CN114350678B - 基因OsLUX在促进水稻抽穗和提高植物抗病性中的应用 - Google Patents
基因OsLUX在促进水稻抽穗和提高植物抗病性中的应用 Download PDFInfo
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Abstract
本发明公开了基因OsLUX在促进水稻抽穗和提高植物抗病性中的应用。OsLUX基因是水稻生物钟的核心组分,本发明通过Mutmap方法鉴定了OsLUX的三个突变体,表现为极端晚抽和晚抽表型。进一步研究发现OsLUX基因表现为昼夜节律表达,定位在细胞核和质膜中并具有转录自激活活性,通过抑制OsGI、Hd1和Ghd7来促进Ehd1、Hd3a和OsMADS14的表达,最终促进抽穗。同时,elh1/oslux‑1接种白叶枯菌相较于野生型表现出更加感病,表明OsLUX对白叶枯菌的抗性起正调控作用。本发明进一步丰富和完善了水稻生物钟基因调控光周期开花的理论基础。
Description
技术领域
本发明属于植物基因工程领域,具体地说,涉及基因OsLUX在促进水稻抽穗和提高植物抗病性中的应用。
背景技术
生物钟被赋予植物“中央处理器”的功能,几乎参与调控了水稻所有的生长发育、物质代谢及应激反应过程。外界环境因子的动态变化经输入途径传递到生物钟系统,使植物感知特定时间的信号变化,进而调控植物相应的生长发育进程以达到与外界环境变化的同步性。在大田环境下,作物的整个生命周期均处于变化的自然条件中(主要是光照和温度),生物钟协调水稻内源时间机制以适应外界环境近24小时的节律变化,此过程对调控作物抽穗和产量的成至关重要,如开花相关的周期性表达以应对环境因素的变化(Nakamichiet al.,2020);生物钟相关基因赋予作物以感知环境信号来直接调控碳水化合的代谢,获得更多能量来源来提高光合作用的利用率;淀粉在白天的合成积累及晚上的降解(Kim etal.,2017)水稻是我国重要的粮食作物,其抽穗期作为最重要的农艺性状之一,严格受控于生物钟的调节。
水稻中,生物钟调节光周期开花的进展主要集中在对OsELF3、OsCCA1/OsLHY、OsGI和OsFKF1等基因的研究上。长日照条件下,OsELF3作为开花激活子调控生物钟相关基因,oself3突变体中,OsCCA1/OsLHY表达量在早上下调,OsPRR95、OsPRR73、OsPRR59、OsPRR37、OsPRR1和OsGI的表达量上调。OsLHY作为生物钟核心组件,通过OsGI-Hd1途径精准调节水稻的抽穗期,在不同日照条件下具有完全依赖于Hd1的提早和延迟抽穗的双重功能(Zhou etal.2021)。OsPRR1和OsGI是生物钟晚间循环中的一环,参与水稻OsGI-Hd1-Hd3a开花途径的调控,可见OsGI在生物钟和水稻开花调节子间起着枢纽作用。此外,OsELF3作为负调节因子对下游Ghd7也有抑制作用,通过OsELF3的两条调控途径,激活了下游Hd3a和RFT1的表达以促进水稻在长日照条件下的开花(Yang et al.,2013)。在拟南芥中,FKF1作为蓝光受体参与CO的调控来诱导长日照下的开花;而在水稻中,OsFKF1可能是独立于光周期的自主开花激活子,通过调控下游Ehd2、Ghd7和Ehd1的表达来诱导开花。根据拟南芥中FKF1蓝光受体的保守性,OsFKF1-OsG1互作可能在蓝光下参与水稻独立于Ehd2和Ghd7途径的Ehd1的调节(Han et al.,2015)。
活性氧(ROS)是植物响应生物和非生物胁迫的重要信号分子,胞内ROS的稳态是由抗氧化系统所调控。研究表明,植物体内H2O2水平在持续光照条件下具有节律振荡。突变ELF3、LUX、TOC1等生物钟相关基因,也均造成H2O2水平和ROS相关基因表达节律性的丧失(Lai et al.,2012)。说明生物钟参与调控了ROS的稳态以维持细胞的正常代谢功能,生物钟相关基因在植物非生物胁迫调控中起作用。在生物胁迫上,植物生物钟也发挥着重要的作用。植株生长过程中,时刻遭受病原菌的侵扰,因此,植物整体上进化出两条防御机制,如PTI过程中的SA、JA和ROS的积累,物理性防御如气孔开张、细胞壁加厚和胼胝质积累等;ETI过程中R基因的特异性表达。研究表明,CCA1和LUX功能缺失引起生物钟节律紊乱而导致对病原菌的防御水平降低(Hevia et al.,2015;Zhang et al.,2013;Korneli et al.,2014)。CCA1至少部分是依赖气孔的调节来参与拟南芥的抗性,GRP7的表达具有节律性,在开花转变和病原菌防御方面发挥作用,GRP7作为CCA1的下游靶基因,调控气孔的开张来抵御病原菌的侵入(Zhang et al.,2013)。除CCA1外,其他生物钟基因如TOC1、ELF3和LUX同样也以依赖气孔的方式参与植物的抗性(Somers et al.,1998;Kinoshita et al.,2011;Zhanget al.,2013)。CCA1过表达植株表现出更感病的表型,然而GRP7过表达植株却没有相应的表型,说明CCA1还有可能通过其他途径参与拟南芥的抗性反应。最新研究表明,LUX能够参与多层次的防御反应,包括气孔的开张、系统获得性抗性、基础抗性和SA及JA信号途径。LUX作为转录抑制子,抑制JA信号途径相关基因JAZ5的表达,并激活下游SA信号相关基因EDS1的表达,SA和JA信号途径的交叉,使得拟南芥获得对不同生活方式病原菌的广谱抗性(Zhang et al.,2019)。
发明内容
本发明的目的是提供基因OsLUX在促进水稻抽穗和提高植物抗病性中的应用。
为了实现本发明目的,第一方面,本发明提供基因OsLUX在促进水稻抽穗中的应用。
本发明中,基因OsLUX为编码如下蛋白质(a)或(b)的基因:
(a)由SEQ ID NO:3所示的氨基酸序列组成的蛋白质;或
(b)SEQ ID NO:3所示序列经取代、缺失或添加一个或几个氨基酸且具有同等功能的由(a)衍生的蛋白质。
第二方面,本发明提供一种促进水稻抽穗的方法,所述方法包括:增强水稻中的基因OsLUX,获得该基因增强的转基因水稻。
所述增强的途径选自以下1)~5),或任选的组合:
1)通过导入具有所述基因的质粒而增强;
2)通过增加染色体上所述基因的拷贝数而增强;
3)通过改变染色体上所述基因的启动子序列而增强;
4)通过将强启动子与所述基因可操作地连接而增强;
5)通过导入增强子而增强。
第三方面,本发明提供基因OsLUX在提高植物抗病性中的应用。
优选地,所述植物为禾本科稻属植物,更优选水稻。
进一步地,所述应用为提高水稻抗病性,所述抗病性包括但不限于抗白叶枯病。
第四方面,本发明提供一种提高水稻抗病性的方法,所述方法包括:增强水稻中的基因OsLUX,获得该基因增强的转基因水稻。
所述增强的途径选自以下1)~5),或任选的组合:
1)通过导入具有所述基因的质粒而增强;
2)通过增加染色体上所述基因的拷贝数而增强;
3)通过改变染色体上所述基因的启动子序列而增强;
4)通过将强启动子与所述基因可操作地连接而增强;
5)通过导入增强子而增强。
所述抗病性包括但不限于抗白叶枯病。
第五方面,本发明提供基因OsLUX的3种突变体,其特征在于,所述突变体为基因oslux-1、oslux-2或oslux-3。
基因oslux-1与基因OsLUX相比,其CDS序列第376位碱基由C突变为T;
基因oslux-2与基因OsLUX相比,其CDS序列第369位碱基由G突变为A;
基因oslux-3与基因OsLUX相比,其CDS序列第469位碱基由C突变为T。
进一步地,基因oslux-1、oslux-2和oslux-3的CDS序列分别如SEQ ID NO:4-6。
第六方面,本发明提供含有所述突变体的生物材料,所述生物材料包括但不限于表达盒、转座子、质粒载体、病毒载体、工程菌或转基因细胞系。
第七方面,本发明提供所述突变体或含有所述突变体的生物材料的以下任一应用:
1)用于延迟水稻抽穗;
2)用于植物育种;
3)用于制备转基因植物。
优选地,所述植物为禾本科稻属植物,更优选水稻。
第八方面,本发明提供一种延迟水稻抽穗的方法,利用基因工程手段,对水稻基因OsLUX进行改造,获得该基因功能缺失的转基因水稻。
第九方面,本发明提供按照上述方法获得的转基因水稻在植物育种中的应用。
育种方法包括但不限于转基因、杂交、回交、自交或无性繁殖。
本发明提供OsLUX基因在控制水稻抽穗期(促进抽穗)和免疫反应(提高水稻抗病性)中的应用,基因OsLUX来自水稻品种日本晴,其cDNA序列如SEQ ID NO:1所示,该基因编码具有238个氨基酸的MYB家族的转录因子,是水稻生物钟的核心组分,通过Mutmap方法鉴定了OsLUX的三个突变体,突变体表现为极端晚抽和晚抽的表型。遗传互补和敲除实验证实了该极端晚抽穗的表型是由OsLUX突变所引起的。进一步研究发现OsLUX基因表现为昼夜节律表达,定位在细胞核和质膜中并具有转录自激活活性,通过抑制OsGI、Hd1和Ghd7来促进Ehd1、Hd3a和OsMADS14的表达,最终促进抽穗。同时,elh1/oslux-1接种白叶枯菌相较于野生型表现出更加感病,表明OsLUX对白叶枯菌的抗性起正调控作用。本发明进一步丰富和完善了水稻生物钟基因调控光周期开花的理论基础。
附图说明
图1为本发明较佳实施例中3个晚抽穗突变体的表型及抽穗期统计。其中,A为elh1、elh2和elh3突变株在自然短日照条件下的表型。B为自然长短日照条件下抽穗期的统计,NLD和NSD分别表示自然长日照和自然短日照条件。***表示不同处理组之间的差异具有统计学意义,***表示P<0.001。
图2为本发明较佳实施例中通过Mutmap方法克隆的OsLUX基因的3种突变类型。
图3为本发明较佳实施例中elh1/oslux-1遗传互补与敲除的表型及抽穗期统计。其中,A、C分别为elh1/oslux-1在自然长短日照条件下(NLDs和NSDs)的互补表型,B、D分别为在NLD和NSD的elh1/oslux-1敲除表型,E为elh1/oslux-1在NLD和NSD的互补和敲除家系的抽穗期统计。***表示不同处理组之间的差异具有统计学意义,***表示P<0.001。
图4为本发明较佳实施例中OsLUX基因的表达模式分析。其中,A是OsLUX在根、茎、叶(剑叶、倒二和倒三叶)、叶鞘和不同发育时期穗的表达量分析,B是OsLUX在不同组织中的GUS染色,C是在正常长日照转到持续光照条件下的OsLUX的表达模式,D是在正常短日照转到持续黑暗条件下的OsLUX的表达模式。LL和DD分别表示持续光照和持续黑暗。
图5为本发明较佳实施例中OsLUX蛋白定位于细胞核和质膜且具有转录自激活活性。其中,A为OsLUX在烟草和水稻原生质体中的亚细胞定位,Bar=10μm,B为OsLUX的转录自激活活性分析。AX表示AbA和X-α-gal。
图6为本发明较佳实施例中在长短日照条件下,开花相关基因Hd1(A)、Ehd1(B)、Hd3a(C)、OsMADS14(D)、Ghd7(E)和OsGI(F)在野生型和oslux-1突变体中的相对表达量分析,浅灰色矩形代表暗期。
图7为本发明较佳实施例中野生型和elh1/oslux-1突变体接种白叶枯菌两周后的表型及病斑长度统计。其中,A和B分别是接种CR1和CR4两个白叶枯菌的发病统计。***表示不同处理组之间的差异具有统计学意义,***表示P<0.001。
具体实施方式
本发明分离克隆了梗稻品种日本晴中携带的一个调控抽穗期和抗病反应的生物钟基因,并鉴定了该基因在水稻抽穗期和抗病性上的作用。
本发明分离得到一种属于MYB转录因子家族的水稻生物钟基因的DNA片段并鉴定其生物学功能。通过氨基酸序列比对,该基因(RAP-DB数据库中登录号为Os01g09718000,RGAP数据库中登录号为LOC_Os01g74020)是拟南芥LUX的同源基因,氨基酸相似度达到58.8%。且GARP结构域序列则高度保守,因此我们把它命名为OsLUX。
本发明提供的调控水稻抽穗期和免疫反应的OsLUX基因,其cDNA序列见SEQ IDNO:1,长度为1243bp,其编码区(CDS)序列见SEQ ID NO:2。该基因编码具有238个氨基酸的GARP结构域蛋白(氨基酸序列见SEQ ID NO:3),包含一个外显子,属于MYB转录因子基因家族,进一步研究发现在短日照和长日照条件下,OsLUX基因通过下调Ghd7、OsGI、Hd1来促进Ehd1、Hd3a和RFT1的表达,最终促进抽穗。
本发明通过Mutmap技术鉴定到了三个OsLUX的突变体,该基因的突变体elh1(extremely late heading 1)/oslux-1,编码区序列第376位C突变为T,造成提前终止,其编码区(CDS)序列见SEQ ID NO:4,在自然长短日照条件下均表现出相较于野生型晚抽两个月的表型,elh2/oslux-2的CDS序列的第369位的G突变为A,造成提前终止,其CDS序列见SEQID NO:5,该突变体的表型与oslux-1的类似,而elh3/oslux-3的CDS序列的第469位的C突变为T,造成氨基酸替换,其CDS序列见SEQ ID NO:6,该突变体的表型在自然长短日照条件下均较野生型晚一月有余。通过节律表达模式分析发现,OsLUX基因呈现明显的昼夜节律表达模式,在光期,OsLUX表达振幅基本不变,但转到暗期时,其表达迅速下降。组织表达分析发现,OsLUX基因主要在叶片中表达,且在倒二叶中表达量较高,GUS染色分析也证实了这个结果。通过亚细胞定位发现,OsLUX-GFP融合蛋白定位在细胞核和细胞质。转录自激活实验发现OsLUX蛋白具有较强的转录自激活活性,并且其C端是转录自激活活性所必须的。qRT-PCR分析发现OsLUX位于水稻开花因子OsGI、Ghd7和Hd1的上游,通过抑制OsGI、Ghd7和Hd1而促进Ehd1/Hd3a的表达,最终促进水稻的抽穗。
以下实施例用于说明本发明,但不用来限制本发明的范围。若未特别指明,实施例均按照常规实验条件,如Sambrook等分子克隆实验手册(Sambrook J&Russell DW,Molecular Cloning:a Laboratory Manual,2001),或按照制造厂商说明书建议的条件。
实施例1 3种晚抽穗突变体的表型
为了鉴定新的调控水稻抽穗期基因,对日本晴品种(Nipponbare)进行了EMS诱变,在突变体库中发现3株晚抽的单株elh1、elh2和elh3,经多年种植,发现突变体在自然长日照和自然短日照条件下表现出稳定的极端晚抽和晚抽的表型(图1)。
实施例2Mutmap方法鉴定3种OsLUX基因的突变体
为了鉴定引起极端晚抽穗的控制基因,将突变体与野生型进行了回交,在回交F2分离群体中利用极端晚抽穗单株进行Mutmap鉴定(图2),根据候选基因,参照水稻全基因组序列数据库的LOC_Os01g74020位点信息[GenBank(http://www.ncbi.nlm.nih.gov)注册号:NC_029256],在OsLUX编序列两侧设计一对扩增引物OsLUX-F和OsLUX-R进行PCR扩增得到包含OsLUX完整编码区序列用于与野生型CDS序列进行序列比对。
实施例3OsLUX基因的功能鉴定
为证实极端晚抽突变体elh1/oslux-1、elh2/oslux-2和elh3/oslux-3的表型是由OsLUX功能缺失引起的,发明人对elh1/oslux-1进行了遗传互补和OsLUX的敲除实验。具体实施如下:以日本晴基因组DNA作为模板,用引物OsLUX-EcoRI-F(SEQ ID NO:7)和OsLUX-EcoRI-R(SEQ ID NO:8)进行PCR扩增得到OsLUX基因的全长序列,随后与EcoRI酶切的pCAMBIA1305.1载体进行同源重组,以elh1/oslux-1为转化受体获得的阳性株,其抽穗期表型恢复到了野生型的表型(图3中A,C,E)。设计OsLUX的靶序列(SEQ ID NO:9)构建到BGK032(购自百格基因科技有限公司)载体上,以Nip为转化受体,获得的阳性敲除突变体的表型表现出类似于与elh1/oslux-1的极端晚抽穗表型(图3中B,D,E)。以上实验结果证实了OsLUX功能丧失造成极端晚抽穗的表型。
实施例4OsLUX基因的时空表达模式分析
为了研究OsLUX在不同组织中的表达模式,利用qRT-PCR检测了OsLUX在水稻中各组织如根、茎、叶(剑叶、倒二叶和倒三叶)、叶鞘、穗(不同发育时期的穗)的表达水平(图4,A)。利用引物OsLUX-qRT-F和OsLUX-qRT-R进行qRT-PCR。结果显示,OsLUX的表达水平在幼嫩的叶片中表达量最高(倒二叶)。同时OsLUX的表达水平也在根和茎中检测到,但在穗中的表达量较低。同时还构建了pOsLUX::GUS表达载体来研究OsLUX的组织表达模式。载体构建方法如下:以日本晴基因组DNA作为模板,用引物OsLUX-GUS-EcoRI-F(SEQ ID NO:10)和OsLUX-GUS-NcoI-R(SEQ ID NO:11)进行PCR扩增得到OsLUX基因启动子序列,然后用EcoRI和NcoI双酶切载体pCAMBIA1305.1将OsLUX基因启动子序列替换切掉的35S promoter序列,然后重组至该位点,用农杆菌侵染的方法转化水稻日本晴品种。不同组织化学染色结果显示,GUS信号在根、茎、叶片、叶鞘和穗中均能检测到,表明OsLUX是组成性表达(图4,B)。
OsLUX是生物钟的核心组分,为了研究其节律表达模式(图4,C和D),我们通过实时荧光定量PCR(qRT-PCR)的方法研究了OsLUX基因的表达水平,引物序列为OsLUX-qRT-F和OsLUX-qRT-R。在光照培养箱长日照(14小时光照,10小时黑暗)和短日照(10小时光照,14小时黑暗)条件下,96小时周期中,第一天均是正常光照条件,随后长日照条件转变为持续光照三天,短日照条件转变为持续黑暗三天,每4小时收集一次叶片。在长短日照条件下,OsLUX的表达水平在暗期前2小时达到峰值,在进入暗期后其表达量迅速降低,同样在持续光照条件下,其表达振幅基本不变,但是在持续暗期中,其表达振幅逐渐变小。这些结果说明OsLUX转录水平表现明显的节律表达且受光照诱导表达。
实施例5OsLUX蛋白的亚细胞定位和转录自激活活性分析
1、OsLUX基因的亚细胞定位
根据NCBI保守结构域预测(https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi)OsLUX编码一个保守的MYB_SHAQKYF转录因子。为了确定OsLUX作为转录因子并且具有核定位功能,我们对其进行亚细胞定位实验,本实施例中构建了一个亚细胞定位载体,载体构建过程如下:用引物OsLUX-GFP-EcoRI-F(SEQ ID NO:12)和OsLUX-GFP-EcoRI-R(SEQ ID NO:13),以OsLUX的全长cDNA,即SEQ ID NO:1所示序列为模板进行扩增,然后通过同源重组的方法将该片段重组至pYBA1132载体(pYBA1132由北京市农林科学院姚磊博士惠赠)中的EcoRI位点,最终形成OsLUX-GFP重组载体。在水稻原生质体和烟草细胞中瞬时表达实验显示OsLUX-GFP融合蛋白定位在细胞核和质膜。
2、OsLUX的转录自激活活性分析
为了进一步分析OsLUX是否具有转录自激活活性,利用pGBKT7载体构建了OsLUX的全长和各种缺失重组载体。首先以OsLUX的全长cDNA,即SEQ ID NO:1所示序列为模板,用引物(表1)扩增OsLUX的CDS和各种缺失片段,然后通过同源重组的方法将这些片段重组至载体pGBKT7的EcoRI和BamHI位点。最终构建得到如下5个载体:BD-OsLUX(AA:1-238),BD-OsLUX-ΔC(AA:1-117),BD-OsLUX-ΔN/ΔMYB(AA:174-238),BD-OsLUX-MYB(AA:118-173)和BD-OsLUX-ΔN(AA:118-238)。空载体BD和5个重组载体转化入酵母菌株Y2H Gold中,随后分别在-Trp和-Trp/-His/-Ade/AbA/X-α-gal的SD培养基上培养。转录自激活活性分析使用Gold Yeast Two-Hybrid System User Manual进行。结果显示,OsLUX具有很强的转录自激活活性,其余4个缺失构建中,除了BD-LUX-ΔC(AA:1-117)的酵母菌株有较强的转录自激活活性外,其余OsLUX的分段构建均无转录自激活活性(图5)。以上实验结果说明OsLUX是一个核转录因子并具有转录自激活活性,且其转录激活域位于OsLUX的N端区域。
表1用于构建OsLUX转录自激活载体的引物
实施例6OsLUX调控开花相关基因来促进水稻抽穗
OsLUX作为生物钟核心组分,且具有光周期响应和节律表达的特性,结合突变的晚抽穗表型,说明该基因是调控水稻抽穗的正调控因子。为了研究OsLUX在水稻光周期开花调控途径中的作用,分别在长日照和短日照条件下,利用qRT-PCR的方法检测了参与水稻光周期途径的相关基因在野生型和elh1/oslux-1突变植株中的表达,引物序列见表2。叶片分别选取自短日照处理40天和长日照处理50天的水稻植株。
首先检测了两个调控水稻抽穗的关键基因Hd1和Ehd1的表达,结果在两天的光周期中,与野生型植株相比,Hd1和Ehd1的转录水平在elh1/oslux-1植株中分别显著地上调和极显著降低。表明OsLUX抑制Hd1和促进Ehd1的表达,随后检测Hd1和Ehd1下游的Hd3a和OsMADS14的表达,它们的表达模式与Ehd1类似,均是极显著的下调,接着分别检测Hd1和Ehd1上游的OsGI和Ghd7,发现OsLUX均抑制两者的表达。以上结果表明OsLUX是通过抑制OsGI、Ghd7、Hd1和促进Ehd1、Hd3a和开花决定基因OsMADS14的表达来调控水稻抽穗的(图6)。
表2 qRT-PCR引物序列
实施例7OsLUX调控水稻的免疫反应
生物钟基因参与植物的生物和非生物胁迫,本发明进一步考察了OsLUX在水稻免疫反应中的作用。在自然长日照的大田环境下,对孕穗期的野生型和elh1/oslux-1突变体的剑叶进行剪叶法接种CR1和CR4(罗利军等,1998;Berchmans等,2003)两个白叶枯菌小种(菌株CR1和CR4由中国水稻研究所吴建利研究员惠赠),两周后观察发病情况,病斑长度代表发病级别。
将实验室保存的CR1和CR4白叶枯菌涂布在培养基上,在培养箱28℃条件下培养一周后,转接到新的培养基上继续生长2天,随后无菌水洗脱菌孢调到OD600=1.0。用沾有CR1和CR4白叶枯菌的剪刀剪掉水稻叶尖部,每个小种接种10个单株,每个单株3个分蘖,接种后14天统计病斑长度发现elh1/oslux-1突变体的病斑长度长于野生型的病斑长度,说明OsLUX正调控水稻抗白叶枯菌的抗性(图7)。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之做一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
参考文献:
1、罗利军梅捍卫赵新华等.水稻白叶枯病抗性基因定位及其小种专化性[J].中国科学C辑,1998,28(6):536-541.
2、Berchmans N.J徐同,宋凤鸣等.哈茨木霉NF9菌株对水稻的诱导抗病性[J]中国生物防治,2003,19(3):111-114.
3、Han S.H.,Yoo S.C.,Lee B.D.,et al.,Rice FLAVIN-BINDING,KELCH REPEAT,F-BOX 1(OsFKF1)promotes flowering independent of photoperiod.Plant CellEnviron,2015;38(12):2527-2540.
4、Hevia M.A.,Canessa P.,Müller-Esparza H.,et al.,A circadianoscillator in the fungus Botrytis cinerea regulates virulence when infectingArabidopsis thaliana.Proc Natl Acad Sci U S A,2015,112(28):8744-8749.
5、Kim J.A.,Kim H.S.,Choi S.H.,et al.,The importance of the circadianclock in regulating plant metabolism.Int J Mol Sci,2017,18(12):2680.
6、Kinoshita T.,Ono N.,Hayashi Y.,et al.,FLOWERING LOCUS T regulatesstomatal opening.Curr Biol,2011;21(14):1232-1238.
7、Korneli C.,Danisman S.,Staiger D.,Differential control of pre-invasive and post-invasive antibacterial defense by the Arabidopsis circadianclock.Plant Cell Physiol,2014;55(9):1613-1622.
8、Lai A.G.,Doherty C.J.,Mueller-Roeber B.,et al.,CIRCADIAN CLOCK-ASSOCIATED 1regulates ROS homeostasis and oxidative stress responses.ProcNatl Acad Sci U S A,2012;109(42):17129-17134.
9、Nakamichi N.,Kudo T.,Makita N.,et al.,Flowering time control inrice by introducing Arabidopsis clock-associated PSEUDO-RESPONSE REGULATOR5.Biosci Biotechnol Biochem,2020;DOI:10.1080/09168451.2020.1719822.
10、Somers D.E.,Webb A.A.,Pearson M.,et al.,The short-period mutant,toc1-1,alters circadian clock regulation of multiple outputs throughoutdevelopment in Arabidopsis thaliana.Development,1998;125(3):485-494.
11、Yang Y.,Peng Q.,Chen G.X.,et al..OsELF3 is involved in circadianclock regulation for promoting flowering under long-day conditions inrice.Mol Plant,2013;6(1):202-215.
12、Zhang C.,Gao M.,Seitz N.C.,et al.,LUX ARRHYTHMO mediates crosstalkbetween the circadian clock and defense in Arabidopsis.Nat Commun,2019;doi.org/10.1038/s41467-019-10485-6.
13、Zhang C.,Xie Q.,Anderson R.G.,et al.,Crosstalk between thecircadian clock and innate immunity in Arabidopsis.PLoS Pathog,2013;9(6):e1003370.
14、Zhou S.,Zhu S.,Cui S.,et al.Transcriptional and post-transcriptional regulation of heading date in rice.New Phytol.2021,230(3):943-956.
序列表
<110> 中国水稻研究所
<120> 基因OsLUX在促进水稻抽穗和提高植物抗病性中的应用
<130> PI202110712
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ccacctcgac tcaactcaac tcgcacaaat atcttctctc ttctcaccca ccgcccgatt 60
cttcgagtcc ccgatttggt tcggcctgaa gaagggtctg ggtctctcta gagagtgagt 120
agccatcgat ttgagatttg attagtaatt cggaggagga ggaacgagtg gtttttttga 180
tttgatttga tttgattgag aagaaaatcc gagtattatg ggcgaggagg cgccggagga 240
gtacgagctg ggcggcgggg aggacgagcg ggtgatggag tgggagacgg ggctgcccgg 300
cgccgacgag ctgaccccgc tgtcgcagcc gctggtgccg gcggggctgg cggcggcgtt 360
ccgcatcccg ccggagcccg ggcgcacgct gctcgacgtg caccgcgcgt cggcggcgac 420
ggtgtcccgg ttgcggcgcg cgtcgtcgtc gtcgtcgagc tcgttcccgg cgttcgcgtc 480
gaagggagcg ggaacgggag cggacgaggc ggagtcaggg ggaggcgcgg atggggggaa 540
cgggaacacc aacaacagca gcagcaagag ggcgcggctg gtgtggacgc cgcagctgca 600
caagaggttc gtggaggtgg tggcgcacct ggggatgaag aacgcggtgc ccaagacgat 660
catgcagctg atgaacgtgg agggcctcac ccgggagaac gtcgccagcc acctccagaa 720
gtatcgcctc tacgtgaagc ggatgcaggg cctctccaac gagggccctt ccccctccga 780
ccacatcttc gcctccaccc ccgtccccca cgcctccctc cacgaccagg ttccttctcc 840
ttaccacccc cacccccacc accactccta caacaacgcc gcctatgccg ccaccgtctc 900
ctcctaccac cactaccacc acgccaacca ctgatccatt ccttctcatc atcatttcat 960
ttccacatct atataccaca tactactcca taacgaaggt gtttcatcaa taagagaaga 1020
gaagagaaga gaatgctccc tcaactgtaa actcctaatt ctaattaagg gcaaagtgta 1080
tccatccatc catccacgct gtttgtttgt agctgttgtt aattgagctg gatgcatgac 1140
atgtatctct atgtacctac ctgcgtgaca aaggcatcat gcatatgtat ggttcccttg 1200
caatttgggg aatccagaaa tgattcatac ttgtgattct tgcaattgtg atgctcacac 1260
tgtactctac acattcagtt ccataaaaaa caaaactaat actacgaat 1309
<210> 2
<211> 717
<212> DNA
<213> Oryza sativa
<400> 2
atgggcgagg aggcgccgga ggagtacgag ctgggcggcg gggaggacga gcgggtgatg 60
gagtgggaga cggggctgcc cggcgccgac gagctgaccc cgctgtcgca gccgctggtg 120
ccggcggggc tggcggcggc gttccgcatc ccgccggagc ccgggcgcac gctgctcgac 180
gtgcaccgcg cgtcggcggc gacggtgtcc cggttgcggc gcgcgtcgtc gtcgtcgtcg 240
agctcgttcc cggcgttcgc gtcgaaggga gcgggaacgg gagcggacga ggcggagtca 300
gggggaggcg cggatggggg gaacgggaac accaacaaca gcagcagcaa gagggcgcgg 360
ctggtgtgga cgccgcagct gcacaagagg ttcgtggagg tggtggcgca cctggggatg 420
aagaacgcgg tgcccaagac gatcatgcag ctgatgaacg tggagggcct cacccgggag 480
aacgtcgcca gccacctcca gaagtatcgc ctctacgtga agcggatgca gggcctctcc 540
aacgagggcc cttccccctc cgaccacatc ttcgcctcca cccccgtccc ccacgcctcc 600
ctccacgacc aggttccttc tccttaccac ccccaccccc accaccactc ctacaacaac 660
gccgcctatg ccgccaccgt ctcctcctac caccactacc accacgccaa ccactga 717
<210> 3
<211> 238
<212> PRT
<213> Oryza sativa
<400> 3
Met Gly Glu Glu Ala Pro Glu Glu Tyr Glu Leu Gly Gly Gly Glu Asp
1 5 10 15
Glu Arg Val Met Glu Trp Glu Thr Gly Leu Pro Gly Ala Asp Glu Leu
20 25 30
Thr Pro Leu Ser Gln Pro Leu Val Pro Ala Gly Leu Ala Ala Ala Phe
35 40 45
Arg Ile Pro Pro Glu Pro Gly Arg Thr Leu Leu Asp Val His Arg Ala
50 55 60
Ser Ala Ala Thr Val Ser Arg Leu Arg Arg Ala Ser Ser Ser Ser Ser
65 70 75 80
Ser Ser Phe Pro Ala Phe Ala Ser Lys Gly Ala Gly Thr Gly Ala Asp
85 90 95
Glu Ala Glu Ser Gly Gly Gly Ala Asp Gly Gly Asn Gly Asn Thr Asn
100 105 110
Asn Ser Ser Ser Lys Arg Ala Arg Leu Val Trp Thr Pro Gln Leu His
115 120 125
Lys Arg Phe Val Glu Val Val Ala His Leu Gly Met Lys Asn Ala Val
130 135 140
Pro Lys Thr Ile Met Gln Leu Met Asn Val Glu Gly Leu Thr Arg Glu
145 150 155 160
Asn Val Ala Ser His Leu Gln Lys Tyr Arg Leu Tyr Val Lys Arg Met
165 170 175
Gln Gly Leu Ser Asn Glu Gly Pro Ser Pro Ser Asp His Ile Phe Ala
180 185 190
Ser Thr Pro Val Pro His Ala Ser Leu His Asp Gln Val Pro Ser Pro
195 200 205
Tyr His Pro His Pro His His His Ser Tyr Asn Asn Ala Ala Tyr Ala
210 215 220
Ala Thr Val Ser Ser Tyr His His Tyr His His Ala Asn His
225 230 235
<210> 4
<211> 717
<212> DNA
<213> Oryza sativa
<400> 4
atgggcgagg aggcgccgga ggagtacgag ctgggcggcg gggaggacga gcgggtgatg 60
gagtgggaga cggggctgcc cggcgccgac gagctgaccc cgctgtcgca gccgctggtg 120
ccggcggggc tggcggcggc gttccgcatc ccgccggagc ccgggcgcac gctgctcgac 180
gtgcaccgcg cgtcggcggc gacggtgtcc cggttgcggc gcgcgtcgtc gtcgtcgtcg 240
agctcgttcc cggcgttcgc gtcgaaggga gcgggaacgg gagcggacga ggcggagtca 300
gggggaggcg cggatggggg gaacgggaac accaacaaca gcagcagcaa gagggcgcgg 360
ctggtgtgga cgccgtagct gcacaagagg ttcgtggagg tggtggcgca cctggggatg 420
aagaacgcgg tgcccaagac gatcatgcag ctgatgaacg tggagggcct cacccgggag 480
aacgtcgcca gccacctcca gaagtatcgc ctctacgtga agcggatgca gggcctctcc 540
aacgagggcc cttccccctc cgaccacatc ttcgcctcca cccccgtccc ccacgcctcc 600
ctccacgacc aggttccttc tccttaccac ccccaccccc accaccactc ctacaacaac 660
gccgcctatg ccgccaccgt ctcctcctac caccactacc accacgccaa ccactga 717
<210> 5
<211> 717
<212> DNA
<213> Oryza sativa
<400> 5
atgggcgagg aggcgccgga ggagtacgag ctgggcggcg gggaggacga gcgggtgatg 60
gagtgggaga cggggctgcc cggcgccgac gagctgaccc cgctgtcgca gccgctggtg 120
ccggcggggc tggcggcggc gttccgcatc ccgccggagc ccgggcgcac gctgctcgac 180
gtgcaccgcg cgtcggcggc gacggtgtcc cggttgcggc gcgcgtcgtc gtcgtcgtcg 240
agctcgttcc cggcgttcgc gtcgaaggga gcgggaacgg gagcggacga ggcggagtca 300
gggggaggcg cggatggggg gaacgggaac accaacaaca gcagcagcaa gagggcgcgg 360
ctggtgtgaa cgccgcagct gcacaagagg ttcgtggagg tggtggcgca cctggggatg 420
aagaacgcgg tgcccaagac gatcatgcag ctgatgaacg tggagggcct cacccgggag 480
aacgtcgcca gccacctcca gaagtatcgc ctctacgtga agcggatgca gggcctctcc 540
aacgagggcc cttccccctc cgaccacatc ttcgcctcca cccccgtccc ccacgcctcc 600
ctccacgacc aggttccttc tccttaccac ccccaccccc accaccactc ctacaacaac 660
gccgcctatg ccgccaccgt ctcctcctac caccactacc accacgccaa ccactga 717
<210> 6
<211> 717
<212> DNA
<213> Oryza sativa
<400> 6
atgggcgagg aggcgccgga ggagtacgag ctgggcggcg gggaggacga gcgggtgatg 60
gagtgggaga cggggctgcc cggcgccgac gagctgaccc cgctgtcgca gccgctggtg 120
ccggcggggc tggcggcggc gttccgcatc ccgccggagc ccgggcgcac gctgctcgac 180
gtgcaccgcg cgtcggcggc gacggtgtcc cggttgcggc gcgcgtcgtc gtcgtcgtcg 240
agctcgttcc cggcgttcgc gtcgaaggga gcgggaacgg gagcggacga ggcggagtca 300
gggggaggcg cggatggggg gaacgggaac accaacaaca gcagcagcaa gagggcgcgg 360
ctggtgtgga cgccgcagct gcacaagagg ttcgtggagg tggtggcgca cctggggatg 420
aagaacgcgg tgcccaagac gatcatgcag ctgatgaacg tggagggctt cacccgggag 480
aacgtcgcca gccacctcca gaagtatcgc ctctacgtga agcggatgca gggcctctcc 540
aacgagggcc cttccccctc cgaccacatc ttcgcctcca cccccgtccc ccacgcctcc 600
ctccacgacc aggttccttc tccttaccac ccccaccccc accaccactc ctacaacaac 660
gccgcctatg ccgccaccgt ctcctcctac caccactacc accacgccaa ccactga 717
<210> 7
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gaccatgatt acgaattctg ctcctgttca gcacctgtc 39
<210> 8
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
ggtaccgagc tcgaattcat ccgtacgaac aaacaagagc a 41
<210> 9
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
ctcccttcga cgcgaacgcc ggg 23
<210> 10
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
gctatgacca tgattacgtg ctcctgttca gcacctgtc 39
<210> 11
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
taccctcaga tctaccataa tactcggatt ttcttctcaa tca 43
<210> 12
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
cccgggctgc aggaattcat gggcgaggag gcgcc 35
<210> 13
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
atcgataagc ttgatatcgt ggttggcgtg gtggta 36
Claims (3)
1.突变体基因oslux-3或含有所述突变体的生物材料在延迟水稻抽穗中的应用;
其中,基因oslux-3的CDS序列如SEQ ID NO:6所示;
所述生物材料为表达盒、转座子、质粒载体、病毒载体或工程菌。
2.一种延迟水稻抽穗的方法,其特征在于,利用基因工程手段,对水稻基因OsLUX进行改造,获得该基因功能缺失的转基因水稻;
改造后的基因的CDS序列如SEQ ID NO:6所示。
3.按照权利要求2所述方法获得的转基因水稻在植物育种中的应用;
育种方法包括转基因、杂交、回交、自交或无性繁殖。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2002272291A (ja) * | 2001-03-21 | 2002-09-24 | Idemitsu Kosan Co Ltd | 病害抵抗性イネ |
CN106047893A (zh) * | 2016-07-20 | 2016-10-26 | 中国水稻研究所 | OsCOL16基因在控制水稻抽穗期中的应用 |
CN107400672A (zh) * | 2017-09-15 | 2017-11-28 | 中国水稻研究所 | OsCOL15基因在调控水稻抽穗期中的应用 |
CN111527101A (zh) * | 2017-12-29 | 2020-08-11 | 合成基因组股份有限公司 | 用于改善生长的光合生物基因调节 |
CN111778262A (zh) * | 2020-08-13 | 2020-10-16 | 浙江省农业科学院 | 一种早熟基因eam.z及其分子标记SNP595和应用 |
CN112457385A (zh) * | 2020-12-22 | 2021-03-09 | 福建农林大学 | 一种控制水稻生育期基因ljp1的应用 |
CN113549632A (zh) * | 2021-09-08 | 2021-10-26 | 广东省农业科学院水稻研究所 | 水稻OsFLZ2基因在调控禾本科植物抽穗期中的应用 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006132389A1 (ja) * | 2005-06-09 | 2006-12-14 | National University Corporation Nagoya University | 核酸、当該核酸をコードするアミノ酸、当該核酸及びアミノ酸からなるプローブ、及び当該プローブを用いたスクリーニング法 |
CN100554424C (zh) * | 2007-09-12 | 2009-10-28 | 华中农业大学 | 一种控制水稻谷粒产量,抽穗期和株高的多效性基因Ghd7的克隆及应用 |
-
2022
- 2022-01-12 CN CN202210032594.9A patent/CN114350678B/zh active Active
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2002272291A (ja) * | 2001-03-21 | 2002-09-24 | Idemitsu Kosan Co Ltd | 病害抵抗性イネ |
CN106047893A (zh) * | 2016-07-20 | 2016-10-26 | 中国水稻研究所 | OsCOL16基因在控制水稻抽穗期中的应用 |
CN107400672A (zh) * | 2017-09-15 | 2017-11-28 | 中国水稻研究所 | OsCOL15基因在调控水稻抽穗期中的应用 |
CN111527101A (zh) * | 2017-12-29 | 2020-08-11 | 合成基因组股份有限公司 | 用于改善生长的光合生物基因调节 |
CN111778262A (zh) * | 2020-08-13 | 2020-10-16 | 浙江省农业科学院 | 一种早熟基因eam.z及其分子标记SNP595和应用 |
CN112457385A (zh) * | 2020-12-22 | 2021-03-09 | 福建农林大学 | 一种控制水稻生育期基因ljp1的应用 |
CN113549632A (zh) * | 2021-09-08 | 2021-10-26 | 广东省农业科学院水稻研究所 | 水稻OsFLZ2基因在调控禾本科植物抽穗期中的应用 |
Non-Patent Citations (4)
Title |
---|
A clock regulatory module is required for salt tolerance and control of heading date in rice.;Xiling Wang等;《Plant Cell Environ》;第44卷;全文 * |
LUX ARRHYTHMO mediates crosstalk between the circadian clock and defense in Arabidopsis.;Chong Zhang等;《nature communications》;第10卷;全文 * |
The clock component OsLUX regulates rice heading through recruiting OsELF3-1 and OsELF4s to repress Hd1 and Ghd7.;Peng Xu等;《Journal of Advanced Research》;全文 * |
The germin-like protein OsGLP2-1 enhances resistance to fungal blast and bacterial blight in rice.;Qing Liu等;《Plant Mol Biol》;第92卷;全文 * |
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