Ca2+在介导MeJA诱导白桦悬浮培养三萜合成中的作用

doi:10.7525/j.issn.1673-5102.2015.01.018

摘要/Abstract

摘要： 以白桦细胞为试材，通过在悬浮培养第七天进行MeJA、CaCl₂及其互作处理，研究Ca²⁺在MeJA诱导白桦三萜合成中的作用。结果表明，MeJA处理后显著促进白桦细胞中总三萜和齐墩果酸的合成，分别于48 h达到最高积累量46.90和1.31 mg·g^-1。MeJA+EGTA和MeJA+LaCl₃·7H₂O处理，均降低了MeJA对三萜合成的促进作用，仍显著高于对照。抗逆酶结果显示，CaCl₂和MeJA+CaCl₂处理均不同程度的提高了CAT、PAL和APX的活性，而SOD的活性则受到Ca²⁺的抑制；两种钙离子抑制剂加入后显著影响抗逆酶的活性。MeJA处理早期（2 h），三萜途径各关键酶基因FPS、SS、SE、BPW、BPY显著上调表达。MeJA+LaCl₃·7H₂O处理早期（2 h）除BPW降低外，BPY、BPX2、SS和SE表达量均高于MeJA单独处理；而同MeJA处理相比，MeJA+EGTA处理2 h，BPW、BPX2、SS、SE和FPS表达量均下调。暗示Ca²⁺参与了MeJA诱导白桦防御系统启动和三萜物质合成，但两种Ca²⁺抑制剂EGTA和LaCl₃在三萜合成中的作用效果和机制存在差异。

关键词: 白桦, 悬浮培养, MeJA, 三萜化合物, Ca²⁺

Abstract: We investigated the role of Ca²⁺ in the MeJA-induced biosynthesis of triterpenoid by treating suspending cells of Betula platyphylla with CaCl₂, MeJA and combining with inhibitors of Ca²⁺. MeJA could promote the synthesis of total triterpenes and oleanolic acid in the cells significantly, which, respectively reached the peak of 46.90 and 1.31 mg·g^-1 after 48 hours’ treatment. The induction effect of MeJA decreased after treating by MeJA+EGTA and MeJA+LaCl₃·7H₂O. The accumulation amount of oleanolic acid were still higher than that of the control. By analyzing the activity of defense enzyme, the activities of CAT, PAL, APX were enhanced after treated by CaCl₂ and MeJA+CaCl₂, while the SOD was inhibited by Ca²⁺. After adding two inhibitors of Ca²⁺, all enzymes were effected significantly. The expression of key enzyme genes in the triterpene pathway such as FPS, SS, SE, BPW, BPY BPX2 was significantly upregulated after two hours’ MeJA treatment. The expression of BPW was reduced after two hours’ treating with MeJA+LaCl₃·7H₂O. The expression of BPY, BPX2, SS and SE regulated by MeJA+LaCl₃·7H₂O were higher than those treated by MeJA. Meanwhile the expression of BPW, BPX2, SS, SE and FPS were down-regulated at two hours after MeJA+EGTA treatment compared with MeJA treatment. Therefore, Ca²⁺ played a role in triterpenoid biosynthesis induced by MeJA, but two Ca²⁺ inhibitors had different effect and mechanism in triterpenoid biosynthesis.

Key words: Betula platyphylla Suk., suspension culture, MeJA, triterpenoid, Ca²⁺

中图分类号:

马泓思1；潘亚婕1；王艳1；尹静1*；詹亚光1,2；赵微1；张梦岩1；梁　甜1. Ca²⁺在介导MeJA诱导白桦悬浮培养三萜合成中的作用[J]. 植物研究, 2015, 35(1): 117-126.

MA Hong-Si1；PAN Ya-Jie1；WAN Yan1；YIN Jing1*；ZHAN Ya-Guan1,2；ZHAO Wei1；ZHANG Meng-Yan1；LIANG Tian1. Effect of Ca²⁺ on Mediating the MeJA-induced Synthesis of Triterpenoid in Suspension Cells of Betula platyphylla Suk.[J]. Bulletin of Botanical Research, 2015, 35(1): 117-126.

参考文献

1.Huha J E,Hongc J M,Baekb Y H,et al.Anti-inflammatory and anti-nociceptive effect of Betula platyphylla var.japonica in human interleukin-1β-stimulated fibroblast-like synoviocytes and in experimental animal models［J］.Journal of Ethnopharmacology,2011,135:126-134.

2.Saxena B B,Zhu L,Hao M,et al.Boc-lysinate-betulonic acid:a potent,anti-prostate cancer agent［J］.Journal of Medicinal Chemistry,2006,14:6249-6258.

3.Huang L,Lee K H,Chen C H.Synthesis and anti-HIV activity of bi-functional BA derivatives［J］.Bioorganic & Medicinal Chemistry,2006,14:2279-2289.

4.Kyoko N G,Koji Y,Masahiko T,et al.Cancer preventive agents.Betulinic acid derivatives as potent cancer chemopreventive agents［J］.Bioorganic & Medicinal Chemistry Letters,2009,19(13):3378-3381.

5.Aharon A,Jongsma M A,Kim T Y,et al.Metabolic engineering of terpenoid biosynthesis in plants［J］.Phytochemistry Reviews,2006,5:49-58.

6.Gallo M B C,Sarachine M J.Biological activities of Lupeol［J］.International Journal of Biomedical and Pharmaceutical Sciences,2009,3(Special Issue 1):46-66.

7.Kim Y S,Han J Y,Lim S,et al.Ginseng metabolic engineering :Regulation of genes related to ginsenoside biosynthesis［J］.Journal of Medicinal Plants Research,2010,3(13):1270-1276.

8.Zhang H,Shibuya M,Yokota S.Oxidosqualene cyclases from cell suspension cultures of Betula platyphylla var.japonica:molecular evolution of oxidosqualene cyclases in higher plants［J］.Biological & Pharmaceutical Bulletin,2003,26(5):642-650.

9.Yin J,Zhan Y G,Ren C L,et al.Distribution and expression characteristics of triterpenoids and OSC genes in white birch(Betula platyphylla suk.)［J］.Molecular Biology Reports,2012,39(3):2321-2328.

10.Turner J G,Ellis C,Devoto A.The jasmonate signal pathway［J］.The Plant Cell,2002,S:153-164.

11.Thaler J S,Owen B,Higgins V J.The role of the jasmonate response in plant susceptibility to diverse pathogens with a range of lifestyles［J］.Plant Physiology,2004,135:530-538.

12.Kim Y S,Cho J H,Park S.Gene regulation patterns in triterpene biosynthetic pathway driven by overexpression of squalene synthase and methyl jasmonate elicitation in Bupleurum falcatum［J］.Planta,2011,233:343-355.

13.Hayashi H,Huang P Y,Inoue K.Up-regulation of soyasaponin biosynthesis by Methyl Jasmonate in cultured cells of Glycyrrhiza glabra［J］.Plant Cell Physiol,2003,44(4):404-411.

14.Martin D M,Gershenzon J,Bohlmann J.Induction of volatile terpene biosynthesis and diurnal emission by Methyl Jasmonate in foliage of Norway spruce［J］.Plant Physiology,2003,132:1586-1599.

15.Haley A,Russell A J,Wood N,et al.Effects of mechanical signaling on plant cell cytosolic calcium［J］.Proceedings of the National Academy of Sciences,1995,92:4124-4128.

16.王俊斌,李明,丁博,等.茉莉酸甲酯诱导保卫细胞气孔关闭的信号转导机制［J］.中国细胞生物学学报,2013,35(2):224-228.

17.王海波,黄椿颖,庞学群,等.茉莉酸甲酯诱导的采后香蕉果实耐冷性与活性氧信号的关系［J］.中国农业科学,2008,41(4):1165-1171.

18.余朝阁,李天来,张亢亢,等.钙对茉莉酸甲酯诱导番茄抗灰霉病及防御酶活性的调控作用［J］.中国蔬菜,2012(18):166-170.

19.Islam M M,Hossain M A,Jannat R,et al.Cytosolic alkalization and cytosolic calcium oscillation in Arabidopsis guard cells response to ABA and MeJA［J］.Plant Cell Physiol,2010,51(10):1721-1730.

20.Chen Y C,Tseng B W,Huang Y L,et al.Expression of the ipomoelin gene from sweet potato is regulated by dephosphorylated proteins,calcium ion and ethylene［J］.Plant,Cell and Environment,2003,26:1373-1383.

21.刘新,石武良,张蜀秋,等.Ca²⁺参与茉莉酸诱导蚕豆气孔关闭的信号转导［J］.实验生物学报,2005,38(4):297-302.

22.孙清鹏,于涌鲲,万善霞,等.胞外及胞内Ca²⁺共同参与拟南芥中茉莉酸诱导的钙动员［J］.中国农业科学,2010,43(5):942-948.

23.Moyen C,Hammond-Kosack K E,Jones J,et al.Systemin triggers an increase of cytoplasmic calcium in tomato mesophyll cells:Ca²⁺ mobilization from intra- and extracellular compartments［J］.Plant Cell Environ,1998,21:1101-1111.

24.Hashimoto K,Kudla J.Calcium decoding mechanisms in plants［J］.Biochimie,2011,93:2054-2059.

25.任春林.诱导子组合对白桦三萜合成调控及MeJA抑制性消减文库分析［D］.哈尔滨:东北林业大学,2012.

26.Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-timequantitative PCR and the 2″AACt method［J］.Methods,2001,25:402-408.

27.蔡昆争,董桃杏,徐涛.茉莉酸类物质（JAs）的生理特性及其在逆境胁迫中的抗性作用［J］.生态环境,2006,15(2):397-404.

28.Reddy A S N,Ali G S,Celesnik H,et al.Coping with stresses:roles of calcium- and calcium/calmodulin-regulated gene expression［J］.The Plant Cell,2011,23:2010-2032.

29.Zhao J,Lawrence C D,Robert V.Elicitor signal transduction leading to production of plant secondary metabolites［J］. Biotechnology Advances,2005,23:283-333.

30.Schneider-Müller S,Kurosaki F,Nishi A.Role of salicylic acid and intracellular Ca²⁺ in the induction of chitinase activity in carrot suspension culture［J］.Physiological and Molecular Plant Pathology,1994,45:101-109.

31.由继红,陆静梅.钙对萝卜幼苗抗寒性及某些生理指标的影响［J］.植物研究,2001,21(3):409-412.

32.李婧男,刘强,李升.生长素和氯化钙对盐胁迫下沙冬青幼苗的缓解作用［J］.植物研究,2010,01:27-31.

33.Raz V,Fluhr R.Calcium requirement for ethylene-dependent responses［J］.The Plant Cell,1992,4:1123-1130.

34.Bush D S.Caleium regulation in plant cells and its role in signaling［J］.Annual Review of Plant Physiology and Plant Molecular Biology,1995,46:95-122.

35.陈军文,曹坤芳.三叶橡胶光合作用能力和抗氧化系统以及单萜类物质对茉莉酸的响应［J］.植物研究,2008,01:47-53.

36.黄永恒,姚青,郭俊,等.钙和钙离子通道阻断剂对丛枝菌根真菌吸收镉的影响［J］.环境科学学报,2010,30(10):2049-2057.

37.Munemasa S,Mori I C,Murata Y.Methyl jasmonate signaling and signal crosstalk between methyl jasmonate and abscisic acid in guard cells［J］.Plant Signal Behav,2011,6(7):939-941.

38.Kenton P,Mur LA J,Atzorn R,et al.(-)-Jasmonicacid accumulation in tobacco hypersensitive response lesions［J］.Molecular Plant-Microbe Interactions,1999,12:74-78.

39.León J,Rojo,E,Sánchez-Serrano J J.Wound signalling in plants［J］.Journal of Experimental Botany,2001,52(354):1-9.

40.Hsu Y Y,Chao Y Y,Kao C H.Methyl Jasmonate-induced lateral root formation in rice:The role of heme oxygenase and calcium［J］.Journal of Plant Physiology,2013,170:63-69.

41.施和平,王云灵,曾宝强,等.外源钙对镉胁迫下南美蟛蜞菊毛状根生长、抗氧化酶活性和镉吸收的缓解效应［J］.生物工程学报,2012,28(6):747-762.

42.王博.促进白桦（Betula platyphylla Suk.）培养物中三萜物质积累的初步研究［D］.哈尔滨:东北林业大学,2008.

43.尹静.白桦三萜合成调控及其关键酶基因表达研究［D］.哈尔滨.东北林业大学，2009.

44.Hashimoto K,Kudla J.Calcium decoding mechanisms in plants［J］.Biochimie,2011,93:2054-2059.

45.Nkembo M K,Kurosaki F,Lee J B,et al.Stimulation of calcium signal transduction results in enhancement of production of scopadulcic acid B by methyl jasmonate in the cultured tissues of Scoparia dulcis［J］.Plant Biotechnology,2005,22:333-337.

46.宗会,李明启.钙信使在植物适应非生物逆境中的作用［J］.植物生理学通讯,2001,37(4):330-335.

47.Boursiac Y,Harper J F.The origin and function of calmodulin regulated Ca²⁺ pumps in plants［J］.Journal of Bioenergetics & Biomembranes,2007,39:409-414.

48.薛鑫,张芊,吴金霞.植物体内活性氧的研究及其在植物抗逆方面的应用［J］.生物技术通报,2013(10):6-11.

49.马君兰,赵越.外源茉莉酸甲酯（MeJA）对大豆异黄酮合成途径的影响［J］.东北农业大学学报,2011,42(5):14-18.

50.Ren A,Qin L,Shi L,et al.Methyl Jasmonate induces ganoderic acid biosynthesis in the basidiomycetous fungus Ganoderma lucidum［J］.Bioresource Technology,2010,101:6785-6790.

[1]	陈柄华, 张杰, 刘桂丰, 李思婷, 高元科, 李慧玉, 李天芳. 白桦半同胞家系纸浆材优良家系选择及选择方法评价[J]. 植物研究, 2023, 43(5): 690-699.
[2]	王景哲, 牛朝奎, 梁馨元, 申晨静, 尹静. 水杨酸在白桦苗期抵御盐碱胁迫中的调控作用[J]. 植物研究, 2023, 43(3): 379-387.
[3]	郝雪峰, 亢春霞, 裴雁曦, 金竹萍. 苜蓿体内H₂S信号与Ca²⁺调节气孔运动的作用机制[J]. 植物研究, 2023, 43(2): 281-287.
[4]	杜金霞, 申婷婷, 王浩然, 林一萍, 李慧玉, 张连飞. 白桦BpSPL9基因抑制表达载体的构建及遗传转化研究[J]. 植物研究, 2023, 43(1): 30-35.
[5]	陈坤, 方功桂, 穆怀志, 姜静. 白桦BpPIN3基因启动子序列及应答特性分析[J]. 植物研究, 2022, 42(4): 592-601.
[6]	陈娇娆, 续旭, 胡章立, 杨爽. 植物感受盐胁迫及相关钙信号的研究进展[J]. 植物研究, 2022, 42(4): 713-720.
[7]	张玉琦, 苏欣, 尤志强, 富金博, 詹亚光, 尹静. 不同激素处理对白桦幼树萌条及三萜合成的影响[J]. 植物研究, 2022, 42(2): 289-298.
[8]	杨蕴力, 渠畅, 王阳, 刘桂丰, 姜静. 白桦BpPIN5基因启动子组织定位及外源激素应答分析[J]. 植物研究, 2022, 42(1): 104-111.
[9]	马庆, 李芳蕊, 刘桂丰, 李慧玉. 航天诱变白桦生长性状分析[J]. 植物研究, 2021, 41(4): 540-546.
[10]	吕东林, 李腾, 郭译文, 姜静, 黄海娇. 转基因白桦杂交子代种子活力及外源基因遗传规律分析[J]. 植物研究, 2021, 41(4): 564-572.
[11]	郭依萍, 刘佳欣, 于颖, 王超, 杨传平. 白桦BpNAC012基因调控白桦木质部发育表达谱分析[J]. 植物研究, 2021, 41(2): 251-261.
[12]	朱畇昊, 张梦佳, 董诚明. 外源MeJA对高温胁迫下半夏抗氧化系统和胁迫基因的影响[J]. 植物研究, 2021, 41(1): 67-73.
[13]	刘佳欣, 刘慧子, 石晶静, 于颖, 王超. 白桦MYB基因响应激素及盐旱处理的表达研究[J]. 植物研究, 2020, 40(5): 743-750.
[14]	王万奇, 齐婉竹, 赵秋爽, 曾栋, 刘轶, 付鹏跃, 曲冠证, 赵曦阳. 白桦BpJMJ18基因启动子克隆及表达分析[J]. 植物研究, 2020, 40(5): 751-759.
[15]	姜骋, 张曦, 田晴, 李莉. 白桦BpbHLH112基因克隆及其启动子表达特性分析[J]. 植物研究, 2020, 40(4): 583-592.

Ca²⁺在介导MeJA诱导白桦悬浮培养三萜合成中的作用

Effect of Ca²⁺ on Mediating the MeJA-induced Synthesis of Triterpenoid in Suspension Cells of Betula platyphylla Suk.

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价