Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.)
Cuticular waxes comprise the hydrophobic layer that protects crops against nonstomatal water loss and biotic and abiotic stresses. Expanding on our current knowledge of the genes that are involved in cuticular wax biosynthesis and regulation plays an important role in dissecting the processes of cut...
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MDPI AG
2022-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/21/13025 |
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| author | Xiuxiu Bian Lirong Yao Erjing Si Yaxiong Meng Baochun Li Xiaole Ma Ke Yang Yong Lai Xunwu Shang Chengdao Li Juncheng Wang Huajun Wang |
| author_facet | Xiuxiu Bian Lirong Yao Erjing Si Yaxiong Meng Baochun Li Xiaole Ma Ke Yang Yong Lai Xunwu Shang Chengdao Li Juncheng Wang Huajun Wang |
| author_sort | Xiuxiu Bian |
| collection | DOAJ |
| description | Cuticular waxes comprise the hydrophobic layer that protects crops against nonstomatal water loss and biotic and abiotic stresses. Expanding on our current knowledge of the genes that are involved in cuticular wax biosynthesis and regulation plays an important role in dissecting the processes of cuticular wax metabolism. In this study, we identified the Cer-GN1 barley (<i>Hordeum vulgare</i> L.) mutant that is generated by ethyl methanesulfonate mutagenesis with a glossy spike phenotype that is controlled by a single recessive nuclear gene. A physiological analysis showed that the total cuticular wax loads of Cer-GN1 were one-third that of the progenitor wild-type (WT), and its water loss rate was significantly accelerated (<i>p</i> < 0.05). In addition, Cer-GN1 was defective in the glume’s cuticle according to the toluidine blue dye test, and it was deficient in the tubule-shaped crystals which were observed on the glume surfaces by scanning electron microscopy. Using metabolomics and transcriptomics, we investigated the impacts of cuticular wax composition and waxy regulatory genes on the loss of the glaucous wax in the spikes of Cer-GN1. Among the differential metabolites, we found that 16-hydroxyhexadecanoic acid, which is one of the predominant C16 and C18 fatty acid-derived cutin monomers, was significantly downregulated in Cer-GN1 when it was compared to that of WT. We identified two novel genes that are located on chromosome 4H and are downregulated in Cer-GN1 (<i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i>) that encode long-chain fatty acid omega-monooxygenase CYP704B1, which regulates the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid. A quantitative real-time PCR revealed that the expression levels of <i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i> were downregulated at 1, 4, 8, 12, and 16 days after the heading stage in Cer-GN1 when it was compared to those of WT. These results suggested that <i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i> have CYP704B1 activity, which could regulate the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid in barley. Their downregulation in Cer-GN1 reduced the synthesis of the cuticular wax components and ultimately caused the loss of the glaucous wax in the spikes. It is necessary to verify whether <i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i> truly encode a CYP704B1 that regulates the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid in barley. |
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| issn | 1661-6596 1422-0067 |
| language | English |
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| spelling | doaj.art-21357989e4a1474ba25c145cab765de62023-11-24T05:01:09ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-10-0123211302510.3390/ijms232113025Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.)Xiuxiu Bian0Lirong Yao1Erjing Si2Yaxiong Meng3Baochun Li4Xiaole Ma5Ke Yang6Yong Lai7Xunwu Shang8Chengdao Li9Juncheng Wang10Huajun Wang11College of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaState Key Lab of Aridland Crop Science/Gansu Key Lab of Crop Improvement and Germplasm Enhancement, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Forestry, Henan Agricultural University, Zhengzhou 450002, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaWestern Crop Genetic Alliance, College of Science, Health, Engineering and Education, Murdoch University, 90 South Street, Murdoch, WA 6150, AustraliaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Agronomy, Gansu Agricultural University, Lanzhou 730070, ChinaCuticular waxes comprise the hydrophobic layer that protects crops against nonstomatal water loss and biotic and abiotic stresses. Expanding on our current knowledge of the genes that are involved in cuticular wax biosynthesis and regulation plays an important role in dissecting the processes of cuticular wax metabolism. In this study, we identified the Cer-GN1 barley (<i>Hordeum vulgare</i> L.) mutant that is generated by ethyl methanesulfonate mutagenesis with a glossy spike phenotype that is controlled by a single recessive nuclear gene. A physiological analysis showed that the total cuticular wax loads of Cer-GN1 were one-third that of the progenitor wild-type (WT), and its water loss rate was significantly accelerated (<i>p</i> < 0.05). In addition, Cer-GN1 was defective in the glume’s cuticle according to the toluidine blue dye test, and it was deficient in the tubule-shaped crystals which were observed on the glume surfaces by scanning electron microscopy. Using metabolomics and transcriptomics, we investigated the impacts of cuticular wax composition and waxy regulatory genes on the loss of the glaucous wax in the spikes of Cer-GN1. Among the differential metabolites, we found that 16-hydroxyhexadecanoic acid, which is one of the predominant C16 and C18 fatty acid-derived cutin monomers, was significantly downregulated in Cer-GN1 when it was compared to that of WT. We identified two novel genes that are located on chromosome 4H and are downregulated in Cer-GN1 (<i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i>) that encode long-chain fatty acid omega-monooxygenase CYP704B1, which regulates the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid. A quantitative real-time PCR revealed that the expression levels of <i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i> were downregulated at 1, 4, 8, 12, and 16 days after the heading stage in Cer-GN1 when it was compared to those of WT. These results suggested that <i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i> have CYP704B1 activity, which could regulate the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid in barley. Their downregulation in Cer-GN1 reduced the synthesis of the cuticular wax components and ultimately caused the loss of the glaucous wax in the spikes. It is necessary to verify whether <i>HvMSTRG.29184</i> and <i>HvMSTRG.29185</i> truly encode a CYP704B1 that regulates the conversion of C16 palmitic acid to 16-hydroxyhexadecanoic acid in barley.https://www.mdpi.com/1422-0067/23/21/13025cuticular waxesbarleygene16-hydroxyhexadecanoic acidsynthesis |
| spellingShingle | Xiuxiu Bian Lirong Yao Erjing Si Yaxiong Meng Baochun Li Xiaole Ma Ke Yang Yong Lai Xunwu Shang Chengdao Li Juncheng Wang Huajun Wang Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.) International Journal of Molecular Sciences cuticular waxes barley gene 16-hydroxyhexadecanoic acid synthesis |
| title | Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.) |
| title_full | Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.) |
| title_fullStr | Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.) |
| title_full_unstemmed | Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.) |
| title_short | Characterization of Glossy Spike Mutants and Identification of Candidate Genes Regulating Cuticular Wax Synthesis in Barley (<i>Hordeum vulgare</i> L.) |
| title_sort | characterization of glossy spike mutants and identification of candidate genes regulating cuticular wax synthesis in barley i hordeum vulgare i l |
| topic | cuticular waxes barley gene 16-hydroxyhexadecanoic acid synthesis |
| url | https://www.mdpi.com/1422-0067/23/21/13025 |
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