中国中药杂志

2020, v.45(02) 321-330

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盐胁迫下外源NO对颠茄氮代谢及次生代谢调控的研究
Nitrogen metabolism and secondary metabolism regulation of Atropa belladonna by exogenous NO under NaCl stress

代欢欢;杨怡;山雨思;何潇;辛正琦;吴能表;
DAI Huan-huan;YANG Yi;SHAN Yu-si;HE Xiao;XIN Zheng-qi;WU Neng-biao;Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, School of Life Science,Southwest University;Deyang City Lushan Road Primary School;

摘要(Abstract):

以颠茄幼苗为实验材料,采用土培法种植,向叶面喷施不同浓度(0,0.05,0.1,0.2,0.5 mmol·L~(-1))NO供体硝普钠(sodium nitroprusside,SNP),研究在100 mmol·L~(-1) NaCl胁迫下不同浓度SNP、不同处理时间(4,8,12,16 d)对颠茄氮代谢、次生代谢产物含量以及托品烷类生物碱合成途径的前体物质含量、关键酶基因表达量的影响。结果表明,随着盐胁迫时间的延长,颠茄的氮代谢及次生代谢均受到了一定程度的抑制。经不同浓度外源SNP处理,铵态氮含量显著降低,硝态氮、游离氨基酸、可溶性蛋白含量和氮代谢关键酶(NR,GS,GDH)活性显著升高,次生代谢途径中的前体氨基酸及多胺含量均有不同程度的上升;qRT-PCR分析显示,外源SNP处理可有效提高颠茄次生代谢途径中的关键酶基因PMT,TRⅠ,H6H的表达量,从而使莨菪碱和东莨菪碱产量增加。综上所述,外施适宜浓度的SNP能够有效缓解盐胁迫对颠茄氮代谢及次生代谢的抑制作用,提高其耐盐性,当SNP浓度为0.1,0.2 mmol·L~(-1)时效果较为显著。
Atropa belladonna seedlings were used as experimental materials and cultivated by soil culture method. Different concentrations(0,0.05,0.1,0.2,0.5 mmol·L~(-1))of NO donor sodium nitroprusside(SNP) were sprayed on the leaves. The effects of different concentrations of SNP and different treatment time(4,8,12,16 d) on nitrogen metabolism, secondary metabolite content, precursor content of tropane alkaloid synthesis pathway and expression of key enzyme genes under 100 mmol·L~(-1) NaCl stress were studied. The results showed that with the prolongation of salt stress, the nitrogen metabolism and the accumulation of secondary metabolites of A. belladonna were inhibited to some extent. After treatment with different concentrations of exogenous SNP, the ammonium nitrogen content decreased dramatically, and the contents of nitrate nitrogen, free amino acid, soluble protein and the activities of key enzymes of nitrogen metabolism(NR, GS, GDH) were all greatly improved; the contents of precursor amino acids(ornithine, arginine) and polyamines(Put, Spd, Spm) in the secondary metabolic pathway have increased to varying degrees. The qRT-PCR analysis showed that exogenous SNP treatment can effectively promote the high expression of key enzyme genes PMT, TRⅠ and H6H in the secondary metabolic pathway of A. belladonna, and the production of hyoscyamine and scopolamine were increased notably. In summary, the application of appropriate concentration of SNP can effectively alleviate the inhibition of salt stress on the nitrogen metabolism and secondary metabolism of Atropa belladonna, and enhance its salt tolerance. Overall, 0.1 mmol·L~(-1) and 0.2 mmol·L~(-1) SNP treatment achieved the most remarkable effect.

关键词(KeyWords): 颠茄;盐胁迫;外源NO;氮代谢;托品烷类生物碱
Atropa belladonna;NaCl stress;exogenous nitric;nitrogen metabolism;tropane alkaloids

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(30500041);; 重庆市科技攻关计划项目(cstc2012gg-yyjs80013)

作者(Author): 代欢欢;杨怡;山雨思;何潇;辛正琦;吴能表;
DAI Huan-huan;YANG Yi;SHAN Yu-si;HE Xiao;XIN Zheng-qi;WU Neng-biao;Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, School of Life Science,Southwest University;Deyang City Lushan Road Primary School;

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