目的 探究NO调控遮光胁迫下长春花体内生物碱积累的分子机制。方法 以长春花幼苗为材料,遮光胁迫下施加外源NO供体(SNP),7 d后采叶样,研究长春花幼苗叶片的生长形态变化,HPLC测定激素和生物碱的含量变化,荧光定量RT-PCR法分析关键基因的表达。结果 在复合处理条件下,长春花的生长指标显著增高,激素和生物碱的含量明显增加,转录因子Myc1和路径中的关键基因G10h,Tdc,Str表达均明显上调。相关性分析表明,6-BA和ABA含量与Myc1表达呈显著正相关(P<0.01);Myc1表达与Tdc、Str、Sgd和Dat表达呈显著正相关(P<0.01);Tdc、Str、Sgd和Dat表达与生物碱含量呈显著正相关(P<0.01)。结论 外源NO供体(SNP)作为信号分子,在遮光胁迫下调节激素合成的增加,调控转录因子的高表达,导致生物碱代谢路径中关键基因表达量显著上调,促进长春花体内长春碱和长春新碱含量的积累。
Abstract
OBJECTIVE To explore the molecular mechanism of no regulating alkaloid accumulation under light- shading stress. METHODS The seedlings of Catharanthus roseus (C. roseus) were used as material, and the leaves were sampled 7 d later. Leaves growth morphology were studied, the contents of alkaloids and hormones were determined by HPLC and the genes expression were analyzed by fluorescence quantitative RT-PCR. RESULTS Under compound treatment (T3), the growth index of leaves from C. roseus significantly increased, and the content of hormones and alkaloids increased significantly. The expression of Myc1 and key genes (G10h, Tdc, Sgd and Str) in the pathway were significantly up-regulated. Correlation analysis showed that the contents of 6-BA and ABA were positively correlated with Myc1 expression (P<0.01); Myc1 expression was positively correlated with the expression of Tdc, Str, Sgd, and Dat (P<0.01); there was a significant positive correlation between the genes expression and alkaloid content (P<0.01). CONCLUSION The exogenous NO donors (SNP) could act as a signaling molecule and regulate the increase of hormones content under light-shading stress, increase the high expression of transcription factors Myc1, activate the expression of key genes in the metabolic pathway of alkaloids, and increase the content of vinblastine and vincristine in C. roseus.
关键词
长春花 /
生物碱 /
内源激素 /
Myc1转录因子 /
外源NO
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Key words
Catharanthus roseus /
alkaloids /
plant hormones /
Myc1 transcription factors /
exogenous NO
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中图分类号：
R281
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参考文献
[1] WU X Z, XIE H Q, LONG X H, et al. Studies on the chemical constituents of Catharanthus roseus[J]. Chin Pharm J(中国药学杂志), 2017, 52(8):631-636.
[2] ZHENG J, ZHAO S J, HU Z B. Advances in metabolism pathways of indole terpenoid alkaloids in Catharanthus roseus [J]. Chin J Pharm Biotechnol(药物生物技术), 2012, 19(3):278-282.
[3] LI C Y, LEOPOLD A L, SANDER G W, et al. CrBPF1 overexpression alters transcript levels of terpenoid indole alkaloid biosynthetic and regulatory genes [J]. Front Plant Sci, 2015, doi:10.3389/fpls. 2015. 00818.
[4] POLLIER J, VANDEN B R, RISCHER H, et al. Selection and validation of reference genes for transcript normalization in gene expression studies in Catharanthus roseus[J]. Plant Physiol Biochem, 2014,83(10):20-25.
[5] CHATEL G, MONTIEL G, PRÉ M, et al. CrMYC1, a Catharanthus roseus elicitor-and jasmonate-responsive b HLH transcription factor that binds the G-box element of the strictosidine synthase gene promoter [J]. J Exp Bot, 2003, 54(392) : 2587.
[6] GUO H X, LI F X, LIU Q Z, et al. Cloning and functional analysis of NtMYC1a transcription factors [J]. Acta Agricul Jiangxi (江西农业学报), 2016, 28(12):80-82,86.
[7] ZHU M Y, YU B F, CHEN H F. Effects of exogenous nitrate levels on physiological metabolism of Catharanthus roseus[J]. Bull Bot Res (植物研究), 2016, 36(4):535-541.
[8] XIANG P P, ZHU Y R, WANG W J, et al. Effects of salt stress on callus growth and amaline accumulation in Catharanthus roseus [J]. Chin Tradit Herb Drugs (中草药), 2011, 42(1):167-170.
[9] LIU Y, ZHAO D M, ZU Y G, et al.Effects of shading on the content of vincristine and the expression of related synthetic genes in the leaves of Catharanthus roseus [J]. J Huazhong Agricul Univ(华中农业大学学报), 2010, 29(5):634-638.
[10] LIU P L, PAN Y Z, YANG R D, et al. Effects of exogenous nitric oxide on ATPase and mineral nutrient uptake of plasma membrane peroxidation in roseus roseus under cadmium stress [J]. J Plant Nutr Fertil(植物营养与肥料学报), 2014, 20(2):445-458.
[11] XU M J, DONG J F. Interaction between nitric oxide and hydrogen peroxide in mediating alkaloid biosynthesis in Catharanthus roseus cells [J]. Progr Nat Sci (自然科学进展), 2008,18(12):1386-1397.
[12] XIAO Y H, ZHAO X L, WANG K C, et al. Effects of different nitrogen forms and concentrations on biomass and alkaloids in leaves of Daqing [J]. China J Chin Mater Med(中国中药杂志), 2013, 38(17):2755-2760.
[13] YANG L, TANG Z H, ZU Y G. Simultaneous determination of vindropin, catharanthine and anhydrovinblastine in Catharanthus roseus by high performance liquid chromatography [J]. Chin J Chromatogr(色谱), 2007,25(4):550-552.
[14] WU C T. Interaction between nitric oxide and phytohormone and its relationship [J]. Genom Appl Biol (基因组学与应用生物学), 2010, 29(6):1169-1176.
[15] WANG Y H, LIU J J, CHEN B L, et al. Physiological mechanism of 6-BA and ABA in alleviating cold stress of cotton fiber development [J]. Chin J Appl Ecol (应用生态学报), 2011, 22(5):1233-1239.
[16] CHEN S J, ZHANG M, JIN S Y, et al. Effects of GA3、6-BA on growth and alkaloid content of dendrobium nobile [J]. J Plant Resour Environ(植物资源与环境学报), 2004,13(3):7-11.
[17] ZHANG M X, WANG Y Y, YU F. Regulation of biosynthesis of monoterpene indole alkaloids in Catharanthus roseus by abscisic acid [J/OL]. Mol Plant Breed(分子植物育种), 2018-03-14. http://kns.cnki.net/kcms/detail/46. 1068. S. 20180212. 1131. 002. html.
[18] GAO K, WANG L, WU S R, et al. Research progress of transcription factors regulating the synthesis of pharmacokinetic components in medicinal plants [J]. Chin Tradit Herb Drugs (中草药), 2015, 46(20):3100-3108.
[19] ZENG J L, LIU W H, LIAO Z H, et al. Difference analysis of gene expression abundance of biosynthesis pathway related genes and gene expression after MeJA treatment in Catharanthus roseus [J]. Genom Appl Biol(基因组学与应用生物学), 2017, 36(3):1009-1020.
[20] LIU Y, LI D W, MENG Q H, et al. Analysis of alkaloid content and related gene expression in leaves of Catharanthus roseus under PEG6000 stress [J]. Bull Botan Res(植物研究), 2015, 35(5):746-750.
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脚注
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基金
十三五国家重点研发计划资助(2017YFD0600706);中央高校基本科研业务费专项资金项目资助( 2572015CA07,2572015CA03);黑龙江省自然科学基金项目资助(QC2017009)
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