Identification and functional analysis of sterol C-24 methyltransferase in rice

ZHI Boyang; LEI Long; HUANG Sishu; LIU Xianqing; LUO Jie

doi:10.15886/j.cnki.rdswxb.20240070

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Volume 16 Issue 1

Mar. 2025

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Article Navigation > Journal of Tropical Biology > 2025 > 16(1): 1-12

ZHI Boyang, LEI Long, HUANG Sishu, LIU Xianqing, LUO Jie. Identification and functional analysis of sterol C-24 methyltransferase in rice[J]. Journal of Tropical Biology, 2025, 16(1): 1-12. doi: 10.15886/j.cnki.rdswxb.20240070

Citation:

ZHI Boyang, LEI Long, HUANG Sishu, LIU Xianqing, LUO Jie. Identification and functional analysis of sterol C-24 methyltransferase in rice[J]. Journal of Tropical Biology, 2025, 16(1): 1-12. doi: 10.15886/j.cnki.rdswxb.20240070

Identification and functional analysis of sterol C-24 methyltransferase in rice

doi: 10.15886/j.cnki.rdswxb.20240070

1. School of Tropical Agriculture and Forestry, Hainan University, Danzhou, Hainan 571737, China;
2. College of Nanfan, Hainan University, Sanya, Hainan 57202, China;
3. Yazhou Bay National Laboratory, Sanya, Hainan 572024, China

Received Date: 2024-04-20
Rev Recd Date: 2024-05-11
Publish Date: 2025-03-15

Abstract

Phytosterols are a class of conserved natural compounds in plants that play a role in the growth and development of plants. They also have pharmacological functions such as lowering cholesterol and preventing cardiovascular diseases, which are of significant value to human health. Currently, the genes related to phytosterol synthesis in rice have not been well elucidated, which has hindered the development of high-phytosterol rice varieties. To further explore the genes associated with phytosterol synthesis in rice, the contents of three major phytosterols（campesterol, stigmasterol, and β-sitosterol） in 533 rice varieties were detected accurately by using liquid chromatography-tandem mass spectrometry. Based on the findings, the OsSMT1 gene in rice was located and cloned by using metabolomic genome-wide association analysis. Further in vitro recombinant protein enzyme activity assays and tobacco transient expression experiments confirmed that the protein encoded by this gene is capable of catalyzing the synthesis of the key intermediate 24-methylenecycloartanol in the biosynthesis of phytosterols.
- rice,
- phytosterols,
- sterol C-24 methyltransferase,
- metabolome Genome-Wide Association Study

References

[1]	HU Z, HE B, MA L, et al. Recent advances in ergosterol biosynthesis and regulation mechanisms in Saccharomyces cerevisiae[J]. Indian Journal of Microbiology, 2017, 57(3):270-277.
[2]	VALITOVA Y N, KOTLOVA E R, NOVIKOV A V, et al.Binding of sterols affects membrane functioning and sphingolipid composition in wheat roots[J]. Biochemistry(Moscow), 2010, 75:554-561.
[3]	WANG H, NAGEGOWDA D A, RAWAT R, et al. Overexpression of Brassica juncea wild-type and mutant HMGCoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance[J]. Plant Biotechnology Journal, 2012, 10(1):31-42.
[4]	YANG Z, XIA J, HONG J, et al. Structural insights into auxin recognition and efflux by Arabidopsis PIN1[J]. Nature, 2022, 609:611-615.
[5]	SCHRICK K, FUJIOKA S, TAKATSUTO S, et al. A link between sterol biosynthesis, the cell wall, and cellulose in Arabidopsis[J]. The Plant Journal:for Cell and Molecular Biology, 2004, 38(2):227-243.
[6]	PEREIRA A M, COIMBRA S. Advances in plant reproduction:from gametes to seeds[J]. Journal of Experimental Botany, 2019, 70(11):2933-2936.
[7]	PALTA J P, WHITAKER B D, WEISS L S. Plasma membrane lipids associated with genetic variability in freezing tolerance and cold acclimation of Solanum species[J]. Plant Physiology, 1993, 103(3):793-803.
[8]	BLITS K C, GALLAGHER J L. Effect of NaCl on lipid content of plasma membranes isolated from roots and cell suspension cultures of the dicot halophyte Kosteletzkya virginica(L.) Presl[J]. Plant Cell Reports, 1990, 9(3):156-159.
[9]	TAKSHAK S, AGRAWAL S B. Alterations in metabolite profile and free radical scavenging activities of Withania somnifera leaf and root extracts under supplemental ultraviolet-B radiation[J]. Acta Physiologiae Plantarum,2015, 37(12):260-271.
[10]	KUMAR M S S, MAWLONG I, ALI K, et al. Regulation of phytosterol biosynthetic pathway during drought stress in rice[J]. Plant Physiology and Biochemistry:PPB, 2018,129:11-20.
[11]	ELKEILSH A, AWAD Y M, SOLIMAN M H, et al. Exogenous application of β-sitosterol mediated growth and yield improvement in water-stressed wheat(Triticum aestivum) involves up-regulated antioxidant system[J]. Journal of Plant Research, 2019, 132(6):881-901.
[12]	VEZZA T, CANET F, DE MARAÑÓN A M, et al. Phytosterols:nutritional health players in the management of obesity and its related disorders[J]. Antioxidants, 2020,9(12):1266.
[13]	CABRAL C E, KLEIN M R S T. Phytosterols in the treatment of hypercholesterolemia and prevention of cardiovascular diseases[J]. Arquivos Brasileiros De Cardiologia, 2017, 109(5):475-482.
[14]	JIANG L, ZHAO X, XU J, et al. The protective effect of dietary phytosterols on cancer risk:a systematic metaanalysis[J]. Journal of Oncology, 2019(6):7479518.
[15]	CABRAL C E, KLEIN M R S T. Phytosterols in the treatment of hypercholesterolemia and prevention of cardiovascular diseases[J]. Arquivos Brasileiros De Cardiologia, 2017, 109(5):475-482.
[16]	余慧,徐宝成,王大红,等.植物甾醇氧化物的形成、摄入及健康相关效应[J]. 食品科学, 2021, 42(23):306-314.
[17]	邱振楠,徐乾坤,王小琦,等.水稻DNA多态标记的开发与验证[J]. 浙江农业学报, 2016, 28(3):361-370.
[18]	KATZEN F. Gateway^®recombinational cloning:a biological operating system[J]. Expert Opinion on Drug Discovery, 2007, 2(4):571-589.
[19]	SAHU P K, CHAKRADHARI S, SIPENIECE E, et al.Fatty acids, tocopherols, tocotrienols, phytosterols, carotenoids, and squalene in seed oils of Hyptis suaveolens,Leonotis nepetifolia, and Ocimum sanctum[J]. European Journal of Lipid Science and Technology, 2020, 122(7):2000053.
[20]	GARRIS A J, TAI T H, COBURN J, et al. Genetic structure and diversity in Oryza sativa L[J]. Genetics, 2005,169(3):1631-1638.
[21]	WANG J, LIU J, SONG Z, et al. Sterol C24-methyltransferase:mechanistic studies of the C-methylation reaction with 24-fluorocycloartenol[J]. Bioorganic&Medicinal Chemistry Letters, 2008, 18(1):232-235.
[22]	周千惠,刘俊霞,刘萍萍,等.不同CCDs基因表达介导藏红花素在烟草中的异源合成[J/OL]. 烟草科技,2024:1-13(2024-04 -19) . https://kns.cnki.net/kcms/detail/41.1137.TS.20240419.1449.002.html.
[23]	Schaller H. The role of sterols in plant growth and development[J]. Progress in lipid research, 2003, 42(3):163-175.
[24]	PAL S, RASTOGI S, NAGEGOWDA D A, et al. RNAi of sterol methyl Transferase1 reveals its direct role in diverting intermediates towards withanolide/phytosterol biosynthesis in Withania somnifera[J]. Plant and Cell Physiology, 2019, 60(3):672-686.
[25]	DIENER A C, LI H, ZHOU W X, et al. STEROL METHYLTRANSFERASE 1 controls the level of cholesterol in plants[J]. The Plant Cell, 2000, 12(6):853-870.
[26]	闫志强,邓茹月,宫彦龙,等.茉莉酸甲酯对籼粳稻农艺性状及品质性状的影响[J]. 杂交水稻, 2022, 37(4):107-114.
[27]	张杰,杨玺,李游山.蛋白酶及其抑制剂关键活性位点研究进展[J]. 生物工程学报, 2021, 37(2):561-579.
[28]	ELSHAHAWY I E, ABD EL-WAHED M S. Suppression of Cephalosporium maydis by the resistance inducer betasitosterol[J]. European Journal of Plant Pathology, 2022,163(3):673-693.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Proportional views

Identification and functional analysis of sterol C-24 methyltransferase in rice

1. School of Tropical Agriculture and Forestry, Hainan University, Danzhou, Hainan 571737, China;

2. College of Nanfan, Hainan University, Sanya, Hainan 57202, China;

3. Yazhou Bay National Laboratory, Sanya, Hainan 572024, China

Received Date: 2024-04-20

Rev Recd Date: 2024-05-11

Publish Date: 2025-03-15

Key words:

rice /
phytosterols /
sterol C-24 methyltransferase /
metabolome Genome-Wide Association Study

Abstract: Phytosterols are a class of conserved natural compounds in plants that play a role in the growth and development of plants. They also have pharmacological functions such as lowering cholesterol and preventing cardiovascular diseases, which are of significant value to human health. Currently, the genes related to phytosterol synthesis in rice have not been well elucidated, which has hindered the development of high-phytosterol rice varieties. To further explore the genes associated with phytosterol synthesis in rice, the contents of three major phytosterols（campesterol, stigmasterol, and β-sitosterol） in 533 rice varieties were detected accurately by using liquid chromatography-tandem mass spectrometry. Based on the findings, the OsSMT1 gene in rice was located and cloned by using metabolomic genome-wide association analysis. Further in vitro recombinant protein enzyme activity assays and tobacco transient expression experiments confirmed that the protein encoded by this gene is capable of catalyzing the synthesis of the key intermediate 24-methylenecycloartanol in the biosynthesis of phytosterols.

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Reference (28)

[1]	HU Z, HE B, MA L, et al. Recent advances in ergosterol biosynthesis and regulation mechanisms in Saccharomyces cerevisiae[J]. Indian Journal of Microbiology, 2017, 57(3):270-277.
[2]	VALITOVA Y N, KOTLOVA E R, NOVIKOV A V, et al.Binding of sterols affects membrane functioning and sphingolipid composition in wheat roots[J]. Biochemistry(Moscow), 2010, 75:554-561.
[3]	WANG H, NAGEGOWDA D A, RAWAT R, et al. Overexpression of Brassica juncea wild-type and mutant HMGCoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance[J]. Plant Biotechnology Journal, 2012, 10(1):31-42.
[4]	YANG Z, XIA J, HONG J, et al. Structural insights into auxin recognition and efflux by Arabidopsis PIN1[J]. Nature, 2022, 609:611-615.
[5]	SCHRICK K, FUJIOKA S, TAKATSUTO S, et al. A link between sterol biosynthesis, the cell wall, and cellulose in Arabidopsis[J]. The Plant Journal:for Cell and Molecular Biology, 2004, 38(2):227-243.
[6]	PEREIRA A M, COIMBRA S. Advances in plant reproduction:from gametes to seeds[J]. Journal of Experimental Botany, 2019, 70(11):2933-2936.
[7]	PALTA J P, WHITAKER B D, WEISS L S. Plasma membrane lipids associated with genetic variability in freezing tolerance and cold acclimation of Solanum species[J]. Plant Physiology, 1993, 103(3):793-803.
[8]	BLITS K C, GALLAGHER J L. Effect of NaCl on lipid content of plasma membranes isolated from roots and cell suspension cultures of the dicot halophyte Kosteletzkya virginica(L.) Presl[J]. Plant Cell Reports, 1990, 9(3):156-159.
[9]	TAKSHAK S, AGRAWAL S B. Alterations in metabolite profile and free radical scavenging activities of Withania somnifera leaf and root extracts under supplemental ultraviolet-B radiation[J]. Acta Physiologiae Plantarum,2015, 37(12):260-271.
[10]	KUMAR M S S, MAWLONG I, ALI K, et al. Regulation of phytosterol biosynthetic pathway during drought stress in rice[J]. Plant Physiology and Biochemistry:PPB, 2018,129:11-20.
[11]	ELKEILSH A, AWAD Y M, SOLIMAN M H, et al. Exogenous application of β-sitosterol mediated growth and yield improvement in water-stressed wheat(Triticum aestivum) involves up-regulated antioxidant system[J]. Journal of Plant Research, 2019, 132(6):881-901.
[12]	VEZZA T, CANET F, DE MARAÑÓN A M, et al. Phytosterols:nutritional health players in the management of obesity and its related disorders[J]. Antioxidants, 2020,9(12):1266.
[13]	CABRAL C E, KLEIN M R S T. Phytosterols in the treatment of hypercholesterolemia and prevention of cardiovascular diseases[J]. Arquivos Brasileiros De Cardiologia, 2017, 109(5):475-482.
[14]	JIANG L, ZHAO X, XU J, et al. The protective effect of dietary phytosterols on cancer risk:a systematic metaanalysis[J]. Journal of Oncology, 2019(6):7479518.
[15]	CABRAL C E, KLEIN M R S T. Phytosterols in the treatment of hypercholesterolemia and prevention of cardiovascular diseases[J]. Arquivos Brasileiros De Cardiologia, 2017, 109(5):475-482.
[16]	余慧,徐宝成,王大红,等.植物甾醇氧化物的形成、摄入及健康相关效应[J]. 食品科学, 2021, 42(23):306-314.
[17]	邱振楠,徐乾坤,王小琦,等.水稻DNA多态标记的开发与验证[J]. 浙江农业学报, 2016, 28(3):361-370.
[18]	KATZEN F. Gateway^®recombinational cloning:a biological operating system[J]. Expert Opinion on Drug Discovery, 2007, 2(4):571-589.
[19]	SAHU P K, CHAKRADHARI S, SIPENIECE E, et al.Fatty acids, tocopherols, tocotrienols, phytosterols, carotenoids, and squalene in seed oils of Hyptis suaveolens,Leonotis nepetifolia, and Ocimum sanctum[J]. European Journal of Lipid Science and Technology, 2020, 122(7):2000053.
[20]	GARRIS A J, TAI T H, COBURN J, et al. Genetic structure and diversity in Oryza sativa L[J]. Genetics, 2005,169(3):1631-1638.
[21]	WANG J, LIU J, SONG Z, et al. Sterol C24-methyltransferase:mechanistic studies of the C-methylation reaction with 24-fluorocycloartenol[J]. Bioorganic&Medicinal Chemistry Letters, 2008, 18(1):232-235.
[22]	周千惠,刘俊霞,刘萍萍,等.不同CCDs基因表达介导藏红花素在烟草中的异源合成[J/OL]. 烟草科技,2024:1-13(2024-04	-19) . https://kns.cnki.net/kcms/detail/41.1137.TS.20240419.1449.002.html.
[23]	Schaller H. The role of sterols in plant growth and development[J]. Progress in lipid research, 2003, 42(3):163-175.
[24]	PAL S, RASTOGI S, NAGEGOWDA D A, et al. RNAi of sterol methyl Transferase1 reveals its direct role in diverting intermediates towards withanolide/phytosterol biosynthesis in Withania somnifera[J]. Plant and Cell Physiology, 2019, 60(3):672-686.
[25]	DIENER A C, LI H, ZHOU W X, et al. STEROL METHYLTRANSFERASE 1 controls the level of cholesterol in plants[J]. The Plant Cell, 2000, 12(6):853-870.
[26]	闫志强,邓茹月,宫彦龙,等.茉莉酸甲酯对籼粳稻农艺性状及品质性状的影响[J]. 杂交水稻, 2022, 37(4):107-114.
[27]	张杰,杨玺,李游山.蛋白酶及其抑制剂关键活性位点研究进展[J]. 生物工程学报, 2021, 37(2):561-579.
[28]	ELSHAHAWY I E, ABD EL-WAHED M S. Suppression of Cephalosporium maydis by the resistance inducer betasitosterol[J]. European Journal of Plant Pathology, 2022,163(3):673-693.

Message Board

Identification and functional analysis of sterol C-24 methyltransferase in rice

doi: 10.15886/j.cnki.rdswxb.20240070

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Proportional views

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