盐胁迫下过表达BoRACK1基因对羽衣甘蓝种子萌发的影响
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摘要
蛋白激酶C1受体(RACK1)是一个高度保守的蛋白,它具有多种功能,对植物生长发育有重要的调控作用。本研究构建过表达BoRACK1转基因羽衣甘蓝植株,以阐明该基因在羽衣甘蓝中对盐胁迫的可能功能。转基因羽衣甘蓝对盐胁迫的耐受力比野生型高,表现在其种子发芽率显著高于野生型;活性氧产生比野生型活跃,H_2O_2也可能参与BoRACK1调控的种子萌发过程。实时定量PCR分析表明,在过表达BoRACK1转基因植株中,BoRbohs基因的转录水平显著增强。这些结果表明,在种子萌发过程中,过表达BoRACK1正调控植物耐盐胁迫。
The receptor for activated C-kinase 1(RACK1) is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic kale plants, in which Bo RACK1 gene was overexpressed, were studied to elucidate the possible functions of Bo RACK1 in response to salt stress in kale. The tolerance to salt stress of the transgenic kale plants was higher as compared with the non-transgenic kale plants in the seed germination. Reactive oxygen species production was more active than in the wild-type. H_2O_2 may also be involved in the regulation of Bo RACK1 during seed germination under salt stress. Quantitative real-time PCR analyses showed that the transcript levels of Bo Rbohs genes were significantly higher in overexpression of Bo RACK1 transgenic lines. These results indicated that overexpression of Bo RACK1 positively regulated kale tolerance to salt stresses.
The receptor for activated C-kinase 1(RACK1) is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic kale plants, in which Bo RACK1 gene was overexpressed, were studied to elucidate the possible functions of Bo RACK1 in response to salt stress in kale. The tolerance to salt stress of the transgenic kale plants was higher as compared with the non-transgenic kale plants in the seed germination. Reactive oxygen species production was more active than in the wild-type. H_2O_2 may also be involved in the regulation of Bo RACK1 during seed germination under salt stress. Quantitative real-time PCR analyses showed that the transcript levels of Bo Rbohs genes were significantly higher in overexpression of Bo RACK1 transgenic lines. These results indicated that overexpression of Bo RACK1 positively regulated kale tolerance to salt stresses.
引文
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Bird RJ,Baillie GS,Yarwood SJ,et al(2016).Interaction with receptor for activated C-kinase 1(RACK1)sensitizes the phosphodiesterase PDE4D5 towards hydrolysis of c AMP and activation by protein kinase C.Biochem J,432:207-216
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Schmitt K,Smolinski N,Neumann P,et al(2016).Asc1p/RACK1 connects ribosomes to eukaryotic phosphosignaling.Mol Cell Biol,37:e00279-16
Wang BS,Yu JJ,Zhu DY,et al(2014).Maize Zm RACK1 is involved in the plant response to fungal phytopathogens.Int J Mol Sci,15:9343-9359
Yan JY,Ma CQ Chang B,et al(2017).A modified CTABmethod for genomic DNA extraction from apple fruit.Mol Plant Breeding,15(9):3610-3615(in Chinese with English abstract)[闫玖英,马长青,常博等(2017).改良CTAB法用于苹果果实基因组DNA的提取.分子植物育种,15(9):3610-3615]
Ye NH,Zhu GH,Liu YG,et al(2012).Ascorbic acid and reactive oxygen species are involved in the inhibition of seed germination by abscisic acid in rice seeds.J Exp Bot,63:1809-1822
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Zhang JX,Wang XL,Vikash V,et al(2016).ROS and ROS-mediated cellular signaling.Oxid Med Cell Longev,2016:4350965
Bird RJ,Baillie GS,Yarwood SJ,et al(2016).Interaction with receptor for activated C-kinase 1(RACK1)sensitizes the phosphodiesterase PDE4D5 towards hydrolysis of c AMP and activation by protein kinase C.Biochem J,432:207-216
Chen JG,Ullah H,Temple B,et al(2006).RACK1 mediates multiple hormone responsiveness and developmental processes in Arabidopsis.J Exp Bot,57:2697-2708
Gao H,Gao YL,Li KH(2015).Optimization of Agrobacterium-mediated transformation system for Brassica oleracea var.acephala with hypocotyls as explants.Biotech Bull,31:111-115(in Chinese with English abstract)[高航,高玉亮,李葵花(2015).羽衣甘蓝下胚轴农杆菌介导遗传转化体系的优化.生物技术通报,31:111-115]
Gao JF(2006).Experimental Instruction of Plant Physiology.Beijing:Higher Education Press,214-216(in Chinese)[高俊凤(2006).植物生理学实验指导.北京:高等教育出版社,214-216]
Guo JJ,Wang JB,Li X,et al(2009).RACK1 is a negative regulator of ABA responses in Arabidopsis.J Exp Bot,60:3819-3833
Guo JJ,Wang SC,Valerius O,et al(2011).Involvement of Arabidopsis RACK1 in protein translation and its regulation by abscisic acid.Plant Physiol,155:370-383
He D,Han C,Yao J,et al(2011).Constructing the metabolic and regulatory pathways in germinating rice seeds through proteomic approach.Proteomics,11:2693-2713
Huang YT,Lin C,He F,et al(2017).Exogenous spermidine improves seed germination of sweet corn via involvement in phytohormone interactions,H2O2,and relevant gene expression.BMC Plant Biol,17:1
Komatsu S,Hiraga S,Nouri MZ(2014).Analysis of flooding-responsive proteins localized in the nucleus of soybean root tips.Mol Biol Rep,41:1127-1139
Kong XQ,Luo Z,Zhang YJ,et al(2017).Soaking in H2O2,regulates ABA biosynthesis and GA catabolism in germinating cotton seeds under salt stress.Acta Physiol Plant,39:12-19
Li DH,Liu H,Yang YL,et al(2009).Down-regulated expression of RACK1 gene by RNA interference enhances drought tolerance in rice.Rice Sci,16(1):14-20
Liu QY,He HH,Hu LF(2013).Research progress in plant Rboh genes function and activity regulation mechanism.Biotech Bull,11:8-13(in Chinese with English abstract)[刘秋圆,贺浩华,胡丽芳(2013).植物Rboh基因功能及其活性调节机制的研究进展.生物技术通报,11:8-13]
Nakashima A,Chen LT,Thao NP,et al(2008).RACK1 functions in rice innate immunity by interacting with the Rac1immune complex.Plant Cell,20:2265-2279
Nielsen MH,Flygaard RK,Jenner LB(2017).Structural analysis of ribosomal RACK1 and its role in translational control.Cell Signal,35:272-281
Panngom K,Chuesaard T,Tamchan N,et al(2018).Comparative assessment for the effects of reactive species on seed germination,growth and metabolisms of vegetables.Sci Hortic,227:85-91
Schmitt K,Smolinski N,Neumann P,et al(2016).Asc1p/RACK1 connects ribosomes to eukaryotic phosphosignaling.Mol Cell Biol,37:e00279-16
Wang BS,Yu JJ,Zhu DY,et al(2014).Maize Zm RACK1 is involved in the plant response to fungal phytopathogens.Int J Mol Sci,15:9343-9359
Yan JY,Ma CQ Chang B,et al(2017).A modified CTABmethod for genomic DNA extraction from apple fruit.Mol Plant Breeding,15(9):3610-3615(in Chinese with English abstract)[闫玖英,马长青,常博等(2017).改良CTAB法用于苹果果实基因组DNA的提取.分子植物育种,15(9):3610-3615]
Ye NH,Zhu GH,Liu YG,et al(2012).Ascorbic acid and reactive oxygen species are involved in the inhibition of seed germination by abscisic acid in rice seeds.J Exp Bot,63:1809-1822
Zhang DP,Chen L,Li DH,et al(2014).Os RACK1 is involved in abscisic acid-and H2O2-mediated signaling to regulate seed germination in rice(Oryza sativa L.).PLo SOne,9:e97120
Zhang JX,Wang XL,Vikash V,et al(2016).ROS and ROS-mediated cellular signaling.Oxid Med Cell Longev,2016:4350965