Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae
Floral nectar plays important roles in the interaction between animal-pollinated plants and pollinators. Its components include water, sugars, amino acids, vitamins, and proteins. Growing empirical evidence shows that most of the proteins secreted in nectar (nectarines) are enzymes that can tailor n...
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Frontiers Media S.A.
2018-07-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fpls.2018.01109/full |
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| author | Hong-Xia Zhou Richard I. Milne Xue-Long Ma Yue-Qin Song Jian-Yu Fang Hang Sun Hong-Guang Zha |
| author_facet | Hong-Xia Zhou Richard I. Milne Xue-Long Ma Yue-Qin Song Jian-Yu Fang Hang Sun Hong-Guang Zha |
| author_sort | Hong-Xia Zhou |
| collection | DOAJ |
| description | Floral nectar plays important roles in the interaction between animal-pollinated plants and pollinators. Its components include water, sugars, amino acids, vitamins, and proteins. Growing empirical evidence shows that most of the proteins secreted in nectar (nectarines) are enzymes that can tailor nectar chemistry for their animal mutualists or reduce the growth of microorganisms in nectar. However, to date, the function of many nectarines remains unknown, and very few plant species have had their nectar proteome thoroughly investigated. Mucuna sempervirens (Fabaceae) is a perennial woody vine native to China. Nectarines from this species were separated using two-dimensional gel electrophoresis, and analyzed using mass spectrometry. A L-gulonolactone oxidase like protein (MsGulLO) was detected, and the full length cDNA was cloned: it codes for a protein of 573 amino acids with a predicted signal peptide. MsGulLO has high similarity to L-gulonolactone oxidase 5 (AtGulLO5) in Arabidopsis thaliana, which was suggested to be involved in the pathway of ascorbate biosynthesis; however, both MsGulLO and AtGulLO5 are divergent from animal L-gulonolactone oxidases. MsGulLO was expressed mainly in flowers, and especially in nectary before blooming. However, cloning and gene expression analysis showed that L-galactonolactone dehydrogenase (MsGLDH), a vital enzyme in plant ascorbate biosynthesis, was expressed in all of flowers, roots, stems, and especially leaves. MsGulLO was purified to near homogeneity from raw MS nectar by gel filtration chromatography. The enzyme was determined to be a neutral monomeric protein with an apparent molecular mass of 70 kDa. MsGulLO is not a flavin-containing protein, and has neither L-galactonolactone dehydrogenase activity, nor the L-gulonolactone activity that is usual in animal GulLOs. However, it has weak oxidase activity with the following substrates: L-gulono-1,4-lactone, L -galactono-1,4-lactone, D-gluconic acid-δ-lactone, glucose, and fructose. MsGulLO is suggested to function in hydrogen peroxide generation in nectar but not in plant ascorbate biosynthesis. |
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| format | Article |
| id | doaj.art-2d45095418b04a1eaf744dca3fe27fb1 |
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| last_indexed | 2024-12-14T01:16:50Z |
| publishDate | 2018-07-01 |
| publisher | Frontiers Media S.A. |
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| spelling | doaj.art-2d45095418b04a1eaf744dca3fe27fb12022-12-21T23:22:33ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2018-07-01910.3389/fpls.2018.01109379222Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, FabaceaeHong-Xia Zhou0Richard I. Milne1Xue-Long Ma2Yue-Qin Song3Jian-Yu Fang4Hang Sun5Hong-Guang Zha6College of Life and Environment Sciences, Huangshan University, Huangshan, ChinaInstitute of Molecular Plant Sciences, University of Edinburgh, Edinburgh, United KingdomCollege of Life and Environment Sciences, Huangshan University, Huangshan, ChinaCollege of Life and Environment Sciences, Huangshan University, Huangshan, ChinaCollege of Life and Environment Sciences, Huangshan University, Huangshan, ChinaKey Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, ChinaCollege of Life and Environment Sciences, Huangshan University, Huangshan, ChinaFloral nectar plays important roles in the interaction between animal-pollinated plants and pollinators. Its components include water, sugars, amino acids, vitamins, and proteins. Growing empirical evidence shows that most of the proteins secreted in nectar (nectarines) are enzymes that can tailor nectar chemistry for their animal mutualists or reduce the growth of microorganisms in nectar. However, to date, the function of many nectarines remains unknown, and very few plant species have had their nectar proteome thoroughly investigated. Mucuna sempervirens (Fabaceae) is a perennial woody vine native to China. Nectarines from this species were separated using two-dimensional gel electrophoresis, and analyzed using mass spectrometry. A L-gulonolactone oxidase like protein (MsGulLO) was detected, and the full length cDNA was cloned: it codes for a protein of 573 amino acids with a predicted signal peptide. MsGulLO has high similarity to L-gulonolactone oxidase 5 (AtGulLO5) in Arabidopsis thaliana, which was suggested to be involved in the pathway of ascorbate biosynthesis; however, both MsGulLO and AtGulLO5 are divergent from animal L-gulonolactone oxidases. MsGulLO was expressed mainly in flowers, and especially in nectary before blooming. However, cloning and gene expression analysis showed that L-galactonolactone dehydrogenase (MsGLDH), a vital enzyme in plant ascorbate biosynthesis, was expressed in all of flowers, roots, stems, and especially leaves. MsGulLO was purified to near homogeneity from raw MS nectar by gel filtration chromatography. The enzyme was determined to be a neutral monomeric protein with an apparent molecular mass of 70 kDa. MsGulLO is not a flavin-containing protein, and has neither L-galactonolactone dehydrogenase activity, nor the L-gulonolactone activity that is usual in animal GulLOs. However, it has weak oxidase activity with the following substrates: L-gulono-1,4-lactone, L -galactono-1,4-lactone, D-gluconic acid-δ-lactone, glucose, and fructose. MsGulLO is suggested to function in hydrogen peroxide generation in nectar but not in plant ascorbate biosynthesis.https://www.frontiersin.org/article/10.3389/fpls.2018.01109/fullL-ascorbate biosynthesisFloral nectarL-gulonolactone oxidase like proteinL-galactonolactone dehydrogenaseMucuna sempervirens HemslNectarin |
| spellingShingle | Hong-Xia Zhou Richard I. Milne Xue-Long Ma Yue-Qin Song Jian-Yu Fang Hang Sun Hong-Guang Zha Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae Frontiers in Plant Science L-ascorbate biosynthesis Floral nectar L-gulonolactone oxidase like protein L-galactonolactone dehydrogenase Mucuna sempervirens Hemsl Nectarin |
| title | Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae |
| title_full | Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae |
| title_fullStr | Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae |
| title_full_unstemmed | Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae |
| title_short | Characterization of a L-Gulono-1,4-Lactone Oxidase Like Protein in the Floral Nectar of Mucuna sempervirens, Fabaceae |
| title_sort | characterization of a l gulono 1 4 lactone oxidase like protein in the floral nectar of mucuna sempervirens fabaceae |
| topic | L-ascorbate biosynthesis Floral nectar L-gulonolactone oxidase like protein L-galactonolactone dehydrogenase Mucuna sempervirens Hemsl Nectarin |
| url | https://www.frontiersin.org/article/10.3389/fpls.2018.01109/full |
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