Effects of trace elements on the expression of LjHCT and LjC3H1 for chlorogenic acid synthesis in Lonicera japonica Thunb.
-
摘要: 绿原酸是金银花（Lonicera japonica Thunb.）入药的主要化学成分，如何稳定和提高绿原酸的含量是近年来金银花研究的热点。本研究通过对金银花分别根施Fe、B和Mo3种微量元素，比较处理前、后金银花中微量元素与绿原酸含量的变化，并定量分析3种微量元素对金银花绿原酸合成关键酶基因LjHCT和LjC3H1表达的影响。研究结果显示，高浓度的Fe处理对LjHCT基因的表达有明显的抑制作用，而中、低浓度的Fe处理可以促进LjHCT基因的表达；随着B和Mo元素浓度的提高，LjHCT基因的表达量也逐渐增加。低浓度的Fe处理可以促进LjC3H1基因的表达，而高浓度的Fe处理对LjC3H1基因的表达具有抑制作用；B元素对LjC3H1基因表达无显著影响，而高浓度的Mo处理可以促进LjC3H1基因的表达。根施中、低浓度的Fe元素，中、高浓度的Mo和B元素后金银花绿原酸含量显著增加；而根施高浓度的Fe元素后金银花中绿原酸含量显著减少。研究结果表明微量元素Fe、B和Mo可通过调节绿原酸生物合成关键酶基因的表达从而有效促进绿原酸的形成和积累。本研究为人工定向调控金银花绿原酸含量、开发人工栽培金银花专用微量元素肥料提供了理论依据。Abstract: Chlorogenic acid is the main chemical constituent of Lonicera japonica Thunb., and is responsible for various pharmacological activities. Therefore, increases in the chlorogenic acid content in L. japonica flowers is economically and medically important. This study aimed to characterize the effects of applying three micro-elements (Fe, B and Mo) on hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (LjHCT) and p-coumaroyl-shikimate 3'-hydroxylase (LjC3H1) genes in L. japonica. Results showed that high Fe concentration had an obvious inhibitory effect on LjHCT gene expression, whereas low and moderate concentrations promoted LjHCT gene expression. With the increase in B and Mo concentrations, the expression of LjHCT also increased significantly. Low Fe concentration promoted LjC3H1 gene expression, whereas high Fe concentration exhibited an inhibitory effect. The effect of B on LjC3H1 expression was not obvious, but high Mo concentration promoted LjC3H1 gene expression. These results showed that chlorogenic acid biosynthesis and accumulation increased with increasing LjHCT and LjC3H1 expressions, which were two key enzymes in the synthesis of chlorogenic acid, and Fe, B and Mo concentrations had significant effects on LjHCT and LjC3H1 expression. Application of moderate or low concentrations of Fe, high or moderate concentrations of Mo, and high or moderate concentrations of B significantly increased the accumulation of chlorogenic acid in L. japonica flowers; however, the application of high concentrations of Fe significantly decreased accumulation. This study provides research-based data for improving the cultivation of L. japonica by increasing the use of trace element fertilizer.
-
Keywords:
- Lonicera japonica Thunb. /
- Trace elements /
- Chlorogenic acid /
- LjHCT /
- LjC3H1
-
-
[1] Lu HX, Zhang L, Huang H. Study on the isolation of active constituents in Lonicera japonica and the mechanism of their anti-upper respiratory tract infection action in children[J]. Afr Health Sci, 2015, 15(4):1295-1301.
[2] Wu J, Wang XC, Liu Y, Du H, Shu QY, Su S, Wang LJ, Li SS, Wang LS. Flavone synthases from Lonicera japonica and L. macranthoides reveal differential flavone accumulation[J]. Sci Rep, 2016, 10(6):1-14.
[3] Lieurance D, Cipollini D. Environmental influences on growth and defence responses of the invasive shrub, Lonicera maackii, to simulated and real herbivory in the juvenile stage[J]. Ann Bot, 2013, 112(4):741-749.
[4] 姜南辉. 金银花化学成分研究[J]. 中药材, 2015, 38(2):315-316. Jiang NH. Study on chemical constituents of Lonicera japonica bud[J]. Journal of Chinese Medicinal Materials, 2015, 38(2):315-316.
[5] Liu ZX, Cheng ZY, He QJ, Lin B, Gao PY, Li LZ, Liu QB, Song SJ. Secondary metabolites from the flower buds of Lonicera japonica and their in vitro anti-diabetic activities[J]. Fitoterapia, 2016, 110:44-51.
[6] 蒋向辉. 金银花绿原酸合成途径关键酶基因克隆与功能分析[D]. 长沙:湖南大学, 2013. Jiang XH. Cloning and function analysis of chlorogenic acids biosynthetic pathway key genes from Lonicera japonica Thunb.[D].Changsha:Hunan university, 2013.
[7] 李津婴. 必需微量金属元素在基因表达中的调控作用[J]. 国外医学卫生学分册, 1996, 23(2):270-274. Li JY. Necessary trace metal elements in regulation of gene expression[J]. Foreign Medical Sciences (Section of Hygiene), 1996, 23(2):270-274.
[8] Lane DJ, Chikhani S, Richardson V, Richardson DR. Transferrin iron uptake is stimulated by ascorbate via an intracellular reductive mechanism[J]. BBA Mol Cell Res, 2013, 1833(6):1527-1541.
[9] 梁新华. 微量元素对甘草中甘草酸形成与积累影响的研究[D]. 北京:北京林业大学, 2010. Liang XH. Effect of micro-elements on biosynthesis and accmulation of glycyrrhizia acid in Glycyrrhiza uralensis Fisch[D]. Beijing:Beijing forestry university, 2010. [10] 李琰, 崔蕾, 雷嘉敏, 李群, 张兴. 不同浓度有机物对雷公藤不定根生长和次生代谢产物含量的影响[J]. 植物科学学报, 2014, 32(2):174-180. Li Y, Cui L, Lei JM, Li Q, Zhang X. Effects of different concentrations of organic affixture on the growth and secondary metabolites contents in adventitious roots of Tripterygium wilfordii[J]. Plant Science Journal, 2014, 32(2):174-180.
[11] 莫尔维德特. 农业中的微量营养元素[M]. 北京:农业出版社, 1984. Mortvedt. Trace Nutrient Elements in Agriculture[M]. Beijing:Agriculture Press, 1984.
[12] 劳家柽. 土壤农化分析手册[M]. 北京:农业出版社, 1998. Lao JC. Analysis for Soil Agro-chemistry Manual[M]. Beijing:Agriculture Press, 1998. [13] 蒋向辉, 苑静, 祝军委, 胡秀虹, 杨承慧. 青钱柳叶片矿质营养元素含量季节动态变化研究[J]. 浙江林业科技, 2015, 35(2):17-21. Jiang XH, Yuan J, Zhu JW, Hu XH, Yang CH. Seasonal variation of mineral nutrient contents in Cyclocarya paliurus leaves[J]. Journal Of Zhejiang for Science & Technology, 2015, 35(2):17-21.
[14] 聂西度, 符靓. 盐酸浸提-电感耦合等离子体发射光谱法测定秋葵中Na、Mg、P、K、Ca含量[J]. 食品科学, 2015, 36(24):101-104. Nie XD, Fu L. Determination of macro elements in okra by inductively doupled plasma optical emission spectrometry with hydrochloric acid extraction[J]. Food Science, 2015, 36(24):101-104.
[15] 郑鹤龄. 微量元素营养诊断[M]. 天津:天津科技翻译出版社, 2010. Zheng HL. Trace Elements Nutrition Diagnosis[M]. Tianjin:Tianjin Science and Technology Translation Publishing Press, 2010.
[16] Campa C, Noirot M, Bourgeois M, Pervent M, Ky CL, Chrestin H, Hamon S, Kochko AD. Genetic mapping of a caffeoyl-coenzyme A 3-O-methyltransferase gene in coffee trees. Impact on chlorogenic acid content[J]. Theor Appl Genet, 2003, 107(4):751-756.
[17] 何柳, 陈士林. 植物中绿原酸合成途径研究进展[J]. 药物生物技术, 2013, 20(5):463-466. He L, Chen SL. Research progresses on synthesis of chlorogenic acid in plants[J]. Pharmaceutical Biotechno-logy, 2013, 20(5):463-466.
[18] Camacho-Cristóbal JJ, Lunar L. Lafont F, Baumert A, González-Fontes A. Boron deficiency causes accumulation of chlorogenic acid and caffeoyl polyamine conjugates in tobacco leaves[J]. J Plant Physiol, 2004, 161(7):879-881.
[19] 刘丽娟, 张卫东, 秦佳梅, 徐鸿雁, 王育民. 返魂草营养元素与绿原酸含量变化规律研究[J]. 河南农业科学, 2010, 10(2):89-91. Liu LJ, Zhang WD, Qin JM, Xu HY, Wang YM. Changes of nutrient elements and chlorogenic acid content in Senecio cannabifolius Less.7-152. Su JR, Deng J, Luo X, Yang WY. Studies on fertilization of young Acer truncatum BungeⅠ. Effect of various nutrition supply conditions on growth and morphology of Acer truncatum Bunge[J]. Forest Research, 2005,18(2):147-152.
[21] Karki P, Kim C, Smith K, Son DS, Aschner M, Lee E. Transcriptional regulation of the astrocytic excitatory amino acid transporter 1(EAAT1) via NF-κB and Yin Yang 1(YY1)[J]. J Biol Chem, 2015, 290(39):23725-23737.
[22] 郭雷, 胡洪, 任列娇, 高士争. 蛋白质翻译起始因子的作用与调控[J]. 云南农业大学学报, 2011, 26(4):554-559. Guo L, Hu H, Ren LJ, Gao SZ. Function and regulation of protein translation initiation factors[J]. Journal of Yunnan Agricultural University, 2011, 26(4):554-559.
[23] 梁新华, 栾维江, 梁军, 张风侠. 硼等4种元素对甘草酸生物合成关键酶基因表达的RT-PCR分析[J]. 时珍国医国药, 2011, 22(2):2351-2353. Liang XH, Luan WJ, Liang J, Zhang FX. RT-PCR analysis on the differential expression of the key enzyme genes involved in glycyrrhizic acid synthetic metabolic pathway under B, Mn, Zn and Mo elements treatment[J]. Lishizhen Medicine and Materia Medica Research, 2011, 22(2):2351-2353.
[24] 陈雅彬, 李永强, 孙琳, 沈妍雯, 陈文荣, 刘霞, 郭卫东. 非酸性根际土壤对蓝莓铁元素吸收及其代谢相关基因表达的影响[J].园艺学报, 2015, 42(2):233-242. Chen YB, Li YQ, Sun L, Shen YW, Chen WR, Liu X, Guo WD. Effects of non-acid rhizosphere pH on the iron elements uptakes and expressions of iron metabolism related genes in blueberry[J]. Acta Horticulturae Sinica, 2015, 42(2):233-242.
计量
- 文章访问数: 964
- HTML全文浏览量: 3
- PDF下载量: 1067
下载:
