CN113801887A - 一种水稻纹枯病菌脂肪基因沉默片段RslipA及其应用 - Google Patents
一种水稻纹枯病菌脂肪基因沉默片段RslipA及其应用 Download PDFInfo
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- CN113801887A CN113801887A CN202110964831.0A CN202110964831A CN113801887A CN 113801887 A CN113801887 A CN 113801887A CN 202110964831 A CN202110964831 A CN 202110964831A CN 113801887 A CN113801887 A CN 113801887A
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- rhizoctonia solani
- rslipa
- gene
- lipase gene
- fragment
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Abstract
本发明公开了一种水稻纹枯病菌脂肪酶基因沉默片段RslipA的应用。本发明提供了一种水稻纹枯病菌脂肪酶基因沉默片段RslipA,其核苷酸序列如SEQ IDNO:5所示。水稻纹枯病菌脂肪酶基因在水稻纹枯病菌侵染水稻的过程中被显著诱导表达,在侵染后48h表达量最高;水稻纹枯病菌脂肪酶基因沉默片段RslipA及dsRNA能够用于制备水稻纹枯病菌防治制剂,且将含有沉默片段RslipA的重组载体转化入植株后,能够得到抗水稻纹枯病菌转基因植物。因此,水稻纹枯病菌脂肪酶基因或水稻纹枯病菌脂肪酶基因沉默片段RslipA在防治水稻纹枯病菌、制备水稻纹枯病菌防治产品、构建抗水稻纹枯病菌转基因植物中均具有广泛的应用前景。
Description
技术领域
本发明属于基因工程技术领域。更具体地,涉及一种水稻纹枯病菌脂肪酶基因沉默片段RslipA的应用。
背景技术
基因沉默(RNAi)是指在真核生物(植物、动物、真菌)由保守的双链RNA(double-strand RNA,dsRNA)诱导鉴定和破坏其细胞质中异常变异或过表达的RNA的一种机制。基因沉默是一种高度保守的、序列特异的RNA降解机制,对于调控发育、维持基因组的稳定性以响应生物和非生物胁迫具有重要作用。RNAi目前已经被广泛运用于植物保护领域,如植物抗病毒、害虫和病害上的防治。
病毒诱导基因沉默技术(Virus-induced gene silencing)简称VIGS技术,基于同源RNA互补结合降解的RNAi原理,是利用植物对病毒的天然防御机制发展的一种瞬时、快速鉴定植物基因功能的技术,是用基因序列验证基因功能的一种反向遗传学方法。该技术在植物病害的应用也取得一系列的成果,可以用于筛选和鉴定病菌的基因功能。
水稻纹枯病是世界性的水稻三大病害之一,给水稻生产造成了严重的经济损失。我国每年纹枯病的发病面积超过2亿亩次,发病程度普遍加重,且逐年呈严重趋势。水稻纹枯病是由立枯丝核菌(Rhizoctonia solani)侵染所引起的,是一种重要的腐生性土传病原真菌。该病原菌主要以菌丝和菌核的形式存在于土壤和病残体中。由于寄主范围广、腐生性强、缺乏高水平的抗源、接种鉴定操作困难等原因,目前能稳定高抗纹枯病的水稻品种比较少。
目前,有关水稻纹枯病菌的脂肪酶基因的研究较少,如专利CN110628813B公开了水稻脂肪酶基因Os07g0586800及其编码蛋白的功能和应用,参与调控水稻对白叶枯病的免疫响应,能够显著提高水稻对白叶枯病的抗性。可通过筛选和克隆水稻纹枯病菌的脂肪酶基因,来研究水稻纹枯病菌的致病性和抗病性;筛选出水稻纹枯病菌的致病基因,对于防治水稻纹枯病菌具有重要的研究意义。
发明内容
本发明要解决的技术问题是通过对水稻纹枯病菌的脂肪酶基因的研究,为水稻纹枯病菌的防治提供一种新的方法思路。
本发明的第一个目的是提供一种水稻纹枯病菌致病基因。
本发明的第二个目的是提供一种水稻纹枯病菌脂肪酶基因的沉默片段RslipA。
本发明的第三个目的是提供一种水稻纹枯病菌致病基因在防治水稻纹枯病菌中的应用。
本发明的第四个目的是提供一种水稻纹枯病菌脂肪酶基因沉默片段RslipA在防治水稻纹枯病菌和/或制备水稻纹枯病菌防治产品中的应用。
本发明的第五个目的是提供一种水稻纹枯病菌致病基因或水稻纹枯病菌脂肪酶基因沉默片段RslipA在构建抗水稻纹枯病菌转基因植物中的应用。
本发明的第六个目的是提供一种水稻纹枯病菌防治制剂。
本发明的第七个目的是提供一种水稻纹枯病菌防治方法。
本发明上述目的通过以下技术方案实现:
本发明首先提供了一种水稻纹枯病菌致病基因,经测序后进行序列对比,将该基因归于脂肪酶基因家族,为水稻纹枯病菌脂肪酶基因,其cDNA序列如SEQ ID NO:1所示,序列长度为909bp。
所述水稻纹枯病菌脂肪酶基因的基因组DNA序列如SEQ ID NO:2所示,序列长度为1417bp,包含9个内含子,分别位于第129~182位、242~304位、374~432位、503~559位、652~713位、822~874位、1003~1055位、1141~1191位和1312~1367位,剪切位置均符合“GT~AG法则”。
所述水稻纹枯病菌脂肪酶基因的编码序列如SEQ ID NO:3所示,序列长度为909bp。
所述水稻纹枯病菌脂肪酶的氨基酸序列如SEQ ID NO:4所示,共302个氨基酸,蛋白分子量为31.56kDa。
本发明提供了一种水稻纹枯病菌脂肪酶基因的沉默片段RslipA,其核苷酸序列如SEQ ID NO:5所示,序列长度为415bp。
所述水稻纹枯病菌脂肪酶基因是水稻纹枯病菌与致病力有关的基因片段,在水稻纹枯病菌侵染水稻过程的显著上调,在接种后基因的表达量逐渐上升,在接种后48h表达量达到峰值,显著高于对照。
因此,本发明提供了一种水稻纹枯病菌致病基因防治水稻纹枯病菌中的应用。
本发明提供了一种水稻纹枯病菌脂肪酶基因沉默片段RslipA在防治水稻纹枯病菌和/或制备水稻纹枯病菌防治产品中的应用。
本发明还提供一种水稻纹枯病菌致病基因或水稻纹枯病菌脂肪酶基因沉默片段RslipA在构建抗水稻纹枯病菌转基因植物中的应用。
优选地,所述构建抗水稻纹枯病菌转基因植物的方法为:将所述基因沉默片段RslipA构建到表达载体中,然后将该载体转化至植物中,获得抗水稻纹枯病菌转基因植物。
更优选地,所述转基因植物为烟草、水稻、玉米、大豆、马铃薯或十字花科蔬菜中的任意一种或几种。
进一步优选地,所述重组载体为TRV2:RslipA载体。
本发明还提供一种水稻纹枯病菌防治制剂,含有能够抑制水稻纹枯病菌致病基因表达的物质。
优选地,能够抑制水稻纹枯病菌致病相关基因表达的物质含有水稻纹枯病菌脂肪酶基因沉默片段RslipA和dsRNA,其中dsRNA的核苷酸序列如SEQ ID NO:6所示。
更优选地,水稻纹枯病菌防治制剂含有水稻纹枯病菌脂肪酶基因沉默片段RslipA的重组载体或重组菌。
本发明还提供一种水稻纹枯病菌的防治方法。
优选地,该水稻纹枯病菌的防治方法采用了水稻纹枯病菌防治制剂。
本发明还提供一种水稻纹枯病菌的防治制剂在防治水稻纹枯病菌和/或制备水稻纹枯病菌防治产品中的应用。
本发明还提供一种水稻纹枯病菌的防治制剂在构建抗水稻纹枯病菌转基因植物中的应用。
除本发明提供的水稻纹枯病菌脂肪酶基因的沉默片段RslipA合成方法外,本领域技术人员还可以通过以下方法获得与该基因等同的基因:(1)通过数据库检索获得;(2)以沉默片段RslipA为探针,筛选其它丝核菌基因组文库或cDNA文库获得;(3)根据沉默片段RslipA的序列信息设计寡核苷酸引物,用PCR扩增的方法从丝核菌属(Rhizoctonia)或其它近缘真菌的基因组、mRNA和cDNA中获取;(4)在沉默片段RslipA的基础上,用基因工程方法改造获得。
本发明具有以下有益效果:
本发明提供了一种水稻纹枯病菌脂肪酶基因,在水稻纹枯病菌侵染水稻过程后期被显著诱导表达,水稻纹枯病菌脂肪酶基因是水稻纹枯病菌与致病力有关的基因,以该基因的片段为靶点可沉默脂肪酶基因从而显著降低水稻纹枯病菌的致病能力。
本发明还采用水稻纹枯病菌脂肪酶基因沉默片段RslipA的重组载体,该重组载体可转化至植物,所得转基因植物受水稻纹枯病菌侵害时,对水稻纹枯病菌具有显著的抗病性。因此,本发明提供的水稻纹枯病菌致病基因和水稻纹枯病菌脂肪酶基因沉默片段RslipA在防治水稻纹枯病菌和/或制备水稻纹枯病菌防治产品,以及在构建抗水稻纹枯病菌转基因植物中,均具有广泛的应用前景,可以解决生产上土传真菌病害的难题,有效地防控土传真菌病害,具有重要的推广应用价值。
附图说明
图1为水稻纹枯病菌GD118菌株RNA电泳图。
图2为水稻纹枯病菌脂肪酶基因的沉默片段RslipA的PCR扩增结果图。
图3为水稻纹枯病菌脂肪酶基因在水稻纹枯病菌侵染水稻过程中的基因表达模式图。
图4为VIGS载体构建示意图。
图5为VIGS载体构建得到的TRV2:RslipA载体图谱。
图6为TRV2:RslipA载体转化至根癌农杆菌GV3101的菌落验证结果图。
图7为转化植株烟草接种纹枯病菌后的发病情况图。
图8为烟草体内真菌生物量分析结果图。
图9为烟草靶标基因沉默效率分析结果图。
具体实施方式
以下结合说明书附图和具体实施例来进一步说明本发明,但实施例并不对本发明做任何形式的限定。除非特别说明,本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。
除非特别说明,以下实施例所用试剂和材料均为市购。
实施例1水稻纹枯病菌RNA的提取及cDNA的获得
1、实验方法
S1.水稻纹枯病菌RNA的提取
水稻纹枯病菌RNA的提取使用TaKaRa总RNA提取试剂盒(货号9769),具体实验步骤如下:
水稻纹枯病病原菌(Rhizoctonia solani AG1-IA)GD118菌株,属于强致病力菌株,为本实验室保存的常用菌株。称取0.1g水稻纹枯病菌菌丝于液氮中迅速研磨成粉末,加入到含有Buffer RL裂解液的1.5mL灭菌离心管中,将裂解液12,000rpm,4℃离心5min;将上清液小心吸取到新的1.5mL灭菌离心管中;加入样品裂解步骤中上清液1/2体积的无水乙醇,立即将混合液(含沉淀)全部转入到RNA Spin Column(含2mL Collection Tube);12,000rpm,离心1min,弃滤;依次将500μL的Buffer RWA,600μL的RWB缓冲液,600μL的BufferRWB加入至RNA Spin Column中,12,000rpm离心30s,弃滤液;然后空管12,000rpm离心1min;将RNA Spin Column安置于1.5mL的无RNase酶收集管,在RNA Spin Column膜中央处加入50~200μL的无RNase酶水(65℃预热),室温静置5min;12,000rpm离心2min洗脱RNA。
S2.cDNA第一链的合成
以步骤S1得到的水稻纹枯病菌的RNA为模板,利用TARAKA PrimeScript反转录试剂盒(货号6210A)合成cDNA,具体实验步骤如下:
在微量离心管中加入2000ng Total RNA、1μL Oligo dT、1μL dNTP(10mM)、补充无RNase酶水至10μL。轻轻混匀,离心数秒后,65℃保温5min,冰上迅速冷却。依次加入4μL 5×PrimeScript II Buffer、0.5μL RNase Inhibitor(40U/μL)、1μL PrimeScript II RTase(200U/μL)、补无RNA酶水至20μL,缓慢混匀。42℃保温50min,95℃保温5min(酶失活),冰上冷却后-20℃保存备用。
2、实验结果
水稻纹枯病菌RNA的琼脂糖凝胶电泳如图1所示,可见28S、18S、5.8S三个条带完整性较好,无蛋白质污染,可作为反转录模板。使用NanoDrop 2000超微量分光光度计测定,样品OD260/280约为2.00,RNA纯度较高。
基于NCBI上已公布的水稻纹枯病菌的基因组与转录组的信息,利用生物信息学软件预测得到可能与致病相关的基因。通过测序分析验证后得到水稻纹枯病菌致病基因,经过测序结果进行序列对比,将该基因归于脂肪酶基因家族;其中,水稻纹枯病菌脂肪酶基因的cDNA序列如SEQ ID NO:1所示,其基因组序列如SEQ ID NO:2所示,其编码序列如SEQ IDNO:3所示,其编码蛋白的氨基酸序列如SEQ ID NO:4所示。
实施例2水稻纹枯病菌脂肪酶基因沉默片段RslipA的合成
1、试验方法
cDNA靶标克隆:以实施例1中步骤S2得到的水稻纹枯病菌cDNA为模板,设计沉默片段RslipA克隆引物5142F/5142R,使用擎科(货号TP001)高保真聚合酶,进行PCR扩增反应。反应条件为:98℃,2min;98℃,10s,58℃,15s,40个循环;最后72℃,10min。
5142F:GCAAGCAAAACTGCGACG;
5142R:GGTGGAGCGACAGATACAGC。
使用TAE缓冲液制作1.0%的琼脂糖凝胶,将PCR产物进行琼脂糖凝胶电泳。使用Axygen PCR纯化试剂盒进行PCR产物纯化、测序。
2、实验结果
水稻纹枯病菌脂肪酶基因的沉默片段的扩增结果如图2所示,可以看出,应用沉默片段RslipA克隆引物5142F/5142R,可以扩增出单一、明亮的条带。经测序分析后得水稻纹枯病菌脂肪酶基因的沉默片段RslipA的核苷酸序列如SEQ ID NO:5所示。
实施例3水稻纹枯病菌脂肪酶基因在水稻纹枯病菌侵染水稻过程中的表达分析
1、试验方法
S1.水稻纹枯病菌接种体的制备
首先使用PDA平板将本实验室保存的水稻纹枯病菌菌株GD118活化,黑暗条件下,28℃培养2d。同时将水稻谷粒浸泡24h后放入250mL锥形瓶中,121℃,30min,高压蒸汽灭菌。将灭菌后的谷粒均匀平铺在培养2d后的PDA平板上,黑暗条件下,28℃培养7d至谷粒表面布满菌丝及菌核,即可用于接种。
S2.水稻的种植和水稻纹枯病菌的接种
分别使用10%的次氯酸钠、30%的双氧水对水稻种子(品种9311)进行消毒,再将种子置于无菌培养皿内,使用无菌水浸泡3d以上直至发芽,然后将发芽的种子转移至含有10cm厚无菌土的方盆(长:80cm;宽:40cm;高:15cm)内。接种选择水稻三叶一心期,在盆内无水干燥条件下,使用步骤S1制备得到的接种体进行接种,在水稻茎基部放置两枚接种体,温度30℃、湿度90%以上利于发病。
S3.样品的荧光定量PCR分析
为了验证水稻纹枯病脂肪酶基因在水稻纹枯病菌侵染水稻过程中的表达情况,选取了接种后的6个时间点10、18、24、32、48和72h后,收集活体接种的水稻茎基部作为样品。使用TaKaRa MiniBEST植物RNA提取试剂盒(货号9769)进行不同时间的水稻样品总RNA的提取,具体方法同“实施例1的步骤S1”。利用TARAKA PrimeScript反转录试剂盒(货号6210A)进行RNA反转录,具体方法同“实施例1的步骤S2”,将反转录所得的cDNA产物稀释10倍后,作为模板,利用Primer Premier 5.0软件设计实施例1中的水稻纹枯病菌脂肪酶基因的荧光定量PCR引物5142QF/R,进行实时荧光定量PCR。
5142QF:ATGGTCGTGTATGCGTCTGG;
5142QR:TGGATCGGTTCCTTGGTTG。
引物的特异性通过凝胶电泳、测序和溶解曲线检验。荧光定量PCR反应体系为20μL,包含10μL Bio-Rad CFX real-time PCR system、0.2μL上/下游引物、2μL cDNA模板和适量的纯水。每个样品进行三次技术重复。
水稻纹枯病菌的内参基因使用GAPDH进行样品间的基因表达的均一化处理,应用引物GAPDH F/R,结果采用2-ΔΔCt法进行分析。
GAPDH F:GGTCGGCAAAGTCATACCAT;
GAPDH R:TCTGCGTCCTTCTTGGAGATA。
2、实验结果
水稻纹枯病菌脂肪酶基因在水稻纹枯病菌侵染水稻过程中的基因表达分析结果如图3所示,由此可知,水稻纹枯病菌脂肪酶基因在水稻纹枯病菌侵染水稻的过程中被显著诱导表达,在侵染后第48h时的表达量显著增高,这表明该基因响应了水稻纹枯病菌的侵染。
实施例4TRV:RslipA载体构建和转化烟草
1、实验方法
S1.TRV2:RslipA载体构建及农杆菌转化
根据实施例1中的水稻纹枯病菌脂肪酶基因的编码序列,设计引物两端包含EcoRI和BamHI酶切位点的特异性引物VIGS-5142F/R引物序列如下所示:
VIGS-5142F:CCGgaattc GCAAGCAAAACTGCGACG;
VIGS-5142R:CGCggatcc GGTGGAGCGACAGATACAGC。
以实施例1中的水稻纹枯病菌cDNA为模板,利用擎科(货号TP001)高保真酶进行PCR扩增,将扩增产物回收后,和载体pYL156同时在37℃下双酶切5h。将酶切后的目的片段和载体利用T4 DNA连接酶,16℃过夜连接,将连接好的产物转化入E.coil DH5α感受态,得到TRV2:RslipA载体。经测序验证确定无误后,扩繁提取质粒,保存备用。
S2.根癌农杆菌感受态的制备及电击转化
将根癌农杆菌GV3101单菌落接于4mL含Rif(25μg/mL)的LB液体培养基中,28℃,200r/min振荡培养2d。取培养液3mL的接种量转接至200mL LB培养液中,200r/min,28℃振荡培养至对数生长期(细胞浓度OD600为0.5~0.6左右)。离心弃去废液,收集菌体。用预冷的无菌双蒸水将菌液重悬洗涤3次,后用2mL无菌预冷的10%(w/v)甘油重悬菌体。取100μL菌悬液于1.5mL离心管,于-70℃保存备用。加3μL质粒于菌液中,轻轻混匀并转移至无菌预冷的电击杯中,电击后迅速加入1mL LB液体培养基,混匀并转移细胞至1.5mL的离心管中,在28℃摇床中200r/min培养2~3h;取100μL菌液涂布于含Rif(25μg/mL)和Kan(50μg/mL)的LB平板中,倒置放于28℃培养箱中培养2天,观察转化子生长情况,取未添加质粒DNA的农杆菌在同样电击条件下的结果作为对照。挑取单菌落使用pYL156载体引物PYL156 F/R进行PCR验证,载体引物PYL156 F/PYL156 R的引物序列如下所示:
PYL156 F:AATTCACTGGGAGATGATACGCTG;
PYL156 R:CCTATGGTAAGACAATGAGTCGGC。
S3.农杆菌培养和烟草瞬时转化
挑取农杆菌单克隆加入5mL的液体LB培养基(25μg/mL Rif和50μg/mL Kan)中,放于28℃摇床中进行200r/min过夜培养,将1mL菌液加入50mL LB(25μg/mL Rif和50μg/mLKan)中,放于28℃摇床中进行200r/min振荡培养。当培养物OD600到0.8~1.0时,以6000rpm低温离心5min收集菌体,弃去废液。将菌体重悬于注射基质(10mM MES、10mM MgCl2、100μM乙酰丁香酮)中,使OD600为0.5~0.6,得到TRV1农杆菌。再把TRV1农杆菌和TRV2:RslipA载体以1:1混合,室温静置3~5h,不要摇动。最后利用注射器将含有注射基质的农杆菌注射于本氏烟草最嫩的叶片中。
S4.烟草致病力检测和生物量测定
通过在植物体内载体表达法来获得dsRNA,从注射后的第10天起,本氏烟草的白化现象就非常明显,这表明了在本氏烟草体内VIGS载体已经产生出大量dsRNA,并发挥着干扰作用,其dsRNA的核苷酸序列如SEQ ID NO:6所示。因此,我们选择了从注射VIGS系列载体后的第14天,进行活体接种。对接菌后的第5天进行本氏烟草的病情指数统计,通过统计叶片病斑和凋萎叶片的数量进行致病力的检测。提取接种水稻纹枯病菌5d后本氏烟草和阴性对照的叶片(营养土以上0~3cm处)的总DNA,以水稻纹枯病菌内部转录间隔区序列为靶标片段,设计特异性引物Rs F/R。同时,以烟草actin基因为内参基因,应用引物EF1a/1b,具体方法同实施例3的步骤S3,结果采用2-ΔΔCt法进行分析。
Rs F:GCCTTTTCTACCTTAATTTGGCAG;
Rs R:GTGTGTAAATTAAGTAGACAGCAAATG;
EF1a:TGGTGTCCTCAAGCCTGGTAT;
EF1b:ACGCTTGAGATCCTTAACCGC。
S5.靶标基因沉默效率测定
提取接菌5d后的烟草叶片总RNA,并进行反转录,获得cDNA。根据实施例1中的水稻纹枯病菌脂肪酶基因设计非沉默片全区域的特异性引物qP-5142-F/R,利用qRT-PCR测定水稻纹枯病菌在侵染烟草过程中靶标基因的转录水平。qRT-PCR采用全式金PerfectStartTMGreen qPCR SuperMix(AQ601-01)。以水稻纹枯病菌甘油醛3-磷酸基因GAPDH基因(GAPDH-F/R)作为内参基因进行样品间的基因表达的均一化处理,使用Bio-Rad CFX real-timePCR system,反应体系为20μL,其中包含10μL Mix、0.2μL上/下游引物、2μL cDNA模板和适量的ddH2O。每个样品进行三次技术重复。结果采用2-ΔΔCt法进行分析。
qP-5142-F:TGTTCCAATTATCCCAGGAAGGG;
qP-5142-R:CAAGGATGTTGGAAACGGTTCC;
GAPDH-F:TACTCCGCAATGCTATCG;
GAPDH-R:TACTCGGTCCCAGTGGT。
2、实验结果
水稻纹枯病菌脂肪酶基因的沉默片段的扩增结果如图2所示,可以看出,应用沉默片段RslipA克隆引物VIGS-5142F/R,可以扩增出单一、明亮的条带。VIGS载体信息图如图4所示,构建得到的TRV2:RslipA载体图谱如图5所示,TRV2:RslipA载体转化至根癌农杆菌GV3101的菌落验证结果如图6所示,可以看出,扩增出单一、明亮的条带,说明TRV2:RslipA载体已转化至根癌农杆菌GV3101。烟草接种水稻纹枯病菌后致病情况如图7所示,对照植株转化GFP载体的叶片都出现病斑,其病情指数为81.2,而转化了TRV2:RslipA载体的烟草植株仅出现个别病斑,其病情指数为22.2。
烟草体内真菌生物量分析结果如图8所示,可以看出,与未接种水稻纹枯病菌的烟草相比,接种水稻纹枯病菌时,烟草体内的真菌生物量显著降低,仅为对照的0.13倍(p<0.01)。烟草体内靶标基因沉默效率分析结果如图9所示,可以看出,与对照组烟草植株中的靶标基因表达量相比,RslipA基因表达水平相较于对照植株也显著下调,沉默效率降低72%(p<0.01)。
由以上结果可知,将含有水稻纹枯病菌脂肪酶基因沉默片段RslipA的重组载体转化入植株后,该片段会伴随烟草脆裂病毒的复制而产生dsRNA,随后被切割成siRNA从而沉默接种时水稻纹枯病菌中脂肪酶基因的表达,表达量降低造成了瞬时转化的烟草抗性提高。由此能够得到抗水稻纹枯病菌转基因植物,与对照组相比其病情指数低,表达水平显著下调,在构建抗水稻纹枯病菌转基因植物:大豆、马铃薯、玉米和十字花科蔬菜等抗纹枯病菌转基因植物中,均具有广泛的应用前景,可以解决生产上土传真菌病害的难题,有效地防控土传真菌病害,具有重要的推广应用价值。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
序列表
<120> 一种水稻纹枯病菌脂肪基因沉默片段RslipA及其应用
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ccctatgctc tactctcgcg agcagggtat tgtcccgcaa gcaaaactgc gacgtggtca 180
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gctgagcgtg gcacttcttc ggttactctt acaggacaca gtctcggtgg tgccatttct 540
ctccttgatg cgctgtatct gtcgctccac ctcccgtcgg ccaaactcaa agtagtcact 600
catggaatgc cgagggttgg aaataccgaa tttgcgactc tcgtcgactc caagatcacc 660
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ccctatgcgt gagtgtctct ccctcggatt gatgtctgct ttaacttatg ctgcccacgt 180
agtctactct cgcgagcagg gtattgtccc gcaagcaaaa ctgcgacgtg gtcatgtgga 240
agtacgttgc cagaggtcca ttcagtgttt cctccaagat actaatacgc tatcacaatt 300
gcagcttcgt gtaacgagct tccaggcatg gtcgtgtatg cgtctggtgg ggacggtgtc 360
gtcactcctt attgtgcgtg ctttctttct cctatatgat tagcctaaat taattggtct 420
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ctcaccaaag tttgattagt gtgccactct tgatcgatgc cgactttagg ctcgactcac 600
ttgataccaa gtttttcccc ggtgtttctt cttcagtcaa gacacacaat ggtacttgtt 660
aaataactct tgtgtccggc acacgtattt atggtgttac gttacttttc caggtttcca 720
agaggcccag aagcgaggcg cccaggccaa acttgcagcc gtcaagaagg ccattgctga 780
gcgtggcact tcttcggtta ctcttacagg acacagtctc ggtacgttag tctatattca 840
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agggttggaa ataccgaatt tgcgactctc gtcgactcca aggtgtgtgt gctctcaacg 1020
aattctggag cagtaaaatt gaaagatgat tatagatcac cgacatttct cgtatcgtca 1080
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gtaataactc ttaaaagttc atgccgacgt atgcttatgc ccagagtcca ggtgaaagac 1200
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gcaccattgg aaccgtttcc aacatccttg tgggagatct caatgaccac ggtaagtttc 1320
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ccctatgctc tactctcgcg agcagggtat tgtcccgcaa gcaaaactgc gacgtggtca 180
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gtcgtcactc cttattggtt cgtcggttac tatcctggac tgaactcggt ggtgatcagc 300
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<211> 302
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<213> 人工序列(Artificial Sequence)
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Met Leu Ala Ser Phe Ala Ala Ala Phe Leu Leu Gly Val Ala Ser Thr
1 5 10 15
Leu Ala Leu Pro Ala Pro Ile Pro Arg Ala Gly Val Thr Thr Leu Thr
20 25 30
Thr Ala Gln Ile Thr Ala Phe Arg Pro Tyr Ala Leu Leu Ser Arg Ala
35 40 45
Gly Tyr Cys Pro Ala Ser Lys Thr Ala Thr Trp Ser Cys Gly Thr Ser
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115 120 125
Pro Gly Val Ser Ser Ser Val Lys Thr His Asn Gly Phe Gln Glu Ala
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Gln Lys Arg Gly Ala Gln Ala Lys Leu Ala Ala Val Lys Lys Ala Ile
145 150 155 160
Ala Glu Arg Gly Thr Ser Ser Val Thr Leu Thr Gly His Ser Leu Gly
165 170 175
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195 200 205
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<212> DNA
<213> 人工序列(Artificial Sequence)
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ttgatcgatg ccgactttag gctcgactca cttgatacca agtttttccc cggtgtttct 240
tcttcagtca agacacacaa tggtttccaa gaggcccaga agcgaggcgc cgaggccaaa 300
cttgcagccg tcaagaaggc cattgctgag cgtggcactt cctcggttac tcttacagga 360
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<210> 6
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<212> RNA
<213> 人工序列(Artificial Sequence)
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gcaagcaaaa cugcgacgug gucaugugga acuucgugua acgagcuucc agguaugguc 60
guguaugcgu cuggugggga cggugucguc acuccuuauu gguucgucgg uuacuauccu 120
ggacugaacu cgguggucau cagcaaccaa ggaaccgauc cauccaaauu ugugccacuc 180
uugaucgaug ccgacuuuag gcucgacuca cuugauacca aguuuuuccc cgguguuucu 240
ucuucaguca agacacacaa ugguuuccaa gaggcccaga agcgaggcgc cgaggccaaa 300
cuugcagccg ucaagaaggc cauugcugag cguggcacuu ccucgguuac ucuuacagga 360
cacagucucg guggugccau uucucuccuu gaugcgcugu aucugucgcu ccacc 415
Claims (10)
1.一种水稻纹枯病菌致病基因,其特征在于,其基因的cDNA序列如SEQ ID NO:1所示;或基因组DNA序列如SEQ ID NO:2所示;或编码序列如SEQ ID NO:3所示。
2.一种水稻纹枯病菌脂肪酶,其特征在于,其氨基酸序列如SEQ ID NO:4所示。
3.一种水稻纹枯病菌脂肪酶基因沉默片段RslipA,其特征在于,其核苷酸序列如SEQID NO:5所示。
4.权利要求1所述水稻纹枯病菌致病基因在防治水稻纹枯病菌中的应用。
5.权利要求3所述水稻纹枯病菌脂肪酶基因沉默片段RslipA在防治水稻纹枯病菌和/或制备水稻纹枯病菌防治产品中的应用。
6.权利要求1所述水稻纹枯病菌致病基因或权利要求3所述水稻纹枯病菌脂肪酶基因沉默片段RslipA在构建抗水稻纹枯病菌转基因植物中的应用。
7.一种水稻纹枯病菌防治制剂,其特征在于,含有能够抑制权利要求1所述水稻纹枯病菌致病基因表达的物质。
8.根据权利要求7所述水稻纹枯病菌防治制剂,其特征在于,所述物质含有权利要求3所述水稻纹枯病菌脂肪酶基因沉默片段RslipA和dsRNA,其dsRNA的核苷酸序列如SEQ IDNO:6所示。
9.一种水稻纹枯病菌的防治方法,其特征在于,采用权利要求7~8任一所述水稻纹枯病菌防治制剂。
10.权利要求7所述水稻纹枯病菌防治制剂在防治水稻纹枯病菌和/或制备水稻纹枯病菌防治产品和/或构建抗水稻纹枯病菌转基因植物中的应用。
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Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07274752A (ja) * | 1994-04-11 | 1995-10-24 | Mitsubishi Corp | 病害抵抗性イネ科植物およびその作出方法 |
JP2003250370A (ja) * | 2002-03-04 | 2003-09-09 | National Institute Of Agrobiological Sciences | 病害抵抗性イネ科植物 |
KR20100130168A (ko) * | 2009-06-02 | 2010-12-10 | 주식회사 바이오애그진앤텍 | 잔디 병원균 검출을 위한 표적 서열 증폭용 프라이머 및 그 용도 |
CN102206674A (zh) * | 2011-04-08 | 2011-10-05 | 扬州大学 | 一种利用RNAi技术培育抗条纹叶枯病水稻的方法 |
CN104788549A (zh) * | 2014-01-16 | 2015-07-22 | 中国科学院微生物研究所 | 水稻抗病性相关蛋白rir1及其编码基因和应用 |
BR112012031221A2 (pt) * | 2010-06-08 | 2015-09-08 | Devgen Private Ltd | método para regular negativamente expressão de genes em fungos |
CN107304422A (zh) * | 2016-04-21 | 2017-10-31 | 浙江省农业科学院 | 控制水稻纹枯病抗性的OsSBR1基因及其RNA干扰片段的应用 |
CN110066811A (zh) * | 2019-03-15 | 2019-07-30 | 四川农业大学 | 一种水稻纹枯病effector基因RsIA_SCR28及其应用 |
CN110358776A (zh) * | 2019-07-09 | 2019-10-22 | 华南农业大学 | 一种水稻纹枯病菌致病相关基因及其应用 |
CN110904139A (zh) * | 2019-12-05 | 2020-03-24 | 沈阳农业大学 | 重组载体、菌株、水稻纹枯病菌效应蛋白的表达纯化方法 |
CN111534501A (zh) * | 2020-04-22 | 2020-08-14 | 华南农业大学 | 一种水稻纹枯病菌MAPK蛋白激酶基因靶标片段Rsmapk及其应用 |
CN111748562A (zh) * | 2020-06-15 | 2020-10-09 | 华南农业大学 | 一种水稻纹枯病菌Atg 22蛋白编码基因及其靶标片段Rsatg22和应用 |
CN111808832A (zh) * | 2020-06-05 | 2020-10-23 | 华南农业大学 | 一种水稻纹枯病菌阳离子转运ATP酶基因及其片段Rscta和应用 |
-
2021
- 2021-08-20 CN CN202110964831.0A patent/CN113801887B/zh active Active
Patent Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07274752A (ja) * | 1994-04-11 | 1995-10-24 | Mitsubishi Corp | 病害抵抗性イネ科植物およびその作出方法 |
JP2003250370A (ja) * | 2002-03-04 | 2003-09-09 | National Institute Of Agrobiological Sciences | 病害抵抗性イネ科植物 |
KR20100130168A (ko) * | 2009-06-02 | 2010-12-10 | 주식회사 바이오애그진앤텍 | 잔디 병원균 검출을 위한 표적 서열 증폭용 프라이머 및 그 용도 |
BR112012031221A2 (pt) * | 2010-06-08 | 2015-09-08 | Devgen Private Ltd | método para regular negativamente expressão de genes em fungos |
CN102206674A (zh) * | 2011-04-08 | 2011-10-05 | 扬州大学 | 一种利用RNAi技术培育抗条纹叶枯病水稻的方法 |
CN104788549A (zh) * | 2014-01-16 | 2015-07-22 | 中国科学院微生物研究所 | 水稻抗病性相关蛋白rir1及其编码基因和应用 |
CN107304422A (zh) * | 2016-04-21 | 2017-10-31 | 浙江省农业科学院 | 控制水稻纹枯病抗性的OsSBR1基因及其RNA干扰片段的应用 |
CN110066811A (zh) * | 2019-03-15 | 2019-07-30 | 四川农业大学 | 一种水稻纹枯病effector基因RsIA_SCR28及其应用 |
CN110358776A (zh) * | 2019-07-09 | 2019-10-22 | 华南农业大学 | 一种水稻纹枯病菌致病相关基因及其应用 |
CN110904139A (zh) * | 2019-12-05 | 2020-03-24 | 沈阳农业大学 | 重组载体、菌株、水稻纹枯病菌效应蛋白的表达纯化方法 |
CN111534501A (zh) * | 2020-04-22 | 2020-08-14 | 华南农业大学 | 一种水稻纹枯病菌MAPK蛋白激酶基因靶标片段Rsmapk及其应用 |
CN111808832A (zh) * | 2020-06-05 | 2020-10-23 | 华南农业大学 | 一种水稻纹枯病菌阳离子转运ATP酶基因及其片段Rscta和应用 |
CN111748562A (zh) * | 2020-06-15 | 2020-10-09 | 华南农业大学 | 一种水稻纹枯病菌Atg 22蛋白编码基因及其靶标片段Rsatg22和应用 |
Non-Patent Citations (3)
Title |
---|
SHUAI LI 等: "The Effector AGLIP1 in Rhizoctonia solani AG1 IA Triggers Cell Death in Plants and Promotes Disease Development Through Inhibiting PAMP-Triggered Immunity in Arabidopsis thaliana", 《FRONT MICROBIOL 》, vol. 10, pages 1 - 12 * |
SRAYAN GHOSH 等: "RS_CRZ1, a C2H2-Type Transcription Factor Is Required for Pathogenesis of Rhizoctonia solani AG1-IA in Tomato", 《MOL PLANT MICROBE INTERACT .》, vol. 34, no. 1, pages 26 - 38 * |
石彦龙;徐国娟;王旭丽;王国梁;: "转录因子OsEIL2正调控水稻对纹枯病的抗性", 植物保护, vol. 48, no. 1, pages 46 - 53 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111748562A (zh) * | 2020-06-15 | 2020-10-09 | 华南农业大学 | 一种水稻纹枯病菌Atg 22蛋白编码基因及其靶标片段Rsatg22和应用 |
CN111748562B (zh) * | 2020-06-15 | 2022-04-29 | 华南农业大学 | 一种水稻纹枯病菌Atg 22蛋白编码基因及其靶标片段Rsatg22和应用 |
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