水稻羧酸酯酶OsCDAP的生物学功能初探
|
摘要
本课题组在对水稻病害的研究中,鉴定并克隆到一个羧酸酯酶基因OsCDAP。为进一步研究该酯酶生物学功能,构建了OsCDAP植物过表达载体,通过农杆菌介导的方法转化水稻"中花11"。转基因水稻表型分析显示,过表达转基因水稻植株株高略高于对照,其第一节间长度明显长于对照,而第五节间长度明显短于对照,其他节间长度差异不明显;OsCDAP过表达株系的剑叶夹角明显大于对照。进一步对转基因水稻GA和BR合成及信号转导相关基因进行分析,发现部分基因的表达发生改变。本试验结果显示OsCDAP可通过调控内源GA和BR的含量及参与相关信号途径调控水稻节间长度及叶夹角大小,进而对水稻的生长发育产生影响。
Our research group cloned and indentified a carboxylesterase gene OsCDAP in the study of rice diseases. In order to study its function, an over-expression vector containing OsCDAP was constructed, and the transgenic lines of rice Zhonghua 11 were harvested through Agrobacterium-mediated transformation. Phenotype analysis showed that the plant height of OsCDAP over-expressed rice was slightly higher than that of control. The length of the first internode of transgenic strains was significantly longer than that of control, while the length in the of the fifth internode was significantly shorter than that of the control. There were almost no significant differences in the other internode length between transgenic strains and control. The leaf angles of flag leaves of transgenic strains were significantly larger than those of control. The expressions of GA and BR synthesis and signal transduction genes were also changed corresponding to the phenotypes variation. The results indicated that OsCDAP might regulate rice internode length and leaf angle by regulating the GA and BR synthesis and participating in signal transduction, which eventually lead to the variation of growth and development of rice.
Our research group cloned and indentified a carboxylesterase gene OsCDAP in the study of rice diseases. In order to study its function, an over-expression vector containing OsCDAP was constructed, and the transgenic lines of rice Zhonghua 11 were harvested through Agrobacterium-mediated transformation. Phenotype analysis showed that the plant height of OsCDAP over-expressed rice was slightly higher than that of control. The length of the first internode of transgenic strains was significantly longer than that of control, while the length in the of the fifth internode was significantly shorter than that of the control. There were almost no significant differences in the other internode length between transgenic strains and control. The leaf angles of flag leaves of transgenic strains were significantly larger than those of control. The expressions of GA and BR synthesis and signal transduction genes were also changed corresponding to the phenotypes variation. The results indicated that OsCDAP might regulate rice internode length and leaf angle by regulating the GA and BR synthesis and participating in signal transduction, which eventually lead to the variation of growth and development of rice.
引文
[1] 洪凯,张斌,高阳,等.水稻剑叶夹角和单株产量的QTL分析[J].分子植物育种,2015,13(4):761-768.
[2] 李臻,王庆国,潘教文,等.水稻细胞色素P450基因OsDWARF48调控株高的功能分析[J].山东农业科学,2018,50(11):1-9.
[3] Sasaki A, Ashikari M, Ueguchi-Tanaka M, et al. Green revolution: a mutant gibberellin-synthesis gene in rice[J]. Nature, 2002, 416(6882):701-702.
[4] Itoh H, Tatsumi T, Sakamoto T, et al. A rice semi-dwarf gene, Tan-Ginbozu(D35), encodes the gibberellin biosynthesis enzyme, ent-kaurene oxidase[J]. Plant Molecular Biology, 2004, 54(4):533-547.
[5] Oikawa T, Koshioka M, Kojima K, et al. A role of OsGA20ox1, encoding an isoform of gibberellin 20-oxidase, for regulation of plant stature in rice[J]. Plant Molecular Biology, 2004, 55(5):687-700.
[6] Tong H, Xiao Y, Liu D, et al.Brassinosteroid regulates cell elongation by modulating gibberellin metabolism in rice[J]. Plant Cell, 2014, 26(11):4376-4393.
[7] Hong Z, Ueguchi-Tanaka M, Umemura K, et al. A rice brassinosteroid-deficient mutant, ebisu dwarf(d2), is caused by a loss of function of a new member of cytochrome P450[J]. Plant Cell, 2003, 15(12):2900-2910.
[8] Tanabe S, Ashikari M, Fujioka S, et al. A novel cytochrome P450 is implicated in brassinosteroid biosynthesis via the characterization of a rice dwarf mutant, dwarf11, with reduced seed length[J]. Plant Cell, 2005, 17(3):776-790.
[9] Mori M, Nomura T, Ooka H, et al.Isolation and characterization of a rice dwarf mutant with a defect in brassinosteroid biosynthesis[J]. Plant Physiology, 2002, 130(3):1152-1161.
[10] Qiao S, Sun S, Wang L, et al. The RLA1/SMOS1 transcription factor functions with OsBZR1 to regulate brassinosteroid signaling and rice architecture[J]. Plant Cell, 2017, 29(2):292-309.
[11] Hirano K, Yoshida H, Aya K, et al.Small organ size 1 and small organ size 2/dwarf and low-tillering form a complex to integrate auxin and brassinosteroid signaling in rice[J]. Molecular Plant, 2017, 10(4):590-604.
[12] Ko M, Cho J H, Seo H H, et al. Constitutive expression of a fungus-inducible carboxylesterase improves disease resistance in transgenic pepper plants[J]. Planta, 2016, 244(2):1-14.
[13] Schmeitzl C, Varga E, Warth B, et al. Identification and characterization of carboxylesterases from brachypodium distachyon deacetylating trichothecene mycotoxins[J]. Toxins, 2015, 8(1):6.
[14] Stuhlfelder C, Lottspeich F, Mueller M J. Purification and partial amino acid sequences of an esterase from tomato[J]. Phytochemistry, 2002, 60(3):233-240.
[15] 刘建光, 赵贵元, 赵俊丽,等. 植物羧酸酯酶的结构、表达调控及生物功能研究进展[J]. 作物杂志, 2018(3): 32-36.
[16] 李臻,潘教文,王庆国,等.水稻羧酸酯酶基因OsCDAP的克隆及表达分析[J].山东农业科学,2018,50(11):10-15.
[17] Fukao T , Bailey-Serres J . Submergence tolerance conferred by Sub1A is mediated by SLR1 and SLRL1 restriction of gibberellin responses in rice[J]. Proceedings of the National Academy of Sciences, 2008, 105(43):16814-16819.
[18] Magome H, Nomura T, Hanada A, et al.CYP714B1 and CYP714B2 encode gibberellin 13-oxidases that reduce gibberellin activity in rice[J]. Proc. Natl. Acad. Sci. U S A,2013, 110 (5):1947-1952.
[19] Luo A, Qian Q, Yin H, et al. EUI1, encoding a putative cytochrome P450 monooxygenase, regulates internode elongation by modulating gibberellin responses in rice.[J].Plant Cell Physiol.,2006, 47(2):181-191.
[20] Qiao F, Zhao K J. The influence of RNAi targeting of OsGA20ox2 gene on plant height in rice[J]. Plant Molecular Biology Reporter, 2011, 29(4):952-960.
[21] De Vleesschauwer D, Seifi S, Filipe O, et al. The DELLA protein SLR1 integrates and amplifies salicylic acid- and jasmonic acid-dependent innate immunity in rice[J]. Plant Physiology, 2016, 170(3):1831-1847
[22] Hirano K, Kouketu E, Katoh H, et al. The suppressive function of the rice DELLA protein SLR1 is dependent on its transcriptional activation activity[J].The Plant Journal, 2012, 71(3):443-453.
[23] Zuo S, Zhou X, Chen M, et al. OsSERK1 regulates rice development but not immunity to Xanthomonas oryzae pv. oryzae or Magnaporthe oryzae[J]. Journal of Integrative Plant Biology, 2014, 56(12):1179-1192.
[2] 李臻,王庆国,潘教文,等.水稻细胞色素P450基因OsDWARF48调控株高的功能分析[J].山东农业科学,2018,50(11):1-9.
[3] Sasaki A, Ashikari M, Ueguchi-Tanaka M, et al. Green revolution: a mutant gibberellin-synthesis gene in rice[J]. Nature, 2002, 416(6882):701-702.
[4] Itoh H, Tatsumi T, Sakamoto T, et al. A rice semi-dwarf gene, Tan-Ginbozu(D35), encodes the gibberellin biosynthesis enzyme, ent-kaurene oxidase[J]. Plant Molecular Biology, 2004, 54(4):533-547.
[5] Oikawa T, Koshioka M, Kojima K, et al. A role of OsGA20ox1, encoding an isoform of gibberellin 20-oxidase, for regulation of plant stature in rice[J]. Plant Molecular Biology, 2004, 55(5):687-700.
[6] Tong H, Xiao Y, Liu D, et al.Brassinosteroid regulates cell elongation by modulating gibberellin metabolism in rice[J]. Plant Cell, 2014, 26(11):4376-4393.
[7] Hong Z, Ueguchi-Tanaka M, Umemura K, et al. A rice brassinosteroid-deficient mutant, ebisu dwarf(d2), is caused by a loss of function of a new member of cytochrome P450[J]. Plant Cell, 2003, 15(12):2900-2910.
[8] Tanabe S, Ashikari M, Fujioka S, et al. A novel cytochrome P450 is implicated in brassinosteroid biosynthesis via the characterization of a rice dwarf mutant, dwarf11, with reduced seed length[J]. Plant Cell, 2005, 17(3):776-790.
[9] Mori M, Nomura T, Ooka H, et al.Isolation and characterization of a rice dwarf mutant with a defect in brassinosteroid biosynthesis[J]. Plant Physiology, 2002, 130(3):1152-1161.
[10] Qiao S, Sun S, Wang L, et al. The RLA1/SMOS1 transcription factor functions with OsBZR1 to regulate brassinosteroid signaling and rice architecture[J]. Plant Cell, 2017, 29(2):292-309.
[11] Hirano K, Yoshida H, Aya K, et al.Small organ size 1 and small organ size 2/dwarf and low-tillering form a complex to integrate auxin and brassinosteroid signaling in rice[J]. Molecular Plant, 2017, 10(4):590-604.
[12] Ko M, Cho J H, Seo H H, et al. Constitutive expression of a fungus-inducible carboxylesterase improves disease resistance in transgenic pepper plants[J]. Planta, 2016, 244(2):1-14.
[13] Schmeitzl C, Varga E, Warth B, et al. Identification and characterization of carboxylesterases from brachypodium distachyon deacetylating trichothecene mycotoxins[J]. Toxins, 2015, 8(1):6.
[14] Stuhlfelder C, Lottspeich F, Mueller M J. Purification and partial amino acid sequences of an esterase from tomato[J]. Phytochemistry, 2002, 60(3):233-240.
[15] 刘建光, 赵贵元, 赵俊丽,等. 植物羧酸酯酶的结构、表达调控及生物功能研究进展[J]. 作物杂志, 2018(3): 32-36.
[16] 李臻,潘教文,王庆国,等.水稻羧酸酯酶基因OsCDAP的克隆及表达分析[J].山东农业科学,2018,50(11):10-15.
[17] Fukao T , Bailey-Serres J . Submergence tolerance conferred by Sub1A is mediated by SLR1 and SLRL1 restriction of gibberellin responses in rice[J]. Proceedings of the National Academy of Sciences, 2008, 105(43):16814-16819.
[18] Magome H, Nomura T, Hanada A, et al.CYP714B1 and CYP714B2 encode gibberellin 13-oxidases that reduce gibberellin activity in rice[J]. Proc. Natl. Acad. Sci. U S A,2013, 110 (5):1947-1952.
[19] Luo A, Qian Q, Yin H, et al. EUI1, encoding a putative cytochrome P450 monooxygenase, regulates internode elongation by modulating gibberellin responses in rice.[J].Plant Cell Physiol.,2006, 47(2):181-191.
[20] Qiao F, Zhao K J. The influence of RNAi targeting of OsGA20ox2 gene on plant height in rice[J]. Plant Molecular Biology Reporter, 2011, 29(4):952-960.
[21] De Vleesschauwer D, Seifi S, Filipe O, et al. The DELLA protein SLR1 integrates and amplifies salicylic acid- and jasmonic acid-dependent innate immunity in rice[J]. Plant Physiology, 2016, 170(3):1831-1847
[22] Hirano K, Kouketu E, Katoh H, et al. The suppressive function of the rice DELLA protein SLR1 is dependent on its transcriptional activation activity[J].The Plant Journal, 2012, 71(3):443-453.
[23] Zuo S, Zhou X, Chen M, et al. OsSERK1 regulates rice development but not immunity to Xanthomonas oryzae pv. oryzae or Magnaporthe oryzae[J]. Journal of Integrative Plant Biology, 2014, 56(12):1179-1192.