低温条件下小麦返白系叶片中H_2O_2累积的原因初探
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摘要
为了研究低温处理下小麦叶色阶段性白化品种返白系与对照品种矮变1号叶片的H_2O_2含量是否存在差异,并初步分析导致差异的原因,采用DAB组织染色法检测返白系和矮变1号在4℃低温处理90 d、25℃下恢复培养10 d时叶片H_2O_2含量的动态变化,测定4℃低温处理下返白系和矮变1号中表征光合电子传递效率的荧光参数F_v/F_m和qP,并采用qRT-PCR方法分析返白系和矮变1号中几个光合电子传递体基因及质体H_2O_2清除相关基因的表达模式。结果显示,低温处理下,矮变1号叶片中H_2O_2含量变化不大,返白系叶片中H_2O_2会大量累积;低温处理下,返白系叶片的叶绿素荧光参数F_v/F_m、qP都明显低于矮变1号。同时,低温下返白系光合电子传递链中部分电子传递体亚基基因petD、petN、ndhB和ndhK,以及质体H_2O_2清除相关基因 APX4和 HO1的表达量低于矮变1号。这些结果表明,低温处理下,相较于矮变1号,返白系叶片中会累积大量的H_2O_2。返白系质体中光合电子传递受阻引起电子泄露,同时质体中清除活性氧的能力下降,可能是导致返白系叶片中积累大量H_2O_2的原因。
Wheat Albinism line,discovered from winter wheat Aibian 1, shows stage albinism phenomena from interior leaves under low temperature(4 ℃) for about 35 to 40 days, and then returns green when temperature rises. In order to study whether there is difference in leaf H_2O_2 content between Albinism line and Aibian 1 under low temperature, and analyze the reasons, the content of H_2O_2 was detected using the diaminobenzidine histochemical staining technique after low temperature treatment, and the chlorophyll fluorescence parameters F_v/F_m and qP of wheat leaves were measured by chlorophyll fluorescence imaging system, and the expression of photosynthetic electron transport related genes and elimination of reactive oxygen species related genes were analyzed. It was indicated that Albinism line accumulated much more H_2O_2 under low temperature compared with that in Aibian 1.The chlorophyll fluorescence parameters F_v/F_m and qP in wheat Albinism line that represent photosynthetic electron transfer capacity were significantly lower than those of Aibian 1 in low temperature. The expression levels of petD, petN, ndhB, and ndhK, the key genes correlated with electron transfer, and APX4 and 1 the key genes related to elimination of excessive H_2O_2 of plastids, were significantly declined in Albinism line compared with those in Aibian 1 under low temperature.To conclude, the electronic leakage caused by the destroy of photosynthetic electron transfer capacity is the main reason for cold-sensitive wheat Albinism line accumulating excess H_2O_2 compared with Aibian 1 under low temperature. This situation was exacerbated by inadequate elimination capacity of excessive H_2O_2 of plastids in Albinism line.
Wheat Albinism line,discovered from winter wheat Aibian 1, shows stage albinism phenomena from interior leaves under low temperature(4 ℃) for about 35 to 40 days, and then returns green when temperature rises. In order to study whether there is difference in leaf H_2O_2 content between Albinism line and Aibian 1 under low temperature, and analyze the reasons, the content of H_2O_2 was detected using the diaminobenzidine histochemical staining technique after low temperature treatment, and the chlorophyll fluorescence parameters F_v/F_m and qP of wheat leaves were measured by chlorophyll fluorescence imaging system, and the expression of photosynthetic electron transport related genes and elimination of reactive oxygen species related genes were analyzed. It was indicated that Albinism line accumulated much more H_2O_2 under low temperature compared with that in Aibian 1.The chlorophyll fluorescence parameters F_v/F_m and qP in wheat Albinism line that represent photosynthetic electron transfer capacity were significantly lower than those of Aibian 1 in low temperature. The expression levels of petD, petN, ndhB, and ndhK, the key genes correlated with electron transfer, and APX4 and 1 the key genes related to elimination of excessive H_2O_2 of plastids, were significantly declined in Albinism line compared with those in Aibian 1 under low temperature.To conclude, the electronic leakage caused by the destroy of photosynthetic electron transfer capacity is the main reason for cold-sensitive wheat Albinism line accumulating excess H_2O_2 compared with Aibian 1 under low temperature. This situation was exacerbated by inadequate elimination capacity of excessive H_2O_2 of plastids in Albinism line.
引文
[1]GABBITA S P,ROBINSON K A,STEWART C A,et al.Redox regulatory mechanisms of cellular signal transduction [J].Archives of Biochemistry and Biophysics,2000,376(1):1.
[2]HANCOCK J T,DESIKAN R,NEILL S J.Role of reactive oxygen species in cell signalling pathways [J].Biochemical Society Transactions,2001,29(2):345.
[3]GECHEV T S,VAN BREUSEGEM F,STONE J M,et al.Reactive oxygen species as signals that modulate plant stress responses and programmed cell death [J].Bioessays,2006,28(11):1092.
[4]WEN F,XING D,ZHANG L R.Hydrogen peroxide is involved in high blue light-induced chloroplast avoidance movements in Arabidopsis [J].Journal of Experimental Botany,2008,59(10):2891.
[5]MANO J.Reactive carbonyl species:Their production from lipid peroxides,action in environmental stress,and the detoxification mechanism [J].Plant Physiology and Biochemistry,2012,59(2):92.
[6]ASADA K.The water-water cycle in chloroplasts:Scavenging of active oxygens and dissipation of excess photons [J].Annual Review of Plant Biology,1999,50(50):602.
[7]QUAN L J,ZHANG B,SHI W W,et al.Hydrogen peroxide in plants:A versatile molecule of the reactive oxygen species network [J].Journal of Integrative Plant Biology,2008,50(1):3.
[8]MELHER A H.Studies on reactions of illuminated chloroplasts.Ⅱ.Stimulation and inhibition of the reaction with molecular oxygen [J].Archives of Biochemistry and Biophysics,1951,34(2):339.
[9]林植芳,李双顺,林桂珠,等.衰老叶片和叶绿体中H2O2的累积与膜脂过氧化的关系[J].植物生理学报,1988,14(1):20. LIN Z F,LI S S,LIN G Z,et al.The accumulation of hydrogen peroxide in senescing leaves and chloroplasts in relation to lipid peroxidation[J].Acta Phytophysiolgica Sinica,1988,14(1):20.
[10]OLSEN L F,ISSINGER O G,GUERRA B.The Yin and Yang of redox regulation[J].Redox Report,2013,18(6):247.
[11]WANG Y Y,HECKER A G,HAUSER B A.The APX4 locus regulates seed vigor and seedling growth in Arabiodopsis thaliana [J].Planta,2014,239(4):909.
[12]LUNDBERG E,STORM P,SCHROER W P,et al.Crystal structure of the TL29 protein from Arabidopsis thaliana:An APX homolog without peroxidase activity[J].Journal of Structural Biology,2011,176(1):24.
[13]CHEN X Y,DING X,XU S,et al.Endogenous hydrogen peroxide plays a positive role in the up-regulation of hemeoxygenase and acclimation to oxidative stress in wheat seedling leaves [J].Journal of Integrative Plant Biology,2009,51(10):952.
[14]XU D K,JIN Q J,XIE Y J,et al.Characterization of a wheat heme oxygenase-1 gene and its responses to different abiotic stresses [J].International Journal of Molecular Sciences,2011,2(11):7693.
[15]苏小静,汪沛洪.小麦突变体返白系返白机理的研究-Ⅰ.返白阶段叶绿体超微结构观察[J].西北农业大学学报,1990,18(2):73. SU X J,WANG P H.Studies on the albescent mechanism in the mutant stage Albinism line of the winter wheat -Ⅰ.Observation of the ultrastructure of chloroplast druingalbescent stage [J].Acta University AgriculturaeBoreali-occidentalis,1990,18(2):73.
[16]赵亚华,郭蔼光,汪沛洪.小麦返白系与不同生态类型小麦品种杂交后代的遗传特性[J].宁夏农学院学报,1995,16(2):50. ZHAO Y H,GUO A G,WANG P H.The genetic feature of the hybrid progenies of the stage Albinism lines and cultivars of wheat of different ecological type [J].Journal of Ningxia Agricultural College,1995,16(2):50.
[17]陈贵松,张静雅,苏适,等.低温、外源ABA和Ca+对小麦返白系质体基因表达的影响[J].麦类作物学报,2014,34(9):1176. CHEN G S,ZHANG J Y,SU S,et al.The effect of low temperature,ABA and Ca+ on the expression of plastid genes in wheat albino line [J].Journal of Triticeae Crops,2014,34(9):1176.
[18]蒋明义,杨文英,徐江,等.渗透胁迫下水稻幼苗中叶绿素降解的活性氧损伤作用[J].植物学报,1994,36(4):291. JIANG M Y,YANG W Y,XU J,et al.Active oxygen damage effect of chlorophyll degradation in rice seedlings under osmotic stress [J].Acta Botanica Sinica,1994,36(4):291.
[19]孙锦,贾永霞,郭世荣,等.海水胁迫对菠菜(Spinaciaolerancea L.)叶绿体活性氧和叶绿素代谢的影响[J].生态学报,2009,29(8):4366. SUN J,JIA Y X,GUO S R,et al.Effects of seawater stress on metabolism of reactive oxygen species and chlorophyll in chloroplasts of spinach(Spinaciaolerancea L.) [J].Acta Ecologica Sinica,2009,29(8):4366.
[20]VAN H P,VAN B H.Photooxidative damage to the photosynthetic apparatus during chilling [J].Physiologia Plantarum,1980,50(1):52.
[21]何洁,刘鸿先,王以柔,等.低温与植物的光合作用[J].植物生理学通讯,1986(2):3. HE J,LIU H X,WANG Y R,et al.Low temperature and photosynthesis of plants [J].Plant Physiology Communications,1986(2):3.
[22]薛娴,许会敏,吴鸿洋,等.植物光合作用循环电子传递的研究进展[J].植物生理学报,2017,53(2):145. XUE X ,XU H M,WU H Y,et al.Research progress of cyclic electron transport in plantphotosynthesis[J].Plant Physiology Journal,2017,53(2):145.
[23]MURGIA I,TARANTINO D,VANNINI C,et al.Arabidopsis thaliana plants overexpressing thylakoidalascorbate peroxidase show increased resistance to paraquat-induced photooxidative stress and to nitric oxide-induced cell death[J].The Plant Journal,2004,38(6):940.
[2]HANCOCK J T,DESIKAN R,NEILL S J.Role of reactive oxygen species in cell signalling pathways [J].Biochemical Society Transactions,2001,29(2):345.
[3]GECHEV T S,VAN BREUSEGEM F,STONE J M,et al.Reactive oxygen species as signals that modulate plant stress responses and programmed cell death [J].Bioessays,2006,28(11):1092.
[4]WEN F,XING D,ZHANG L R.Hydrogen peroxide is involved in high blue light-induced chloroplast avoidance movements in Arabidopsis [J].Journal of Experimental Botany,2008,59(10):2891.
[5]MANO J.Reactive carbonyl species:Their production from lipid peroxides,action in environmental stress,and the detoxification mechanism [J].Plant Physiology and Biochemistry,2012,59(2):92.
[6]ASADA K.The water-water cycle in chloroplasts:Scavenging of active oxygens and dissipation of excess photons [J].Annual Review of Plant Biology,1999,50(50):602.
[7]QUAN L J,ZHANG B,SHI W W,et al.Hydrogen peroxide in plants:A versatile molecule of the reactive oxygen species network [J].Journal of Integrative Plant Biology,2008,50(1):3.
[8]MELHER A H.Studies on reactions of illuminated chloroplasts.Ⅱ.Stimulation and inhibition of the reaction with molecular oxygen [J].Archives of Biochemistry and Biophysics,1951,34(2):339.
[9]林植芳,李双顺,林桂珠,等.衰老叶片和叶绿体中H2O2的累积与膜脂过氧化的关系[J].植物生理学报,1988,14(1):20. LIN Z F,LI S S,LIN G Z,et al.The accumulation of hydrogen peroxide in senescing leaves and chloroplasts in relation to lipid peroxidation[J].Acta Phytophysiolgica Sinica,1988,14(1):20.
[10]OLSEN L F,ISSINGER O G,GUERRA B.The Yin and Yang of redox regulation[J].Redox Report,2013,18(6):247.
[11]WANG Y Y,HECKER A G,HAUSER B A.The APX4 locus regulates seed vigor and seedling growth in Arabiodopsis thaliana [J].Planta,2014,239(4):909.
[12]LUNDBERG E,STORM P,SCHROER W P,et al.Crystal structure of the TL29 protein from Arabidopsis thaliana:An APX homolog without peroxidase activity[J].Journal of Structural Biology,2011,176(1):24.
[13]CHEN X Y,DING X,XU S,et al.Endogenous hydrogen peroxide plays a positive role in the up-regulation of hemeoxygenase and acclimation to oxidative stress in wheat seedling leaves [J].Journal of Integrative Plant Biology,2009,51(10):952.
[14]XU D K,JIN Q J,XIE Y J,et al.Characterization of a wheat heme oxygenase-1 gene and its responses to different abiotic stresses [J].International Journal of Molecular Sciences,2011,2(11):7693.
[15]苏小静,汪沛洪.小麦突变体返白系返白机理的研究-Ⅰ.返白阶段叶绿体超微结构观察[J].西北农业大学学报,1990,18(2):73. SU X J,WANG P H.Studies on the albescent mechanism in the mutant stage Albinism line of the winter wheat -Ⅰ.Observation of the ultrastructure of chloroplast druingalbescent stage [J].Acta University AgriculturaeBoreali-occidentalis,1990,18(2):73.
[16]赵亚华,郭蔼光,汪沛洪.小麦返白系与不同生态类型小麦品种杂交后代的遗传特性[J].宁夏农学院学报,1995,16(2):50. ZHAO Y H,GUO A G,WANG P H.The genetic feature of the hybrid progenies of the stage Albinism lines and cultivars of wheat of different ecological type [J].Journal of Ningxia Agricultural College,1995,16(2):50.
[17]陈贵松,张静雅,苏适,等.低温、外源ABA和Ca+对小麦返白系质体基因表达的影响[J].麦类作物学报,2014,34(9):1176. CHEN G S,ZHANG J Y,SU S,et al.The effect of low temperature,ABA and Ca+ on the expression of plastid genes in wheat albino line [J].Journal of Triticeae Crops,2014,34(9):1176.
[18]蒋明义,杨文英,徐江,等.渗透胁迫下水稻幼苗中叶绿素降解的活性氧损伤作用[J].植物学报,1994,36(4):291. JIANG M Y,YANG W Y,XU J,et al.Active oxygen damage effect of chlorophyll degradation in rice seedlings under osmotic stress [J].Acta Botanica Sinica,1994,36(4):291.
[19]孙锦,贾永霞,郭世荣,等.海水胁迫对菠菜(Spinaciaolerancea L.)叶绿体活性氧和叶绿素代谢的影响[J].生态学报,2009,29(8):4366. SUN J,JIA Y X,GUO S R,et al.Effects of seawater stress on metabolism of reactive oxygen species and chlorophyll in chloroplasts of spinach(Spinaciaolerancea L.) [J].Acta Ecologica Sinica,2009,29(8):4366.
[20]VAN H P,VAN B H.Photooxidative damage to the photosynthetic apparatus during chilling [J].Physiologia Plantarum,1980,50(1):52.
[21]何洁,刘鸿先,王以柔,等.低温与植物的光合作用[J].植物生理学通讯,1986(2):3. HE J,LIU H X,WANG Y R,et al.Low temperature and photosynthesis of plants [J].Plant Physiology Communications,1986(2):3.
[22]薛娴,许会敏,吴鸿洋,等.植物光合作用循环电子传递的研究进展[J].植物生理学报,2017,53(2):145. XUE X ,XU H M,WU H Y,et al.Research progress of cyclic electron transport in plantphotosynthesis[J].Plant Physiology Journal,2017,53(2):145.
[23]MURGIA I,TARANTINO D,VANNINI C,et al.Arabidopsis thaliana plants overexpressing thylakoidalascorbate peroxidase show increased resistance to paraquat-induced photooxidative stress and to nitric oxide-induced cell death[J].The Plant Journal,2004,38(6):940.