茶氨酸诱导烟草抵御低钾胁迫的研究
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摘要
通过施用茶氨酸来减轻低钾胁迫对烟草幼苗的毒害,并从抗氧化系统、钾离子含量及相关基因来探讨其作用机理。结果表明,在低钾胁迫下,0.1和0.2 mmol·L-1茶氨酸均能促进烟苗根长、鲜重及叶绿素含量的显著增加;外源茶氨酸预处理不仅能显著提高低钾胁迫下烟草幼苗脯氨酸、谷胱甘肽(GSH)含量,还能增强烟草幼苗抗坏血酸还原酶(APX)、过氧化物酶(POD)和过氧化氢酶(CAT)活性,降低过氧化氢(H2O2)和丙二醛(MDA)含量。此外,茶氨酸还能促进烟草幼苗对钾离子的吸收。半定量PCR结果表明,茶氨酸预处理可以诱导低钾胁迫下烟草幼苗Nt LKS1、Nt KC和Nt KT1上调。综上,茶氨酸缓解烟草幼苗低钾胁迫是通过激发植物体抗氧化系统、促进钾离子的吸收及相关基因上调的综合结果。
To study the role of theanine in protecting tobacco seedlings against low potassium stress, the antioxidant system, the potassium content and the transcription of potassium uptake-related genes of tobacco seedlings were investigated. The results showed that both 0.1 and 0.2 mmol·L-1 theanine could effectively improve root length, fresh weight and the chlorophyll content of tobacco under low potassium stress. Meanwhile, physiological-biochemical indices analysis indicated that theanine enhanced the contents of glutathione(GSH), proline as well as the activities of antioxidant enzymes, such as ascorbate peroxidase(APX), peroxidase(POD) and catalase(CAT), but theanine lowered the content of malondialdehyde(MDA) and the hydrogen peroxide(H2O2) level than that under low potassium stress alone. Moreover, theanine significantly increased the potassium content under low potassium stress. Furthermore, semi-quantitative RT-PCR(Reverse transcription-PCR) analysis demonstrated that the transcript levels of Nt LKS1, Nt KCand Nt KT1 were up-regulated by theanine treatment under low potassium stress, while Nt HAK1 displayed no difference between the low potassium-untreated and treated groups. Taken together, it could be concluded that theanine is beneficial to relieving low potassium stress on tobacco seedlings, which is comprehensive consequence of stimulating antioxidant system, increasing the content of potassium and up-regulating the expression of Nt LKS1, Nt KC and Nt KT1.
To study the role of theanine in protecting tobacco seedlings against low potassium stress, the antioxidant system, the potassium content and the transcription of potassium uptake-related genes of tobacco seedlings were investigated. The results showed that both 0.1 and 0.2 mmol·L-1 theanine could effectively improve root length, fresh weight and the chlorophyll content of tobacco under low potassium stress. Meanwhile, physiological-biochemical indices analysis indicated that theanine enhanced the contents of glutathione(GSH), proline as well as the activities of antioxidant enzymes, such as ascorbate peroxidase(APX), peroxidase(POD) and catalase(CAT), but theanine lowered the content of malondialdehyde(MDA) and the hydrogen peroxide(H2O2) level than that under low potassium stress alone. Moreover, theanine significantly increased the potassium content under low potassium stress. Furthermore, semi-quantitative RT-PCR(Reverse transcription-PCR) analysis demonstrated that the transcript levels of Nt LKS1, Nt KCand Nt KT1 were up-regulated by theanine treatment under low potassium stress, while Nt HAK1 displayed no difference between the low potassium-untreated and treated groups. Taken together, it could be concluded that theanine is beneficial to relieving low potassium stress on tobacco seedlings, which is comprehensive consequence of stimulating antioxidant system, increasing the content of potassium and up-regulating the expression of Nt LKS1, Nt KC and Nt KT1.
引文
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[23]梁艳荣,胡晓红,张颍力,等.植物过氧化物酶生理功能研究进展[J].内蒙古农业大学学报(自然科学版),2003,24(2):110-113.
[24]LIN Y F,AARTS M G.The molecular mechanism of zinc and cadmium stress response in plants[J].Cellular and molecular life sciences:CMLS,2012,69(19):3187-3206.
[25]GUAN Z,CHAI T,ZHANG Y,et al.Enhancement of Cd tolerance in transgenic tobacco plants overexpressing a Cd-induced catalase c DNA[J].Chemosphere,2009,76(5):623-630.
[26]AHMAD P,SARWAT M,SHARMA S.Reactive oxygen species,antioxidants and signaling in plants[J].Journal of Plant Biology,2008,51(3):167-173.
[27]MARTINEZ-CORDERO M A,MARTINEZ V,RUBIO F.Cloning and functional characterization of the high-affinity K+transporter HAK1 of pepper[J].Plant Molecular Biology,2004,56:413-421.
[28]NIEVES CORDONES M,MILLER A J,ALEMAN F,et al.A putative role for the plasma membrane potential[J].Plant Mol Biol,2008,68:521-532.
[29]卢云峰.β-氨基丁酸诱导烟草抗低钾胁迫和抗TMV侵染的研究[D].合肥:合肥工业大学,2012.
[2]MAATHUIS F J.Physiological functions of mineral macronutrients[J].Current Opinion in Plant Biology,2009,12(3):250-258.
[3]彭冠云,周清明,易克,等.烤烟钾素营养研究进展[J].作物研究,2005(5):386-389
[4]汪波,翟曙光.钾盐在烟草燃烧过程中的作用[J].中国科学技术大学学报,2002,32(4):433-439.
[5]MENGEL K,KIRKBY E A.Potassium.Principles of plant nutrition[R].Kluwer,Norwell,2001:503-509
[6]LIU Q,DUAN H,LUAN J,et al.Effects of theanine on growth of human lung cancer and leukemia cells as well as migration and invasion of human lung cancer cells[J].Cytotechnology,2009,59(3):211-217.
[7]YOKOGOSHI H,KATO Y,SAGESAKA Y M,et al.Reduction effect of theanine on blood pressure and brain5-hydroxyindoles in spontaneously hypertensive rats[J].Bioscience,Biotechnology and Biochemistry,1995,59(4):615-618.
[8]ZHENG G,SAYAMA K,OKUBO T,et al.Anti-obesity effects of three major components of green tea,catechins,caffeine and theanine,in mice[J].In Vivo,2004,18(1):55-62.
[9]MIYAGAWA K,HAYASHI Y,KURIHARA S,et al.Co-administration of l-cystine and l-theanine enhances efficacy of influenza vaccination in elderly persons:Nutritional status-dependent immunogenicity[J].Geriatrics&Gerontology International,2008,8(4):243-250.
[10]HIRSCH R E.A role for the AKT1 potassium channel in plant nutrition[J].Science,1998,280(5365):918-921
[11]朱广廉.植物生理学[M].北京:北京大学出版社,1990.
[12]MICHAEL P I,KRISHNASWAMY M.The effect of zinc stress combined with high irradiance stress on membrane damage and antioxidative response in bean seedlings[J].Environmental and Experimental Botany,2011,74:171-177.
[13]STEWART R R C,BEWLEY J D.Lipid peroxidation associated with accelerated aging of soybean axes[J].Plant physiology,1980,65(2):245-248
[14]邹琦.植物生理学实验指导[M].北京:中国农业出版社,2000
[15]RAMAKRISHNA B,RAO S S R.24-Epibrassinolide alleviated zinc-induced oxidative stress in radish(Raphanus sativus L.)seedlings by enhancing antioxidative system[J].Plant Growth Regul,2012,68(2):249-259.
[16]BEWLEY J D.Physiological aspects of desiccation tolerance-a retrospect[J].Int J Plant Sci,1995:393-403.
[17]NAKANO Y,ASADA K.Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts[J].Plant Cell Physiol,1981,22(5):867-880.
[18]APEL K,HIRT H.Reactive oxygen species:metabolism,oxidative stress,and signal transduction[J].Annu Rev Plant Biol,2004,55:373-399.
[19]NOCTOR G,FOYER C H.Ascorbate and glutathione:keeping active oxygen under control[J].Annual Review of Plant Biology,1998,49(1):249-279.
[20]SIRIPORNADULSIL S,TRAINA S,VERMA D P S,et al.Molecular mechanisms of proline-mediated tolerance to toxic heavy metals in transgenic microalgae[J].The Plant Cell,2002,14(11):2837-2847.
[21]王艺霖.提高烟草氮钾营养利用率的遗传操作及烟草低钾响应基因的鉴定[D].昆明:昆明理工大学,2013.
[22]薛艳茹,曹敏建,王晓光,等.低钾胁迫下玉米逆境反应机制初探[J].玉米科学,2008,16(2):101-103.
[23]梁艳荣,胡晓红,张颍力,等.植物过氧化物酶生理功能研究进展[J].内蒙古农业大学学报(自然科学版),2003,24(2):110-113.
[24]LIN Y F,AARTS M G.The molecular mechanism of zinc and cadmium stress response in plants[J].Cellular and molecular life sciences:CMLS,2012,69(19):3187-3206.
[25]GUAN Z,CHAI T,ZHANG Y,et al.Enhancement of Cd tolerance in transgenic tobacco plants overexpressing a Cd-induced catalase c DNA[J].Chemosphere,2009,76(5):623-630.
[26]AHMAD P,SARWAT M,SHARMA S.Reactive oxygen species,antioxidants and signaling in plants[J].Journal of Plant Biology,2008,51(3):167-173.
[27]MARTINEZ-CORDERO M A,MARTINEZ V,RUBIO F.Cloning and functional characterization of the high-affinity K+transporter HAK1 of pepper[J].Plant Molecular Biology,2004,56:413-421.
[28]NIEVES CORDONES M,MILLER A J,ALEMAN F,et al.A putative role for the plasma membrane potential[J].Plant Mol Biol,2008,68:521-532.
[29]卢云峰.β-氨基丁酸诱导烟草抗低钾胁迫和抗TMV侵染的研究[D].合肥:合肥工业大学,2012.