CN111286505A - 水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用 - Google Patents

水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用 Download PDF

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CN111286505A
CN111286505A CN201910408254.XA CN201910408254A CN111286505A CN 111286505 A CN111286505 A CN 111286505A CN 201910408254 A CN201910408254 A CN 201910408254A CN 111286505 A CN111286505 A CN 111286505A
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王石平
张标明
张海涛
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Abstract

本发明属于植物基因工程技术领域。具体涉及水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用。本发明提供了一种水稻抗白叶枯病基因Xa14,其具有SEQ ID NO:1所示的核苷酸序列,该基因的cDNA序列如SEQ ID NO:2所示,其编码的蛋白质序列如SEQ ID NO:3所示。Xa14基因属于BED‑NB‑LRR基因家族,该基因编码的蛋白质与植物抗病性相关。通过遗传转化,将Xa14基因转入宿主普通水稻品种,能够提高水稻对白叶枯病的抗性,从而减轻白叶枯病产生的危害,以达到增产的目的。

Description

水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用
技术领域
本发明属于植物基因工程技术领域,具体涉及水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用。 本发明Xa14基因属于BED-NB-LRR基因家族,编码的蛋白质与植物抗病性相关。通过遗传转化将Xa14 基因转入普通水稻品种,来提高水稻对白叶枯病的抗性。
背景技术
植物的抗病反应是由多基因调控的复杂信号传递过程。参与抗病反应的基因主要分为两类:主效抗病 (major resistance,MR)基因,它们大多直接参与植物和病原菌的识别反应,介导不同的抗病信号路径; 抗病相关基因,它们往往是抗病信号中处于下游的重要成员。根据植物抗病反应的特点和抗病程度,植物 的抗性又可分为质量抗性和数量抗性。通常情况下,对于质量抗性,植物仅对病原菌中某些特定致病小种 有专化性抗性,而对其他的的致病小种没有抗性。这类抗病反应可以由单个基因调控且抗病程度高,也被 称为主效抗病基因调控的抗性。这类抗性的不足是抗病性可能会因为致病小种的进化突变而丢失(Kou和 Wang 2010;Zhang和Wang 2013)。数量抗性的特点是植物与病原菌没有专化性的抗病性,不存在致病小 种的特异性。这类抗性一般由多个基因调控且具有广谱和持久抗病性,不足的是抗病水平较低,所以也可 以称为数量性状位点(quantitative trait locus,QTL)调控的抗性(Kou和Wang 2010)。
截至目前,水稻中已经有超过41个抗白叶枯病主效基因被鉴定(Zhang等2016)。其中11个基因被 克隆,分别是7个显性基因Xa1、Xa3/Xa26、Xa4、Xa10、Xa21、Xa23、Xa27(Song等1995;Yoshimura 等1998;Sun等2004;Gu等2005;Tian等2014;;Wang等2015;Hu等2017);4个隐性基因xa5、 xa13、xa25、xa41(Iyer和McCouch 2004;Chu等2006;Antony等2010;Liu等2011)。它们编码的蛋 白类型不尽相同,只有Xa1编码典型的NB-LRR类抗性蛋白,其它基因的编码产物均是非典型的抗性蛋白 (张海涛和王石平2016)。这些非典型的蛋白通过不同的方式调节水稻的抗病性,其中有多个抗性蛋白与 病原菌中的类转录激活效应子(transcription activator like effector,TAL effector)有关(Zhang和Wang 2013)。病原菌中TAL效应子可以靶定在宿主的启动子上,诱导植物抗病基因或感病基因表达,使植物表现出抗病 或者感病反应(Zhang等2015)。
水稻是单子叶植物研究的模式植物,同时也是世界上最主要的粮食作物之一。但是水稻在实际生产中 却面临着很多挑战,病害和非生物环境的影响常常造成产量和品质的下降,从而对粮食的安全形势造成非 常严重的影响。白叶枯病是水稻生产过程中最重要的三大病害之一,其侵染的方式是通过水孔或伤口进入 水稻叶片,靠吸取水稻的营养物质供自己繁殖,在维管束细胞传播,最后导致水稻叶片形成长条枯死状病 斑的症状。白叶枯病菌侵染水稻,往往会使水稻枯死,从而导致大幅度减产,严重时使水稻减产20%-30%(Kou和Wang 2010)。实践和研究表明,发掘和利用优良抗性基因资源改良水稻是控制病害发生、减少或 避免病害所带来的损失最经济有效的措施。
发明内容
本发明的目的是提供一种水稻抗白叶枯病基因Xa14,其具有SEQ ID NO:1所示的核苷酸序列。
本发明另一目的是提供一种利用抗白叶枯病基因Xa14提高水稻对白叶枯病抗性的应用。
本发明的技术方案如下所述:
本发明采用遗传学的方法,构建水稻抗白叶枯病菌的分离群体,利用图位克隆的方法,分离到水稻抗 白叶枯病基因Xa14。通过共分离标记检测表明该基因与白叶枯病抗性是共分离的。通过遗传转化Xa14基 因,使原感病水稻品种对白叶枯病菌产生抗性,证实了该基因的功能。
本发明Xa14基因的核苷酸序列如SEQ ID NO:1所示,该基因全长6311bp,具有2个内含子和3个 外显子,其CDS分别为199-505bp、601-913bp和1563-5922bp,cDNA全长为4980bp,编码1659个氨基 酸,其蛋白质序列如序列表SEQ ID NO:3所示。该蛋白属于BED-NB-LRR家族,活性中心301-592区段 为保守的NB-ARC区,包括ATP结合区等。应当理解,在不影响Xa14蛋白活性的前提下,本领域技术人 员可对SEQ ID NO:3所示的蛋白质序列进行各种替换、添加或/和缺失一个或几个氨基酸获得具有同等功 能的氨基酸序列。
此外,考虑到密码子的简并性,例如可在其编码区或非编码区不影响蛋白表达的条件下,对SEQ ID NO:3所示蛋白质序列进行修改。因此,本发明还包括对编码上述蛋白的基因序列进行的替换、添加或/ 和缺失一个或多个核苷酸,与上述编码蛋白基因具有相同或相似功能的核苷酸序列。本发明还包括基于所 述基因的正义或反义序列,包括含有所述核苷酸序列或其片段的克隆载体或表达载体,含有所述载体的宿 主细胞以及含有所述核苷酸序列或其片段的转化植物细胞和转基因植物。
附图说明
图1:本发明鉴定和分离克隆水稻抗病基因Xa14以及验证Xa14基因功能的流程图。
图2:感病品种IR24和抗病品种IRBB14杂交F2群体在孕穗期接种PXO112后病斑长度分布图。
图3:候选基因与Xa1的同源性分析。
图4:.Xa14的定位。A:分子标记下方的数值表示标记与Xa14重组单数,Xa14位于分子标记SNP1 和RM5473之间;B:SNP1和RM5473之间的物理图谱。
图5:本发明起始转化载体pCAMBIA1301的图谱。
图6:本发明所用遗传转化载体D261O的结构示意图。附图标记说明:RB和LB分别表示T-DNA的 右边界和左边界,GUS表示β-葡糖苷酸酶基因,Hpt表示潮霉素磷酸转移酶基因,NOS表示胭脂碱合成 酶基因的多聚腺苷酸化信号,PXa14表示Xa14自身启动子。
图7:T0代遗传转化植株(D261OIR)中基因型与抗性检测。附图标记说明:图7中的A图为接种白 叶枯病菌后叶片的发病情况,图7中的B图为用Xa14基因内部特异引物Xa14-6F/Xa14-6R鉴定T0代转基 因植株(1~20),图7中的C图为接种白叶枯病菌后叶片的病斑长度调查分析,对照为水稻感病品种IR24 (遗传转化的受体)。星号(**)表示与对照相比,转化植株的病斑长度显著(P<0.01)减小。
图8:T1代遗传转化植株(D261OIR)对白叶枯病菌PXO112的抗性增强表型与Xa14基因型共分离。 附图标记说明:图7中的A图和图7中的B图为1号遗传转化植株(D261OIR-1)的T1代接种白叶枯病 菌的病斑长度和Xa14基因型,图7中的C图和D图为3号遗传转化植株(D261OIR-3)的T1代接种白叶 枯病菌的病斑长度和Xa14基因型,图7中的E图和F图为8号遗传转化植株(D261OIR-8)的T1代接种 白叶枯病菌的病斑长度和Xa14基因型。对照是遗传转化受体IR24。病斑长度为接种白叶枯病菌两周后的 数据,每个数据是平均值(4个重复)±标准差。星号(**)表示与对照相比,转化植株的病斑长度显著 (P<0.01)减小。
具体实施方式
对序列表的说明:
序列表SEQ ID NO:1是本发明分离克隆的Xa14基因的核苷酸序列。其中该序列的199-5922位核苷酸 序列是该基因的编码区。
序列表SEQ ID NO:2是Xa14基因的cDNA序列。
序列表SEQ ID NO:3是Xa14基因编码的蛋白质序列。
以下实施例进一步定义本发明。但不应理解为对本发明的限制。根据以下的描述和这些实施例,本领 域技术人员可以确定本发明的基本特征,并且在不偏离本发明精神和范围的情况下,可以对本发明做出各 种修改,以使其适用各种用途和条件。
实施例1:Xa14基因的定位克隆
1.精细定位显性基因Xa14
将抗白叶枯病水稻品种IRBB14(国际水稻研究所)和感白叶枯病水稻品种IR24(国际水稻研究所) 杂交,构建了含Xa14的F2群体。为了鉴定该基因的显、隐性,在大田种植了F2群体并从中随机选择315 个单株组成随机群体。根据接种后调查的病斑长度,判定Xa14为显性抗病基因(图1)。用PCR和聚丙烯 酰胺凝胶电泳,以及SSR和SNP分子标记的方法,检测F2各个单株分子标记的分离情况。根据F2的分 子标记分型,应用JoinMap3.0软件,构建水稻染色体的分子标记遗传连锁图。再根据前期的研究结果,用 PCR和聚丙烯酰胺凝胶电泳的方法,以RM17487 (5′-CGGAGCATGTGGAGAGGAACTCG-3′/5′-GGAGAGGGCAAGGGCTTCTTCG-3′)和RM17524 (5′-TCCAGCTAGTTTGGTTGCATACG-3′/5′-GGACAATGAACTACAGTAGCACACG-3′)分子标记筛选F2群体得到 了13株重组单株(表1)。在此基础上,通过设计SNP和In/Del分子标记,进一步将Xa14基因定位于SNP1 (5′-ATTTCACGATGCCCCAAGC-3′/5′-GGAGCCCTGTGGGAAGATCT-3′)和RM5473 (5′-GGAGATAAGACACGAGGGAATTATGC-3′/5′-AGATTAACTACGCGCGCTCATCC-3′)之间。在定位期间还用到 三个中间分子标记,分别为RM17497 (5′-AGGCTCACAGCAATTAGAGGGAGAGG-3′/5′-CACTTGAAGGTTCAGAGGAGCACACC-3′)、RM17501 (5′-CTGGAGAGGGTGGTTGGGATCG-3′/5′-CAGCAGCAGCACGCGAAACC-3′)、RM3843 (5′-CCAGATCATCCAGGCATAACATCACC-3′/5′-CGGCGCTGGTAAACTCCATTCC-3′)。
表1 从599个单株中筛选到13个重组单株
Figure BDA0002061950730000041
2.Xa14候选基因的确定
根据Gramene数据库的信息(http://archive.gramene.org/markers/),分子标记SNP1与RM5473之间的 物理距离大约为173kb。根据水稻基因组注释数据库(RICE GenomeAnnotation Project; http://rice.plantbiology.msu.edu/)的信息,在该区域有20个蛋白编码位点,其中7个预测编码NB-LRR类 蛋白(表2)。
表2 编码NB-LRR蛋白的7个基因
Figure BDA0002061950730000042
通过测序,发现RGAb和RGAe编码的蛋白质只包含LRR区域,RGAg因为核苷酸的插入导致翻译提 前终止。其余4个基因编码完整的NB-LRR蛋白。RGAa、RGAc和RGAd分别编码了常见的NB-LRR蛋白, 而只有RGAf编码了含有锌指结构的蛋白。用生物信息学方法分析这些基因,结果发现RGAf与Xa1基因 高度同源,Xa1与RGAf的一致性为86.9%(图2)。因此,将RGAf基因命名为Xa14。
实施例2:Xa14候选基因的结构和编码产物分析
1.基因末端序列分析
按常规方法抽取水稻IRBB14总RNA,随后采用Invitrogen公司的Micro-FastTrackTM 2.0Kit (Invitrogen K1520-02)抽提mRNA,再采用Invitrogen公司的GeneRacer Core Kit试剂盒,通过 5’-RACE(rapid amplification of cDNA end)和3’-RACE分析方法,确定水稻家系IRBB14中的Xa14候选基 因的5’和3’末端序列,操作按Invitrogen公司提供的试剂盒说明书进行。进行5’-RACE第一轮扩增的引 物是Xa14-1R(5′-CAAAGCTGATGTCCCGTTCTTAGATGTGC-3′)和UPM(试剂盒自带);第二轮扩增引物是 Xa14-2R(5′-ATGCCTTGGACCGTTTCTTG-3′)和NUP(试剂盒自带);进行3’-RACE第一轮扩增的引物是Xa14-1F(5′-CAGGGCTATGAGCTATGCCC-3′)和UPM;第二轮扩增引物是Xa14-2F (5′-GAAATGGAAAGACTAACGGACG-3′)和NUP;然后通过电泳回收目标大小的片段,用pGEM-TVector System I试剂盒(美国Promega公司)对回收片段进行克隆。最后对克隆进行测序分析。
2.基因末端序列分析
采用GenScan(http://genes.mit.edu/GENSCAN.html)软件对包含Xa14基因区段的基因组DNA序列进 行分析,预测存在2个内含子和3个外显子,采用跨越内含子的三对PCR引物Xa14-3F (5′-CGAGAAGGTGGATGGGGT-3′)和Xa14-2R、Xa14-4F(5′-ACGGTTTGAGGCTGAAGACAC-3′)和 Xa14-4R(5′-GGGTGCGTGCTAACTGCCTCGTGGTGTG-3′)扩增基因组DNA和RNA反转录产物,RT-PCR 产物Xa14-3F/Xa14-2R和Xa14-4F/Xa14-4R均比从基因组中扩增出的产物小,说明确实存在2个内含子。 将RT-PCR产物克隆、测序、与基因组DNA序列比较,确定了各内含子长度及的剪切位点。采用Xa14基 因区段的其它引物进行RT-PCR、5’RACE和3’RACE分析,没有再发现其它内含子的存在。
比较分析Xa14基因的基因组序列和cDNA序列,确定Xa14基因由6311个核苷酸组成,包含3个外 显子和2个内含子。第一个外显子由505个核苷酸组成,它包括由198个核苷酸组成的5′端非翻译区 (UTR)(位于序列表SEQ ID NO:1的1-198bp处)和由306个核苷酸组成的编码区(位于序列表SEQ ID NO: 1的179-485bp处);第一个内含子由94个核苷酸组成(位于序列表SEQ ID NO:1的506-600bp处);第二 个外显子(即第二段编码区)由312个核苷酸组成(位于序列表SEQ ID NO:1的601-913bp处);第二个内含 子由648个核苷酸组成(位于序列表SEQ ID NO:1的914-1562bp处);第三个外显子由4748个核苷酸组 成,它包括由4359个核苷酸组成的第四段编码区(位于序列表SEQ ID NO:1的1563-5922bp处)和由388个核苷酸组成的3′端UTR(位于序列表SEQ ID NO:1的5923-6311bp处)。
3.基因编码产物分析
Xa14基因编码一个长度为1659个氨基酸的蛋白质。用Blastp方法分析,推测该基因编码一个 BED-NB-LRR类蛋白质,包含核苷酸结合位点和富亮氨酸拉链结构域。预测显示该蛋白质的氨基末端有一 个保守的BED锌指结构域。
实施例3:Xa14基因的功能验证与应用
本发明采用农杆菌介导的遗传转化方法,通过将Xa14基因转入水稻感病品种IR24(国际水稻所)中, 进一步验证该基因的功能。
1.遗传转化载体的构建
本发明所用的起始载体是常用载体pCAMBIA1301(图5,Cao等,2007)。根据基因组测序的结果, 设计引物(5'-GGTACCGGCATAAGAAATCATAGCAGTGG-3', 5'-CCAAGCTTGCATGCCTGCAGGTCGACGGTGTAAGGCCTCCATTCTAACAG-3'),其中包括HindIII和 BamHI的接头,扩增抗白叶枯病水稻IRBB14的基因组。PCR反应体系总体积50μl:1μl DNA,10×buffer 5μl,10mMdNTPs 5μl,10μM引物各1μl,高保真Taq酶0.5μl;灭菌的双蒸水37.5μl;反应条件:94℃2min,94℃15s,58℃30s,72℃7min,共30个循环;产物用TAE胶分离,过回收柱纯化。所得序列包 括Xa14上游2000bp的启动子、5724bp的基因组序列和下游的460bp的3'非编码区。用限制性内切酶 HindIII和BamHI组合酶切,酶切体系总体积50μl:PCR产物约15μl,10×反应buffer 5μl,酶各2μl, 混匀后37℃酶切过夜,试剂盒纯化回收所需片段。pCAMBIA1301载体酶切体系同上,试剂盒纯化回收。 连接反应:基因组片段3.8μl,载体0.5μl,T4ligase 0.2μl,10×反应buffer 0.5μl,4℃,连接过夜;连接 产物与大肠杆菌DH10B混匀,电转化后涂布于含卡那霉素的LB平板,37℃,过夜培养。挑单克隆,扩大 培养,抽质粒,酶切验证。把构建好的载体电转化农杆菌EHA105,抽质粒,PCR验证正确,加入等体积 的50%的甘油混匀,置于-70℃冰箱保存。
2.遗传转化水稻和T0代基因型检测
采用农杆菌EHA105介导的遗传转化方法(Lin和Zhang,2005)将上述Xa14基因组转化载体(D261O) 转入感病水稻品种IR24中。具体的操作步骤在上述参考方法的基础上改良如下:
(1)诱导愈伤组织:将水稻品种IR24的成熟种子去壳后,用0.15%的氯化汞浸泡消毒20分钟,每隔 5分钟剧烈震荡一次。之后在无菌操作台上用灭菌蒸馏水清洗种子5次。将种子放置在诱导培养基上(成 分见后),置于暗室培养一个月,培养温度为25±1℃。
(2)愈伤组织继代:在无菌操作台上挑取亮黄色、质地紧密且干燥的愈伤组织,放置在继代培养基 上(成分见后),置于暗室培养两周,培养温度为25±1℃。
(3)愈伤组织预培养:在无菌操作台上挑取亮黄色、质地紧密且干燥的愈伤组织,放置在预培养基 上(成分见后),置于暗室培养两周,培养温度为25±1℃。
(4)农杆菌培养:在带有对应抗性选择的LA培养基上(成分见后)预培养含有D261O载体的农杆 菌EHA105(来源于澳大利亚CAMBIA实验室商用菌株),培养时间为两天,培养温度为28℃;之后将上 述农杆菌转移至悬浮培养基(成分见后),于28℃摇床上培养2-3小时。
(5)农杆菌侵染愈伤组织:调节农杆菌的悬浮液至OD600数值为0.8-1.0,将预培养后的愈伤组织在 农杆菌悬浮液中浸泡30分钟,之后转移至无菌滤纸上吸干,然后放置在共培养基(成分见后)上于黑暗 处培养3天,培养温度为19℃。
(6)愈伤组织选择培养:用灭菌蒸馏水彻底清洗愈伤,之后转移愈伤至无菌滤纸上吸干,将愈伤转 移至选择培养基(成分见后)上培养2-3次,每次2周。
(7)愈伤组织分化:将抗性愈伤转移至预分化培养基(成分见后)上于黑暗处培养5-7天,之后转移 至分化培养基上(成分见后),于光照培养室内培养,温度为26℃。
(8)遗传转化苗生根处理:剪去分化得到的遗传转化苗的一部分根,将其转移至生根培养基中,于 光照培养室内培养2-3周,温度为26℃。之后将遗传转化苗移栽在土壤中。
遗传转化中所用的培养基组分及其配方如下:
(1)试剂和溶液的缩写
本发明中培养基所用到的植物激素的缩写表示如下:6-BA(6-BenzylaminoPurine,6-苄基氨基嘌呤); CN(Carbenicillin,羧苄青霉素);KT(Kinetin,激动素);NAA(Napthalene acetic acid,萘乙酸);IAA (Indole-3-acetic acid,吲哚乙酸);2,4-D(2,4-Dichlorophenoxyacetic acid,2,4-二氯苯氧乙酸);AS (Acetosringone,乙酰丁香酮);CH(Casein Enzymatic Hydrolysate,水解酪蛋白);HN(Hygromycin B, 潮霉素);DMSO(Dimethyl Sulfoxide,二甲基亚砜);N6max(N6培养基大量成分溶液);N6mix(N6培 养基微量成分溶液);MSmax(MS基本培养基大量成分溶液);MSmix(MS基本培养基微量成分溶液)。
(2)主要溶液配方
1)N6培养基大量元素母液[10倍浓缩液(10X)]的配制:
Figure BDA0002061950730000071
2)N6培养基微量元素母液[100倍浓缩液(100X)]的配制:
Figure BDA0002061950730000072
3)铁盐(Fe2EDTA)贮存液(100X)的配制:
准备800ml的去离子水并加热至70℃,加入3.73克的乙二铵四乙酸二钠(Na2EDTA·2H2O),充分 溶解后在70℃水浴中保持2小时,最后定容至1000ml。
4)维生素贮存液(100X)的配制:
Figure BDA0002061950730000073
Figure BDA0002061950730000081
5)MS基本培养基大量元素母液(10X)的配制:
Figure BDA0002061950730000082
6)MS基本培养基微量元素母液(100X)的配制:
Figure BDA0002061950730000083
7)2,4-D贮存液(1mg/ml)的配制:
称取2,4-D 100mg,用1ml 1N氢氧化钾溶解5分钟,然后加10ml蒸馏水溶解完全后定容至100ml。
8)6-BA贮存液(1mg/ml)的配制:
称取6-BA 100mg,用1ml 1N氢氧化钾溶解5分钟,然后加10ml蒸馏水溶解完全后定容至100ml。
9)萘乙酸(NAA)贮存液(1mg/ml)的配制:
称取NAA 100mg,用1ml 1N氢氧化钾溶解5分钟,然后加10ml蒸馏水溶解完全后定容至100ml。
10)吲哚乙酸(IAA)贮存液(1mg/ml)的配制:
称取IAA 100mg,用1ml 1N氢氧化钾溶解5分钟,然后加10ml蒸馏水溶解完全后定容至100ml。
11)葡萄糖贮存液(0.5g/ml)的配制:
称取葡萄糖125g,然后用蒸馏水溶解定容至250ml,灭菌后4℃保存备用。
12)AS贮存液的配制:
称取AS 0.392g,加入DMSO 10ml,溶解后分装至1.5ml离心管内,4℃保存备用。
13)1N氢氧化钾贮存液的配制:
称取氢氧化钾5.6g,用蒸馏水溶解并定容至100ml,室温保存备用。
(3)用于水稻遗传转化的培养基配方
1)诱导培养基
Figure BDA0002061950730000084
Figure BDA0002061950730000091
加蒸馏水至900ml,1N氢氧化钾调节pH值到5.9,煮沸并定容至1000ml,分装到50ml三角瓶(25 ml/瓶),封口灭菌。
2)继代培养基
Figure BDA0002061950730000092
加蒸馏水至900ml,1N氢氧化钾调节pH值到5.9,煮沸并定容至1000ml,分装到50ml三角瓶 (25ml/瓶),封口灭菌。
3)预培养基
Figure BDA0002061950730000093
加蒸馏水至250ml,1N氢氧化钾调节pH值到5.6,封口灭菌。使用前加热溶解并加入5ml葡萄糖 贮存液和250μl AS贮存液,分装倒入培养皿中(25ml/皿)。
4)共培养基
Figure BDA0002061950730000094
Figure BDA0002061950730000101
加蒸馏水至250ml,1N氢氧化钾调节pH值到5.6,封口灭菌。使用前加热溶解并加入5ml葡萄糖 贮存液和250μl AS贮存液,分装倒入培养皿中(25ml/每皿)。
5)悬浮培养基
Figure BDA0002061950730000102
加蒸馏水至100ml,调节pH值到5.4,分装到两个100ml的三角瓶中,封口灭菌。使用前加入1ml 葡萄糖贮存液和100μl AS贮存液。
6)选择培养基
Figure BDA0002061950730000103
加蒸馏水至250ml,调节pH值到6.0,封口灭菌。使用前溶解培养基,加入对应抗生素,分装倒入 培养皿中(25ml/皿)。
7)预分化培养基
Figure BDA0002061950730000104
Figure BDA0002061950730000111
加蒸馏水至250ml,1N氢氧化钾调节pH值到5.9,封口灭菌。使用前溶解培养基,加入对应抗生素, 分装倒入培养皿中(25ml/皿)。
8)分化培养基
Figure BDA0002061950730000112
加蒸馏水至900ml,1N氢氧化钾调节pH值到6.0。煮沸并定容至1000ml,分装到50ml三角瓶(50 ml/瓶),封口灭菌。
9)生根培养基
Figure BDA0002061950730000113
加蒸馏水至900ml,1N氢氧化钾调节pH值到5.8。煮沸并定容至1000ml,分装到生根管中(25ml /管),封口灭菌。
最终获得的遗传转化植株被命名为D261OIR(其中前面部分为遗传转化载体名称,IR代表水稻品种 IR24)。本发明共获得独立转化植株20株。在抽穗期阶段取植株的剑叶,抽提DNA,采用PCR技术检测 转化植株中Xa14基因型。结果显示,与对照IR24相比,所有的阳性遗传转化植株中的DNA带型都是杂 合的两条带。
3.T1代遗传转化植株的抗病性分析
为检测Xa14基因遗传转化植株是否对白叶枯病表现出抗性,申请人选取T0代3株遗传转化植株 (D261OIR-1,D261OIR-3和D261OIR-8)的T1代家系进行基因型和抗病性分析。在孕穗期接种白叶枯病 菌PXO112,接种两周后调查,同时取样抽提DNA,采用PCR技术检测转化植株的基因型。结果显示, T1代遗传转化家系中,与对照IR24相比,所有携带Xa14基因的阳性植株对白叶枯病菌PXO112的抗性显 著增强(P<0.01),而转基因阴性植株对白叶枯病菌的抗性没有变化(图7)。这些结果说明,Xa14基 因能够显著增强水稻对白叶枯病的抗性。
参考文献
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Chu Z,Fu B,Yang H,Xu C,Li Z,Sanchez A,Park YJ,Bennetzen JL,Zhang Q,Wang S(2006)Targeting Xa13,a recessive gene for bacterial blight resistancein rice.Theor Appl Genet,112:455-461;
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序列表
<110> 华中农业大学
<120> 水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用
<141> 2019-05-10
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 6311
<212> DNA
<213> 水稻(Oryza sativa)
<220>
<221> gene
<222> (1)..(6311)
<220>
<221> 3'UTR
<222> (5923)..(6311)
<220>
<221> exon
<222> (1563)..(5922)
<220>
<221> Intron
<222> (914)..(1562)
<220>
<221> exon
<222> (601)..(913)
<220>
<221> Intron
<222> (506)..(600)
<220>
<221> exon
<222> (199)..(505)
<220>
<221> 5'UTR
<222> (1)..(198)
<400> 1
catcagacga ttaatccacg acgaccagga gcgctgcccc gagccacacg cccacacgca 60
ctggtgaatc ttctcctttc cacctagttt tcactctgca gtctcctctc ctagctacta 120
ctctgccttc ccatcctcgc ttgttgcagc cctcttgcac acgccattgg ccacagtcca 180
aggactgctc ctgttccgat ggaggaggtg gaagccggtt tgctggaggg cgggatcagg 240
tggctggcgg agaccatcct ggataacctg gatgccgaca agctggatga gtggattcgc 300
cagattaggc tcgccgctga caccgagaag ctacgggctg agatcgagaa ggtggatggg 360
gtggtcgctg ccgtgaaggg gagggcgatc gggaacaggt cgctggcccg atctctcggc 420
cgtctcaggg ggttgctgta cgacgccgac gatgcggtcg acgagctcga ctacttcagg 480
ctccagcagc aggtcgaggg aggaggtact gtctttgcat atccgtgcca ttttaattaa 540
gtttgcaagc tgcgtgcctg caacaatggc gtcagtttcc aacccatgct tgtgctacag 600
ttattacacg gtttgaggct gaagacacgg ttggagatgg agcagaggac gaaggcgata 660
ttccgatgga caatactgat gtaccggcgg cggctgcggc aggccgcagc agcaagaaac 720
ggtccaaggc atgggaacac tttactcccg tagagttcac tgctgatggg aaggcttcta 780
aagcacggtg caagtactgc cacaaggacc tatgttgcac atctaagaac gggacatcag 840
ctttgcgcaa ccatctcaat gtttgcaaga ggaaacgtgt aacaagtact gaccaaccgg 900
taaatccatc aaggtaatgc taatggagtt ctgaatttag tgtaaatccg ttgaagtgta 960
aatttggccc gttacatctg cttaagatct cattctgtct ctaatcttct aatagccaac 1020
tcatggtcat tttttttcct aatatatagt accggtgatg gtgcaccaaa tgtaattaga 1080
tgcaaggaaa caaaagtgaa caattgtata tatcaaatat aattatatct aaaacatgag 1140
tagtgtatca aatccaattc tttcaaaaat ctactatgca aaattgagtg acaaaatctg 1200
ctgccttttt tttttacaga aagcaaccaa ttaatataag tcaaatataa aaacgctttg 1260
tagtctccaa taaaatagct cattgtttcg tttatactta tgtttataaa tttaaattta 1320
aaacttaatt ttggagttga ttttgtggtt ttcttttcat cctattttat tttacaacat 1380
ttgattttga atagttaaga atgcgtatat aaaaatttta cccataagtt attttttaaa 1440
ttgttaataa atcgtaagga taatcataag tataagtgaa acgattcgct cttcatctac 1500
ttaagattgc gttatattgc tgacctttct aatcgcctaa ccacgatcac atgctcttcc 1560
agtgccggtg agggtgcatc aaatgcaact ggtaattcag taggcagaaa aaggatgaga 1620
atggatggga cttcaacaca ccacgaggca gttagcacgc acccttggaa caaggctgaa 1680
ctttccaaca ggatccaatg catgactcat cagttagaag aggctgtaaa tgaggttatg 1740
aggctatgtc gatcctcaag ttcaaaccag agtcgacagg gtacatcacc ggccacaaat 1800
gcaacaacat cgtcttatct tccggagccc atagtgtatg ggagggctgc agagatggaa 1860
accatcaaac agctgatcat gagcaataga tctaatggca taaccgtcct gccaattgta 1920
ggcaatggag ggataggaaa aaccactttg gcgcaactgg tctgcaaaga tctggtaatt 1980
aaaagtcagt ttaatgttaa gatatgggtg tatgtatctg ataaatttga tgtagttaag 2040
attacaaggc agattttgga tcatgtctcc aaccagagcc acgaaggaat aagcaacctt 2100
gatacgcttc agcaggatct tgaggaacaa atgaaatcta agaagttcct cattgtctta 2160
gatgatgtgt gggaaatccg tacagatgac tggaaaaaac tactggctcc tttaagacct 2220
aatgatcagg tgaattcgtc acaggaagag gcaacaggta atatgataat tttgacaact 2280
cgtatacaga gtattgccaa aagtcttgga acagtacaat caattaagtt agaagctctg 2340
aaagatgacg atatatggtc actatttaaa gtgcatgctt ttggtaatga taaacatgat 2400
agtagtccag gcttacaggt tcttgggaag caaattgcta gcgagctaaa aggcaaccca 2460
ctggcagcaa aaactgtggg ttcactatta ggaacgaatc ttaccatcga tcattgggat 2520
agcattataa agagtgaaga atggaaatcc ctgcaacaag cttatggcat catgcaagcg 2580
ctgaagttga gctatgatca tctatccaac cccttacagc aatgcgtctc ttattgttct 2640
cttttcccca agggttattc tttcagcaaa gcacaactaa tacaaatatg gattgctcaa 2700
ggatttgtgg aagaatccag tgagaagttg gagcagaaag gatggaaata tctagctgag 2760
ttggtaaatt cgggtttcct tcagcaagtt gaaagcacac ggttttcatc agaatatttt 2820
gttatgcacg atcttatgca tgatttagcg caaaaggttt cacaaacaga atatgcaact 2880
atagatggct cagagtgcac agagttagcc ccaagtatac gccatttgtc aatagtaact 2940
gattctgcat accgcaagga gaaatataga aacatatctc gtaatgaggt gtttgagaaa 3000
aggttgatga aagttaagtc aaggagtaag ttgaggtcac tggtattaat tgggcaatat 3060
gattctcatt tttttaaata tttcaaagat gctttcaagg aagcacaaca tctgcgactg 3120
ctgcagatca ctgcaactta tgctgattct gattcatttc tctccagttt ggtaaattct 3180
acacatctcc ggtatctgaa aattgtgacc gaagaatccg gcagaacttt gccccgatct 3240
ctaaggaagt attaccatct tcaagtacta gatattggct atagatttgg aattccccgt 3300
atatctaatg atataaataa tcttctcagc ctgcggcatc ttgttgcata tgatgaagtg 3360
tgttcttcca ttgctaacat tggtaaaatg acctcacttc aggaactagg caattttatt 3420
gttcagaata atttaagtgg ttttgaggtg acacaattga aatccatgaa caagcttgta 3480
caacttagtg tgtctcaact tggaaatgtt agaactcagg aggaggcatg tggggcaaaa 3540
ctgaaagaca aacaacactt agaaaagcta catttgtcct ggaaggatgc atggaatgga 3600
tatgacagtg acgaaagcta tgaagatgaa tacggcagtg atatgaatat agaaacagaa 3660
ggggaggaac tgtcagttgg tgatgccaat ggtgcccaaa gcttacaaca tcacagtaat 3720
ataagctctg aacttgcttc aagtgaggtg ctcgaaggtc ttgaaccaca tcacggcctc 3780
aagtatctac ggatatctgg gtataatgga tctacctccc caacttggct tccttcttca 3840
cttacctgtc tgcaaacact tcatctagaa aaatgtggaa aatggcaaat acttccttta 3900
gaaaggctag ggttacttgt aaagctcgtg ttgatcaaaa tgaggaatgc aacagaactc 3960
tcaatccctt cactggagga gcttgtgtta attgcattgc caagcttgaa cacatgctcc 4020
tgcacttcca tcaggaactt gaactccagt ttaaaggttc tgaaaattaa gaattgccct 4080
gtactgaagg tatttccctt gtttgagatt tgccagaaat ttgaaatcga gcggacgtcg 4140
tcatggttgc cccatcttag caagcttacc atctataatt gtcctctttc ctgtgtgcac 4200
agttctctgc caccttcatc tattacatat tgctatgatt taacattcta cggcagcaag 4260
gtagatttcg ctggatttac ctcccttgag gagttagtga tttcacgatg ccccaagctg 4320
gtgtcgttct tggcgcataa cgacggaaat gatgaacagt cgaatggaag atggctccta 4380
ccgctatcac ttggaaaact tgagattaac tatgttgatt ccctaaaaac gctgcagctc 4440
tgctttccgg ggaacctcac ccgcctgaaa aaactagtag tgttgggaaa ccaaagttta 4500
acatctctgc agctccattc ctgcacagca ctccaagagt tgataattcg aagctgtgag 4560
tcgcttaatt ctctggaagg cttgcaattg ctcggcaatc tcaggttgct gtgtgcacac 4620
agatgcctca gcggccatga agaagatgga atgtgtatcc ttccgcaatc acttgaggaa 4680
attgacatct gggagtactc tcaagaaacg ctgcagccct gctttccagg gaacctcacc 4740
agcttgaaaa aactagtagt acagggaagc caaaagttaa tatctctgca gctgtattcc 4800
tgcacagcac tccaagagtt gatgattgaa agttgtgtgt cgcttaattc tctggaaggc 4860
ctgcaatggc tcgtcaacct caggttgctg cgggcacaca gatgcctcag tggttatgga 4920
gaaaatggaa ggtgtatcct tccacaatca cttgaggaac tttacatcag agagtattct 4980
caagaaacgc tgcagccctg ctttccaggg aacctcacca gcttgaaaaa actagtagta 5040
cagggaagcc aaaagttaat atctctgcag ctgtattcct gcacagcact ccaagagttg 5100
atgattggaa gttgtgtgtc gcttaattct ctggaaggcc tgcaatggct cgtcaacctc 5160
aggttgctgc gggcacacag atgcctcagt ggttatggag aaaatggaag gtgtatcctt 5220
ccacaatcac ttgagggtct ttacatcaga gagtattctc aagaaattct acagccctgc 5280
ttccagacga atctcacttg cttaaaaaga ttagaggtat caggcactgg aagtttcaaa 5340
tctctgaagt tgcaatcatg cactgcactc gaacatttga ggattgaatg gtgtgcgtcg 5400
cttgctacat tagagggctt gcaattcctc catgccctta agcatttgga agtattcaga 5460
tgccctggct tgcctccata tttggggagt ttatcagggc agggctatga gctatgccca 5520
ctactggaaa ggctcgagat cgatgacccc tctatcctta ccacgtcgtt ttgcaagaac 5580
ctcacctctc tccaatacct agagctttgc agtcacggat tggaaatgga aagactaacg 5640
gacgaggaag agagagcgct tcaactcctc acttccctgc aagagctccg atttaactgt 5700
tgctacaatc tcgtagatct tcccacaggg ctccacaacc ttctctccct caagaggttg 5760
gagatctgga attgcgggag catcgcgagg ccgctggaaa agggtctccc accttcgttg 5820
gaagaactgg atatcctcgg ttgtagtaaa gagctagttc agcagtgcag aactctagca 5880
agcaagctga aggtcgaaat taatggggaa tatgtgaact gattactcgg tggcttgtta 5940
ggcgcacctt tttccacctg tccaactggc gtgggctcgt tcaggcgttc aagctgctgt 6000
aaattccatt gccgcaatga cgaccttcag aaccgttaca caatacaaag gacatatgat 6060
ggctagatca actgtcgcag agctagcttt ggttcacctg aaaacataag gccaaacgcg 6120
tggttctttt aatcagaagt atcaaaaatt ggttttggtt tttaacgaaa agatgttaga 6180
aattggtttt aatgtgccag attctgtacc agaattctgt attttgctcc gtaattctgt 6240
gctacgactg atgttgtatt taactgatag caatcgaccg gcaaaaccaa tccgctccaa 6300
tgcattgtca c 6311
<210> 2
<211> 4980
<212> DNA
<213> 水稻(Oryza sativa)
<220>
<221> gene
<222> (1)..(4980)
<220>
<221> CDS
<222> (1)..(4980)
<400> 2
atg gag gag gtg gaa gcc ggt ttg ctg gag ggc ggg atc agg tgg ctg 48
Met Glu Glu Val Glu Ala Gly Leu Leu Glu Gly Gly Ile Arg Trp Leu
1 5 10 15
gcg gag acc atc ctg gat aac ctg gat gcc gac aag ctg gat gag tgg 96
Ala Glu Thr Ile Leu Asp Asn Leu Asp Ala Asp Lys Leu Asp Glu Trp
20 25 30
att cgc cag att agg ctc gcc gct gac acc gag aag cta cgg gct gag 144
Ile Arg Gln Ile Arg Leu Ala Ala Asp Thr Glu Lys Leu Arg Ala Glu
35 40 45
atc gag aag gtg gat ggg gtg gtc gct gcc gtg aag ggg agg gcg atc 192
Ile Glu Lys Val Asp Gly Val Val Ala Ala Val Lys Gly Arg Ala Ile
50 55 60
ggg aac agg tcg ctg gcc cga tct ctc ggc cgt ctc agg ggg ttg ctg 240
Gly Asn Arg Ser Leu Ala Arg Ser Leu Gly Arg Leu Arg Gly Leu Leu
65 70 75 80
tac gac gcc gac gat gcg gtc gac gag ctc gac tac ttc agg ctc cag 288
Tyr Asp Ala Asp Asp Ala Val Asp Glu Leu Asp Tyr Phe Arg Leu Gln
85 90 95
cag cag gtc gag gga gga gtt att aca cgg ttt gag gct gaa gac acg 336
Gln Gln Val Glu Gly Gly Val Ile Thr Arg Phe Glu Ala Glu Asp Thr
100 105 110
gtt gga gat gga gca gag gac gaa ggc gat att ccg atg gac aat act 384
Val Gly Asp Gly Ala Glu Asp Glu Gly Asp Ile Pro Met Asp Asn Thr
115 120 125
gat gta ccg gcg gcg gct gcg gca ggc cgc agc agc aag aaa cgg tcc 432
Asp Val Pro Ala Ala Ala Ala Ala Gly Arg Ser Ser Lys Lys Arg Ser
130 135 140
aag gca tgg gaa cac ttt act ccc gta gag ttc act gct gat ggg aag 480
Lys Ala Trp Glu His Phe Thr Pro Val Glu Phe Thr Ala Asp Gly Lys
145 150 155 160
gct tct aaa gca cgg tgc aag tac tgc cac aag gac cta tgt tgc aca 528
Ala Ser Lys Ala Arg Cys Lys Tyr Cys His Lys Asp Leu Cys Cys Thr
165 170 175
tct aag aac ggg aca tca gct ttg cgc aac cat ctc aat gtt tgc aag 576
Ser Lys Asn Gly Thr Ser Ala Leu Arg Asn His Leu Asn Val Cys Lys
180 185 190
agg aaa cgt gta aca agt act gac caa ccg gta aat cca tca agt gcc 624
Arg Lys Arg Val Thr Ser Thr Asp Gln Pro Val Asn Pro Ser Ser Ala
195 200 205
ggt gag ggt gca tca aat gca act ggt aat tca gta ggc aga aaa agg 672
Gly Glu Gly Ala Ser Asn Ala Thr Gly Asn Ser Val Gly Arg Lys Arg
210 215 220
atg aga atg gat ggg act tca aca cac cac gag gca gtt agc acg cac 720
Met Arg Met Asp Gly Thr Ser Thr His His Glu Ala Val Ser Thr His
225 230 235 240
cct tgg aac aag gct gaa ctt tcc aac agg atc caa tgc atg act cat 768
Pro Trp Asn Lys Ala Glu Leu Ser Asn Arg Ile Gln Cys Met Thr His
245 250 255
cag tta gaa gag gct gta aat gag gtt atg agg cta tgt cga tcc tca 816
Gln Leu Glu Glu Ala Val Asn Glu Val Met Arg Leu Cys Arg Ser Ser
260 265 270
agt tca aac cag agt cga cag ggt aca tca ccg gcc aca aat gca aca 864
Ser Ser Asn Gln Ser Arg Gln Gly Thr Ser Pro Ala Thr Asn Ala Thr
275 280 285
aca tcg tct tat ctt ccg gag ccc ata gtg tat ggg agg gct gca gag 912
Thr Ser Ser Tyr Leu Pro Glu Pro Ile Val Tyr Gly Arg Ala Ala Glu
290 295 300
atg gaa acc atc aaa cag ctg atc atg agc aat aga tct aat ggc ata 960
Met Glu Thr Ile Lys Gln Leu Ile Met Ser Asn Arg Ser Asn Gly Ile
305 310 315 320
acc gtc ctg cca att gta ggc aat gga ggg ata gga aaa acc act ttg 1008
Thr Val Leu Pro Ile Val Gly Asn Gly Gly Ile Gly Lys Thr Thr Leu
325 330 335
gcg caa ctg gtc tgc aaa gat ctg gta att aaa agt cag ttt aat gtt 1056
Ala Gln Leu Val Cys Lys Asp Leu Val Ile Lys Ser Gln Phe Asn Val
340 345 350
aag ata tgg gtg tat gta tct gat aaa ttt gat gta gtt aag att aca 1104
Lys Ile Trp Val Tyr Val Ser Asp Lys Phe Asp Val Val Lys Ile Thr
355 360 365
agg cag att ttg gat cat gtc tcc aac cag agc cac gaa gga ata agc 1152
Arg Gln Ile Leu Asp His Val Ser Asn Gln Ser His Glu Gly Ile Ser
370 375 380
aac ctt gat acg ctt cag cag gat ctt gag gaa caa atg aaa tct aag 1200
Asn Leu Asp Thr Leu Gln Gln Asp Leu Glu Glu Gln Met Lys Ser Lys
385 390 395 400
aag ttc ctc att gtc tta gat gat gtg tgg gaa atc cgt aca gat gac 1248
Lys Phe Leu Ile Val Leu Asp Asp Val Trp Glu Ile Arg Thr Asp Asp
405 410 415
tgg aaa aaa cta ctg gct cct tta aga cct aat gat cag gtg aat tcg 1296
Trp Lys Lys Leu Leu Ala Pro Leu Arg Pro Asn Asp Gln Val Asn Ser
420 425 430
tca cag gaa gag gca aca ggt aat atg ata att ttg aca act cgt ata 1344
Ser Gln Glu Glu Ala Thr Gly Asn Met Ile Ile Leu Thr Thr Arg Ile
435 440 445
cag agt att gcc aaa agt ctt gga aca gta caa tca att aag tta gaa 1392
Gln Ser Ile Ala Lys Ser Leu Gly Thr Val Gln Ser Ile Lys Leu Glu
450 455 460
gct ctg aaa gat gac gat ata tgg tca cta ttt aaa gtg cat gct ttt 1440
Ala Leu Lys Asp Asp Asp Ile Trp Ser Leu Phe Lys Val His Ala Phe
465 470 475 480
ggt aat gat aaa cat gat agt agt cca ggc tta cag gtt ctt ggg aag 1488
Gly Asn Asp Lys His Asp Ser Ser Pro Gly Leu Gln Val Leu Gly Lys
485 490 495
caa att gct agc gag cta aaa ggc aac cca ctg gca gca aaa act gtg 1536
Gln Ile Ala Ser Glu Leu Lys Gly Asn Pro Leu Ala Ala Lys Thr Val
500 505 510
ggt tca cta tta gga acg aat ctt acc atc gat cat tgg gat agc att 1584
Gly Ser Leu Leu Gly Thr Asn Leu Thr Ile Asp His Trp Asp Ser Ile
515 520 525
ata aag agt gaa gaa tgg aaa tcc ctg caa caa gct tat ggc atc atg 1632
Ile Lys Ser Glu Glu Trp Lys Ser Leu Gln Gln Ala Tyr Gly Ile Met
530 535 540
caa gcg ctg aag ttg agc tat gat cat cta tcc aac ccc tta cag caa 1680
Gln Ala Leu Lys Leu Ser Tyr Asp His Leu Ser Asn Pro Leu Gln Gln
545 550 555 560
tgc gtc tct tat tgt tct ctt ttc ccc aag ggt tat tct ttc agc aaa 1728
Cys Val Ser Tyr Cys Ser Leu Phe Pro Lys Gly Tyr Ser Phe Ser Lys
565 570 575
gca caa cta ata caa ata tgg att gct caa gga ttt gtg gaa gaa tcc 1776
Ala Gln Leu Ile Gln Ile Trp Ile Ala Gln Gly Phe Val Glu Glu Ser
580 585 590
agt gag aag ttg gag cag aaa gga tgg aaa tat cta gct gag ttg gta 1824
Ser Glu Lys Leu Glu Gln Lys Gly Trp Lys Tyr Leu Ala Glu Leu Val
595 600 605
aat tcg ggt ttc ctt cag caa gtt gaa agc aca cgg ttt tca tca gaa 1872
Asn Ser Gly Phe Leu Gln Gln Val Glu Ser Thr Arg Phe Ser Ser Glu
610 615 620
tat ttt gtt atg cac gat ctt atg cat gat tta gcg caa aag gtt tca 1920
Tyr Phe Val Met His Asp Leu Met His Asp Leu Ala Gln Lys Val Ser
625 630 635 640
caa aca gaa tat gca act ata gat ggc tca gag tgc aca gag tta gcc 1968
Gln Thr Glu Tyr Ala Thr Ile Asp Gly Ser Glu Cys Thr Glu Leu Ala
645 650 655
cca agt ata cgc cat ttg tca ata gta act gat tct gca tac cgc aag 2016
Pro Ser Ile Arg His Leu Ser Ile Val Thr Asp Ser Ala Tyr Arg Lys
660 665 670
gag aaa tat aga aac ata tct cgt aat gag gtg ttt gag aaa agg ttg 2064
Glu Lys Tyr Arg Asn Ile Ser Arg Asn Glu Val Phe Glu Lys Arg Leu
675 680 685
atg aaa gtt aag tca agg agt aag ttg agg tca ctg gta tta att ggg 2112
Met Lys Val Lys Ser Arg Ser Lys Leu Arg Ser Leu Val Leu Ile Gly
690 695 700
caa tat gat tct cat ttt ttt aaa tat ttc aaa gat gct ttc aag gaa 2160
Gln Tyr Asp Ser His Phe Phe Lys Tyr Phe Lys Asp Ala Phe Lys Glu
705 710 715 720
gca caa cat ctg cga ctg ctg cag atc act gca act tat gct gat tct 2208
Ala Gln His Leu Arg Leu Leu Gln Ile Thr Ala Thr Tyr Ala Asp Ser
725 730 735
gat tca ttt ctc tcc agt ttg gta aat tct aca cat ctc cgg tat ctg 2256
Asp Ser Phe Leu Ser Ser Leu Val Asn Ser Thr His Leu Arg Tyr Leu
740 745 750
aaa att gtg acc gaa gaa tcc ggc aga act ttg ccc cga tct cta agg 2304
Lys Ile Val Thr Glu Glu Ser Gly Arg Thr Leu Pro Arg Ser Leu Arg
755 760 765
aag tat tac cat ctt caa gta cta gat att ggc tat aga ttt gga att 2352
Lys Tyr Tyr His Leu Gln Val Leu Asp Ile Gly Tyr Arg Phe Gly Ile
770 775 780
ccc cgt ata tct aat gat ata aat aat ctt ctc agc ctg cgg cat ctt 2400
Pro Arg Ile Ser Asn Asp Ile Asn Asn Leu Leu Ser Leu Arg His Leu
785 790 795 800
gtt gca tat gat gaa gtg tgt tct tcc att gct aac att ggt aaa atg 2448
Val Ala Tyr Asp Glu Val Cys Ser Ser Ile Ala Asn Ile Gly Lys Met
805 810 815
acc tca ctt cag gaa cta ggc aat ttt att gtt cag aat aat tta agt 2496
Thr Ser Leu Gln Glu Leu Gly Asn Phe Ile Val Gln Asn Asn Leu Ser
820 825 830
ggt ttt gag gtg aca caa ttg aaa tcc atg aac aag ctt gta caa ctt 2544
Gly Phe Glu Val Thr Gln Leu Lys Ser Met Asn Lys Leu Val Gln Leu
835 840 845
agt gtg tct caa ctt gga aat gtt aga act cag gag gag gca tgt ggg 2592
Ser Val Ser Gln Leu Gly Asn Val Arg Thr Gln Glu Glu Ala Cys Gly
850 855 860
gca aaa ctg aaa gac aaa caa cac tta gaa aag cta cat ttg tcc tgg 2640
Ala Lys Leu Lys Asp Lys Gln His Leu Glu Lys Leu His Leu Ser Trp
865 870 875 880
aag gat gca tgg aat gga tat gac agt gac gaa agc tat gaa gat gaa 2688
Lys Asp Ala Trp Asn Gly Tyr Asp Ser Asp Glu Ser Tyr Glu Asp Glu
885 890 895
tac ggc agt gat atg aat ata gaa aca gaa ggg gag gaa ctg tca gtt 2736
Tyr Gly Ser Asp Met Asn Ile Glu Thr Glu Gly Glu Glu Leu Ser Val
900 905 910
ggt gat gcc aat ggt gcc caa agc tta caa cat cac agt aat ata agc 2784
Gly Asp Ala Asn Gly Ala Gln Ser Leu Gln His His Ser Asn Ile Ser
915 920 925
tct gaa ctt gct tca agt gag gtg ctc gaa ggt ctt gaa cca cat cac 2832
Ser Glu Leu Ala Ser Ser Glu Val Leu Glu Gly Leu Glu Pro His His
930 935 940
ggc ctc aag tat cta cgg ata tct ggg tat aat gga tct acc tcc cca 2880
Gly Leu Lys Tyr Leu Arg Ile Ser Gly Tyr Asn Gly Ser Thr Ser Pro
945 950 955 960
act tgg ctt cct tct tca ctt acc tgt ctg caa aca ctt cat cta gaa 2928
Thr Trp Leu Pro Ser Ser Leu Thr Cys Leu Gln Thr Leu His Leu Glu
965 970 975
aaa tgt gga aaa tgg caa ata ctt cct tta gaa agg cta ggg tta ctt 2976
Lys Cys Gly Lys Trp Gln Ile Leu Pro Leu Glu Arg Leu Gly Leu Leu
980 985 990
gta aag ctc gtg ttg atc aaa atg agg aat gca aca gaa ctc tca atc 3024
Val Lys Leu Val Leu Ile Lys Met Arg Asn Ala Thr Glu Leu Ser Ile
995 1000 1005
cct tca ctg gag gag ctt gtg tta att gca ttg cca agc ttg aac aca 3072
Pro Ser Leu Glu Glu Leu Val Leu Ile Ala Leu Pro Ser Leu Asn Thr
1010 1015 1020
tgc tcc tgc act tcc atc agg aac ttg aac tcc agt tta aag gtt ctg 3120
Cys Ser Cys Thr Ser Ile Arg Asn Leu Asn Ser Ser Leu Lys Val Leu
1025 1030 1035 1040
aaa att aag aat tgc cct gta ctg aag gta ttt ccc ttg ttt gag att 3168
Lys Ile Lys Asn Cys Pro Val Leu Lys Val Phe Pro Leu Phe Glu Ile
1045 1050 1055
tgc cag aaa ttt gaa atc gag cgg acg tcg tca tgg ttg ccc cat ctt 3216
Cys Gln Lys Phe Glu Ile Glu Arg Thr Ser Ser Trp Leu Pro His Leu
1060 1065 1070
agc aag ctt acc atc tat aat tgt cct ctt tcc tgt gtg cac agt tct 3264
Ser Lys Leu Thr Ile Tyr Asn Cys Pro Leu Ser Cys Val His Ser Ser
1075 1080 1085
ctg cca cct tca tct att aca tat tgc tat gat tta aca ttc tac ggc 3312
Leu Pro Pro Ser Ser Ile Thr Tyr Cys Tyr Asp Leu Thr Phe Tyr Gly
1090 1095 1100
agc aag gta gat ttc gct gga ttt acc tcc ctt gag gag tta gtg att 3360
Ser Lys Val Asp Phe Ala Gly Phe Thr Ser Leu Glu Glu Leu Val Ile
1105 1110 1115 1120
tca cga tgc ccc aag ctg gtg tcg ttc ttg gcg cat aac gac gga aat 3408
Ser Arg Cys Pro Lys Leu Val Ser Phe Leu Ala His Asn Asp Gly Asn
1125 1130 1135
gat gaa cag tcg aat gga aga tgg ctc cta ccg cta tca ctt gga aaa 3456
Asp Glu Gln Ser Asn Gly Arg Trp Leu Leu Pro Leu Ser Leu Gly Lys
1140 1145 1150
ctt gag att aac tat gtt gat tcc cta aaa acg ctg cag ctc tgc ttt 3504
Leu Glu Ile Asn Tyr Val Asp Ser Leu Lys Thr Leu Gln Leu Cys Phe
1155 1160 1165
ccg ggg aac ctc acc cgc ctg aaa aaa cta gta gtg ttg gga aac caa 3552
Pro Gly Asn Leu Thr Arg Leu Lys Lys Leu Val Val Leu Gly Asn Gln
1170 1175 1180
agt tta aca tct ctg cag ctc cat tcc tgc aca gca ctc caa gag ttg 3600
Ser Leu Thr Ser Leu Gln Leu His Ser Cys Thr Ala Leu Gln Glu Leu
1185 1190 1195 1200
ata att cga agc tgt gag tcg ctt aat tct ctg gaa ggc ttg caa ttg 3648
Ile Ile Arg Ser Cys Glu Ser Leu Asn Ser Leu Glu Gly Leu Gln Leu
1205 1210 1215
ctc ggc aat ctc agg ttg ctg tgt gca cac aga tgc ctc agc ggc cat 3696
Leu Gly Asn Leu Arg Leu Leu Cys Ala His Arg Cys Leu Ser Gly His
1220 1225 1230
gaa gaa gat gga atg tgt atc ctt ccg caa tca ctt gag gaa att gac 3744
Glu Glu Asp Gly Met Cys Ile Leu Pro Gln Ser Leu Glu Glu Ile Asp
1235 1240 1245
atc tgg gag tac tct caa gaa acg ctg cag ccc tgc ttt cca ggg aac 3792
Ile Trp Glu Tyr Ser Gln Glu Thr Leu Gln Pro Cys Phe Pro Gly Asn
1250 1255 1260
ctc acc agc ttg aaa aaa cta gta gta cag gga agc caa aag tta ata 3840
Leu Thr Ser Leu Lys Lys Leu Val Val Gln Gly Ser Gln Lys Leu Ile
1265 1270 1275 1280
tct ctg cag ctg tat tcc tgc aca gca ctc caa gag ttg atg att gaa 3888
Ser Leu Gln Leu Tyr Ser Cys Thr Ala Leu Gln Glu Leu Met Ile Glu
1285 1290 1295
agt tgt gtg tcg ctt aat tct ctg gaa ggc ctg caa tgg ctc gtc aac 3936
Ser Cys Val Ser Leu Asn Ser Leu Glu Gly Leu Gln Trp Leu Val Asn
1300 1305 1310
ctc agg ttg ctg cgg gca cac aga tgc ctc agt ggt tat gga gaa aat 3984
Leu Arg Leu Leu Arg Ala His Arg Cys Leu Ser Gly Tyr Gly Glu Asn
1315 1320 1325
gga agg tgt atc ctt cca caa tca ctt gag gaa ctt tac atc aga gag 4032
Gly Arg Cys Ile Leu Pro Gln Ser Leu Glu Glu Leu Tyr Ile Arg Glu
1330 1335 1340
tat tct caa gaa acg ctg cag ccc tgc ttt cca ggg aac ctc acc agc 4080
Tyr Ser Gln Glu Thr Leu Gln Pro Cys Phe Pro Gly Asn Leu Thr Ser
1345 1350 1355 1360
ttg aaa aaa cta gta gta cag gga agc caa aag tta ata tct ctg cag 4128
Leu Lys Lys Leu Val Val Gln Gly Ser Gln Lys Leu Ile Ser Leu Gln
1365 1370 1375
ctg tat tcc tgc aca gca ctc caa gag ttg atg att gga agt tgt gtg 4176
Leu Tyr Ser Cys Thr Ala Leu Gln Glu Leu Met Ile Gly Ser Cys Val
1380 1385 1390
tcg ctt aat tct ctg gaa ggc ctg caa tgg ctc gtc aac ctc agg ttg 4224
Ser Leu Asn Ser Leu Glu Gly Leu Gln Trp Leu Val Asn Leu Arg Leu
1395 1400 1405
ctg cgg gca cac aga tgc ctc agt ggt tat gga gaa aat gga agg tgt 4272
Leu Arg Ala His Arg Cys Leu Ser Gly Tyr Gly Glu Asn Gly Arg Cys
1410 1415 1420
atc ctt cca caa tca ctt gag ggt ctt tac atc aga gag tat tct caa 4320
Ile Leu Pro Gln Ser Leu Glu Gly Leu Tyr Ile Arg Glu Tyr Ser Gln
1425 1430 1435 1440
gaa att cta cag ccc tgc ttc cag acg aat ctc act tgc tta aaa aga 4368
Glu Ile Leu Gln Pro Cys Phe Gln Thr Asn Leu Thr Cys Leu Lys Arg
1445 1450 1455
tta gag gta tca ggc act gga agt ttc aaa tct ctg aag ttg caa tca 4416
Leu Glu Val Ser Gly Thr Gly Ser Phe Lys Ser Leu Lys Leu Gln Ser
1460 1465 1470
tgc act gca ctc gaa cat ttg agg att gaa tgg tgt gcg tcg ctt gct 4464
Cys Thr Ala Leu Glu His Leu Arg Ile Glu Trp Cys Ala Ser Leu Ala
1475 1480 1485
aca tta gag ggc ttg caa ttc ctc cat gcc ctt aag cat ttg gaa gta 4512
Thr Leu Glu Gly Leu Gln Phe Leu His Ala Leu Lys His Leu Glu Val
1490 1495 1500
ttc aga tgc cct ggc ttg cct cca tat ttg ggg agt tta tca ggg cag 4560
Phe Arg Cys Pro Gly Leu Pro Pro Tyr Leu Gly Ser Leu Ser Gly Gln
1505 1510 1515 1520
ggc tat gag cta tgc cca cta ctg gaa agg ctc gag atc gat gac ccc 4608
Gly Tyr Glu Leu Cys Pro Leu Leu Glu Arg Leu Glu Ile Asp Asp Pro
1525 1530 1535
tct atc ctt acc acg tcg ttt tgc aag aac ctc acc tct ctc caa tac 4656
Ser Ile Leu Thr Thr Ser Phe Cys Lys Asn Leu Thr Ser Leu Gln Tyr
1540 1545 1550
cta gag ctt tgc agt cac gga ttg gaa atg gaa aga cta acg gac gag 4704
Leu Glu Leu Cys Ser His Gly Leu Glu Met Glu Arg Leu Thr Asp Glu
1555 1560 1565
gaa gag aga gcg ctt caa ctc ctc act tcc ctg caa gag ctc cga ttt 4752
Glu Glu Arg Ala Leu Gln Leu Leu Thr Ser Leu Gln Glu Leu Arg Phe
1570 1575 1580
aac tgt tgc tac aat ctc gta gat ctt ccc aca ggg ctc cac aac ctt 4800
Asn Cys Cys Tyr Asn Leu Val Asp Leu Pro Thr Gly Leu His Asn Leu
1585 1590 1595 1600
ctc tcc ctc aag agg ttg gag atc tgg aat tgc ggg agc atc gcg agg 4848
Leu Ser Leu Lys Arg Leu Glu Ile Trp Asn Cys Gly Ser Ile Ala Arg
1605 1610 1615
ccg ctg gaa aag ggt ctc cca cct tcg ttg gaa gaa ctg gat atc ctc 4896
Pro Leu Glu Lys Gly Leu Pro Pro Ser Leu Glu Glu Leu Asp Ile Leu
1620 1625 1630
ggt tgt agt aaa gag cta gtt cag cag tgc aga act cta gca agc aag 4944
Gly Cys Ser Lys Glu Leu Val Gln Gln Cys Arg Thr Leu Ala Ser Lys
1635 1640 1645
ctg aag gtc gaa att aat ggg gaa tat gtg aac tga 4980
Leu Lys Val Glu Ile Asn Gly Glu Tyr Val Asn
1650 1655
<210> 3
<211> 1659
<212> PRT
<213> 水稻(Oryza sativa)
<400> 3
Met Glu Glu Val Glu Ala Gly Leu Leu Glu Gly Gly Ile Arg Trp Leu
1 5 10 15
Ala Glu Thr Ile Leu Asp Asn Leu Asp Ala Asp Lys Leu Asp Glu Trp
20 25 30
Ile Arg Gln Ile Arg Leu Ala Ala Asp Thr Glu Lys Leu Arg Ala Glu
35 40 45
Ile Glu Lys Val Asp Gly Val Val Ala Ala Val Lys Gly Arg Ala Ile
50 55 60
Gly Asn Arg Ser Leu Ala Arg Ser Leu Gly Arg Leu Arg Gly Leu Leu
65 70 75 80
Tyr Asp Ala Asp Asp Ala Val Asp Glu Leu Asp Tyr Phe Arg Leu Gln
85 90 95
Gln Gln Val Glu Gly Gly Val Ile Thr Arg Phe Glu Ala Glu Asp Thr
100 105 110
Val Gly Asp Gly Ala Glu Asp Glu Gly Asp Ile Pro Met Asp Asn Thr
115 120 125
Asp Val Pro Ala Ala Ala Ala Ala Gly Arg Ser Ser Lys Lys Arg Ser
130 135 140
Lys Ala Trp Glu His Phe Thr Pro Val Glu Phe Thr Ala Asp Gly Lys
145 150 155 160
Ala Ser Lys Ala Arg Cys Lys Tyr Cys His Lys Asp Leu Cys Cys Thr
165 170 175
Ser Lys Asn Gly Thr Ser Ala Leu Arg Asn His Leu Asn Val Cys Lys
180 185 190
Arg Lys Arg Val Thr Ser Thr Asp Gln Pro Val Asn Pro Ser Ser Ala
195 200 205
Gly Glu Gly Ala Ser Asn Ala Thr Gly Asn Ser Val Gly Arg Lys Arg
210 215 220
Met Arg Met Asp Gly Thr Ser Thr His His Glu Ala Val Ser Thr His
225 230 235 240
Pro Trp Asn Lys Ala Glu Leu Ser Asn Arg Ile Gln Cys Met Thr His
245 250 255
Gln Leu Glu Glu Ala Val Asn Glu Val Met Arg Leu Cys Arg Ser Ser
260 265 270
Ser Ser Asn Gln Ser Arg Gln Gly Thr Ser Pro Ala Thr Asn Ala Thr
275 280 285
Thr Ser Ser Tyr Leu Pro Glu Pro Ile Val Tyr Gly Arg Ala Ala Glu
290 295 300
Met Glu Thr Ile Lys Gln Leu Ile Met Ser Asn Arg Ser Asn Gly Ile
305 310 315 320
Thr Val Leu Pro Ile Val Gly Asn Gly Gly Ile Gly Lys Thr Thr Leu
325 330 335
Ala Gln Leu Val Cys Lys Asp Leu Val Ile Lys Ser Gln Phe Asn Val
340 345 350
Lys Ile Trp Val Tyr Val Ser Asp Lys Phe Asp Val Val Lys Ile Thr
355 360 365
Arg Gln Ile Leu Asp His Val Ser Asn Gln Ser His Glu Gly Ile Ser
370 375 380
Asn Leu Asp Thr Leu Gln Gln Asp Leu Glu Glu Gln Met Lys Ser Lys
385 390 395 400
Lys Phe Leu Ile Val Leu Asp Asp Val Trp Glu Ile Arg Thr Asp Asp
405 410 415
Trp Lys Lys Leu Leu Ala Pro Leu Arg Pro Asn Asp Gln Val Asn Ser
420 425 430
Ser Gln Glu Glu Ala Thr Gly Asn Met Ile Ile Leu Thr Thr Arg Ile
435 440 445
Gln Ser Ile Ala Lys Ser Leu Gly Thr Val Gln Ser Ile Lys Leu Glu
450 455 460
Ala Leu Lys Asp Asp Asp Ile Trp Ser Leu Phe Lys Val His Ala Phe
465 470 475 480
Gly Asn Asp Lys His Asp Ser Ser Pro Gly Leu Gln Val Leu Gly Lys
485 490 495
Gln Ile Ala Ser Glu Leu Lys Gly Asn Pro Leu Ala Ala Lys Thr Val
500 505 510
Gly Ser Leu Leu Gly Thr Asn Leu Thr Ile Asp His Trp Asp Ser Ile
515 520 525
Ile Lys Ser Glu Glu Trp Lys Ser Leu Gln Gln Ala Tyr Gly Ile Met
530 535 540
Gln Ala Leu Lys Leu Ser Tyr Asp His Leu Ser Asn Pro Leu Gln Gln
545 550 555 560
Cys Val Ser Tyr Cys Ser Leu Phe Pro Lys Gly Tyr Ser Phe Ser Lys
565 570 575
Ala Gln Leu Ile Gln Ile Trp Ile Ala Gln Gly Phe Val Glu Glu Ser
580 585 590
Ser Glu Lys Leu Glu Gln Lys Gly Trp Lys Tyr Leu Ala Glu Leu Val
595 600 605
Asn Ser Gly Phe Leu Gln Gln Val Glu Ser Thr Arg Phe Ser Ser Glu
610 615 620
Tyr Phe Val Met His Asp Leu Met His Asp Leu Ala Gln Lys Val Ser
625 630 635 640
Gln Thr Glu Tyr Ala Thr Ile Asp Gly Ser Glu Cys Thr Glu Leu Ala
645 650 655
Pro Ser Ile Arg His Leu Ser Ile Val Thr Asp Ser Ala Tyr Arg Lys
660 665 670
Glu Lys Tyr Arg Asn Ile Ser Arg Asn Glu Val Phe Glu Lys Arg Leu
675 680 685
Met Lys Val Lys Ser Arg Ser Lys Leu Arg Ser Leu Val Leu Ile Gly
690 695 700
Gln Tyr Asp Ser His Phe Phe Lys Tyr Phe Lys Asp Ala Phe Lys Glu
705 710 715 720
Ala Gln His Leu Arg Leu Leu Gln Ile Thr Ala Thr Tyr Ala Asp Ser
725 730 735
Asp Ser Phe Leu Ser Ser Leu Val Asn Ser Thr His Leu Arg Tyr Leu
740 745 750
Lys Ile Val Thr Glu Glu Ser Gly Arg Thr Leu Pro Arg Ser Leu Arg
755 760 765
Lys Tyr Tyr His Leu Gln Val Leu Asp Ile Gly Tyr Arg Phe Gly Ile
770 775 780
Pro Arg Ile Ser Asn Asp Ile Asn Asn Leu Leu Ser Leu Arg His Leu
785 790 795 800
Val Ala Tyr Asp Glu Val Cys Ser Ser Ile Ala Asn Ile Gly Lys Met
805 810 815
Thr Ser Leu Gln Glu Leu Gly Asn Phe Ile Val Gln Asn Asn Leu Ser
820 825 830
Gly Phe Glu Val Thr Gln Leu Lys Ser Met Asn Lys Leu Val Gln Leu
835 840 845
Ser Val Ser Gln Leu Gly Asn Val Arg Thr Gln Glu Glu Ala Cys Gly
850 855 860
Ala Lys Leu Lys Asp Lys Gln His Leu Glu Lys Leu His Leu Ser Trp
865 870 875 880
Lys Asp Ala Trp Asn Gly Tyr Asp Ser Asp Glu Ser Tyr Glu Asp Glu
885 890 895
Tyr Gly Ser Asp Met Asn Ile Glu Thr Glu Gly Glu Glu Leu Ser Val
900 905 910
Gly Asp Ala Asn Gly Ala Gln Ser Leu Gln His His Ser Asn Ile Ser
915 920 925
Ser Glu Leu Ala Ser Ser Glu Val Leu Glu Gly Leu Glu Pro His His
930 935 940
Gly Leu Lys Tyr Leu Arg Ile Ser Gly Tyr Asn Gly Ser Thr Ser Pro
945 950 955 960
Thr Trp Leu Pro Ser Ser Leu Thr Cys Leu Gln Thr Leu His Leu Glu
965 970 975
Lys Cys Gly Lys Trp Gln Ile Leu Pro Leu Glu Arg Leu Gly Leu Leu
980 985 990
Val Lys Leu Val Leu Ile Lys Met Arg Asn Ala Thr Glu Leu Ser Ile
995 1000 1005
Pro Ser Leu Glu Glu Leu Val Leu Ile Ala Leu Pro Ser Leu Asn Thr
1010 1015 1020
Cys Ser Cys Thr Ser Ile Arg Asn Leu Asn Ser Ser Leu Lys Val Leu
1025 1030 1035 1040
Lys Ile Lys Asn Cys Pro Val Leu Lys Val Phe Pro Leu Phe Glu Ile
1045 1050 1055
Cys Gln Lys Phe Glu Ile Glu Arg Thr Ser Ser Trp Leu Pro His Leu
1060 1065 1070
Ser Lys Leu Thr Ile Tyr Asn Cys Pro Leu Ser Cys Val His Ser Ser
1075 1080 1085
Leu Pro Pro Ser Ser Ile Thr Tyr Cys Tyr Asp Leu Thr Phe Tyr Gly
1090 1095 1100
Ser Lys Val Asp Phe Ala Gly Phe Thr Ser Leu Glu Glu Leu Val Ile
1105 1110 1115 1120
Ser Arg Cys Pro Lys Leu Val Ser Phe Leu Ala His Asn Asp Gly Asn
1125 1130 1135
Asp Glu Gln Ser Asn Gly Arg Trp Leu Leu Pro Leu Ser Leu Gly Lys
1140 1145 1150
Leu Glu Ile Asn Tyr Val Asp Ser Leu Lys Thr Leu Gln Leu Cys Phe
1155 1160 1165
Pro Gly Asn Leu Thr Arg Leu Lys Lys Leu Val Val Leu Gly Asn Gln
1170 1175 1180
Ser Leu Thr Ser Leu Gln Leu His Ser Cys Thr Ala Leu Gln Glu Leu
1185 1190 1195 1200
Ile Ile Arg Ser Cys Glu Ser Leu Asn Ser Leu Glu Gly Leu Gln Leu
1205 1210 1215
Leu Gly Asn Leu Arg Leu Leu Cys Ala His Arg Cys Leu Ser Gly His
1220 1225 1230
Glu Glu Asp Gly Met Cys Ile Leu Pro Gln Ser Leu Glu Glu Ile Asp
1235 1240 1245
Ile Trp Glu Tyr Ser Gln Glu Thr Leu Gln Pro Cys Phe Pro Gly Asn
1250 1255 1260
Leu Thr Ser Leu Lys Lys Leu Val Val Gln Gly Ser Gln Lys Leu Ile
1265 1270 1275 1280
Ser Leu Gln Leu Tyr Ser Cys Thr Ala Leu Gln Glu Leu Met Ile Glu
1285 1290 1295
Ser Cys Val Ser Leu Asn Ser Leu Glu Gly Leu Gln Trp Leu Val Asn
1300 1305 1310
Leu Arg Leu Leu Arg Ala His Arg Cys Leu Ser Gly Tyr Gly Glu Asn
1315 1320 1325
Gly Arg Cys Ile Leu Pro Gln Ser Leu Glu Glu Leu Tyr Ile Arg Glu
1330 1335 1340
Tyr Ser Gln Glu Thr Leu Gln Pro Cys Phe Pro Gly Asn Leu Thr Ser
1345 1350 1355 1360
Leu Lys Lys Leu Val Val Gln Gly Ser Gln Lys Leu Ile Ser Leu Gln
1365 1370 1375
Leu Tyr Ser Cys Thr Ala Leu Gln Glu Leu Met Ile Gly Ser Cys Val
1380 1385 1390
Ser Leu Asn Ser Leu Glu Gly Leu Gln Trp Leu Val Asn Leu Arg Leu
1395 1400 1405
Leu Arg Ala His Arg Cys Leu Ser Gly Tyr Gly Glu Asn Gly Arg Cys
1410 1415 1420
Ile Leu Pro Gln Ser Leu Glu Gly Leu Tyr Ile Arg Glu Tyr Ser Gln
1425 1430 1435 1440
Glu Ile Leu Gln Pro Cys Phe Gln Thr Asn Leu Thr Cys Leu Lys Arg
1445 1450 1455
Leu Glu Val Ser Gly Thr Gly Ser Phe Lys Ser Leu Lys Leu Gln Ser
1460 1465 1470
Cys Thr Ala Leu Glu His Leu Arg Ile Glu Trp Cys Ala Ser Leu Ala
1475 1480 1485
Thr Leu Glu Gly Leu Gln Phe Leu His Ala Leu Lys His Leu Glu Val
1490 1495 1500
Phe Arg Cys Pro Gly Leu Pro Pro Tyr Leu Gly Ser Leu Ser Gly Gln
1505 1510 1515 1520
Gly Tyr Glu Leu Cys Pro Leu Leu Glu Arg Leu Glu Ile Asp Asp Pro
1525 1530 1535
Ser Ile Leu Thr Thr Ser Phe Cys Lys Asn Leu Thr Ser Leu Gln Tyr
1540 1545 1550
Leu Glu Leu Cys Ser His Gly Leu Glu Met Glu Arg Leu Thr Asp Glu
1555 1560 1565
Glu Glu Arg Ala Leu Gln Leu Leu Thr Ser Leu Gln Glu Leu Arg Phe
1570 1575 1580
Asn Cys Cys Tyr Asn Leu Val Asp Leu Pro Thr Gly Leu His Asn Leu
1585 1590 1595 1600
Leu Ser Leu Lys Arg Leu Glu Ile Trp Asn Cys Gly Ser Ile Ala Arg
1605 1610 1615
Pro Leu Glu Lys Gly Leu Pro Pro Ser Leu Glu Glu Leu Asp Ile Leu
1620 1625 1630
Gly Cys Ser Lys Glu Leu Val Gln Gln Cys Arg Thr Leu Ala Ser Lys
1635 1640 1645
Leu Lys Val Glu Ile Asn Gly Glu Tyr Val Asn
1650 1655

Claims (2)

1.Xa14基因在改良水稻对白叶枯病抗性中的应用,其特征在于,所述的Xa14基因的核苷酸序列如序列表SEQ ID NO:1所示。
2.Xa14基因在改良水稻对白叶枯病抗性中的应用,其特征在于,所述的Xa14基因编码的蛋白质序列如序列表SEQ ID NO:3所示。
CN201910408254.XA 2019-05-16 2019-05-16 水稻抗白叶枯病基因Xa14在改良水稻抗病性中的应用 Pending CN111286505A (zh)

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