药学学报

丹参酮和丹酚酸类化合物的生物合成及其转录调控机制

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詹忠根 ^* , 李杏

药学学报 | 综述 2020,55(9): 2110-2121

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药学学报 | 综述 2020, 55(9): 2110-2121

丹参酮和丹酚酸类化合物的生物合成及其转录调控机制

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詹忠根^*, 李杏

作者信息

浙江经贸职业技术学院生物制药教研室, 浙江杭州 310018

通讯作者:

*詹忠根,Tel: 86-571-86929836, E-mail:zhzg9321@163.com

Biosynthesis and transcriptional regulation of tanshinones and salvianolic acids

Zhong-gen ZHAN^*, Xing LI

Affiliations

Biophamaceutical Laboratory, Zhejiang Institute of Economics and Trade, Hangzhou 310018, China

出版时间: 2020-09-12 doi: 10.16438/j.0513-4870.2020-0429

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摘要

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丹参酮和丹酚酸类化合物是治疗冠心病、心肌梗死、高血压、高血脂、脑卒中等疾病的重要物质基础。近年来，随着基因组学、转录组学、代谢组学和生物信息学的快速发展，其生物合成途径和转录调控机制研究取得较大进展。文章在总结丹参酮和丹酚酸类化合物生物合成最新研究的基础上，着重综述了转录因子对其生物合成的调控作用，认为进一步厘清丹参酮和丹酚酸类化合物的生物合成途径（尤其是下游合成途径的研究）和不同家族转录因子的相互作用、信号响应和协同调控机制，对于挖掘其合成、转运、调节、修饰相关的新基因及揭示调控网络的分子机制十分必要；同时，根据丹参活性成分生物合成规律，人工设计并构建新的、具有特定生理功能的生物系统，从而大幅度提高合成酶基因的表达及活性物质的产量，是有待进一步深入研究的方向之一。

关键词

丹参酮类化合物 / 丹酚酸类化合物 / 生物合成途径 / 转录因子

Abstract

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Tanshinones and salvianolic acids are important materials in the treatment of coronary heart disease, myocardial infarction, hypertension, hyperlipidemia, stroke and others illnesses. In recent years, with the development of genomics, transcriptome, metabolomics and bioinformatics, many advances have been made in the biosynthesis and transcriptional regulation of tanshinones and salvianolic acids. This paper summarizes these advances and suggests further study on the downstream synthesis pathways and transcriptional regulatory mechanisms to reveal new molecular mechanism of synthesis, transport, regulation and modification. Additionally, we discuss the design and construction of new biological pathways to increase the expression of biosynthesis genes and the production of secondary components, is a newly developing research field.

Key words

tanshinone / salvianolic acid / biosynthesis pathway / transcription factor

引用本文

詹忠根, 李杏. 丹参酮和丹酚酸类化合物的生物合成及其转录调控机制. 药学学报, 2020 , 55 (9) : 2110 -2121 . DOI: 10.16438/j.0513-4870.2020-0429

Zhong-gen ZHAN, Xing LI. Biosynthesis and transcriptional regulation of tanshinones and salvianolic acids[J]. Acta Pharmaceutica Sinica, 2020 , 55 (9) : 2110 -2121 . DOI: 10.16438/j.0513-4870.2020-0429

正文

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中药丹参为唇形科(Labiatae)鼠尾草属(Salvia)植物丹参(Salvia miltiorrhiza Bge.)的干燥根及茎, 含有丰富的丹参酮类化合物, 如丹参酮Ⅰ (tanshinone Ⅰ)、丹参酮ⅡA (tanshinone ⅡA)、丹参酮ⅡB (tanshinone ⅡB)、二氢丹参酮Ⅰ (dihydrotanshinone Ⅰ)、隐丹参酮(cryptotanshinone)等(图 1)和丹酚酸类化合物, 如丹参素(3, 4-dihydroxyphenyllactic acid)、咖啡酸(caffeic acid)、迷迭香酸(rosmarinic acid)、丹酚酸A (salvianolic acid A)和丹酚酸B (salvianolic acid B)等(图 2), 具有活血化瘀、消肿止痛、养心安神之功效, 是治疗冠心病、心肌梗死、高血压、高血脂等疾病的主要物质基础^[1]。然而天然资源中丹参酮和丹酚酸类化合物含量极低, 且其累积与地理位置、生态环境及遗传背景有关, 也受诱导子、关键酶、调节基因或转录因子的调控。为充分利用生物技术方法, 有效提高丹参酮和丹酚酸等药效成分含量, 近年来, 测序和解析了丹参全基因组、叶绿体和线粒体^{[2, 3]}, 发表了20多个来自丹参不同器官及生长阶段的转录组数据^{[4, 5]}, 以及克隆与鉴定了大部分关键酶基因^{[6, 7]}, 为阐明丹参酮和丹酚酸类化合物的生物合成途径, 揭示转录因子单独或协同调控机制作出了极大贡献。前人已经对丹参酮和丹酚酸类化合物的生物合成、诱导子的诱导与调控等进行了详细的总结^[6-8], 本文主要介绍该类化合物生物合成的最新研究, 着重综述转录因子的调控作用。

1 丹参主要药效成分的生物合成研究

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1.1 丹参酮类化合物的生物合成

丹参酮类化合物属于二萜类, 主要在丹参根部周皮合成和积累, 迄今已分离到50余种。其生物合成包括牻牛儿基牻牛儿基焦磷酸(GGPP)合成、GGPP环化和次丹参酮二烯修饰等3个阶段(图 3)。GGPP的合成与其他天然二萜类基本一致, 即甲羟戊酸途径(MVA)合成的异戊烯基焦磷酸(IPP)与甲基赤鲜醇4-磷酸途径(MEP)合成的二甲基烯丙基焦磷酸(DMAPP)在法呢基焦磷酸合酶(SmFPPS)、牻牛儿基牻牛儿基焦磷酸环化酶(SmGGPPS)作用下生成GGPP。但不同于其他天然二萜类的是丹参酮生物合成中, GGPP的合成前体主要来源于MEP途径^[9]。GGPP环化阶段, 先由柯巴基焦磷酸合酶(SmCPS)催化GGPP转化成柯巴基焦磷酸(CPP), 再由类贝壳衫烯合酶(SmKSL)将CPP环化成次丹参酮二烯(miltiradiene)^{[10, 11]}。在此过程中, SmCPS的作用非常重要, 直接决定次丹参酮二烯碳架结构的形成。抑制SmCPS1的表达, 丹参酮ⅡA和丹参酮Ⅰ的合成显著下降^[10]。而SmKSL之所以能催化CPP生成次丹参酮二烯, 这一有别于裸子植物中紫杉醇、银杏内酯等生物合成过程的新功能, 得益于其编码基因SmKSL在进化过程中缺失了‘internal/γ’结构域^[11]。次丹参酮二烯的修饰, 主要包括细胞色素P450家族蛋白SmCYP76AH1催化次丹参酮二烯生成铁锈醇(ferruginol)^[12], SmCYP76AH3羟化铁锈醇的C-7和C-11位生成11-羟基铁锈醇(11-hydroxy ferruginol)、柳杉酚(sugiol)和11-羟基柳杉酚(11-hydroxyl sugiol), 再由SmCYP76AK1羟化11-羟基铁锈醇和11-羟基柳杉酚的C-20生成11, 20-二羟基铁锈醇(11, 20-hydroxy ferruginol)和11, 20-二羟基柳杉酚(11, 20-hydroxyl sugiol), 由于11, 20-二羟基柳杉酚不稳定, 可自氧化成10-羟甲基四氢丹参新酮(10-hydroxymerhyl tetrahydromiltirone)^[13], 最后在其他CYP450氧化酶、脱羧酶、脱氢酶、还原酶的催化下形成丹参酮ⅡA、隐丹参酮等活性成分^[14]。CYP450s是萜类骨架物质氧化修饰的主要酶类, 能将C-H键氧化, 使之成为可进一步转化的羟基。前期通过cDNA芯片^[15]、基因组^[2]和转录组^[5]等筛选或预测到SmCYP450s 500余个, 其中33个在丹参根周皮显著高表达, 尤其是SmCYP76亚家族成员与丹参酮的下游合成途径关系密切^[16]。目前, SmCYP76AH1、SmCYP76AH3、SmCYP76AK1已得到功能验证, 推测SmCYP71D375在丹参新酮合成的上游区域起作用, 分别从不同通路合成隐丹参酮和二氢丹参酮Ⅰ^[17]。

1.2 丹酚酸类化合物的生物合成

丹酚酸类化合物属于酚酸类, 主要在丹参的韧皮部和木质部中合成和积累, 迄今已分离到30余种。从结构上看, 大多数丹酚酸类化合物可视为咖啡酸的衍生物, 如迷迭香酸(rosmarinic acid, RA)为咖啡酸与丹参素的二聚物, 丹酚酸B为迷迭香酸二聚体, 丹酚酸A由1分子丹参素与2分子咖啡酸缩合而成(图 2)^[18]。因此, RA可能是大多数丹酚酸类化合物生物合成的共同前体。目前认为, 丹参中RA的合成有3种可能, 但均起源于苯丙烷代谢途径的两条平行支路。传统认为, RA由苯丙氨酸(L-phe)支路生成的4-香豆酰CoA (4-coumaroyl-CoA)和由酪氨酸(L-tyr)支路生成的4-羟基苯乳酸(4-hydroxyphenyllactic acid)在迷迭香酸合成酶(SmRAS)催化下生成2-氧-(4-香豆酰)-3-(4-羟基苯)-乳酸(4-coumaroyl-4'-hydroxyphenyllactic acid), 然后由细胞色素P450蛋白CYP98A14催化生成RA^[19]。但Di等^{[20, 21]}认为, 苯丙氨酸支路中4-香豆酸(4-coumaric acid)先被转化为咖啡酸(caffeic acid), 然后咖啡酰CoA (caffeoyl CoA)与4-羟基苯乳酸在SmRAS催化下生成咖啡酸-4-羟基苯乳酸(caffeoyl-4'-hydroxyphenyllactic acid), 再经CYP98A14转化成RA, 才是RA合成的主要路线。而酪氨酸支路的4-羟基苯乳酸经丹参素与4-香豆酰CoA在SmRAS催化下生成4-香豆酰-3', 4'-二羟基苯乳酸(4-coumaroyl-3', 4'-dihydroxyphenyllactic acid)后, 由CYP98A14催化形成RA是一条衍生路线(图 4)。相对于RA的合成, 丹酚酸下游生源途径还没有完全明确。Di等^[21]、Li等^{[22, 23]}认为丹酚酸B是由两分子RA在漆酶(lactase)等氧化酶的作用下生成苯环2位自由基和α位自由基, 然后二聚化反应直接生成或经丹酚酸E (salvianolic acid E)转化而成。然而, 综合Ag⁺和茉莉酸甲酯(MeJA)诱导后丹参毛状根中RA和丹酚酸B含量的变化情况^[24], 过表达SmRAS仅能提高迷迭香酸的含量却无法提高丹酚酸B的含量, 且丹酚酸B的主要降解产物是丹参素, 并测得合成过程中间体—紫草酸(lithospermicacid)^[25]; 以及MeJA和真菌提取物(YE)处理后酪氨酸支路上关键酶基因的响应与丹酚酸B合成的相关性等研究^{[26, 27]}表明, 丹酚酸B也有可能由咖啡酸与RA结合后生成紫草酸, 然后由紫草酸与丹参素自发酯化而成(图 4)。

2 丹参主要药效成分生物合成的转录调控研究

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丹参酮和丹酚酸类化合物的生物合成受YE、Ag⁺、MeJA、水杨酸(SA)、脱落酸(ABA)、赤酶素(GA)和乙烯(Eth)等生物及非生物诱导子的诱导^{[1, 28]}, 其机制之一就是通过转录因子激活或抑制合成途径关键酶基因的表达以调控次生代谢产物的合成。

2.1 JAZ蛋白与bHLH转录因子家族

茉莉酸类物质(JAs)通过调控JA信号通路相应的转录因子进而调控丹参酮和丹酚酸类化合物的合成^[29], 其介导的转录调控核心模型是SCF^COI1-JAZs-MYC2复合体。即正常生长的植物内源JAs水平较低, JAZ (jasmonate-ZIM domain)蛋白与MYC2等转录因子结合, 抑制茉莉酸早期响应基因的表达。当植物内源JAs或JA-异亮氨酸复合体(JA-Ile)含量增高时, JAZ蛋白与E3泛素连接酶SCF^COI1复合物相互作用, 并导致其自身被降解, 释放出MYC2等转录因子激活下游JAs应答基因的表达^[30]。

从丹参中分离的9条SmJAZ基因均受MeJA诱导, 过表达SmJAZs, 则SmRAS1、SmCYP98A14等基因的表达量下降, 负调控丹酚酸B的合成。与丹酸酚不同的是, 仅SmJAZ3/4/8负调控丹参酮的合成, 而SmJAZ1/2/5/6/9则起正调控作用, 其中SmJAZ2/4/9通过调控SmCPS1、SmCYP76AH1的表达来实现调控功能^[31]。SmJAZs转录因子家族中, SmJAZ8的作用较为特殊, 因其缺失JAs结构域而无法与COI1蛋白结合, 主要通过N端的两性抑制基序(EAR)起稳定抑制转录因子的活性, 抑制SmCPS1、SmCYP76AH1及处于SmCYP98A14下游酶基因的表达, 并参与JAs信号诱导的丹参初生代谢和次生代谢的负调控作用^[31]。过表达SmJAZ8, 能显著抑制SmJAZ1/4/5/6/10 的表达, 甚至产生对JA不敏感的丹参表型, 而SmJAZ8-RNAi则提升家族其他成员的表达量^[31], 是丹酚酸合成过程中较为重要的负调控因子。

SmJAZ作为JA信号通路中负调控茉莉酸信号从SCF^COI1复合体向下游应答基因传导的核心蛋白, 其启动子区域含有能与SmMYC2转录因子结合的元件^{[32, 33]}。MYC2属于bHLH (basic helix-loop-helix transcription factors)转录因子家族, 含有高度保守的碱性螺旋-环-螺旋结构域, 其N端的碱性区域能识别DNA上的E-box和G-box位点, C端HLH结构域形成的二聚体或四聚体能与靶基因的启动子结合, 是调控JA信号通路基因表达的另一核心成员。研究表明:正常情况下, SmMYC2a/2b与JAZ1/2结合, 调控功能被抑制。当内源JAs或JA-Ile含量升高时, 释放出的SmMYC2a激活SmHCT6、SmCYPA14; SmMYC2b激活SmCYPA14; SmMYC2激活SmTAT1、SmPAL1、SmC4H和SmCYP98A14等基因的转录表达^{[34, 35]}, 正向调控丹酚酸的生物合成。但在已注释的131条SmbHLH转录因子中^[36-38], 也有负调控丹酚酸合成的转录因子。如SmbHLH7能与JAZ3L结合, 负调控SmRAS1、SmCYP98A14、SmTAT1等基因表达^[38]; SmbHLH37能与SmJAZ3/8结合, 负调控SmTAT1、SmPAL1基因^[39]; 而SmbHLH51与R2R3-MYB类转录因子AtPAP1形成调控转录复合体后, 正向调控丹酚酸类物质的合成(图 5)^[40]。

除调控丹酚酸合成外, bHLH转录因子家族还对丹参酮的生物合成起调控作用。如SmMYC2可通过抑制SmbHLH74的转录, 削弱SmbHLH74对丹参酮合成关键酶基因SmHMGR1、SmGGPPS1、SmCYP76AH1的负调控^[41]。JAMs在结构上比MYC2转录因子在N端少一个酸性激活结构域, 起负调控作用。拟南芥异源表达研究表明, SmJAM3能通过负调控AtMYC2/3/4和正调控AtJAM1以进一步调控MYC2下游靶基因AtERF1、AtMYB75、AtPDF1.2、AtVSP2的表达。而在丹参中, SmJAM3、SmMYC2-likes分别与SmJAZ3、SmJAZ9蛋白互作, 上调表达SmGGPPS等基因的转录, 促进丹参酮合成^[42]。此外, SmbHLH7能正调控SmHMGS、SmGGPPS1、SmCYP76AH1、SmKSL1等丹参酮合成基因^[38], SmbHLH10能与丹参酮合成关键酶SmDXS2、SmCPS1、SmCPS5基因启动子的G-box位点结合, 激活转录^[43], 而SmbHLH148几乎能激活所有丹酚酸和丹参酮生物合成途径上的基因表达(图 6)^[44]。

需要指出的是, JAZs作为一种阻遏蛋白, 通常起抑制作用^[45]。但过表达SmJAZ1/2/5/6/9却显著提高转基因毛状根中丹参酮的含量, 且SmJAZs能与SmMYC2a、SmMYB39、SmMYC2b、SmPAP1等转录因子互作^{[32, 33]}, 促进SmMYC2a、SmMYC2b和SmPAP1表达, 抑制SmMYB39表达^[33]。因此, SmJAZs蛋白在调控JA诱导的丹参次生代谢物合成中发挥中心枢纽作用, 存在复杂的共调控网络, 这种多个SmJAZs共同调控一种或一类次生代谢产物合成的机制还有待进一步阐明。此外, 由于采样或命名等原因而导致的不同研究者研究同一名称的转录因子(如SmbHLH7)却得到不一致结论的情况, 也应引起重视^{[38, 42]}。

2.2 MYB转录因子家族

MYB (v-myb avian myeloblastosis viral oncogene homolog)是植物中最大的转录因子家族, 根据所含串联重复序列(repeat, R)数量, 可将MYB转录因子分为4大类。其中, 参与植物次生代谢产物合成调控的转录因子主要来自R2R3-MYB亚家族, 可通过蛋白质互作、转录酶调控、氧化还原反应、ABA信号系统或参与JA信号系统行使功能。

Li等^[46]从丹参全基因组中筛选出110个R2R3-MYB转录因子, 将其划分为37个亚族, 并预测S3、S4、S5、S6、S7、S13、S20、S21亚族对丹参酮和丹酚酸类化合物的合成具有调控作用。研究表明, S4亚族的R2R3-MYB转录因子主要通过乙烯响应因子相关的EAR对苯丙烷代谢产物的合成起抑制作用^[47], 如SmMYB4通过抑制酚酸类上游苯丙烷代谢途径的SmPAL1和SmC4H基因和丹参酮合成途径下游关键酶基因SmGGGPPS3, 负调控两类化合物的积累^[48]; SmMYB39作为负调控因子, 除直接抑制迷迭香酸代谢途径关键酶基因SmTAT、SmC4H和丹参酮代谢途径SmDXS2、SmDXR、SmHMGR1、SmGGPPS、SmKSL1的转录外, 还能通过抑制SmbHLH7转录或干扰SmbHLH7与其他转录因子形成正调控MBW复合体等机制行使负调控功能^{[49, 50]}。S6亚族转录因子SmPAP1与拟南芥中的PAP1蛋白的同源性高达75%, 在丹参中过表达拟南芥AtPAP1、AtPAP2, 能不同程度正向调控苯丙烷类代谢途径上SmPAL、SmC4H、SmRAS等多种酶基因的表达^{[51, 52]}。SmPAP1基因启动子区域上含有多种响应元件和bHLH结合位点^[53], 可与SmMYC2转录因子协作, 共同激活迷迭香酸合成途径SmTAT、SmPAL1和SmC4H等关键酶基因表达, 同时抑制丹参酮类生物合成关键酶基因SmCPS转录^[54]。S7亚族的转录因子SmMYB111, 基因启动子区含有多种激素响应元件和MYB结合位点, 受MeJA、SA、GA和光等诱导, 能与SmTTG1、SMBHH51形成SmTTG1-SmMYB111-SmbHLH51三元转录复合物, 共同参与丹酚酸生物合成的正向调节^[55]。SmMYB36位于S5和S15之间, 能与SmCPS1、SmCYP76AH1和SmKSL1等靶基因启动子区的MBS1、MBSⅠ、MBSⅡ响应元件及SmbHLH128发生互作, 促进丹参酮的积累和降低丹酚酸类物质的含量^[56]。转录组和代谢组分析表明, SmMYB36对丹参酮类化合物合成的调控是全方位的, 苯丙烷途径、酪氨酸衍生途径、甲基赤藓糖醇磷酸途径和下游丹参酮生物合成途径的基因转录水平均发生了显著变化^[57]。

另外, S20亚族转录因子SmMYB98b、SmMYB9a和SmMYB9b, 均能激活MEP途径中SmDXS2、SmDXR、SmGGPPS和SmKSL1等关键酶基因的转录, 显著提高丹参毛状根中丹参酮的含量^{[58, 59]}, 而SmMYB98则激活SmGGPPS1、SmPAL1、SmRAS1等基因的转录, 对丹参酮和丹酚酸的合成起正向调控作用^[60]。而且, SmMYB98b和SmMYB9a还参与低磷对丹参酮的调控, 低磷诱导的丹参酮积累能被SmMYB98b和SmMYB9a基因的过表达所抑制^[61]。但这种调控方式有别于AtMYB2、AtMYB62对低磷的响应机制, AtMYB2、AtMYB62分别通过活化miR399的转录和调节GA信号及其代谢, 进而调节磷饥饿响应^{[62, 63]}, 而SmMYB98b、SmMYB9a则与低磷响应基因SmSPX2和SmSPX4互作, 介导SPX对下游低磷响应基因(PSI)的调节作用(图 7)^[61]。

2.3 WRKY转录因子家族

WRKY转录因子家族的DNA结合区包含1~2个WRKY核心结构域, C端为锌指结构, 主要通过ABA信号通路参与植物的生长发育、响应逆境胁迫以及参与次生代谢的调控, 也能与SA、JA等信号通路协同, 间接调控应答基因^{[64, 65]}。丹参中共注释到WRKY转录因子69个, 其中从基因组数据中发现61个^[66], 从转录组数据中筛选出8个^[67]。Li等^[66]对其中61条SmWRKYs基因进行研究, 发现42条能响应MeJA和Ag⁺胁迫, 推测SmWRKY3/9正向调控丹参酮的合成。Yu等^[67]进一步推测SmWRKY19正向调控丹参酮、SmWRKY45/68正向调控丹酚酸、SmWRKY1/7/29/52/56/58对丹参酮和丹酚酸类化合物的生物合成均有正向调控作用。其中, Ⅲ类转录因子SmWRKY1, 已被证实能通过结合蛋白的方式对SmDXR进行正调控, 使转基因品系中丹参酮产量增加5倍^[68]。Ⅰ类转录因子SmWRKY2, 能与SmCPS启动子区的W-box结合以激活转录, 正调控丹参酮的生物合成^[69]。SmWRKY54^[70]、SmWRKY3和SmWRKY70^[71]等转录因子也能正向调控代谢途径上关键酶基因的转录, 促进丹参酮的合成。而受MeJA和YE强烈诱导的Ⅱa类转录因子SmWRKY9, 则正调控SmRAS1和SmCYP98A14的转录(SmPAL3、Sm4CL2、Sm4CL3等酶基因可能也参与), 促进迷迭香酸的生物合成(图 8)^[72]。

2.4 EIL转录因子家族

JAs与乙烯都能促进植物次生代谢产物的积累, 两者在拟南芥中对基因的表达具有协同诱导、相互依赖作用, 任一激素信号通路被阻断, 对基因的诱导表达作用即消失。研究表明, EIN3 (ethyleneinsensitive factor 3)/EILs (EIN3-likes)转录因子位于乙烯与JA这两条信号通路的交叉点。正常情况下, 乙烯能促进EIN3/EILs的积累, 而JAZ蛋白可通过组蛋白去乙酰化酶(HDA6)直接抑制EIN3/EILs转录因子的活性, 当JAs与其受体SCF^COI1-JAZ结合后, JAZ蛋白随之降解, 从而解除HDA6对EIN3/EIL1的抑制, 启动EIN3/EIL1调控的基因表达^[73]。丹参SmEIN3转录因子响应ABA、YE、MeJA和SA的诱导, 但GA抑制其表达。在转录水平上丹参SmEIN3不响应乙烯诱导, 乙烯调节的是转录后水平的EIN3/EIL蛋白及其活性, 以激活丹酚酸合成关键酶基因Sm4CL1、SmCYP98A14、SmHPPR、SmRAS和丹参酮合成关键酶基因SmDXSⅡ、SmDXR、SmCYP76AH1、SmKSL、SmGGPPS、SmHMGS、SmAACT、SmHMGR、SmCPS的转录。从SmEIN3与SmMYC2、SmJAZs存在互作现象分析, 丹参酮和丹酚酸类化合物的生物合成调节中可能存在乙烯信号通路与JA信号通路的交流, 两者起共同调控作用(图 9)^[74]。

2.5 GRAS转录因子家族

GA3处理后, 能激活SmCPS1、SmKSL1、SmPAL和SmTAT等关键酶基因的表达, 促进丹参酮和丹参酚类物质的合成^[75]。DELLA是GRAS转录因子超家族中的重要成员, 其N端具有GA受体GID1结合域, 为GA信号途径的负调控因子。GA可通过调控SLEEPY1等蛋白以促进DELLA降解, 而DELLA的降解则促进JAs的合成与JAZ降解, 从而进一步促进了DELLA的去抑制化^[76]。同时, DELLA蛋白也可通过结合不同的转录因子介导GA信号调控下游靶基因^[77], 如以DELLA-JAZ蛋白互作的形式达到GA-JA途径调控网络的实现^[78]; 在缺氮时, DELLA通过与PAP1蛋白相互作用, 增强PAP1对靶基因的调控(图 9)^[79]。Bai等^[80]从丹参中克隆出5个能响应GA或乙烯诱导的SmGRAS转录因子, 通过激活SmCPS1和SmKSL1的转录, 正向调控丹参次生代谢的合成。Wang^[81]从丹参中筛选出同时响应GA3、水杨酸(SA)和JAs信号途径的SmGRAS转录因子5个, 并鉴定出11个亚家族共34个SmGRAS转录因子, 其中4个转录因子属于SmDELLA亚家族, 都能与SmJAZ1、SmJAZ3、SmMYB36、SmMYC2等蛋白互作, 推测对丹参次生代谢产物的合成具有调控作用。

2.6 ERF类转录因子家族

ERF类转录因子属于AP2/ERF家族成员, 在AP2结构域的N-端具有特异识别顺式作用元件的β-折叠结构, 而C-端的α-螺旋结构则用于与其他转录因子或DNA的GCC-box相互作用。AP2/ERF类转录因子共有5个亚族, ERF (ethylene response factors)亚族种类最多, 大多数成员响应乙烯信号通路(也可响应JA、SA信号通路), 在植物生长、发育和次生代谢中扮演重要作用^[82]。Ji等^[83]从丹参基因组数据中注释到79个ERF转录因子, 并根据丹参酮的分布、转录因子的表达模式及启动子顺式元件等推测SmERF128/152与丹参酮生物合成相关, SmERF8/166与丹酚酸合成相关。SmERF2和SmO3L3通过结合(A/G/C) GCCGCC型的GCC-box, 分别激活丹参酮合成途径的SmDXR、SmDXS2、SmHMGS、SmKSL和丹酚酸合成途径的Sm4CL1、SmRAS等酶基因表达^[84]。SmERF6和SmERF8可识别SmCPS1和SmKSL1启动子区的乙烯反应元件GCC-box, 正向调控丹参酮的生物合成^{[85, 86]}。SmERF115受MeJA诱导, 但不受YE、SA、ET诱导, 能直接结合SmRAS1启动子激活表达(可能也激活SmPAL3、SmTAT3、Sm4CL5等), 促进丹酚酸类化合物的生成^[87]。而SmERF1L1则对MeJA、YE、SA、ET的诱导均有响应, 能激活SmDXR、SmDXS2、SmHMGS、SmKSL的转录, 促进丹参酮的合成^[88]。SmERF128可结合靶基因的GCC盒、CRTDREHBCF2 (CBF2)基序和RAV1AAT (RAA)基序, 激活SmCPS1、SmKSL1和SmCYP76AH1的表达(图 9)^[89]。

2.7 LBD转录因子家族

LBD (lateral organ boundaries domain)是一类植物特有的转录因子家族。具有完整亮氨酸拉链基序(Class Ⅰ)的成员, 主要参与组织发育的调控, 而不具有完整亮氨酸拉链基序的成员(Class Ⅱ), 主要与花青素生物合成、氮代谢的调控相关。从丹参基因组中鉴定出51个SmLBD, 90%的SmLBD具有保守的CX2CX6CX3C结构, 74%的SmLBD在叶片表达, 推测SmLBD9/13/21/50等与丹酚酸化合物的合成关系密切, SmLBD44与丹参酮的合成调控相关^[90]。研究表明, LBD转录因子的保守区段即LOB结构域可与bHLH、MYB转录因子相互作用, 参与JA信号下游途径调控^[91], 且同类LBD转录因子的功能相对保守, 但作用方式有所区别^[92]。如SmLBD16/23/50均能负向调控丹酚酸的合成, 且与SmJAZ1、SmMYB36/97、SmbHLH37和SmMYC2a/2b等转录因子存在蛋白互作, 但SmLBD16在调控苯丙烷途径关键酶基因表达时需SmPAP1参与, 而SmLBD23则不需要^[93]。

3 总结与展望

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近年来, 丹参酮和丹酚酸类化合物的生物合成及其转录因子调控机制研究取得长足进展, 一方面得益于丹参全基因组图谱的完成, 不同组织、器官的转录组测序, 使其能在基因组和转录组层面整体分析与丹参酮和丹酚酸类化合物生物合成相关的差异基因和转录因子^[94], 另一方面得益于拟南芥、烟草、水稻等模式植物研究中所建立的高效成熟的转化系统。但是, 丹参作为药用模式植物, 其次生代谢产物的生物合成及调控机制研究, 还需继续深入, 主要体现在4个方面: ①丹参酮和丹酚酸类化合物的生物合成途径还未被完全阐明, 急需明确参与丹参酮后期修饰及迷迭香酸下游途径的关键酶, 并进一步揭示其合成、转运、调节、修饰的新基因及作用机制。②丹参中转录因子众多, 家族成员庞大, 同一家族(亚家族)成员功能相似等, 大大增加了转录因子调控机制研究的难度, 但是家族成员之间及不同家族转录因子之间的相互作用、响应的信号通路、参与的合成途径等错综复杂的调控关系, 仍是今后的研究重点。③部分转录因子的研究, 如SPL (SQUAMOSA promoter binding protein-like)转录因子^[95]、bZIP (basic region/leucine zipper motif)转录因子^[96], 还停留在功能预测层面, 急需进一步深入。同时, 转录因子自身的转录也受到其他基因的调控, 如microRNAs、ncRNAs等^[97], 但这方面的研究较少。④人工设计并构建新的、具有特定生理功能的生物系统, 是一种极具潜力的丹参活性成分资源获取方法。如通过构建SmHMGR催化活性域-SmGGPPS, SmFPS融合蛋白-SmKSL, SmCPS融合蛋白酵母工程菌, 使次丹参酮二烯的产量达到了365 mg·L^-1 ^[98]; 组合改造FPP合成酶和GGPP合成酶, 将次丹参酮二烯的产量提高到488 mg·L^-1^[99]; 引入CYP76AH1、SmCPR1基因, 将铁锈醇的最高产量提高到10.5 mg·L^-1^[12]。而转录因子能同时诱导一个或多个基因的协同表达, 调控效果优于单纯提高单个或多个结构基因的表达, 是另一大幅提高合成酶基因表达及活性物质产量的可能方法, 值得进一步深入研究。

作者贡献:詹忠根主要负责选题, 文献检索和初稿写作, 李杏主要负责文稿修改与校对。

利益冲突:无利益冲突。

参考文献

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参考文献引证文献

排序方式：

[1]

Wang

, Dai

, Xiang

et al . Advances in the molecular mechanism of salvianolic acid and tanshinone for intervention of diabetic kidney disease[J]. Acta Pharm Sin (药学学报), 2019, 54: 1356-1363. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=yxxb201908003

[2]

, Song

, Lou

et al . Analysis of the genome sequence of the medicinal plant Salvia miltiorrhiza[J]. Mol Plant, 2016, 9: 949-952.

https://doi.org/10.1016/j.molp.2016.03.010

[3]

Qian

. Study on Chloroplast and Mitochondrial Genomes of Salvia miltiorrhiza (丹参的叶绿体和线粒体基因组研究)[D]. Beijing: Chinese Academy of Medical Sciences·and Peking Union Medical College, 2014.

[4]

Luo

, Zhu

, Song

et al . Transcriptional data mining of Salvia miltiorrhiza in response to methyl jasmonate to examine the mechanism of bioactive compound biosynthesis and regulation[J]. Physiol Plant, 2014, 152: 241-255.

https://doi.org/10.1111/ppl.12193

[5]

, Peters

, Weirather

et al . Full-length transcriptome sequences and splice variants obtained by a combination of sequencing platforms applied to different root tissues of Salvia miltiorrhiza and tanshinone biosynthesis[J]. Plant J, 2015, 82: 951-961.

https://doi.org/10.1111/tpj.12865

[6]

, Ji

, Zhang

et al . Biosynthesis and regulation of active compounds in medicinal model plant Salvia miltiorrhiza[J]. Chin Herb Med, 2016, 8: 3-11.

https://doi.org/10.1016/S1674-6384(16)60002-3

[7]

Wang

, Xu

, Gong

et al . Biosynthesis, chemistry, and pharmacology of polyphenols from Chinese Salvia species:a review[J]. Molecules, 2019, 24: 155-177.

https://doi.org/10.3390/molecules24010155

[8]

, Gao

, Shao

et al . Effect of elicitors on induction and manipulation of secondary metabolic effective ingredients in Salvia miltiorrhiza[J]. China J Chin Mater Med (中国中药杂志), 2011, 36: 258-262. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgzyzz201103007

[9]

Gao

, Sun

, Xiao

et al . Combining metabolomics and transcriptomics to characterize tanshinone biosynthesis in Salvia miltiorrhiza[J]. BMC Genomics, 2014, 15: 73-86.

https://doi.org/10.1186/1471-2164-15-73

[10]

Cheng

, Su

, Hu

et al . RNA interference-mediated repression of SmCPS (copalyldiphosphate synthase) expression in hairy roots of Salvia miltiorrhiza causes a decrease of tanshinones and sheds light on the functional role of SmCPS[J]. Biotechnol Lett, 2014, 36: 363-369.

https://doi.org/10.1007/s10529-013-1358-4

[11]

Hillwig

, Xu

, Toyomasu

et al . Domain loss has independently occurred multiple times in plant terpene synthase evolution[J]. Plant J Cell Mol Biol, 2011, 68: 1051-1060.

https://doi.org/10.1111/j.1365-313X.2011.04756.x

[12]

Guo

, Zhou

, Hillwig

et al . CYP76AH1 catalyzes turnover of miltiradiene in tanshinones biosynthesis and enables heterologous production of ferruginol in yeasts[J]. Proc Natl Acad Sci U S A, 2013, 110: 12108-12113.

https://doi.org/10.1073/pnas.1218061110

[13]

Guo

, Ma

, Cai

et al . Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones[J]. New Phytologist, 2016, 210: 525-534.

https://doi.org/10.1111/nph.13790

[14]

Yang

, Ding

, Lin

et al . Transcriptome analysis of medicinal plant Salvia miltiorrhiza and identification of genes related to tanshinone biosynthesis[J]. PLoS One, 2013, 8:e80464

https://doi.org/10.1371/journal.pone.0080464

[15]

Cui

, Huang

, Tang

et al . Candidate genes involved in tanshinone biosynthesis in hairy roots of Salvia miltiorrhiza revealed by cDNA microarray[J]. Mol Biol Rep, 2011, 38: 2471-2478.

https://doi.org/10.1007/s11033-010-0383-9

[16]

Chang

, Wang

, Li

. Transcriptomic analysis reveals potential genes involved in tanshinone biosynthesis in Salvia miltiorrhiza[J]. Sci Rep, 2019, 9: 14929-14940.

https://doi.org/10.1038/s41598-019-51535-9

[17]

, Xu

, Song

. Cloning and functional identification of novel CYP71D375 related to tanshinone biosynthesis in Salvia miltiorrhiza[J]. Sci Sin Vit (中国科学:生命科学), 2018, 48: 390-398.

https://doi.org/10.1360/N052017-00196

[18]

Shi

, Huang

, Deng

et al . Bioactivities, biosynthesis and biotechnological production of phenolic acids in Salvia miltiorrhiza[J]. Crit Rev Food Sci Nutr, 2019, 59: 953-964.

https://doi.org/10.1080/10408398.2018.1474170

[19]

Petersen

, H usler

, Karwatzki

et al . Proposed biosynthetic pathway for rosmarinic acid in cell cultures of Coleus blumei Benth[J]. Planta, 1993, 189: 10-14.

https://doi.org/10.1007/BF00201337

[20]

. Exploring the Biosynthetic Pathway of Phenolic Acids and Functional Study of the Involved Genes in Salvia miltiorrhiza Bunge (丹参酚酸类成分生源途径的探索及相关基因的克隆与功能研究)[D]. Shanghai: Second Military Medical University, 2012.

[21]

, Zhang

, Chen

et al . C¹³ tracer reveals phenolic acids biosynthesis in hairy root cultures of Salvia miltiorrhiza[J]. ACS Chem Biol, 2013, 8: 1537-1548.

https://doi.org/10.1021/cb3006962

[22]

, Li

, Zhou

et al . Analysis of the laccase gene family and miR397-/miR408-mediated posttranscriptional regulation in Salvia miltiorrhiza[J]. Peer J, 2019, 7:e7605

https://doi.org/10.7717/peerj.7605

[23]

, Feng

, Chen

et al . Genome-wide identification and characterization of Salvia miltiorrhiza laccases reveal potential targets for salvianolic acid B biosynthesis[J]. Front Plant Sci, 2019, 10: 435-449.

https://doi.org/10.3389/fpls.2019.00435

[24]

Xiao

, Gao

, Di

et al . Lithospermic acid B is more responsive to silver ions (Ag⁺) than rosmarinic acid in Salvia miltiorrhiza hairy root cultures[J]. Biosci Rep, 2010, 30: 33-40.

https://doi.org/10.1042/BSR20080124

[25]

Xing

, Yang

, Guo

et al . Ag⁺ as a more effective elicitor for production of tanshinones than phenolic acids in Salvia miltiorrhiza hairy roots[J]. Molecules, 2015, 20: 309-324. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=9fd7ca326e14eff6cc9add7868808c93

[26]

Guo

, Zhang

, Wang

et al . Hydrolytic kinetics of lithospermic acid B extracted from roots of Salvia miltiorrhiza[J]. J Pharm Biomed Anal, 2007, 43: 435-439.

https://doi.org/10.1016/j.jpba.2006.07.046

[27]

Zhang

, Yan

, Wang

et al . Selective responses of enzymes in the two parallel pathways of rosmarinic acid biosynthetic pathway to elicitors in Salvia miltiorrhiza hairy root cultures[J]. J Biosci Bioeng, 2014, 117: 645-651.

https://doi.org/10.1016/j.jbiosc.2013.10.013

[28]

Liang

, Ma

, Xu

et al . Effects of abscisic acid, gibberellin, ethylene and their interactions on production of phenolic acids in Salvia miltiorrhiza Bunge hairy roots[J]. PLoS One, 2013, 8:e72806

https://doi.org/10.1371/journal.pone.0072806

[29]

Zhou

, Huang

, Wu

et al . Comprehensive transcriptome profiling of Salvia miltiorrhiza for discovery of genes associated with the biosynthesis of tanshinones and phenolic acids[J]. Sci Rep, 2017, 7: 10554-10565.

https://doi.org/10.1038/s41598-017-10215-2

[30]

Chini

, Boter

, Solano

. Plant oxylipins:COl1/JAZs/MYC2 as the core jasmonic acid-signalling module[J]. FEBS J, 2009, 76: 4682-4692.

https://doi.org/10.1111/j.1742-4658.2009.07194.x

[31]

Pei

, Ma

, Ding

et al . SmJAZ8 acts as a core repressor regulating JA-induced biosynthesis of salvianolic acids and tanshinones in Salvia miltiorrhiza hairy roots[J]. J Exp Bot, 2018, 69: 1663-1678.

https://doi.org/10.1093/jxb/erx484

[32]

, Zhang

, Hua

et al . Combination of transcriptomic and metabolomic analyses reveals a JAZ repressor in the jasmonate signaling pathway of Salvia miltiorrhiza[J]. Sci Rep, 2015, 5: 14048-14061.

https://doi.org/10.1038/srep14048

[33]

Shi

, Zhou

, Zhang

et al . Methyl jasmonate induction of tanshinone biosynthesis in Salvia miltiorrhiza hairy roots is mediated by jasmonate zim-domain repressor proteins[J]. Sci Rep, 2016, 6: 20919-20929.

https://doi.org/10.1038/srep20919

[34]

Zhou

, Sun

, Chen

et al . SmMYC2a and SmMYC2b played similar but irreplaceable roles in regulating the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza[J]. Sci Rep, 2016, 6: 22852-22863.

https://doi.org/10.1038/srep22852

[35]

Yang

, zhou

, Su

et al . Overexpression of SmMYC2 increases the production of phenolic acids in Salvia miltiorrhiza[J]. Front Plant Sci, 2017, 8: 1804-1815.

https://doi.org/10.3389/fpls.2017.01804

[36]

Hua

, Zhang

, Song

et al . De novo transcriptome sequencing in Salvia miltiorrhiza to identify genes involved in the biosynthesis of active ingredients[J]. Genomics, 2011, 98: 272-279.

https://doi.org/10.1016/j.ygeno.2011.03.012

[37]

Zhang

, Luo

, Xu

et al . Genome-wide characterisation and analysis of bHLH transcription factors related to tanshinone biosynthesis in Salvia miltiorrhiza[J]. Sci Rep, 2015, 5: 11244.

https://doi.org/10.1038/srep11244

[38]

Cao

. The Functional Study of SmbHLH7 Transcription Factor in Salvia miltiorrhiza (丹参SmbHLH7转录因子的克隆与功能初探)[D]. Shanghai: Shanghai Normal University, 2018.

[39]

, Niu

, Su

et al . SmbHLH37 functions antagonistically with SmMYC2 in regulating jasmonate-mediated biosynthesis of phenolic acids in Salvia miltiorrhiza[J]. Front Plant Sci, 2018, 9: 1720-1732.

https://doi.org/10.3389/fpls.2018.01720

[40]

, Zhang

, Li

et al . AtPAP1 interacts with and activates SmbHLH51, a positive regulator to phenolic acids biosynthesis in Salvia miltiorrhiza[J]. Front Plant Sci, 2018, 9: 1687-1689.

https://doi.org/10.3389/fpls.2018.01687

[41]

Zhang

. Molecular Mechanism of SmbHLH74 in Regulation of Tanshinone Biosynthesis in Salvia miltiorrhiza Bunge (丹参转录因子SmbHLH74调控丹参酮生物合成的分子机制)[D]. Shenyang: Shenyang Agricultural University, 2019.

[42]

Wang

. Isolation and Function Analysis of bHLH Translation Factor in Salvia miltiorrhiza (丹参bHLH类转录因子的基因克隆及功能初步研究)[D]. Shanghai: Shanghai Normal University, 2015.

[43]

Xing

, Yang

, Yu

et al . Overexpression of SmbHLH10 enhances tanshinones biosynthesis in Salvia miltiorrhiza hairy roots[J]. Plant Sci, 2018, 276: 229-238.

https://doi.org/10.1016/j.plantsci.2018.07.016

[44]

Xing

, Liang

, Liu

et al . Overexpression of SmbHLH148 induced biosynthesis of tanshinones as well as phenolic acids in Salvia miltiorrhiza hairy roots[J]. Plant Cell Rep, 2018, 37: 1681-1692.

https://doi.org/10.1007/s00299-018-2339-9

[45]

Goossens

, Mertens

, Goossens

. Role and functioning of bHLH transcription factors in jasmonate signaling[J]. J Exp Bot, 2017, 68: 1333-1347.

https://doi.org/10.1093/jxb/erw440

[46]

, Lu

. Genome-wide characterization and comparative analysis of R2R3-MYB transcription factors shows the complexity of MYB-associated regulatory networks in Salvia miltiorrhiza[J]. BMC Genomics, 2014, 15: 277-288.

https://doi.org/10.1186/1471-2164-15-277

[47]

Liu

, Osboum

, Ma

. MYB transcription factors as regulators of phenylpropanoid metabolism in plants[J]. Mol Plant, 2015, 8: 689-708.

https://doi.org/10.1016/j.molp.2015.03.012

[48]

Zhu

. Study on the Regulation of SmMYB4 on the Synthesis of Secondary Metabolites in Salvia miltiorrhiza (SmMYB4对丹参次生代谢产物合成的调节作用研究)[D]. Xi'an: Shaanxi Normal University (陕西师范大学), 2016.

[49]

Zhang

, Ma

, Yang

et al . Cloning and characterization of a putative R2R3 MYB transcriptional repressor of the rosmarinic acid biosynthetic pathway from Salvia miltiorrhiza[J]. PLoS One, 2013, 8:e73259

[50]

Xing

. Molecular Mechanism of bHLH7 Interacting with MYB39 to Regulate Phenolic Acids and Tanshinones Biosynthesis in Salvia miltiorrhiza (丹参转录因子bHLH7与MYB39互作调控酚酸类及丹参酮类物质代谢分子机制研究)[D]. Beijing: University of Chinese Academy of Sciences, 2019.

[51]

Zhang

, Yan

, Wang

. The Arabidopsis PAP1 transcription factor plays an important role in the enrichment of phenolic acids in Salvia miltiorrhiza[J]. J Agric Food Chem, 2010, 58: 12168-12175.

https://doi.org/10.1021/jf103203e

[52]

Liu

. Effect of Transcription Factor PAP2 on the Synthesis of Salvianolic Acids (转录因子PAP2对丹参酚酸类产物合成的影响)[D]. Xi'an: Shaanxi Normal University, 2011.

[53]

. Analysis of the Expression and Regulation Mode of SmPAP1 Gene in Salvia miltiorrhiza (丹参SmPAP1基因的表达调控模式分析)[D]. Xi'an: Shaanxi Normal University, 2013.

[54]

Hao

, Jiang

, Feng

et al . Cloning, molecular characterization and functional analysis of a putative R2R3-MYB transcription factor of the phenolic acid biosynthetic pathway in S. miltiorrhiza Bge. f. alba[J]. Plant Cell Tissue Organ Cult, 2016, 124: 151-168.

https://doi.org/10.1007/s11240-015-0883-3

[55]

, Wu

, Kuang

et al . SmMYB111 Is a key factor to phenolic acid biosynthesis and interacts with both SmTTG1 and SmbHLH51 in Salvia miltiorrhiza[J]. J Agric Food Chem, 2018, 66: 8069-8078.

https://doi.org/10.1021/acs.jafc.8b02548

[56]

Ding

, Pei

, Bai

et al . SmMYB36, a novel R2R3-MYB transcription factor, enhances tanshinone accumulation and decreases phenolic acid content in Salvia miltiorrhiza hairy roots[J]. Sci Rep, 2017, 7: 5104.

https://doi.org/10.1038/s41598-017-04909-w

[57]

Zhao

. Terpenoids and Phenylpropanoids Metabolic Regulation by SmMYB36 in Salvia miltiorrhiza Bunge (丹参SmMYB36基因对萜类和苯丙烷代谢途径调控的研究)[D]. Yangling: Northwest A & F University, 2019.

[58]

Liu

, Yang

, Xing

et al . SmMYB98b positive regulation to tanshinones in Salvia miltiorrhiza Bunge hairy roots[J]. Plant Cell Tissue Organ Cult, 2020, 140: 459-467.

https://doi.org/10.1007/s11240-019-01716-1

[59]

Zhang

, Zhou

, Zheng

et al . Overexpression of SmMYB9b enhances tanshinone concentration in Salvia miltiorrhiza hairy roots[J]. Plant Cell Rep, 2017, 36: 1297-1309.

https://doi.org/10.1007/s00299-017-2154-8

[60]

Hao

, Pu

, Cao

et al . Tanshinone and salvianolic acid biosynthesis are regulated by SmMYB98 in Salvia miltiorrhiza hairy roots[J]. J Adv Res, 2020, 23: 1-12.

https://doi.org/10.1016/j.jare.2020.01.012

[61]

Liu

. Study on the Physiological Response to Low Phosphate, and the Function of Genes SmMYB98b and SmMYB9a in Salvia miltiorrhiza (丹参对低磷胁迫的生理响应及基因SmMYB98b和SmMYB9a的功能研究)[D]. Yangling: Northwest A & F University, 2018.

[62]

Baek

, Kim

, Chun

et al . Regulation of miR399f transcription by AtMYB2 affects phosphate starvation responses in Arabidopsis[J]. Plant Physiol, 2013, 161: 362-373.

https://doi.org/10.1104/pp.112.205922

[63]

Devaiah

, Madhuvanthi

, Karthikeyan

et al . Phosphate starvation responses and gibberellic acid biosynthesis are regulated by the MYB62 transcription factor in Arabidopsis[J]. Mol Plant, 2009, 2: 43-58.

https://doi.org/10.1093/mp/ssn081

[64]

Rushton

, Tripathi

, Rabara

et al . WRKY transcription factors:key components in abscisic acid signalling[J]. Plant Biotechnol J, 2012, 10: 2-11.

https://doi.org/10.1111/j.1467-7652.2011.00634.x

[65]

Schluttenhofer

, Pattanaik

, Patra

et al . Analyses of catharanthus roseus and Arabidopsis thaliana WRKY transcription factors reveal involvement in jasmonate signaling[J]. BMC Genomics, 2014, 15: 502-521.

https://doi.org/10.1186/1471-2164-15-502

[66]

, Li

, Shao

et al . Molecular cloning and expression analysis of WRKY transcription factor genes in Salvia miltiorrhiza[J]. BMC Genomics, 2015, 16: 200-220.

https://doi.org/10.1186/s12864-015-1411-x

[67]

, Guo

, Yang

et al . Transcriptional profiles of SmWRKY family genes and their putative roles in the biosynthesis of tanshinone and phenolic acids in Salvia miltiorrhiza[J]. Int J Mol Sci, 2018, 19: 1593-1609.

https://doi.org/10.3390/ijms19061593

[68]

Cao

, Wang

, Shi

et al . Transcription factor SmWRKY1 positively promote the biosynthesis of tanshinones in Salvia miltiorrhiza[J]. Front Plant Sci, 2018, 9: 554-563.

https://doi.org/10.3389/fpls.2018.00554

[69]

Deng

, Hao

, Shi

et al . Tanshinone production could be increased by the expression of SmWRKY2 in Salvia miltiorrhiza hairy roots[J]. Plant Sci, 2019, 284: 1-8.

https://doi.org/10.1016/j.plantsci.2019.03.007

[70]

. Preliminary Study on the Function of SmWRKY54 Transcription Factor in Salvia miltiorrhiza (丹参SmWRKY54转录因子功能初步研究)[D]. Shanghai: Shanghai Normal University, 2016.

[71]

Hao

. The Function Study of SmWRKY3 and SmWRKY70 Transcription Factors in Salvia miltiorrhiza (丹参SmWRKY3和SmWRKY70转录因子功能研究)[D]. Shanghai: Shanghai Normal University, 2014.

[72]

Zhang

. The Functional Study of SmWRKY9 Transcription Factor in Salvia miltiorrhiza (丹参SmWRKY9转录因子功能研究)[D]. Shanghai: Shanghai Normal University, 2019.

[73]

Zhu

, An

, Feng

et al . Derepression of ethylene-stabilized transcription factors (EIN3/EIL1) mediates jasmonate and ethylene signaling synergy in Arabidopsis[J]. Proc Natl Acad Sci U S A, 2011, 108: 12539-12544.

https://doi.org/10.1073/pnas.1103959108

[74]

Wang

. Cloning and Functional Analysis of eILtranscription Factor Gene EIN3 from Salvia miltiorrhiza (丹参EIL类转录因子EIN3基因的克隆与功能分析)[D]. Shanghai: Shanghai Normal University, 2018.

[75]

, Li

et al . Genome-wide analysis, molecular cloning and expression profiling reveal tissue-specifically expressed, feedback-regulated, stress-responsive and alternatively spliced novel genes involved in gibberellin metabolism in Salvia miltiorrhiza[J]. BMC Genomics, 2015, 16: 1087-1108.

https://doi.org/10.1186/s12864-015-2315-5

[76]

Hou

, Lee

LYC

, Xia

et al . DELLAs modulate jasmonate signaling via competitive binding to JAZs[J]. Devel Cell, 2010, 19: 884-894.

https://doi.org/10.1016/j.devcel.2010.10.024

[77]

Locascio

, Blázquez

, Alabadí

et al . Genomic analysis of DELLA protein activity[J]. Plant Cell Physiol, 2013, 54: 1229-1237.

https://doi.org/10.1093/pcp/pct082

[78]

Chini

, Fonseca

, Fernandez

et al . The JAZ family of repressors is the missing link in jasmonate signalling[J]. Nature, 2007, 448: 666-671.

https://doi.org/10.1038/nature06006

[79]

Weiss

, Ori

. Mechanisms of cross talk between gibberellin and other hormones[J]. Plant Physiol, 2007, 144: 1240-1246.

https://doi.org/10.1104/pp.107.100370

[80]

Bai

, Xia

, Wang

et al . Molecular cloning and characterization of five SmGRAS genes associated with tanshinone biosynthesis in Salvia miltiorrhiza hairy roots[J]. PLoS One, 2017, 12:e0185322

[81]

Wang

. Identification of GRAS Transcription Factor Family and Study on the Function of DELLA Subfamily Gene in Salvia miltiorrhiza (丹参GRAS转录因子家族鉴定及DELLA亚家族基因功能研究)[D]. Xi'an: Shaanxi Normal University, 2018.

[82]

Scarpeci

, Frea

, Zanor

et al . Overexpression of AtERF019 delays plant growth and senescence and improves drought tolerance in Arabidopsis[J]. J Exp Bot, 2017, 68: 673-685. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=ea7c79ff5c967ace76255bc5df7085b5

[83]

, Luo

, Xu

et al . Genome-wide identification of the AP2/ERF gene family involved in active constituent biosynthesis in Salvia miltiorrhiza[J]. Plant Gen, 2016, 9: 77-87.

https://doi.org/10.3835/plantgenome2015.08.0077

[84]

Huang

. Isolation and Functional Analysis of Two ERF Transcription Factors in Salvia miltiorrhiza (丹参两个ERF类转录因子的克隆与功能分析)[D]. Shanghai: Shanghai Normal University, 2015.

[85]

Bai

, Li

, Jia

et al . The ethylene response factor SmERF6 co-regulates the transcription of SmCPS1 and SmKSL1 and is involved in tanshinone biosynthesis in Salvia miltiorrhiza hairy roots[J]. Planta, 2018, 248: 243-255.

https://doi.org/10.1007/s00425-018-2884-z

[86]

Bai

. Study on the Transcriptional Regulation of SmCPS1 and SmKSL1, are Involved in Tanshinone Biosynthesis (丹参酮合成关键基因SmCPS1与SmKSL1转录调控的研究)[D]. Yangling: Northwest A&FUniversity, 2019.

[87]

Sun

, Shi

, Wang

et al . The biosynthesis of phenolic acids is positively regulated by the JA-responsive transcription factor ERF115 in Salvia miltiorrhiza[J]. J Exp Bot, 2019, 70: 243-254.

https://doi.org/10.1093/jxb/ery349

[88]

Huang

, Sun

, Yuan

et al . The AP2/ERF transcription factor SmERF1L1 regulates the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza[J]. Food Chem, 2019, 274: 368-375.

https://doi.org/10.1016/j.foodchem.2018.08.119

[89]

Zhang

, Ji

, Xu

et al . The AP2/ERF transcription factor SmERF128 positively regulates diterpenoid biosynthesis in Salvia miltiorrhiza[J]. Plant Mol Biol, 2019, 100: 83-93.

https://doi.org/10.1007/s11103-019-00845-7

[90]

, Liang

, Li

et al . Genome-wide characterisation and expression profiling of the LBD family in Salvia miltiorrhiza reveals the function of LBD50 in jasmonate signaling and phenolic biosynthesis[J]. Ind Crops Products, 2020, 144: 112006-112019.

https://doi.org/10.1016/j.indcrop.2019.112006

[91]

Thatcher

, Powell

, Aitken

EAB

et al . The lateral organ boundaries domain transcription factor LBD20 functions in fusarium wilt susceptibility and jasmonate signaling in Arabidopsis[J]. Plant Physiol, 2012, 160: 407-418.

https://doi.org/10.1104/pp.112.199067

[92]

, Liu

, An

et al . Cloning and elucidation of the functional role of apple MdLBD13 in anthocyanin biosynthesis and nitrate assimilation[J]. Plant Cell Tissue Organ Cult, 2017, 130: 47-59.

https://doi.org/10.1007/s11240-017-1203-x

[93]

Feng

. Cloning and Function of Transcription Factors SmLBD23 and SmLBD16 in Salvia miltiorrhiza (丹参转录因子SmLBD23和SmLBD16的克隆及功能研究)[D]. Xi'an: Shaanxi Normal University, 2018.

[94]

Chen

, Song

, Sun

et al . Herbal genomics:examining the biology of traditional medicines[J]. Science, 2015, 347:S27-S29 https://www.researchgate.net/profile/Qihe_Xu/publication/270960764_Evaluating_the_safety_of_herbal_medicines_Integrated_toxicological_approaches/links/54baf28a0cf29e0cb04bd73c.pdf

[95]

Zhang

, Wu

, Zhao

et al . Genome-wide analysis and molecular dissection of the SPL gene family in Salvia miltiorrhiza[J]. J Integr Plant Biol, 2014, 56: 38-50.

https://doi.org/10.1111/jipb.12111

[96]

Zhang

, Xu

, Ji

et al . Genomic survey of bZIP transcription factor genes related to tanshinone biosynthesis in Salvia miltiorrhiz[J]. Acta Pharm Sin B, 2018, 8: 295-305.

https://doi.org/10.1016/j.apsb.2017.09.002

[97]

, Shao

, Lu

. Identification and characterization of mRNA-like noncoding RNAs in Salvia miltiorrhiza[J]. Planta, 2015, 241: 1131-1143.

https://doi.org/10.1007/s00425-015-2246-z

[98]

Zhou

, Gao

, Rong

et al . Modular pathway engineering of diterpenoid synthases and the mevalonic acid pathway for miltiradiene production[J]. J Am Chem Soc, 2012, 134: 3234-3241.

https://doi.org/10.1021/ja2114486

[99]

Dai

, Liu

, Huang

et al . Production of miltiradiene by metabolically engineered Saccharomyces cerevisiae[J]. Biotechnol Bioeng, 2012, 109: 2845-2853

https://doi.org/10.1002/bit.24547

2020年第55卷第9期

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doi: 10.16438/j.0513-4870.2020-0429

接收时间：2020-03-30
首发时间：2026-01-23
出版时间：2020-09-12

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收稿日期：2020-03-30
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浙江经贸职业技术学院生物制药教研室, 浙江杭州 310018

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*詹忠根,Tel: 86-571-86929836, E-mail:zhzg9321@163.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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