番茄红素ε-环化酶调节烟草抗旱性的研究
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摘要
类胡萝卜素是一类重要的天然色素,在植物生长发育和抵御逆境的过程中起着重要作用.番茄红素ε-环化酶是类胡萝卜素合成的关键酶,催化番茄红素向α类胡萝卜素的转化.本文以烟草为材料,通过转基因手段分别获得了烟草Ntε-LCY基因的过量表达和基因沉默株系.表型分析结果显示:与野生型植株相比,Ntε-LCY过量表达植株的抗旱能力显著降低,而Ntε-LCY表达沉默株系的抗旱能力明显增强.生理指标测定结果显示:Ntε-LCY过量表达植株叶片的失水效率、活性氧(ROS)含量显著高于野生型植株,而Ntε-LCY沉默株系的生理指标值明显低于野生型植株.这些生理指标值的改变趋势与植株ABA含量的变化趋势相反,表明Ntε-LCY基因可能通过调控ABA含量,影响植株的失水效率和ROS累积,进而调控植物的抗旱性.
Carotenoids are crucial pigments functioning in the growth and adaption against environmental stresses in plants. Lycopene ε-cyclase(ε-LCY) is a key enzyme catalyzing the synthesis of α carotenoid. In this study, the authors generated Ntε-LCY overproducing and expressing silencing lines in tobacco. Phenotypic analysis shows that Ntε-LCY overproducing plants are more sensitive to drought stress compared with wild-type plants, whereas Ntε-LCY silencing plants are more tolerant. Consistently, the water loss rate and reactive oxygen species(ROS) content are significantly higher in Ntε-LCY overproducing plants and lower in Ntε-LCY silencing plants than wild-type plants. These physiological parameters changes seem to be related to the altered abscisic acid(ABA) by Ntε-LCY manipulation in plants. A significant increase is in Ntε-LCY overproducing plants and an observed decrease is in Ntε-LCY silencing plants compared with wild-type plants. Taken together, the results indicate that Ntε-LCY may affect ABA synthesis and in turn influence water loss and ROS accumulation, finally modulating the drought tolerance in tobacco.
Carotenoids are crucial pigments functioning in the growth and adaption against environmental stresses in plants. Lycopene ε-cyclase(ε-LCY) is a key enzyme catalyzing the synthesis of α carotenoid. In this study, the authors generated Ntε-LCY overproducing and expressing silencing lines in tobacco. Phenotypic analysis shows that Ntε-LCY overproducing plants are more sensitive to drought stress compared with wild-type plants, whereas Ntε-LCY silencing plants are more tolerant. Consistently, the water loss rate and reactive oxygen species(ROS) content are significantly higher in Ntε-LCY overproducing plants and lower in Ntε-LCY silencing plants than wild-type plants. These physiological parameters changes seem to be related to the altered abscisic acid(ABA) by Ntε-LCY manipulation in plants. A significant increase is in Ntε-LCY overproducing plants and an observed decrease is in Ntε-LCY silencing plants compared with wild-type plants. Taken together, the results indicate that Ntε-LCY may affect ABA synthesis and in turn influence water loss and ROS accumulation, finally modulating the drought tolerance in tobacco.
引文
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[2] WELSCH R,ZHOU X,YUAN H,et al.Clp protease and OR directly control the proteostasis of phytoene synthase,the crucial enzyme for carotenoid biosynthesis in Arabidopsis[J].Molecular Plant,2018,11:149-162.
[3] NISAR N,LI L,LU S,et al.Carotenoid metabolism in plants[J].Molecular Plant,2015,8:68-82.
[4] LIU L,SHAO Z,ZHANG M,et al.Regulation of carotenoid metabolism in tomato[J].Molecular Plant,2015,8:28-39.
[5] WALTER M H,STRACK D.Carotenoids and their cleavage products:Biosynthesis and functions[J].Natural Product Reports,2011,28:663-692.
[6] 李锋,王燃,武明珠,等.基于过表达技术的烟草NtGGPPS1基因功能的研究[J].河南大学学报(自然科学版),2019,49(2):180-185.LI F,WANG R,WU M Z,et al.Study on NtGGPPS1 function in Nicotiana tabacum based on overexpression technology[J].Journal of Henan University(Natural Science),2019,49(2):180-185.
[7] CAZZONELLI C.Carotenoids in nature:Insights from plants and beyond[J].Functional Plant Biology,2011,38:833-847.
[8] YOUNG A J.The photoprotective role of carotenoids in higher plants[J].Physiologia Plantarum,1991,83:702-708.
[9] POGSON B J,NIYOGI K K,BJORKMAN O,et al.Altered xanthophyll compositions adversely affect chlorophyll accumulation and nonphotochemical quenching in Arabidopsis mutants[J].Proceedings of the National Academy of Sciences of the United States of America USA,1998,95:13324-13329.
[10] NIYOGI K K,GROSSMAN A R,BJORKMAN O.Arabidopsis mutants define a central role for the xanthophyll cycle in the regulation of photosynthetic energy conversion[J].Plant Cell,1998,10:1121-1134.
[11] POGSON B J,RISSLER H M.Genetic manipulation of carotenoid biosynthesis and photoprotection[J].Philosophical Transactions of the Royal Society B:Biological Sciences,2000,355:1395-1403.
[12] NAMBARA E,MARION P A.Abscisic acid biosynthesis and catabolism[J].Annual Review of Plant Biology,2005,56:165-185.
[13] SHI Y,GUO J,ZHANG W,et al.Cloning of the lycopene beta-cyclase gene in Nicotiana tabacum and its overexpression confers salt and drought tolerance[J].International Journal of Molecular Sciences,2015,16:30438-30457.
[14] MERLOT S,MUSTILLI A C,GENTY B,et al.Use of infrared thermal imaging to isolate Arabidopsis mutants defective in stomatal regulation[J].The Plant Journal,2002,30:601-609.
[15] DAUDI A,O'BRIEN J A.Detection of hydrogen peroxide by DAB staining in Arabidopsis leaves[J].Bio-protocol,2012,2(18):1-4.
[16] KUMAR D,YUSUF M A,SINGH P,et al.Histochemical detection of superoxide and H2O2 accumulation in Brassica juncea seedlings[J].Bio-protocol,2014,4:1108.
[17] MENG Z,ZHANG B,LIU B,et al.High carotenoids content can enhance resistance of selected Pinctada fucata families to high temperature stress[J].Fish & Shellfish Immunology,2017,61:211-218.
[18] SHI Y,LIU P,XIA Y,et al.Downregulation of the lycopene ε-cyclase gene confers tolerance to salt and drought stress in Nicotiana tabacum[J].Acta Physiologiae Plantarum,2015,37:210.
[19] QUAN W,HU Y,MU Z,et al.Overexpression of AtPYL5 under the control of guard cell specific promoter improves drought stress tolerance in Arabidopsis[J].Plant Physiology and Biochemistry,2018,129:150-157.
[20] ZOU J J,LI X D,RATNASEKERA D,et al.Arabidopsis calcium-dependent protein kinase8 and catalase3 function in abscisic acid-mediated signaling and H2O2 homeostasis in stomatal guard cells under drought stress[J].Plant Cell,2015,27:1445-1460.
[21] SAH S K,REDDY K R,LI J.Abscisic acid and abiotic stress tolerance in crop plants[J].Frontiers in Plant Science,2016,7:571.
[22] RAGHAVENDRA A S,GONUGUNTA V K,CHTISTMANN A,et al.ABA perception and signalling[J].Trends in Plant Science,2010,15:395-401.