水稻OsFAD2、OsFAD6的克隆及其家族成员对非生物胁迫的响应

doi:10.3724/SP.J.1005.2010.00839

遗传 ›› 2010, Vol. 32 ›› Issue (8): 839-847.doi: 10.3724/SP.J.1005.2010.00839

水稻OsFAD2、OsFAD6的克隆及其家族成员对非生物胁迫的响应

曹英萍, 石金磊, 李钟, 明凤

复旦大学生命科学学院植物科学研究所, 遗传工程国家重点实验室, 上海 200433

收稿日期:2009-10-23 修回日期:2010-01-08 出版日期:2010-08-20 发布日期:2010-08-23
通讯作者: 明凤 E-mail:fming@fudan.edu.cn
基金资助:
国家863项目(编号：2008AA10Z116), 转基因生物新品种培育重大专项(编号：#2008ZX08009-001), 上海市青年科技启明星计划 (编号：08QH14003), 上海市科委重点科技攻关项目(编号：08391910400)和国家基础科学人才培养基金项目(编号：J0630643)资助。

Isolation of OsFAD2, OsFAD6 and FAD family members response to abiotic stresses in Oryza sativa L.

CAO Ying-Ping, SHI Jin-Lei, LI Zhong, MING Feng

State Key Laboratory of Genetic Engineering, Institute of Plant Biology, School of Life Science, Fudan University, Shanghai 200433, China

Received:2009-10-23 Revised:2010-01-08 Online:2010-08-20 Published:2010-08-23
Contact: MING Feng E-mail:fming@fudan.edu.cn

摘要/Abstract

摘要： 植物中的不饱和脂肪酸由脂肪酸去饱和酶(Fatty acid desaturase, FAD)合成, 它在植物的生长发育以及植物非生物胁迫方面起着重要的作用。文章采用RT-PCR方法, 从水稻日本晴(Oryza sativa L.)中克隆了分别与FAD2、FAD6同源的脂肪酸脱氢酶序列, 命名为OsFAD2和OsFAD6。OsFAD2的ORF为1 167 bp, 推测其编码蛋白含有388个氨基酸, 等电点为8.17, 分子量为52.24 kDa, C端有内质网定位序列; OsFAD6的ORF长度为1 365 bp, 推测编码454个氨基酸的蛋白质序列, 分子量44.35 kDa, 等电点为9.24, 推测N端38个氨基酸为叶绿体导肽。两者都具有膜整合脂肪酸去饱和酶特有的3个组氨酸簇。RT-PCR分析表明, OsFAD2和OsFAD6在水稻所有器官中都表达, 在叶中表达量为最高。在水稻FAD基因家族中, 叶中OsFAD2、OsFAD6 的mRNA对低温不响应, 而OsFAD7和OsFAD8的 mRNA在低温下上升。水稻叶中OsFAD2、OsFAD6、OsFAD3和OsFAD7的mRNA表达具有昼夜节律性, 在光照下表达量低, 而在随后的黑暗中表达量高, OsFAD6和OsFAD7 mRNA表达的昼夜节律性可能与水稻幼苗叶片中NADPH量的改变有关。

关键词: 水稻, 脂肪酸去饱和酶, 非生物胁迫, 昼夜节律, 表达谱

Abstract: Unsaturated fatty acids are synthesized by fatty acid desaturase in plant, which play an important role in plant development and defense to abiotic stresses. The key enzymes for the conversion of oleic into linoleic acid were isolated from rice (Oryza sativa L.) and were designated OsFAD2 and OsFAD6, respectively. The open reading frame (ORF) of OsFAD2 was 1 167 bp in length, which encoded a 388 amino acids sequence with the isoelectric point of 8.17 and molecular mass of 52.24 kDa, the OsFAD2 protein contained a C-terminal ER retrieval motif. The ORF of OsFAD6 was 1 365 bp in length and the predicted OsFAD6 protein has 454 amino acids with an estimated molecular mass of 44.35 kDa and an isoelectric point of 9.24, the predicted OsFAD6 protein possessed a putative N-terminal plastidial signal peptide. Both of them had three his-boxes, which were peculiar to membrane integrated fatty acid desaturase by Clustal X analysis. RT-PCR analysis showed that both genes were expressed in all tissues rice seedlings, with the maximum transcript accumulation in leaves. Among FAD family members of Oryza sativa L., the mRNA abundance of OsFAD2 and OsFAD6 in leaves did not change under cold stress; however, the mRNA abundance of OsFAD7 and OsFAD8 increased in the same condition. It was also found that the expression of FAD family members had diurnal rhythm phenomena. Based on the results of this study, it suggested that diurnal rhythm expression of OsFAD6 and OsFAD7 was related to the change of NADPH abundance.

Key words: expression pattern, Oryza sativa L., fatty acid desaturase, abiotic stress, diurnal rhythm

曹英萍，石金磊，李钟，明凤. 水稻OsFAD2、OsFAD6的克隆及其家族成员对非生物胁迫的响应[J]. 遗传, 2010, 32(8): 839-847.

CAO Yang-Ping, DAN Jin-Lei, LI Zhong, MENG Feng. Isolation of OsFAD2, OsFAD6 and FAD family members response to abiotic stresses in Oryza sativa L.[J]. HEREDITAS, 2010, 32(8): 839-847.

参考文献

[1] Dai XF, Xiao L, Wu YH, Wu G, Lu GM. An overview of plant fatty acid desaturases and the coding genes. Chinese Bull Bot, 2007, 24(1): 105–113. [2] Los DA, Murata N. Structure and expression of fatty acid desaturases. Biochim Biophys Acta-Lipids Lipid Metabol, 1998, 1394(1): 3–15. [3] Somerville C, Browse J. Dissecting desaturation: plants prove advantageous. Trends Cell Biol, 1996, 6(4): 148–153. [4] Iba K, Gibson S, Nishiuchi T, Fuse T, Nishimura M, Arondel V, Hugly S, Somerville C. A gene encoding a chloroplast omega-3 fatty acid desaturase complements alterations in fatty acid desaturation and chloroplast copy number of the fad7 mutant of Arabidopsis thaliana. J Biol Chem, 1993, 268(32): 24099–24105. [5] Gibson S, Arondel V, Iba K, Somerville C. Cloning of a temperature-regulated gene encoding a chloroplast omega-3 desaturase from Arabidopsis thaliana. Plant Physiol, 1994, 106(4): 1615–1621. [6] Okuley J, Lightner J, Feldmann K, Yadav N, Lark E, Browse J. Arabidopsis Fad2 gene encodes the enzyme that is essential for polyunsaturated lipid-synthesis. Plant Cell, 1994, 6(1): 147–158. [7] Falcone DL, Gibson S, Lemieux B, Somerville C. Identi-fication of a gene that complements an Arabidopsis mutant deficient in chloroplast ω-6 desaturase activity. Plant Physiol, 1994, 106(4): 1452–1459. [8] Hita WD, Carlson TJ, Booth JR, Jr, Kinney AJ, Stecca Kland Yada NS. Cloning of a higher-plant plastid ω-6 fatty acid desaturase cDNA and its expression in a Cyanobacterium. Plant Physiol, 1994, 105(2): 634–641. [9] Banilas G, Moressis A, Nikoloudakis N, Hatzopoulos P. Spatial and temporal expressions of two distinct oleate desaturases from olive (Olea europaea L.). Plant Sci, 2005, 168(2): 547–555. [10] Teixeira MC, Coelho N, Olsson ME, Brodelius PE, Car-valho IS, Brodelius M. Molecular cloning and expression analysis of three omega-6 desaturase genes from purslane (Portulaca oleracea L.). Biotechnol Lett, 2009, 31(7): 1089–1101. [11] Mikkilineni V, Rocheford TR. Sequence variation and genomic organization of fatty acid desaturase-2 (fad2) and fatty acid desaturase-6 (fad6) cDNAs in maize. Theor Appl Genet, 2003, 106(7): 1326–1332. [12] Collados R, Andreu V, Picorel R, Alfonso M. A light- sensitive mechanism differently regulates transcription and transcript stability of ω-3 fatty-acid desaturases (FAD3, FAD7 and FAD8) in soybean photosynthetic cell suspensions. FEBS Lett, 2006, 580(20): 4934–4940. [13] Kis M, Zsiros O, Farkas T, Wada H, Nagy F, Gombos Z. Light-induced expression of fatty acid desaturase genes. Biochemistry, 1998, 95(8): 4209–4214. [14] Klyachko-Gurvicha GL, Tsoglina LN, Douchab J, Kopet-skiib J, Shebalina (Ryabykh) IB, Semenenkoa VE. De-saturation of fatty acids as an adaptive response to shifts in light intensity. Physiol Plant, 1999, 107(2): 240–249. [15] Neidleman SL. Effects of temperature on lipid unsatura-tion. Biotechnol Genet Eng Rev, 1987, 5: 245–268. [16] Wang JW, Ming F, Pittman J, Han YY, Hu J, Guo B, Shen DL. Characterization of a rice (Oryza sativa L.) gene en-coding a temperature-dependent chloroplast ω-3 fatty acid desaturase. Bichem Biophys Res Commun, 2002, 340(4): 1209–1216. [17] Gibson S, Arondel V, Iba K, Somerville C. Cloning of a temperature-regulated gene encoding a chloroplast ω-3 desaturase from Arabidopsis thaliana. Plant Physiol, 1994, 106(4): 1615–1621. [18] Berberich T, Harad M, Sugawara K, Kodama H, Iba K, Kusano T. Two maize genes encoding ω-3 fatty-acid de-saturase and their differential expression to temperature. Plant Mol Biol, 1998, 36(2): 297–306. [19] Kargiotidou A, Deli D, Galanopoulou D, Tsaftaris A, Farmaki T. Low temperature and light regulate delta 12 fatty acid desaturases (FAD2) at a transcriptional level in cotton (Gossypium hirsutum). J Exp Bot, 2008, 59(8): 2043–2056. [20] Kodama H, Hamada T, Horiguchi G, Nishimura M, Iba K. Genetic enhancement of cold tolerance by expression of a gene for chloroplast Omega-3 fatty-acid desaturase in transgenic tobacco. Plant Physiol, 1994, 105(2): 601–605. [21] 刘立侠, 柳青, 许守民. 基因工程在改善植物油营养价值中的应用. 植物学通报, 2005, 22(5): 623–631. [22] Yara A, Yaeno T, Hasegawa M, Seto H, Montillet JL, Ku-sumi K, Seo S, Iba K. Disease resistance against Mag-naporthe grisea is enhanced in transgenic rice with sup-pression of ω-3 fatty acid desaturases. Plant Cell Physiol, 2007, 48(9): 1263–1274. [23] Kodama H, Akagi H, Kusumi K, Fujimura T, Iba K. Structure, chromosomal location and expression of a rice gene encoding the microsome omega-3 fatty acid desatu-rase. Plant Mol Biol, 1993, 33(3): 493–502. [24] Yoshida S, Forno DA, Cock JH, Gomez KA. Laboratory manual for physiological studies of rice. International Rice Research Institute, Manila, The Philippines, 1976. [25] Thompson JD, Higgins DG, Gibson TJ. Clustal W: im-proving the sensitivity of progressive multiple sequence alignment through sequence weighting matrix choice. Nucleic Acids Res, 1994, 22(22): 4673–4680. [26] Kumar S, Tamura K, Jakobsen IB, Nei M. MEGA2: mo-lecular evolutionary genetics analysis software. Arizona State University, 2001, Tempe, AZ. [27] Saitou N, Nei M. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol, 1987, 4: 406–425. [28] Terry R r. Metabolic Pathways of Agrochemicals. Cam-bridge: The Royal Society of Chemistry, 1998, 735. [29] McConn M, Browse J. The critical requirement for linolenic acid is pollen development, not photosynthesis, in an Arabi-dopsis mutant. Plant Cell, 1996, 8(3): 403–416. [30] McConn M, Creelman RA, Bell E, Mullet JE, Browse J. Jasmonate is essential for insect defense in Arabidopsis. Proc Natl Acad Sci USA, 1997, 94(10): 5473–5477. [31] Pearcy RW. Effect of growth temperature on the fatty acid composition of the leaf lipids in Atriplex lentiformis (Torr.) Wats. Plant Physiol, 1978, 61(4): 484–486. [32] Huang J, Wang JF, Zhang HS. Advance on plant pentose phosphate pathway and its key enzymes. Chinese Bull Bot, 2004, 21(2): 139–145.

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水稻OsFAD2、OsFAD6的克隆及其家族成员对非生物胁迫的响应

Isolation of OsFAD2, OsFAD6 and FAD family members response to abiotic stresses in Oryza sativa L.

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