CN110747202A - 一种岷江百合WRKY转录因子基因LrWRKY11及应用 - Google Patents
一种岷江百合WRKY转录因子基因LrWRKY11及应用 Download PDFInfo
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Abstract
本发明公开了一种岷江百合WRKY转录因子基因LrWRKY11，其核苷酸序列如SEQ ID NO:1所述，编码如SEQ ID NO:2所示氨基酸序列的蛋白质，本发明通过功能基因组学相关技术研究证实LrWRKY11基因具有提高植物抗真菌的功能，将本发明抗真菌LrWRKY11基因构建到植物表达载体上并转入烟草中过量表达，转基因烟草具有很强的对真菌病原菌的抗性，实验结果显示超表达LrWRKY11的转基因烟草对稻黑孢霉、轮枝镰刀菌、葡萄座腔菌、茄腐镰刀菌、人参链格孢的侵染具有很高水平的抗性。
Description
技术领域
本发明涉及分子生物学以及基因工程相关技术研究领域，特别是一种岷江百合WRKY转录因子基因LrWRKY11及应用。
背景技术
植物在生物或非生物胁迫因子的影响下，会发生一系列形态、生理和生化上的病理变化，并阻碍其正常生长，危害其经济价值。生物胁迫因子包括真菌、细菌等，产生植物病害的症状类型主要为变色、坏死、腐烂、萎蔫、畸形，其中以坏死、腐烂、萎蔫为主。由真菌引起的植物病害约占70~80%，病症常表现为各种霉、点、粒、粉等，严重影响植物的生长发育和产量。由于真菌性病害容易通过气流和水流传播，一般多采用化学药剂进行防治，或通过改进栽培管理措施加以防治。化学药剂如杀菌剂，虽见效极快，却存在滥用杀菌剂等问题，使得化学物残留于植株或土壤，影响环境，危害人类健康。改进栽培管理措施如农作物轮作，虽有一定效果，但耗时长，见效慢。随着基因工程技术的迅速发展，通过DNA重组技术导入抗真菌基因，培育抗病植物新品种有望从根本上解决真菌病害问题。
WRKY转录因子是植物防御信号的调节因子，在受到病原体攻击后调节下游靶基因的转录，从而调控宿主植物的免疫应答。先天免疫系统在识别病原体相关的分子模式或病原体通过植物的特定受体释放的效应分子后被激活（Jones JDG, Dangl JL. The plantimmune system. Nature, 2006, 444:323-329）。WRKY 转录因子不仅在植物对生物和非生物胁迫的应答中起重要作用，而且还参与碳水化合物的合成、衰老、发育和次级代谢产物的合成。WRKY转录因子的DNA 结合结构域被命名为WRKY 结构域，其具有不变的WRKYGQK 序列和CX4-5CX22-23HXH 锌指模体（Bakshi M, Oelmüller R. WRKY transcription factors.Plant Signal Behav, 2014, 9:247-258）。基于WRKY结构域的数目和锌指模体的类型将WRKY转录因子分为3类。第I类的成员通常含有两个WRKY结构域，而具有一个WRKY结构域的大多数蛋白质属于第II类。第III类蛋白质也只有一个WRKY结构域，但锌指基序的模式不同。第I和第II类成员所具有的锌指模体为C2-H2（C-X4-5-CX22-23-H-X1-H），其中X是任意氨基酸。第III 类WRKY 蛋白所含的锌指模体为C2-HC（C-X7-C-X23-H-X-C）。第I类的WRKY蛋白随后被分为2个亚族，I a含有C2H2锌指基序，I b含有C2HC锌指基序。基于WRKY结构域的系统发育分析，进一步将第II类分为5个亚组（Li H, Yan X, Yu X, et al. Expressionand functional analysis of two genes encoding transcription factors, VpWRKY1,and VpWRKY2, isolated from Chinese wild Vitis pseudoreticulata. Planta, 2010,232:1325-1337）。
目前已经从多种植物中分离了大量WRKY转录因子基因。酵母双杂交和EMSA试验表明拟南芥（Arabidopsis thaliana）的WRKY50转录因子通过其DNA结合域与PR1的两个启动子元件结合，验证了AtWRKY50在水杨酸诱导的系统性获得抗性中发挥重要作用（HussainRMF, Sheikh AH, Imran H, et al. Arabidopsis WRKY50 and TGA transcriptionfactors synergistically activate expression of PR1. Front Plant Sci, 2018, 9:930-946）。野草莓（Fragaria vesca）中的WRKY42转录因子基因在拟南芥中过表达，增强了转基因拟南芥对白粉菌（Golovinomyces cichoracearum）的抗性，并且也提高了对盐和干旱胁迫的耐受性（Wei W, Meng-Yuan C, Hu Y, et al. Ectopic expression of,FvWRKY42, a WRKY transcription factor from the diploid woodland strawberry ( Fragaria vesca ), enhances resistance to powdery mildew, improves osmoticstress resistance, and increases abscisic acid sensitivity in Arabidopsis.Plant Sci, 2018, 275:60-74）。在对水稻立枯病和叶鞘腐败病具有抗性的水稻（Oryza sativa）品种中检测到WRKY12、WRKY13以及抗病相关基因TIFY9和PR2的高表达；将OsWRKY13基因在敏感性的水稻品种中过表达后，其对立枯丝核菌（Rhizoctonia solani）和稻帚枝霉（Sarocladium oryzae）的抗性增强（Jimmy JL, Babu S. Gene network mediated byWRKY13 to regulate resistance against sheath infecting fungi in rice (Oryza sativa L.). Plant Sci, 2019, 280:269-282）。病毒诱导的基因沉默引起的辣椒(Capsicum annuum) WRKY22基因功能丧失，增强了辣椒对青枯雷尔氏菌（Ralstonia solanacearum）的敏感性，并下调防御相关基因CaPO2、CaPR4、CaACC、CaBPR1、CaDEF1、CaHIR1、CaWRKY40的表达，引起细胞死亡；相反，在辣椒叶片中CaWRKY22的瞬时过表达上调了防御相关基因的表达，对青枯雷尔氏菌抗性增强（Hussain A, Li X, Weng Y, et al.CaWRKY22 acts as a positive regulator in pepper response to Ralstonia Solanacearum by constituting networks with CaWRKY6, CaWRKY27, CaWRKY40, andCaWRKY58. Int J Mol Sci, 2018, 19(5):1426-1445）。将大麦WRKY6，WRKY40和WRKY70在小麦（Triticum aestivum）叶片中瞬时表达，提高了小麦对叶锈菌（Puccinia triticina）的抗性（Gao J, Bi W, Li H, et al. WRKY transcription factors associated withNPR1-mediated acquired resistance in barley are potential resources toimprove wheat resistance to Puccinia triticina. Front Plant Sci, 2018, 9:1486-1500）。
不同植物的WRKY转录因子除了显示正调节作用外，也有负调节作用。来自葡萄（Vitis quinquangularis）的WRKY52转录因子在水杨酸信号转导途径中起着至关重要的作用，在拟南芥中VqWRKY52的异位表达，与野生型相比，转基因拟南芥增强了对白粉病和丁香假单胞菌（Pseudomonas syringae）的抗性，同时增加了对灰葡萄孢菌（Botrytis cinerea）的敏感性（Wang X, Guo R, Tu M, et al. Ectopic expression of the wild grapeWRKY transcription factor VqWRKY52 in Arabidopsis thaliana enhancesresistance to the biotrophic pathogen powdery mildew but not to thenecrotrophic pathogen Botrytis cinerea. Front Plant Sci, 2017, 8:97-110）。甘蔗（Saccharum）ScWRKY3蛋白定位中位于烟草叶片的细胞核中，接种黑穗病病原菌（Sporisorium scitamineum）后，ScWRKY3在抗黑穗病品种中稳定表达，在易染黑穗病品种中转录水平被下调（Wang L, Liu F, Zhang X, et al. Expression characteristicsand functional analysis of the ScWRKY3 gene from sugarcane. Int J Mol Sci,2018, 19:4059-4081）。此外，ScWRKY3基因在烟草中的瞬时过表达，然后别接种茄腐镰刀菌（Fusarium solani）和青枯雷尔氏菌，转基因叶片的枯萎病症状比对照组更明显，表明ScWRKY3在防卫反应中起负调节作用。通过病毒诱导的基因沉默产生GbWRKY1敲除的棉花(Gossypium barbadense)植株，结果对灰葡萄孢和大丽轮枝菌（Verticillium dahliae）具有更强的抗性；而在棉花和拟南芥中过表达GbWRKY1基因，转基因植株减弱了对灰葡萄孢和大丽轮枝菌的抗性（Li C, He X, Luo X, et al. Cotton WRKY1 mediates the plantdefense-to-development transition during infection of cotton by Verticillium dahliae by activating JASMONATE ZIM-DOMAIN1 expression. Plant Physiol, 2014,166:2179-2194）。显然，GbWRKY1在防卫反应中负调控作用。
在种球繁殖及鲜切花生产过程中，百合易受到真菌、病毒、细菌等多种病原菌的危害。目前发现的百合病害达几十种之多，主要由尖孢镰刀菌(F.oxysporum)引起的枯萎病（又称为基腐病、茎腐病）是百合生产中危害最严重的病害。镰刀菌侵染百合种球后引起基盘坏死、鳞片腐烂脱落，造成种球质量下降；植株感染镰刀菌后叶片变黄、萎蔫下垂，植株提早枯萎死亡，严重影响百合切花的产量和品质。岷江百合(Lilium regale Wilson)为我国特有品种，仅分布于岷江流域海拔800～2700m 的河谷到山腰的岩石缝中，具有很强的枯萎病抗性，是现代百合育种的重要种质资源。WRKYs参与应答多种生物和非生物胁迫，是植物防御系统的重要组成部分。从岷江百合中分离参与抗病防卫反应的WRKY转录因子基因，并分析其功能，有助于深入发掘岷江百合的珍稀基因资源，为植物抗病基因工程提供候选基因。
发明内容
本发明的目的是提供一种岷江百合WRKY转录因子基因LrWRKY11及应用，即其在提高烟草对稻黑孢霉（Nigrospora oryzae）、轮枝镰刀菌（F. verticillioides）、葡萄座腔菌（Botryosphaeria dothidea）、茄腐镰刀菌、人参链格孢（Alternaria panax）抗性中的应用。
本发明从岷江百合中克隆获得的具有抗真菌活性的岷江百合WRKY转录因子基因LrWRKY11的全长基因，LrWRKY11的核苷酸序列如SEQ ID NO:1所示，该基因全长为931 bp，包含一个528 bp的开放阅读框、185 bp的5′非翻译区(untranslated region, UTR)及218bp的3′UTR，编码如SEQ ID NO:2所示氨基酸序列的蛋白质。
本发明分离克隆岷江百合的一个抗真菌相关转录因子基因的完整cDNA片段，通过根癌农杆菌(Agrobacterium tumefaciens)介导将目的基因转入受体植物中过量表达，并通过进一步实验验证该基因是否具有抗真菌的活性，为后期利用该基因改良烟草及其他植物抵御真菌病害的能力奠定基础；发明人将这个基因命名为LrWRKY11。
本发明所述WRKY转录因子基因LrWRKY11的编码区是序列表SEQ ID NO:1中第186-713位所示的核苷酸序列。
本发明将岷江百合WRKY转录因子基因LrWRKY11应用在提高烟草对稻黑孢霉、轮枝镰刀菌、葡萄座腔菌、茄腐镰刀菌、人参链格孢的抗性中，具体操作如下：
(1)采用扩增LrWRKY11的特异引物，从接种尖孢镰刀菌后的岷江百合根中提取总RNA，通过逆转录-聚合酶链式反应(reverse transcription-polymerase chain reaction，RT-PCR)扩增出LrWRKY11的全长编码区，然后将其连接到pGEM-T载体上，经测序获得具有目的基因的克隆；
(2)用限制性内切酶BamHI和XbaI酶切pGEM-T-LrWRKY11载体和植物表达载体pCAMBIA2300S，通过胶回收得到目的基因片段和载体大片段；再将所获得LrWRKY11基因片段与pCAMBIA2300S载体片段连接，构建植物超表达载体；之后将所构建的重组载体通过根癌农杆菌介导转入烟草中表达；
(3)重组载体T-DNA上具有卡那霉素抗性基因，用添加卡那霉素的分化培养基筛选转化子，并通过PCR以及RT-PCR检测得到真正的转基因植株，分析转基因植株对于植物病原真菌的抗性，最后筛选出对真菌抗性明显增强的转基因植株。
本发明为提高植物对真菌病害的抗性提供了一种新的方法，通过基因工程手段培育抗病植物可以克服传统育种的不足，不仅育种周期缩短，而且操作简单，容易获得高抗材料。本发明来自岷江百合的LrWRKY11转录因子能增强植物对几种病原真菌的抗性，将该基因导入烟草中，可以产生具有真菌抗性的新品种和新材料。利用基因工程技术培育抗性植物品种和材料具有明显的优势和不可取代的重要性。它不仅可以为大规模生产农作物、花卉、药用植物、林木等提供方便，大量减少化学农药的使用，还可以为农业生产节约成本、减小环境污染，因此本发明具有广阔的市场应用前景。
附图说明
图1是本发明中部分LrWRKY11转基因烟草基因组DNA的PCR检测结果，其中Marker：DL2000 DNA Marker (大连宝生物)；阳性对照：质粒pGEM-T-LrWRKY11为模板的PCR反应；WT：非转基因烟草(野生型)总DNA为模板进行的PCR；
图2是本发明中部分阳性LrWRKY11转基因烟草中LrWRKY11转录水平的表达分析结果图，其中Marker：DL2000 DNA Marker (大连宝生物)；WT：非转基因烟草总RNA逆转录cDNA为模板的PCR产物；阳性对照：质粒pGEM-T-LrWRKY11为模板的PCR产物；
图3是本发明中LrWRKY11转基因烟草抗病性分析的结果图；其中a、b、c、d、e图示中接种的真菌分别是稻黑孢霉、轮枝镰刀菌、葡萄座腔菌、茄腐镰刀菌、人参链格孢；WT为野生型烟草，3、4、5、14为LrWRKY11转基因烟草。
具体实施方式
下面通过附图和实施例对本发明作进一步详细说明，但本发明保护范围不局限于所述内容，实施例中方法如无特殊说明均为常规方法，使用的试剂如无特殊说明均为常规市售试剂或按常规方法配制的试剂。
实施例1：LrWRKY11全长基因克隆以及序列分析
用尖孢镰刀菌接种百合的根，用接种后24 h的根提取总RNA，用液氮将处理过的百合的根研磨成粉末，然后转入离心管中，采用异硫氰酸胍法提取总RNA；采用M-MLV逆转录酶(promega)以总RNA为模板合成cDNA第一链，反应体系和操作过程为：取5 μg Total RNA，依次加入50 ng oligo (dT)、2 μL dNTP Mix (2.5mM each)，用DEPC水将反应体积补齐至14.5 μL；混匀后，70℃加热变性5 min后迅速在冰上冷却5 min，然后依次加入4 μL 5×First-stand buffer、0.5 μL RNasin (200U)、1 μL M-MLV (200U)，混匀并简短离心，42℃温浴1.5 h，取出后70℃加热10 min，终止反应；cDNA第一链合成后置于-20℃保存备用。
以合成的第一链cDNA为模板，扩增目的基因LrWRKY11，所用上下游引物序列分别为5’ATGGCGGCTGTAGGAGTT3’及5’GCAGCTTTCAATTACTCAAAAG3’。采用AdvantageTM 2 PCREnzyme (Clontech)扩增出目的基因。PCR反应条件：95℃ 1 min；94℃ 30 s，60℃ 30 s，72℃ 32 s，32个循环；72℃ 7 min。反应体系(20 μL)为1 μL cDNA、2 μL 10×Advantage 2PCR Buffer、1.8 μL dNTP Mix (10 mM each)、0.2μL 正向引物(10 μM)、0.2 μL 反向引物(10 μM)、0.2 μL Advantage 2 PCR Polymerase Mix、14.6 μL PCR-Grade water。PCR结束后，取10 μL进行琼脂糖凝胶电泳，用以检测扩增产物的特异性以及大小。
对PCR产物进行TA克隆，使用的试剂盒为pGEM-T vector kit（Promega），反应体系和操作过程为：取1.5 μL PCR产物，依次加入1 μL pGEM-T vector（50 ng/μL）和2.5 μL 2×Ligation solution I，混匀后置于16 ℃过夜反应。采用热激转化法将连接产物转入大肠杆菌DH5α中。使用含有氨苄青霉素（ampicillin，Amp）的LB固体培养基筛选阳性克隆，挑选若干个单菌落，摇菌后用扩增LrWRKY11的特异引物鉴定出多克隆位点插入LrWRKY11的克隆，将所鉴定的克隆进行测序，最终获得的LrWRKY11全长cDNA为931 bp，通过NCBI ORFfinder （http://www.ncbi.nlm.nih.gov/gorf/gorf.html）分析发现其包含一个528 bp的开放读码框（见序列表），LrWRKY11编码一个含175个氨基酸的蛋白质LrWRKY11，其分子量约为19.6 KDa，等电点约为5.92。
实施例2：植物超表达载体构建
采用SanPrep柱式质粒DNA小量抽提试剂盒（上海生工）提取插入LrWRKY11的大肠杆菌质粒pGEM-T-LrWRKY11以及植物表达载体pCAMBIA2300S的质粒，取1 μL用于琼脂糖凝胶电泳以检测所提取质粒的完整性及浓度高低；用限制性内切酶BamHI和XbaI分别对质粒pGEM-T-LrWRKY11和pCAMBIA2300S进行双酶切(100 μL体系)，反应体系和操作过程为：分别取20μL pGEM-T-LrWRKY11和pCAMBIA2300S质粒、依次加入10 μL 10×K buffer、5 μLBamHI、5 μL XbaI、60 μL ddH2O，混匀后短时离心，置于37℃过夜反应。将所有酶切产物进行琼脂糖凝胶电泳，然后使用试剂盒对LrWRKY11片段和pCAMBIA2300S载体大片段分别进行胶回收，取1μL回收产物通过琼脂糖凝胶电泳检测回收片段的大小以及浓度，置于-20℃保存备用。
利用T4 DNA Ligase (TaKaRa)，将回收的LrWRKY11DNA片段和pCAMBIA2300S载体片段连接起来，反应体系(20 μL)和操作过程为：取10 μL LrWRKY11DNA片段依次加入2 μLpCAMBIA2300S载体DNA、2 μL 10×T4 DNA Ligase Buffer、1 μL T4 DNA Ligase、5 μLddH2O，混匀后短时离心，然后16℃水浴过夜反应。接着采用热激转化法将连接产物转入大肠杆菌DH5α中，用含有50 mg/L卡那霉素(kanamycin，Km)的固体培养基筛选阳性克隆。挑选单菌落摇菌，以菌液为模板用扩增LrWRKY11的特异引物进行PCR，挑选出LrWRKY11与pCAMBIA2300S成功连接的克隆，在得到的阳性菌株中加入甘油并置于-80℃保存备用。
采用试剂盒提取并纯化上述大肠杆菌DH5α中的pCAMBIA2300S-LrWRKY11质粒。随后用液氮冻融法将上述构建的植物表达载体pCAMBIA2300S-LrWRKY11转入所制备的根癌农杆菌LBA4404感受态细胞中。操作步骤为：取0.2 μg pCAMBIA2300S-LrWRKY11，质粒加入含有100 μL感受态细胞的离心管中，轻轻混匀后冰浴5 min，随后转入液氮中冷冻1 min，然后迅速置于37℃水浴5 min，再冰浴2 min，之后加入500 μL LB液体培养基于28℃振荡培养4h。将活化后的农杆菌涂于含有50 mg/L Km的LB固体培养基上，28℃倒置培养。挑选单菌落摇菌，再用扩增LrWRKY11的特异性引物进行PCR反应，检测pCAMBIA2300S-LrWRKY11是否转入农杆菌中；对于阳性克隆，加入甘油后置于-80℃保存备用。
实施例3：农杆菌介导的植物遗传转化以及转基因植物筛选
本实验的转基因受体是烟草(Nicotiana tabacum)，将烟草种子用75%的酒精浸泡30s，用无菌水洗涤后用0.1 %的HgCl2浸泡8 min，然后再用无菌水洗涤若干次，播种于1/2 MS培养基上，28℃暗培养5-8d，发芽后转至光照培养箱（25℃，16h/d光照），以后每月用1/2MS培养基继代一次。
从-80℃冰箱中取出保存的含有pCAMBIA2300S-LrWRKY11质粒的农杆菌LBA4404菌种，取20 μL接种于5 mL含有50 mg/L Km和20 mg/L利福平的LB液体培养基中，28℃培养至培养基浑浊。吸取1 mL浑浊的菌液至含有50 mg/L Km的LB固体培养基上，28℃培养48 h；随后将LB固体培养基上的农杆菌刮下适量接种于附加有20 mg/L的乙酰丁香酮的MGL液体培养基中，28℃振荡培养5-8 h以活化农杆菌。
取烟草无菌苗叶片切成1 cm2左右的叶盘，完全浸泡于上述含有活化农杆菌的MGL液体培养基中，浸染时间为15 min，用无菌滤纸吸干叶片表面的菌液，将叶盘置于共培养基上进行室温培养，烟草转化的共培养基为MS+0.02 mg/L 6-BA+2.1 mg/L NAA+30 g/Lsucrose+6 g/L琼脂，22℃无光条件下共培养2天。
将共培养后的叶盘转到加有抗生素的MS筛选培养基中分化成苗，同时筛选转基因植株。烟草筛选培养基为MS+0.5 mg/L 6-BA+0.1 mg/L NAA+30 g/L sucrose+6 g/L琼脂+50 mg/L Km+200 mg/L 头孢霉素（cefotaxime sodium salt，Cef）；筛选培养时将培养瓶转移至光照培养箱培养（25℃，16h/d光照，8h/d黑暗），待烟草长出芽后用含有50 mg/L Km和200 mg/L Cef的MS培养基继代培养。
采用CTAB法提取转基因烟草植株叶片的基因组DNA，将提取的基因组DNA取1 μL通过琼脂糖凝胶电泳检测其完整性和浓度，以转基因植株的基因组DNA为模板用扩增LrWRKY11的特异引物进行PCR，PCR结束后，取8 μL产物用于琼脂糖凝胶电泳以检测阳性转基因植株，部分烟草转基因植株的扩增结果如图1所示，LrWRKY11转基因烟草共筛选到45株阳性转基因植株。
实施例4：转基因烟草中LrWRKY11的表达分析以及转基因植株抗病性分析
取阳性转基因单株以及非转基因烟草（野生型）的嫩叶提取总RNA，逆转录生成cDNA第一链，并以此为模板用扩增LrWRKY11的特异引物进行PCR，根据PCR结果分析各转基因单株中LrWRKY11转录水平的表达，总RNA提取以及RT-PCR的方法与实施例1中相同，PCR结束之后，取10μL用于琼脂糖凝胶电泳，部分单株的检测结果如图2所示，共检测到32个转基因单株中LrWRKY11在转录水平大量表达，这些单株的编号为1～32。
将实验室保存的几种真菌接种于PDA固体培养基（200 g/L马铃薯，15 g/L琼脂，20g/L葡萄糖）上活化，28℃暗培养。以温室栽培的LrWRKY11转基因烟草和野生型烟草为试材，用砂纸在叶片上轻轻摩擦出同等规格的伤口，接上大小相等的新鲜菌丝块，然后用保鲜膜包裹叶片保持湿度。气候培养箱内培养一段时间后，收集接种叶片，观察LrWRKY11转基因烟草叶片和野生型烟草叶片受真菌侵染的发病情况，结果如图3所示，野生型烟草接种稻黑孢霉、轮枝镰刀菌、葡萄座腔菌、茄腐镰刀菌、人参链格孢等几种病原真菌后，发病症状明显，病斑面积大。然而，LrWRKY11转基因烟草对上述五种病原真菌的侵染具有很强的抗性，发病的症状很轻微，发病面积也明显小于转基因烟草。
序列表
<110> 昆明理工大学
<120> 一种岷江百合WRKY转录因子基因LrWRKY11及应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 931
<212> DNA
<213> 岷江百合(Lilium regale Wilson)
<400> 1
atttagttga agcactacac agaaacaaca gaaccaccca agaaatttcc ccagtcgagt 60
aaggaaactt acaacgaagt tatccctcgc tataaaaact acacattctc ccctatatcc 120
atcatcaata gtgctcgaga taaagtcata taaacccgta cgttcattac aatcctcact 180
agatcatggc ggctgtagga gttccctcat tccatcccgc catgtgcagc tcccactccc 240
agcagacctc tgcagtggac atttcggctc cattcgatct ctccgattgc atcttgttgg 300
acgatgatct gctcgttgag tccccgccta ctgcacagac actgattcca aatctccctt 360
acagcagcag aagggacgac gttaagaaag agacagcagg agataaccat aggatcgcat 420
tccgaacaag gtcggaggtc gatatcatgg atgatgggta cagatggaga aagtacggta 480
agaagaccgt caagaacagc ccaaatccaa ggaactacta ccgatgctcg acagatggtt 540
gctcggtgaa gaagagggtg gagagggcca gagatgatcc gagctacgtg atcaccactt 600
atgagggaac ccataatcat acaagcccga gtgttgttta ctacacagct caggatgatg 660
actcgggtcg gttcttcgtc tccgggtgcc aagatttacc tcaaagctca tgaaacctaa 720
cttttgagta attgaaagct gcactaccgt atttttatga gttgtatctg ataatgaact 780
catttgagct catcttgttg aagcttatag ggatggtgtg taccttattt tcttgctaaa 840
ttttgatgta tatatatgtc ttatatgttt aaccagcttt ttcgtaaaag ttgattaatg 900
caattagaat tctcgagtgg gaagtttgtc g 931
<210> 2
<211> 175
<212> PRT
<213> 岷江百合(Lilium regale Wilson)
<400> 2
Met Ala Ala Val Gly Val Pro Ser Phe His Pro Ala Met Cys Ser Ser
1 5 10 15
His Ser Gln Gln Thr Ser Ala Val Asp Ile Ser Ala Pro Phe Asp Leu
20 25 30
Ser Asp Cys Ile Leu Leu Asp Asp Asp Leu Leu Val Glu Ser Pro Pro
35 40 45
Thr Ala Gln Thr Leu Ile Pro Asn Leu Pro Tyr Ser Ser Arg Arg Asp
50 55 60
Asp Val Lys Lys Glu Thr Ala Gly Asp Asn His Arg Ile Ala Phe Arg
65 70 75 80
Thr Arg Ser Glu Val Asp Ile Met Asp Asp Gly Tyr Arg Trp Arg Lys
85 90 95
Tyr Gly Lys Lys Thr Val Lys Asn Ser Pro Asn Pro Arg Asn Tyr Tyr
100 105 110
Arg Cys Ser Thr Asp Gly Cys Ser Val Lys Lys Arg Val Glu Arg Ala
115 120 125
Arg Asp Asp Pro Ser Tyr Val Ile Thr Thr Tyr Glu Gly Thr His Asn
130 135 140
His Thr Ser Pro Ser Val Val Tyr Tyr Thr Ala Gln Asp Asp Asp Ser
145 150 155 160
Gly Arg Phe Phe Val Ser Gly Cys Gln Asp Leu Pro Gln Ser Ser
165 170 175
<210> 3
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<212> DNA
<213> 人工序列(Artificial)
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atggcggctg taggagtt 18
<210> 4
<211> 22
<212> DNA
<213> 人工序列(Artificial)
<400> 4
gcagctttca attactcaaa ag 22
Claims (2)
1.一种岷江百合WRKY转录因子基因LrWRKY11，其特征在于：其核苷酸序列如SEQ IDNO:1所示，编码如SEQ ID NO:2所示氨基酸序列的蛋白质。
2.权利要求1所述的岷江百合WRKY转录因子基因LrWRKY11在提高烟草对稻黑孢霉（Nigrospora oryzae）、轮枝镰刀菌（Fusarium verticillioides）、葡萄座腔菌（Botryosphaeria dothidea）、茄腐镰刀菌（Fusarium solani）、人参链格孢（Alternaria panax）抗性中的应用。
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111217896A (zh) * | 2020-02-28 | 2020-06-02 | 天津大学 | 人参PgWRKY4X转录因子在调控人参中人参皂苷类化合物含量的应用 |
| CN111675755A (zh) * | 2020-06-05 | 2020-09-18 | 中国科学院武汉植物园 | 调控植物耐盐性的狗牙根转录因子CdWRKY50及其应用 |
| CN112481294A (zh) * | 2020-12-10 | 2021-03-12 | 山东省烟台市农业科学研究院 | 一种柿wrky转录因子基因在提高柿炭疽病抗性中的应用 |
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Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020076775A1 (en) * | 2000-03-17 | 2002-06-20 | Crane Virginia C. | WRKY transcription factors and methods of use |
| US6664446B2 (en) * | 1999-03-23 | 2003-12-16 | Mendel Biotechnology, Inc. | Transgenic plants comprising polynucleotides encoding transcription factors that confer disease tolerance |
| CN101423549A (zh) * | 2007-10-30 | 2009-05-06 | 韩国生命工学研究院 | 来源于甘薯根的IbTIP1基因及其启动子 |
| CN101883783A (zh) * | 2007-11-26 | 2010-11-10 | 巴斯夫植物科学有限公司 | 具有增强的产量相关性状的植物及其制备方法 |
| CN101979550A (zh) * | 2010-10-09 | 2011-02-23 | 山东大学 | 玉米磷酸烯醇式丙酮酸羧激酶基因启动子克隆和应用 |
| CN103320448A (zh) * | 2013-06-26 | 2013-09-25 | 昆明理工大学 | 一种岷江百合bZIP转录因子基因LrbZIP1及应用 |
| CN104946668A (zh) * | 2015-06-29 | 2015-09-30 | 湖南省植物保护研究所 | cry1Ia基因及应用、由其编码的Cry1Ia蛋白及制备方法和应用 |
| CN105087601A (zh) * | 2015-09-07 | 2015-11-25 | 昆明理工大学 | 一种珠子参转录因子基因PjWRKY1的应用 |
| CN105441460A (zh) * | 2016-01-06 | 2016-03-30 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY1及应用 |
| CN109825510A (zh) * | 2019-03-19 | 2019-05-31 | 长江师范学院 | 一种岷江百合LrWRKY2基因及应用 |
| CN110151806A (zh) * | 2018-02-13 | 2019-08-23 | 昆明理工大学 | 一种熟三七及其制备方法和补血用途 |
| US10398100B1 (en) * | 2015-11-13 | 2019-09-03 | Syngenta Participations Ag | Markers associated with sudden death syndrome tolerance in soybeans |
| CN110734482A (zh) * | 2019-11-13 | 2020-01-31 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY4及应用 |
| CN110818782A (zh) * | 2019-11-13 | 2020-02-21 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY3及应用 |
| CN110818783A (zh) * | 2019-11-13 | 2020-02-21 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY2及应用 |
| CN111909941A (zh) * | 2020-09-04 | 2020-11-10 | 长江师范学院 | 一种百合转录因子基因LrWRKY-L1及应用 |
| CN112359049A (zh) * | 2020-12-10 | 2021-02-12 | 昆明理工大学 | 一种岷江百合几丁质酶基因LrCHI2及其应用 |
| CN112831504A (zh) * | 2021-03-16 | 2021-05-25 | 昆明理工大学 | 三七WRKY转录因子基因PnWRKY9及其应用 |
-
2019
- 2019-11-13 CN CN201911105582.9A patent/CN110747202B/zh active Active
Patent Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6664446B2 (en) * | 1999-03-23 | 2003-12-16 | Mendel Biotechnology, Inc. | Transgenic plants comprising polynucleotides encoding transcription factors that confer disease tolerance |
| US20020076775A1 (en) * | 2000-03-17 | 2002-06-20 | Crane Virginia C. | WRKY transcription factors and methods of use |
| CN101423549A (zh) * | 2007-10-30 | 2009-05-06 | 韩国生命工学研究院 | 来源于甘薯根的IbTIP1基因及其启动子 |
| CN101883783A (zh) * | 2007-11-26 | 2010-11-10 | 巴斯夫植物科学有限公司 | 具有增强的产量相关性状的植物及其制备方法 |
| CN101979550A (zh) * | 2010-10-09 | 2011-02-23 | 山东大学 | 玉米磷酸烯醇式丙酮酸羧激酶基因启动子克隆和应用 |
| CN103320448A (zh) * | 2013-06-26 | 2013-09-25 | 昆明理工大学 | 一种岷江百合bZIP转录因子基因LrbZIP1及应用 |
| CN104946668A (zh) * | 2015-06-29 | 2015-09-30 | 湖南省植物保护研究所 | cry1Ia基因及应用、由其编码的Cry1Ia蛋白及制备方法和应用 |
| CN105087601A (zh) * | 2015-09-07 | 2015-11-25 | 昆明理工大学 | 一种珠子参转录因子基因PjWRKY1的应用 |
| US10398100B1 (en) * | 2015-11-13 | 2019-09-03 | Syngenta Participations Ag | Markers associated with sudden death syndrome tolerance in soybeans |
| CN105441460A (zh) * | 2016-01-06 | 2016-03-30 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY1及应用 |
| CN110151806A (zh) * | 2018-02-13 | 2019-08-23 | 昆明理工大学 | 一种熟三七及其制备方法和补血用途 |
| CN109825510A (zh) * | 2019-03-19 | 2019-05-31 | 长江师范学院 | 一种岷江百合LrWRKY2基因及应用 |
| CN110734482A (zh) * | 2019-11-13 | 2020-01-31 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY4及应用 |
| CN110818782A (zh) * | 2019-11-13 | 2020-02-21 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY3及应用 |
| CN110818783A (zh) * | 2019-11-13 | 2020-02-21 | 昆明理工大学 | 一种岷江百合WRKY转录因子基因LrWRKY2及应用 |
| CN111909941A (zh) * | 2020-09-04 | 2020-11-10 | 长江师范学院 | 一种百合转录因子基因LrWRKY-L1及应用 |
| CN112359049A (zh) * | 2020-12-10 | 2021-02-12 | 昆明理工大学 | 一种岷江百合几丁质酶基因LrCHI2及其应用 |
| CN112831504A (zh) * | 2021-03-16 | 2021-05-25 | 昆明理工大学 | 三七WRKY转录因子基因PnWRKY9及其应用 |
Non-Patent Citations (5)
| Title |
|---|
| CUI,Q.: ""Lilium regale WRKY51 (WRKY51) mRNA, partial cds"", 《GENBANK DATABASE》 * |
| QI CUI等: ""Analysis of WRKY transcription factors and characterization of two Botrytis cinerea-responsive LrWRKY genes from Lilium regale"", 《PLANT PHYSIOLOGY AND BIOCHEMISTRY》 * |
| 周俐宏等: ""基于转录组百合WRKY 转录因子的挖掘与分析"", 《分子植物育种》 * |
| 普丽梅: ""岷江百合WRKY转录因子基因家族的发掘及分析"", 《中国优秀硕士学位论文全文数据库 基础科学辑》 * |
| 王国东等: ""漾濞大泡核桃JsWＲKY1过表达增强转基因烟草对胶孢炭疽菌的抗性"", 《华北农学报》 * |
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| CN112831504B (zh) * | 2021-03-16 | 2023-03-24 | 昆明理工大学 | 三七WRKY转录因子基因PnWRKY9及其应用 |
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