本文主要研究内容
作者李慧,阚家亮,王影,蔺经,杨青松,常有宏(2019)在《豆梨中谷胱甘肽还原酶基因的分离、表达特点及酶活性分析》一文中研究指出:【目的】测定谷胱甘肽还原酶活性变化,结合其催化产物含量、编码基因表达特点分析,明确该酶是否参与豆梨应对镉胁迫的调控过程。【方法】采用紫外/可见分光光度法检测谷胱甘肽还原酶(glutathione reductase,GR)活性、谷胱甘肽池组成及过氧化氢(hydrogen peroxide,H2O2)含量,RT-PCR和PCR克隆PcGRchl和PcGRcyt的cDNA和DNA序列,利用生物信息学方法进行序列比较分析,荧光定量PCR检测它们在镉胁迫下转录水平变化。【结果】镉胁迫情况下,豆梨叶片GR活性上升,谷胱甘肽池中总谷胱甘肽(total glutathione,T-GSH)和还原型谷胱甘肽(reduced glutathione,GSH)减少、氧化型谷胱甘肽(oxidized glutathione,GSSG)上升,H2O2积累增加。豆梨叶绿体PcGRchl和胞质PcGRcyt的序列长度、基因结构及所编码蛋白特征各不相同,它们在豆梨叶片中的表达量上调以响应镉胁迫信号,以PcGRchl的转录占主导。外源GSH预处理有助于豆梨预先储存GSH,减缓镉胁迫下H2O2的积累,与Cd(2 mmol·L-1CdCl2·2.5H2O+Hoagland营养液)组相比较,GC(2 mmol·L-1,GSH预处理12 h转入2 mmol·L-1CdCl2·2.5H2O+Hoagland营养液)组PcGRchl和PcGRcyt的表达水平没有太大变化,但GR活性部分受抑制;外源BSO预处理抑制植株叶片GSH合成,镉胁迫下BC(2 mmol·L-1丁硫氨酸-亚砜亚胺预处理12 h转入2 mmol·L-1CdCl2·2.5H2O+Hoagland营养液)组H2O2产生加剧,GR活性上升幅度变大,但PcGRchl和PcGRcyt的转录不受影响。上述结果表明,GSH预处理或BSO预处理,可改变豆梨叶片GSH池的组成,从而在蛋白质水平上反馈调节镉胁迫情况下GR活性。【结论】豆梨GR参与应对镉胁迫的调控过程。镉处理后,豆梨叶片中GSH减少,促使GR酶活性上升,以补充植株应对逆境所需GSH,这一过程主要通过叶绿体PcGRchl的转录上调来实现;GSH或BSO预处理,改变植株GSH含量,从而影响GR活性,减缓或加剧镉胁迫下H2O2的产生,这一过程主要是在GR基因翻译后的蛋白水平上进行反馈调节。
Abstract
【mu de 】ce ding gu guang gan tai hai yuan mei huo xing bian hua ,jie ge ji cui hua chan wu han liang 、bian ma ji yin biao da te dian fen xi ,ming que gai mei shi fou can yu dou li ying dui ge xie pai de diao kong guo cheng 。【fang fa 】cai yong zi wai /ke jian fen guang guang du fa jian ce gu guang gan tai hai yuan mei (glutathione reductase,GR)huo xing 、gu guang gan tai chi zu cheng ji guo yang hua qing (hydrogen peroxide,H2O2)han liang ,RT-PCRhe PCRke long PcGRchlhe PcGRcytde cDNAhe DNAxu lie ,li yong sheng wu xin xi xue fang fa jin hang xu lie bi jiao fen xi ,ying guang ding liang PCRjian ce ta men zai ge xie pai xia zhuai lu shui ping bian hua 。【jie guo 】ge xie pai qing kuang xia ,dou li xie pian GRhuo xing shang sheng ,gu guang gan tai chi zhong zong gu guang gan tai (total glutathione,T-GSH)he hai yuan xing gu guang gan tai (reduced glutathione,GSH)jian shao 、yang hua xing gu guang gan tai (oxidized glutathione,GSSG)shang sheng ,H2O2ji lei zeng jia 。dou li xie lu ti PcGRchlhe bao zhi PcGRcytde xu lie chang du 、ji yin jie gou ji suo bian ma dan bai te zheng ge bu xiang tong ,ta men zai dou li xie pian zhong de biao da liang shang diao yi xiang ying ge xie pai xin hao ,yi PcGRchlde zhuai lu zhan zhu dao 。wai yuan GSHyu chu li you zhu yu dou li yu xian chu cun GSH,jian huan ge xie pai xia H2O2de ji lei ,yu Cd(2 mmol·L-1CdCl2·2.5H2O+Hoaglandying yang ye )zu xiang bi jiao ,GC(2 mmol·L-1,GSHyu chu li 12 hzhuai ru 2 mmol·L-1CdCl2·2.5H2O+Hoaglandying yang ye )zu PcGRchlhe PcGRcytde biao da shui ping mei you tai da bian hua ,dan GRhuo xing bu fen shou yi zhi ;wai yuan BSOyu chu li yi zhi zhi zhu xie pian GSHge cheng ,ge xie pai xia BC(2 mmol·L-1ding liu an suan -ya feng ya an yu chu li 12 hzhuai ru 2 mmol·L-1CdCl2·2.5H2O+Hoaglandying yang ye )zu H2O2chan sheng jia ju ,GRhuo xing shang sheng fu du bian da ,dan PcGRchlhe PcGRcytde zhuai lu bu shou ying xiang 。shang shu jie guo biao ming ,GSHyu chu li huo BSOyu chu li ,ke gai bian dou li xie pian GSHchi de zu cheng ,cong er zai dan bai zhi shui ping shang fan kui diao jie ge xie pai qing kuang xia GRhuo xing 。【jie lun 】dou li GRcan yu ying dui ge xie pai de diao kong guo cheng 。ge chu li hou ,dou li xie pian zhong GSHjian shao ,cu shi GRmei huo xing shang sheng ,yi bu chong zhi zhu ying dui ni jing suo xu GSH,zhe yi guo cheng zhu yao tong guo xie lu ti PcGRchlde zhuai lu shang diao lai shi xian ;GSHhuo BSOyu chu li ,gai bian zhi zhu GSHhan liang ,cong er ying xiang GRhuo xing ,jian huan huo jia ju ge xie pai xia H2O2de chan sheng ,zhe yi guo cheng zhu yao shi zai GRji yin fan yi hou de dan bai shui ping shang jin hang fan kui diao jie 。
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论文作者分别是来自果树学报的李慧,阚家亮,王影,蔺经,杨青松,常有宏,发表于刊物果树学报2019年01期论文,是一篇关于豆梨论文,镉胁迫论文,谷胱甘肽还原酶论文,活性调节论文,表达特点论文,果树学报2019年01期论文的文章。本文可供学术参考使用,各位学者可以免费参考阅读下载,文章观点不代表本站观点,资料来自果树学报2019年01期论文网站,若本站收录的文献无意侵犯了您的著作版权,请联系我们删除。