfaces. Biochemical analysis showed strong reductions of the cuticular wax load on leaves of both eceriferum lines. The cer-za.227 mutant was particularly affected in primary alcohols and esters while in cer-ye.267, the amounts of all cuticular wax substances were reduced. A bulked segregant RNA-sequencing approach led to the identification of the two genes HORVU5Hr1G089230 and HORVU4Hr1G063420 which carry mutations in the lines cer-za.227 and cer-ye.267, respectively. Consideration of additional allelic cer-za and cer-ye lines revealed that these allelic mutants carry mutations in the same two genes. The gene product of HORVU5Hr1G089230, CER-ZA, was confirmed to harbour fatty acyl-CoA reductase (FAR) activity. Its heterologous expression in E. coli, yeast and the wax-deficient A. thaliana cer4-3 mutant resulted in the accumulation of primary alcohols. CER-ZA is the first protein of the FAR family that has been characterised in H. vulgare. Accordingly, CER-ZA was named HvFAR1. The CER-ZA enzyme catalyses the reduction of very long chain acyl-CoAs to primary alcohols and contributes crucially to the biosynthesis of cuticular waxes in barley. The protein was localised to the ER. The cer-ye.267 mutant carries a mutation in the gene HORVU4Hr1G063420. This gene codes for a protein with sequence similarity to for a β-ketoacyl-CoA synthase (KCS). KCS enzymes generate very long chain fatty acids as precursors for the synthesis of different wax lipids. The same gene has previously been described as Cer-zh. Within this study, it was confirmed that cer-ye.267 and cer-zh.54 are allelic mutants. The protein affected in cer-ye.267 and cer-zh.54 was named HvKCS1 and it catalyses the elongation of C16 and C18 acyl-CoAs to very long-chain fatty acids as part of a fatty acid elongation complex. Comparisons of the barrier properties of cer-za.227 and cer-ye.267 with Bowman indicated that the dominant compounds in barley, i.e. primary alcohols, do not contribute to the formation of a functional water-loss and penetration barrier, but that this barrier function rather depends on the ratio of polar and non-polar compounds.Changing environmental conditions demand the expansion of the genetic resources to enable the rapid adaption and development of novel crop species. The cuticle as direct interface between plants and their environment is thereby of a special interest. This study aimed to contribute to the knowledge about the genetic background of the cuticular wax biosynthesis in barley (Hordeum vulgare) with the characterisation of the two wax-deficient eceriferum mutants cer-za.227 and cer-ye.267. Both lines showed reduced water-repellence properties due to the reduction of epicuticular wax crystals on the leaf surfaces. Biochemical analysis showed strong reductions of the cuticular wax load on leaves of both eceriferum lines. The cer-za.227 mutant was particularly affected in primary alcohols and esters while in cer-ye.267, the amounts of all cuticular wax substances were reduced. A bulked segregant RNA-sequencing approach led to the identification of the two genes HORVU5Hr1G089230 and HORVU4Hr1G063420 which carry mutations in the lines cer-za.227 and cer-ye.267, respectively. Consideration of additional allelic cer-za and cer-ye lines revealed that these allelic mutants carry mutations in the same two genes. The gene product of HORVU5Hr1G089230, CER-ZA, was confirmed to harbour fatty acyl-CoA reductase (FAR) activity. Its heterologous expression in E. coli, yeast and the wax-deficient A. thaliana cer4-3 mutant resulted in the accumulation of primary alcohols. CER-ZA is the first protein of the FAR family that has been characterised in H. vulgare. Accordingly, CER-ZA was named HvFAR1. The CER-ZA enzyme catalyses the reduction of very long chain acyl-CoAs to primary alcohols and contributes crucially to the biosynthesis of cuticular waxes in barley. The protein was localised to the ER. The cer-ye.267 mutant carries a mutation in the gene HORVU4Hr1G063420. This gene codes for a protein with sequence similarity to for a β-ketoacyl-CoA synthase (KCS). KCS enzymes generate very long chain fatty acids as precursors for the synthesis of different wax lipids. The same gene has previously been described as Cer-zh. Within this study, it was confirmed that cer-ye.267 and cer-zh.54 are allelic mutants. The protein affected in cer-ye.267 and cer-zh.54 was named HvKCS1 and it catalyses the elongation of C16 and C18 acyl-CoAs to very long-chain fatty acids as part of a fatty acid elongation complex. Comparisons of the barrier properties of cer-za.227 and cer-ye.267 with Bowman indicated that the dominant compounds in barley, i.e. primary alcohols, do not contribute to the formation of a functional water-loss and penetration barrier, but that this barrier function rather depends on the ratio of polar and non-polar compounds.
Schlagwörter
Kutikel, Wachse, Apoplastische Barriere, Cuticle, cuticular waxes, apoplastic barrier
Klassifikation (DDC)
580 Pflanzen (Botanik)Müller, Yannic: The Genomic Loci Cer-za and Cer-ye Contribute to Cuticular Wax Biosynthesis in Hordeum vulgare. - Bonn, 2022. - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn.
Online-Ausgabe in bonndoc: https://nbn-resolving.org/urn:nbn:de:hbz:5-67127
Online-Ausgabe in bonndoc: https://nbn-resolving.org/urn:nbn:de:hbz:5-67127
@phdthesis{handle:20.500.11811/9980,
urn: https://nbn-resolving.org/urn:nbn:de:hbz:5-67127,
author = {{Yannic Müller}},
title = {The Genomic Loci Cer-za and Cer-ye Contribute to Cuticular Wax Biosynthesis in Hordeum vulgare},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2022,
month = jun,
note = {Changing environmental conditions demand the expansion of the genetic resources to enable the rapid adaption and development of novel crop species. The cuticle as direct interface between plants and their environment is thereby of a special interest. This study aimed to contribute to the knowledge about the genetic background of the cuticular wax biosynthesis in barley (Hordeum vulgare) with the characterisation of the two wax-deficient eceriferum mutants cer-za.227 and cer-ye.267. Both lines showed reduced water-repellence properties due to the reduction of epicuticular wax crystals on the leaf surfaces. Biochemical analysis showed strong reductions of the cuticular wax load on leaves of both eceriferum lines. The cer-za.227 mutant was particularly affected in primary alcohols and esters while in cer-ye.267, the amounts of all cuticular wax substances were reduced. A bulked segregant RNA-sequencing approach led to the identification of the two genes HORVU5Hr1G089230 and HORVU4Hr1G063420 which carry mutations in the lines cer-za.227 and cer-ye.267, respectively. Consideration of additional allelic cer-za and cer-ye lines revealed that these allelic mutants carry mutations in the same two genes. The gene product of HORVU5Hr1G089230, CER-ZA, was confirmed to harbour fatty acyl-CoA reductase (FAR) activity. Its heterologous expression in E. coli, yeast and the wax-deficient A. thaliana cer4-3 mutant resulted in the accumulation of primary alcohols. CER-ZA is the first protein of the FAR family that has been characterised in H. vulgare. Accordingly, CER-ZA was named HvFAR1. The CER-ZA enzyme catalyses the reduction of very long chain acyl-CoAs to primary alcohols and contributes crucially to the biosynthesis of cuticular waxes in barley. The protein was localised to the ER. The cer-ye.267 mutant carries a mutation in the gene HORVU4Hr1G063420. This gene codes for a protein with sequence similarity to for a β-ketoacyl-CoA synthase (KCS). KCS enzymes generate very long chain fatty acids as precursors for the synthesis of different wax lipids. The same gene has previously been described as Cer-zh. Within this study, it was confirmed that cer-ye.267 and cer-zh.54 are allelic mutants. The protein affected in cer-ye.267 and cer-zh.54 was named HvKCS1 and it catalyses the elongation of C16 and C18 acyl-CoAs to very long-chain fatty acids as part of a fatty acid elongation complex. Comparisons of the barrier properties of cer-za.227 and cer-ye.267 with Bowman indicated that the dominant compounds in barley, i.e. primary alcohols, do not contribute to the formation of a functional water-loss and penetration barrier, but that this barrier function rather depends on the ratio of polar and non-polar compounds.Changing environmental conditions demand the expansion of the genetic resources to enable the rapid adaption and development of novel crop species. The cuticle as direct interface between plants and their environment is thereby of a special interest. This study aimed to contribute to the knowledge about the genetic background of the cuticular wax biosynthesis in barley (Hordeum vulgare) with the characterisation of the two wax-deficient eceriferum mutants cer-za.227 and cer-ye.267. Both lines showed reduced water-repellence properties due to the reduction of epicuticular wax crystals on the leaf surfaces. Biochemical analysis showed strong reductions of the cuticular wax load on leaves of both eceriferum lines. The cer-za.227 mutant was particularly affected in primary alcohols and esters while in cer-ye.267, the amounts of all cuticular wax substances were reduced. A bulked segregant RNA-sequencing approach led to the identification of the two genes HORVU5Hr1G089230 and HORVU4Hr1G063420 which carry mutations in the lines cer-za.227 and cer-ye.267, respectively. Consideration of additional allelic cer-za and cer-ye lines revealed that these allelic mutants carry mutations in the same two genes. The gene product of HORVU5Hr1G089230, CER-ZA, was confirmed to harbour fatty acyl-CoA reductase (FAR) activity. Its heterologous expression in E. coli, yeast and the wax-deficient A. thaliana cer4-3 mutant resulted in the accumulation of primary alcohols. CER-ZA is the first protein of the FAR family that has been characterised in H. vulgare. Accordingly, CER-ZA was named HvFAR1. The CER-ZA enzyme catalyses the reduction of very long chain acyl-CoAs to primary alcohols and contributes crucially to the biosynthesis of cuticular waxes in barley. The protein was localised to the ER. The cer-ye.267 mutant carries a mutation in the gene HORVU4Hr1G063420. This gene codes for a protein with sequence similarity to for a β-ketoacyl-CoA synthase (KCS). KCS enzymes generate very long chain fatty acids as precursors for the synthesis of different wax lipids. The same gene has previously been described as Cer-zh. Within this study, it was confirmed that cer-ye.267 and cer-zh.54 are allelic mutants. The protein affected in cer-ye.267 and cer-zh.54 was named HvKCS1 and it catalyses the elongation of C16 and C18 acyl-CoAs to very long-chain fatty acids as part of a fatty acid elongation complex. Comparisons of the barrier properties of cer-za.227 and cer-ye.267 with Bowman indicated that the dominant compounds in barley, i.e. primary alcohols, do not contribute to the formation of a functional water-loss and penetration barrier, but that this barrier function rather depends on the ratio of polar and non-polar compounds.},
url = {https://hdl.handle.net/20.500.11811/9980}
}
urn: https://nbn-resolving.org/urn:nbn:de:hbz:5-67127,
author = {{Yannic Müller}},
title = {The Genomic Loci Cer-za and Cer-ye Contribute to Cuticular Wax Biosynthesis in Hordeum vulgare},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2022,
month = jun,
note = {Changing environmental conditions demand the expansion of the genetic resources to enable the rapid adaption and development of novel crop species. The cuticle as direct interface between plants and their environment is thereby of a special interest. This study aimed to contribute to the knowledge about the genetic background of the cuticular wax biosynthesis in barley (Hordeum vulgare) with the characterisation of the two wax-deficient eceriferum mutants cer-za.227 and cer-ye.267. Both lines showed reduced water-repellence properties due to the reduction of epicuticular wax crystals on the leaf surfaces. Biochemical analysis showed strong reductions of the cuticular wax load on leaves of both eceriferum lines. The cer-za.227 mutant was particularly affected in primary alcohols and esters while in cer-ye.267, the amounts of all cuticular wax substances were reduced. A bulked segregant RNA-sequencing approach led to the identification of the two genes HORVU5Hr1G089230 and HORVU4Hr1G063420 which carry mutations in the lines cer-za.227 and cer-ye.267, respectively. Consideration of additional allelic cer-za and cer-ye lines revealed that these allelic mutants carry mutations in the same two genes. The gene product of HORVU5Hr1G089230, CER-ZA, was confirmed to harbour fatty acyl-CoA reductase (FAR) activity. Its heterologous expression in E. coli, yeast and the wax-deficient A. thaliana cer4-3 mutant resulted in the accumulation of primary alcohols. CER-ZA is the first protein of the FAR family that has been characterised in H. vulgare. Accordingly, CER-ZA was named HvFAR1. The CER-ZA enzyme catalyses the reduction of very long chain acyl-CoAs to primary alcohols and contributes crucially to the biosynthesis of cuticular waxes in barley. The protein was localised to the ER. The cer-ye.267 mutant carries a mutation in the gene HORVU4Hr1G063420. This gene codes for a protein with sequence similarity to for a β-ketoacyl-CoA synthase (KCS). KCS enzymes generate very long chain fatty acids as precursors for the synthesis of different wax lipids. The same gene has previously been described as Cer-zh. Within this study, it was confirmed that cer-ye.267 and cer-zh.54 are allelic mutants. The protein affected in cer-ye.267 and cer-zh.54 was named HvKCS1 and it catalyses the elongation of C16 and C18 acyl-CoAs to very long-chain fatty acids as part of a fatty acid elongation complex. Comparisons of the barrier properties of cer-za.227 and cer-ye.267 with Bowman indicated that the dominant compounds in barley, i.e. primary alcohols, do not contribute to the formation of a functional water-loss and penetration barrier, but that this barrier function rather depends on the ratio of polar and non-polar compounds.Changing environmental conditions demand the expansion of the genetic resources to enable the rapid adaption and development of novel crop species. The cuticle as direct interface between plants and their environment is thereby of a special interest. This study aimed to contribute to the knowledge about the genetic background of the cuticular wax biosynthesis in barley (Hordeum vulgare) with the characterisation of the two wax-deficient eceriferum mutants cer-za.227 and cer-ye.267. Both lines showed reduced water-repellence properties due to the reduction of epicuticular wax crystals on the leaf surfaces. Biochemical analysis showed strong reductions of the cuticular wax load on leaves of both eceriferum lines. The cer-za.227 mutant was particularly affected in primary alcohols and esters while in cer-ye.267, the amounts of all cuticular wax substances were reduced. A bulked segregant RNA-sequencing approach led to the identification of the two genes HORVU5Hr1G089230 and HORVU4Hr1G063420 which carry mutations in the lines cer-za.227 and cer-ye.267, respectively. Consideration of additional allelic cer-za and cer-ye lines revealed that these allelic mutants carry mutations in the same two genes. The gene product of HORVU5Hr1G089230, CER-ZA, was confirmed to harbour fatty acyl-CoA reductase (FAR) activity. Its heterologous expression in E. coli, yeast and the wax-deficient A. thaliana cer4-3 mutant resulted in the accumulation of primary alcohols. CER-ZA is the first protein of the FAR family that has been characterised in H. vulgare. Accordingly, CER-ZA was named HvFAR1. The CER-ZA enzyme catalyses the reduction of very long chain acyl-CoAs to primary alcohols and contributes crucially to the biosynthesis of cuticular waxes in barley. The protein was localised to the ER. The cer-ye.267 mutant carries a mutation in the gene HORVU4Hr1G063420. This gene codes for a protein with sequence similarity to for a β-ketoacyl-CoA synthase (KCS). KCS enzymes generate very long chain fatty acids as precursors for the synthesis of different wax lipids. The same gene has previously been described as Cer-zh. Within this study, it was confirmed that cer-ye.267 and cer-zh.54 are allelic mutants. The protein affected in cer-ye.267 and cer-zh.54 was named HvKCS1 and it catalyses the elongation of C16 and C18 acyl-CoAs to very long-chain fatty acids as part of a fatty acid elongation complex. Comparisons of the barrier properties of cer-za.227 and cer-ye.267 with Bowman indicated that the dominant compounds in barley, i.e. primary alcohols, do not contribute to the formation of a functional water-loss and penetration barrier, but that this barrier function rather depends on the ratio of polar and non-polar compounds.},
url = {https://hdl.handle.net/20.500.11811/9980}
}
Die folgenden Nutzungsbestimmungen sind mit dieser Ressource verbunden:
