CN112410308A - 水稻ACCase突变型基因及其蛋白在植物抗除草剂中的应用 - Google Patents

水稻ACCase突变型基因及其蛋白在植物抗除草剂中的应用 Download PDF

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CN112410308A
CN112410308A CN202011316935.2A CN202011316935A CN112410308A CN 112410308 A CN112410308 A CN 112410308A CN 202011316935 A CN202011316935 A CN 202011316935A CN 112410308 A CN112410308 A CN 112410308A
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张保龙
王金彦
凌溪铁
王凤斌
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Jiangsu Academy of Agricultural Sciences
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Abstract

本发明公开了水稻ACCase突变型基因及其蛋白在植物抗除草剂中的应用，所述水稻ACCase突变型蛋白的氨基酸序列存在如下突变：其对应于野生型水稻ACCase的氨基酸序列的第2010位和/或2099发生突变。通过转基因方法将ACCase突变型基因导入对除草剂敏感的水稻基因组并使其表达，受体植物可以具有对乙酰辅酶A羧化酶抑制剂类除草剂的抗（耐）性。该ACCase突变型基因及其蛋白可广泛应用于培育抗除草剂的植物育种。

Description

水稻ACCase突变型基因及其蛋白在植物抗除草剂中的应用
技术领域
本发明涉及水稻ACCase突变型基因及其蛋白在植物抗除草剂中的应用，属于植物基因工程和植物抗除草剂育种领域。
背景技术
乙酰辅酶A羧化酶(ACCase)是植物中脂肪酸从头合成途径中的限速酶，在质体内参与以乙酰辅酶A为前体生成丙二酰辅酶A的过程。ACCase由三部分组成，即生物素羧基载体蛋白(BCCP)、生物素羧化酶(BC)和羧基转移酶(CT)。由于ACCase对植物的正常生长发育具有关键作用，因此，农药公司以ACCase为靶标，开发研制了一系列除草剂，在市场推广至今。
ACCase是植物脂肪酸生物合成的关键酶，它催化ATP依赖乙酰辅酶A羧化作用诱导脂肪酸与类黄酮生物合成中的一种关键代谢产物——丙二酸单酰辅酶A，这是植物中脂肪酸生物合成的首要关键步骤，然后在脂肪酸合成酶作用下形成脂肪酸。若丙二酸单酰辅酶A的合成停止，则对ACCase的抑制将使植物丧失这种中间产物，导致膜结构的破坏，透性增强，细胞破坏。
ACCase抑制剂类除草剂对谷类作物和双子叶作物田中的禾本科杂草均有极强的防效，在全世界广泛使用，从而导致了抗ACCase抑制剂杂草的发生。由于化学除草剂的广泛使用，杂草抗性发生速度如此之快、范围如此之广，不但增加了除草成本，也增加了环境压力。
ACCase同乙酰乳酸合成酶(ALS)和多酚氧化酶(PPO)一样是高效除草剂的靶标酶，针对该靶标的除草剂主要有芳氧苯氧基丙酸类(AOPP)和环己烯酮类(CHD)除草剂，它们对双子叶植物有高度的选择性，而且具有低的哺乳动物毒性和良好的环境特性，受到越来越多的关注。AOPP类除草剂是一类著名的防除禾本科杂草的高活性除草剂，也是一类发展比较迅速、不断开发出新品种的除草剂类型。该类除草剂最早是在20世纪60年代开始研究。目前商品化的AOPP类除草剂有禾草灵、稳杀得、高效盖草能、精禾草克、恶草醚、精喹禾灵等。该类除草剂存在立体异构，而且这种异构体中只有右旋体对ACCase有显著活性。
由于化学除草剂的广泛使用，杂草抗性发生的速度如此之快、范围如此之广，不但增加了除草成本，也增加了环境压力。目前，对抗性杂草种群形成的原因有两种观点：一种认为杂草长期与除草剂接触，而使除草剂作用部位的编码基因发生突变或者基因表达发生改变，结果导致杂草对除草剂的解毒能力提高或者对除草剂具有耐药性或抗药性的种群。总的来说，杂草抗性的形成，基本上是一个抗性基因被选择的过程，抗药性基因的遗传是杂草产生抗药性的基础。之前的报道认为，抗药性的产生不仅是由于植物改变了对除草剂的吸收与转运或植物加速了除草剂的代谢，且主要是因为除草剂的作用部位发生了改变。相当一部分杂草通过靶标酶ACCase的变化来解除或减少其与除草剂的亲和性而获得抗性。瑞士黑麦草对禾草灵的抗性是由于部分ACCase基因突变造成。
目前，还没有研究报道水稻中的ACCase位点突变后可以用于抗精喹禾灵和盖草能等除草剂。
发明内容
发明目的：本发明所要解决的技术问题是提供了一种可以赋予植物抗除草剂抗性的突变型蛋白。进一步地，本发明所要解决的技术问题是提供一种可以赋予植物抗(耐)乙酰辅酶A羧化酶类除草剂抗性的水稻ACCase突变型蛋白。
本发明还要解决的技术问题是提供一种编码水稻ACCase突变型蛋白的核酸或基因。
本发明还要解决的技术问题是提供了表达盒、重组载体或细胞，其含有所述的突变型基因或核酸。
本发明还要解决的技术问题是提供了所述的蛋白，所述的突变型基因或核酸，所述的表达盒、重组载体或细胞在植物抗除草剂方面的应用。
本发明还要解决的技术问题是提供了获得具有除草剂抗性的植物的方法。
本发明最后要解决的技术问题是提供了鉴定植物的方法。
技术方案：本发明提供了水稻ACCase突变型蛋白，所述突变型蛋白的氨基酸序列对应于野生型水稻ACCase的氨基酸序列的第2010位和/或2099发生突变。
作为优选，所述突变型蛋白的氨基酸序列的第2010位由色氨酸突变为亮氨酸和/或2099位由半胱氨酸突变为精氨酸。
优选地，本发明的水稻ACCase的氨基酸序列第2010位氨基酸突变为除了亮氨酸之外的其它氨基酸并且第2099位氨基酸突变为除了精氨酸之外的其它氨基酸。
优选地，本发明的ACCase突变型蛋白发生突变的氨基酸包括但不仅限于上述几种突变，在上述位点突变成其他氨基酸的情况也在本发明的保护范围之内，例如：第2010位氨基酸由色氨酸突变为丙氨酸、甘氨酸、苯丙氨酸、丝氨酸、酪氨酸、脯氨酸、苏氨酸、异亮氨酸、天冬氨酸、谷氨酰胺、赖氨酸、组氨酸、甲硫氨酸、亮氨酸、缬氨酸、谷氨酸、精氨酸；第2099位氨基酸由半胱氨酸突变为丙氨酸、甘氨酸、苯丙氨酸、丝氨酸、酪氨酸、脯氨酸、苏氨酸、异亮氨酸、天冬氨酸、谷氨酰胺、赖氨酸、组氨酸、甲硫氨酸、亮氨酸、缬氨酸、谷氨酸、精氨酸。
本发明所述的水稻ACCase突变型蛋白，包括：
(a)其氨基酸序列如SEQ ID NO.2和SEQ ID NO.4所示；或
(b)在(a)中的氨基酸序列经过取代和/或缺失和/或添加一个或多个氨基酸，且具有乙酰辅酶A羧化酶活性的由(a)衍生的蛋白质。
本发明内容还包括水稻ACCase突变型基因，其编码所述的突变型蛋白。
进一步地，本发明的水稻ACCase突变型基因在水稻ACCase基因的第6029位核苷酸发生突变和/或第6295位核苷酸发生突变。
作为优选，本发明的水稻ACCase突变型基因，所述水稻ACCase基因在水稻ACCase基因的第6029位核苷酸由G突变为T和/或第6295位核苷酸由T突变为C。
进一步地，本发明的水稻ACCase突变型基因的核苷酸序列如SEQ ID NO：1和SEQID NO：3所示。
进一步地，本发明的水稻ACCase突变型基因还包括在严格条件下与限定的核苷酸序列杂交且编码具有乙酰辅酶A羧化酶活性的蛋白质的核苷酸序列；
本发明的ACCase突变型基因发生突变的核苷酸包括但不仅限于上述突变，在上述位点突变成其他核苷酸的情况也在本发明的保护范围之内，例如：G突变为C、A等均在本发明的保护范围之内。
本发明还提供一种表达盒、重组载体或细胞，其含有所述的核酸或基因。
本发明还提供所述的水稻ACCase突变型蛋白、核酸或基因，表达盒、重组载体或细胞在植物抗除草剂方面的应用。
进一步地，本发明的除草剂包括但不仅限于盖草能或精喹禾灵。
其中，本发明的除草剂还包括烯草酮、氰氟草酯、禾草灵、精氟吡禾草灵、禾草灭、烯禾啶、笨草酮、芳氧苯基环己二酮、三酮类环己二酮、拿扑净中的一种或多种。
本发明还提供获得具有除草剂抗性的植物的方法，包括如下步骤：
1)使植物包含所述核酸或基因；或
2)使植物表达所述水稻ACCase突变型蛋白。
进一步地，所述获得具有除草剂抗性的植物的方法包括转基因、杂交、或回交。
本发明还提供一种鉴定植物的方法，其中所述植物是包含所述的核酸或基因的植物、表达所述的蛋白的植物或由所述的方法获得的植物，包括以下步骤：
1)测定所述植物是否包含所述的核酸或基因；或，
2)测定所述植物是否表达所述的突变型蛋白。
本发明还提供一种除草剂的防护剂，该防护剂包括所述的水稻ACCase突变型蛋白。
本发明还提供一种控制杂草的方法，包括：对种植作物的大田施用有效剂量的除草剂，所述植物包含所述的核酸或基因或所述的表达盒、重组载体或细胞，所述除草剂为精喹禾灵和盖草能中的一种或两种。
本发明还提供一种用于保护植物免受由除草剂引起的损伤的方法，包括：对种植作物的大田施用有效剂量的除草剂，所述植物包含所述的核酸或基因或将所述的表达盒、重组载体导入植物，导入后的植物产生除草剂抗性蛋白，所述除草剂为精喹禾灵和盖草能中的一种或两种。
进一步地，本发明的野生型基因来源于水稻的禾广丝苗。
进一步地，本发明的禾广丝苗的野生型植株不具有盖草能或精喹禾灵除草剂抗性，本发明的含有ACCase突变位点的转基因植株能赋予水稻植物增加对乙酰辅酶A羧化酶类除草剂的耐受性。
其中，本发明的水稻禾广丝苗的ACCase突变型突变型基因序列的编号及其对应的核苷酸和氨基酸序列分别是：禾广丝苗的ACCase基因的第6029位核苷酸由G突变为T，2010位氨基酸由色氨酸突变为亮氨酸，其核苷酸序列如SEQ ID No：1所示，其氨基酸序列如SEQID No：2所示；
此外，另一个禾广丝苗的ACCase突变基因的第6029位核苷酸由G突变为T、第6295位核苷酸由T突变为C，其蛋白在2010位氨基酸由色氨酸突变为亮氨酸，2099位氨基酸由半胱氨酸突变为精氨酸，其核苷酸序列如SEQ ID No：3所示，其氨基酸序列如SEQ IDNo：4所示；
本发明还提供了获得具有除草剂抗性的植物的方法。进一步地，本发明提供了用于增强植物、植物组织、植物细胞对于至少一种除草剂的耐受性的方法，所述除草剂干扰ACCase酶的活性。因此，本发明中除草剂抗性指的是抗(耐)乙酰辅酶A羧化酶类除草剂。
本发明所述的增强植物、植物组织或者植物细胞对除草剂的抗性方法可以通过转化或者杂交、自交和无性繁殖的方法实施，也可通过基因定点突变的方式获得，使改变的植株包括本发明的核苷酸序列SEQ ID No：1和SEQ ID No：3中的一种。
本发明所述的增强植物、植物组织或者植物细胞对除草剂的抗性方法可以通过转化或者杂交、自交和无性繁殖的方法实施，也可通过基因定点突变的方式获得，使改变的植株表达本发明的氨基酸序列SEQ ID No：2和SEQ ID No：4中的一种。
本发明还提供了耐受除草剂的水稻植物，其表达含有抗除草剂突变位点的ACCase基因，该基因序列不同于野生型水稻植物的ACCase基因序列，是ACCase基因含有抗除草剂突变位点的序列。
通过该方法获得的上述携带有突变位点的水稻ACCase突变序列的方法也在本发明的保护范围之内，将所述水稻ACCase突变序列转入到野生型水稻品种，发现携带有所述ACCase突变位点的转基因水稻可以抗ACCase类除草剂，而野生型水稻对ACCase类除草剂表现敏感。
本发明还提供了所述核酸或基因及蛋白在植物育种中的应用，用于培育具有除草剂抗性的植物，尤其是农作物，还提供了这些蛋白及其编码基因在转基因或者非转基因如水稻等植物中的应用。
本发明所述的农作物包括但不仅限于水稻，还可以包含其他的农作物。
有益效果：相对于现有技术，本发明具备以下优点：
1)本发明首次通过实验研究得出野生型水稻为籼型常规水稻种子禾广丝苗的水稻ACCase的氨基酸序列的第2010位发生突变，该基因突变得到的突变型蛋白对除草剂具有显著的抗性功能，特别是乙酰辅酶A羧化酶类除草剂。
2)本发明通过实验研究得出野生型水稻为籼型常规水稻种子禾广丝苗的水稻ACCase的氨基酸序列的第2010位和第2099位发生突变，该基因突变得到的突变型蛋白对除草剂具有显著的抗性功能，特别是乙酰辅酶A羧化酶类除草剂。
3)将ACCase突变序列转入到野生型水稻中，转基因水稻获得了ACCase酶类除草剂抗性。在田间喷施ACCase抑制剂类除草剂盖草能的实验结果表明，含有本发明的水稻ACCase突变蛋白的转基因水稻在施用4倍推荐使用浓度的乙酰辅酶A羧化酶抑制剂类除草剂后，植株仍然正常生长发育和结实，而野生型水稻幼苗在施用1倍推荐使用浓度的乙酰辅酶A羧化酶抑制剂类除草剂后，植株生长逐渐停止，叶片逐渐失绿、变浅黄色，植株不能拔节，最终整株死亡。
4)在田间喷施ACCase抑制剂类除草剂精喹禾灵的实验结果表明，含有本发明的水稻ACCase突变蛋白的转基因水稻正常生长，而野生型水稻则死亡。
附图说明
图1、抗乙酰辅酶A羧化酶抑制剂类除草剂突变体HGJK-3植株；
图2、抗乙酰辅酶A羧化酶抑制剂类除草剂突变体HGJK-4植株；
图3、PCR扩增含有水稻ACCase突变位点的ACCase基因全长结果图；第1泳道为Marker；Marker分子量从上到下依次为15kb、8kb、5kb、3kb、2kb、1.5kb、1kb、750bp、500bp，第2泳道为野生型禾广丝苗的ACCase的DNA，第3泳道为突变体HGJK-3植株的ACCase的DNA，第4泳道为突变体HGJK-4植株的ACCase的DNA；
图4、突变型ACCase基因水稻PCR检测图；总共有5个泳道，第1泳道为Marker；Marker分子量从上到下依次为15kb、8kb、5kb、3kb、2kb、1.5kb、1kb、750bp、500bp；第2泳道为野生型禾广丝苗的ACCase的DNA，第3泳道为重组质粒的PCR扩增产物，第4泳道为转化含有ACCase-6029抗除草剂突变体ACCase基因的转基因植株的PCR扩增产物，第5泳道为转化含有ACCase-6029和ACCase-6295抗除草剂突变体ACCase基因的转基因植株的PCR扩增产物。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述，但是本领域技术人员将会理解，下列实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
实施例1水稻M3突变体抗乙酰辅酶A羧化酶类除草剂(精喹禾灵)
将粳型常规水稻种子禾广丝苗(江苏省农业种质资源保护与利用平台惠赠，此为M0，用清水浸泡2小时)150kg分6次用0.5-1.0％(w/w)甲磺酸乙酯(EMS)室温下浸泡6-9小时，期间每1小时摇动一次种子；弃去EMS溶液，自来水翻动浸泡种子5次，每次5分钟，然后用自来水冲洗种子过夜，次日进行田间播种，并进行常规肥水管理(此为M1)。植株成熟后，种子混收、晾干，过冬保存。次年播种田间。待水稻(此为M2)幼苗长至3-4叶期时，喷施1mL精喹禾灵/L水，30天后还呈正常绿色植株即为抗乙酰辅酶A羧化酶类除草剂的水稻突变体(图1)。将禾广丝苗抗除草剂突变体收获的种子播种出苗(此为M3)，待M3水稻幼苗长至3-4叶期时，喷施1mL精喹禾灵/L水(4倍推荐浓度)。喷施除草剂15天后，抗性苗M3呈正常绿色、能继续长高至20-30cm，而非抗性苗叶片失去绿色甚至部分枯黄，植株不长高，只有5-9cm。30天后，M3抗性株均呈正常绿色植株，而喷施同样浓度除草剂的野生型水稻都已全部枯死，而突变体的存活率在90.8％，表明突变水稻能够抗4倍推荐浓度的精喹禾灵除草剂。伤害比率为1-存活率。同时，我们又对ACCase类除草剂盖草能(1mL盖草能/L水)敏感性进行实验，发现突变体也可以抗盖草能除草剂(表1)。
表1除草剂对野生型和HGJK-3突变体的伤害比率
Figure BDA0002789305650000071
实施例2：水稻镇糯19号野生型Accase全长基因的获得
选取禾广丝苗野生型植株的叶片，提取基因组DNA，根据NCBI中日本晴ACCase基因(NCBI：XM_015783727)设计引物进行扩增，采用KOD DNApolymerase聚合酶(购自Toyobo公司)扩增ACCase基因，其反应体系如下：
Figure BDA0002789305650000072
PCR扩增反应程序采用两步法，退火和延伸合为一起，采用68度。
程序如下：预变性：94℃3min；35个循环：变性94℃10sec；延伸68℃10min；保温：72℃10min。
取2μl PCR产物经1％琼脂糖凝胶电泳检测，发现有预期大小的片段后(图3)，剩余的PCR产物经PCR清洁试剂盒(购自Axygen公司)清洁回收后，克隆至pMD19-T载体(购自Takara公司)，然后转化大肠杆菌。每个转化随机挑取12个大肠杆菌单克隆进行PCR检测，取PCR结果呈阳性的6个单克隆，送南京一道生物科技有限公司测序，获得野生型ACCase基因序列，该基因片段长度为6984bp，禾广丝苗野生型ACCase序列全长基因片段的测序结果如下：
Figure BDA0002789305650000081
Figure BDA0002789305650000091
Figure BDA0002789305650000101
Figure BDA0002789305650000111
实施例3：水稻抗乙酰辅酶A羧化酶类除草剂突变体HGJK-3突变位点分析
将获得的抗除草剂水稻突变植株HGJK-3选取叶片并提取基因组DNA，分别将野生型水稻禾广丝苗和水稻突变植株HGJK-3的基因组DNA作为模板进行PCR扩增，扩增体系和扩增条件同实施例2，扩增结果如图3。同时送南京一道生物科技有限公司进行测序。测序结果与禾广丝苗野生型ACCase基因比较，发现在ACCase基因上发生了1个位点的突变，第6029位点碱基发生突变，由G变成T，导致其相应编码的氨基酸序列的第2010位点由色氨酸变为亮氨酸，即抗除草剂突变体的ACCase基因的核苷酸序列如SEQ ID NO.1所示，其编码的ACCase蛋白的氨基酸序列如SEQ ID NO.2所示。此水稻突变体其分类命名为水稻种子HGJK-3(Oryza sativa Indica Group HGJK-3)，该菌株已于2020年9月16日保藏于中国典型培养物保藏中心(CCTCC)，地址：湖北省武汉市武昌区武汉大学保藏中心(武汉大学第一附属小学对面)，邮编：430072，保藏编号为CCTCC No：P202012。
实施例4：水稻抗乙酰辅酶A羧化酶类除草剂突变体HGJK-4突变位点分析
将上述抗乙酰辅酶A羧化酶类突变体HGJK-3进行二次诱变，用0.5-1.0％(w/w)甲磺酸乙酯(EMS)室温下浸泡6-9小时，期间每1小时摇动一次种子；弃去EMS溶液，自来水翻动浸泡种子5次，每次5分钟，然后用自来水冲洗种子过夜，次日进行田间播种，并进行常规肥水管理(此为M1)。植株成熟后，种子混收、晾干，过冬保存。次年播种田间。待水稻(此为M2)幼苗长至3-4叶期时，喷施1mL精喹禾灵/L水，30天后还呈正常绿色植株即为抗乙酰辅酶A羧化酶类除草剂的水稻突变体HGJK-4(图2)。将HGJK-4抗除草剂突变体收获的种子播种出苗(此为M3)，待M3水稻幼苗长至3-4叶期时，喷施1mL精喹禾灵/L水(4倍推荐浓度)。喷施除草剂15天后，抗性苗HGJK-4的存活率要比HGJK-3高2.8％。同时，我们对突变体HGJK-4对盖草能的敏感性进行实验，发现HGJK-4的存活率要比HGJK-3突变体高2.7％。
通过提取HGJK-4的基因组DNA，分别将野生型水稻禾广丝苗和水稻突变植株HGJK-3的基因组DNA以及HGJK-4的基因组DNA作为模板进行PCR扩增，扩增体系和扩增条件同实施例2，扩增结果如图3，同时送南京一道生物科技有限公司进行测序，测序结果与HGJK-3的ACCase基因比较，发现除了在第6029位点碱基发生突变由G变成T之外，在第6295位点碱基由T变为C，导致其相应编码的氨基酸序列第2010位点由色氨酸变为亮氨酸和第2099位点由半胱氨酸变为精氨酸，即抗除草剂突变体的ACCase基因的核苷酸序列如SEQ ID NO.3所示，其编码的ACCase蛋白的氨基酸序列如SEQ ID NO.4所示。此水稻突变体其分类命名为水稻种子HGJK-04(Oryza sativa Indica Group HGJK-04)，该菌株已于2020年11月6日保藏于中国典型培养物保藏中心(CCTCC)，地址：湖北省武汉市武昌区武汉大学保藏中心(武汉大学第一附属小学对面)，邮编：430072，保藏编号为CCTCC NO：P202013。
表2除草剂对野生型和HGJK-3突变体和HGJK-4突变体的伤害比率
Figure BDA0002789305650000131
实施例5：转基因ACCase水稻抗精喹禾灵除草剂
设计特异引物5’-CCCGGGATGACATCCACACATGTGGCGACAT-3’和5’-GGGTACCCTCAACCAAGGACCTTCCTGATTTCT-3’，其5’分别加入SmaI和KpnI酶切修饰位点。通过PCR从上述水稻突变体禾广丝苗抗除草剂突变体的基因组DNA中扩增出突变ACCase基因(SEG ID NO：1)，此外，通过定向突变的方法获得的ACCase基因(SEG ID NO：3)，用SmaI和KpnI分别双酶切突变ACCase基因片段和植物表达载体pCAMBIA1301质粒(购自pcambia公司)，酶切产物用T4-DNA酶(购自TaKaRa公司)连接，连接产物转化大肠杆菌。重组质粒提取DNA，用SmaI和KpnI双酶切验证，可以产生大的质粒片段和小的基因片段，将基因片段进行电泳验证，结果如图4，证明已分别将核苷酸序列如SEQ ID NO.1所示和SEQ ID NO.3所示的ACCase基因克隆至植物表达载体pCAMBIA1301质粒(购自pcambia公司)中。将构建好的质粒载体转化农杆菌EHA105，培养菌体。采用常规的农杆菌介导法转化粳型水稻日本晴(购自江苏省农业种质资源保护与利用平台)，获得转基因植株收种后，后代植株长至3-4叶期时，PCR检测转基因植株(图4)。PCR检测引物为正向引物35SF 5’-ATGGTTAGAGAGGCTTACGC-3’，反向引物5R 5’-AGCAACAGGTCAGCCTTATCCAC-3’，扩增片段包括CaMV35S启动子和ACCase基因的5’端序列，大小约2kb。PCR扩增反应程序采用两步法，退火和延伸合为一起，采用68℃。扩增程序如下：预变性：98℃3min；30个循环：变性98℃10sec；延伸68℃2min；保温：72℃10min。经PCR鉴定呈阳性后，对转基因水稻和野生型水稻幼苗喷施1ml精喹禾灵/L水(4倍推荐使用浓度)，30天后发现转基因水稻生长状态良好，而非转基因日本晴水稻则全部枯死(表3)。
表3乙酰辅酶A羧化酶类除草剂对野生型和转基因水稻的伤害比率
Figure BDA0002789305650000141
序列表
<110> 江苏省农业科学院 江苏中旗作物保护科技有限公司
<120> 水稻ACCase突变型基因及其蛋白在植物抗除草剂中的应用
<160> 11
<170> SIPOSequenceListing 1.0
<210> 1
<211> 6984
<212> DNA
<213> 水稻禾广丝苗ACCase-6029突变核苷酸序列(Oryza sativa)
<400> 1
atgacatcca cacatgtggc gacattggga gttggtgccc aggcacctcc tcgtcaccag 60
aaaaagtcag ctggcactgc atttgtatca tctgggtcat caagaccctc ataccgaaag 120
aatggtcagc gtactcggtc acttagggaa gaaagcaatg gaggagtgtc tgattccaaa 180
aagcttaacc actctattcg ccaaggtctt gctggcatca ttgacctccc aaatgacgca 240
gcttcagaag ttgatatttc acatggttcc gaagatccca gggggcctac ggtcccaggt 300
tcctaccaaa tgaatgggat tatcaatgaa acacataatg ggaggcatgc ttcagtctcc 360
aaggttgttg agttttgtac ggcacttggt ggcaaaacac caattcacag tgtattagtg 420
gccaacaatg gaatggcagc agctaagttc atgcggagtg tccgaacatg ggctaatgat 480
acttttggat cagagaaggc aattcagctg atagctatgg caactccgga ggatctgagg 540
ataaatgcag agcacatcag aattgccgat caatttgtag aggtacctgg tggaacaaac 600
aacaacaact atgcaaatgt ccaactcata gtggagatag cagagagaac aggtgtttct 660
gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720
gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780
gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840
catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900
gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960
atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020
gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080
atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140
aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200
gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260
cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320
atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380
gtcactgagt ggatagctga agtaaatttg cctgcggctc aagttgctgt tggaatgggt 1440
ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500
tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560
tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620
gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680
tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740
ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800
ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860
ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920
gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980
tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040
ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100
gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160
aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220
ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280
attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340
acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400
ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460
gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520
ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580
caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640
aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700
ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760
gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820
tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880
ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940
gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000
tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060
attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120
gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180
atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240
tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300
acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360
actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420
gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480
agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540
catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600
gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660
tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720
ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780
ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840
aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900
ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960
tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020
tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080
ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140
ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200
tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260
accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320
actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380
ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440
ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500
gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560
aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620
gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680
catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740
tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800
gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860
tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920
ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980
aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040
acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100
ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160
ggcatagcag atgaagtgaa atcttgcttc cgtgttgggt ggtctgatga tggcagccct 5220
gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280
gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340
gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400
gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460
ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520
attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580
cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640
tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700
attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760
attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820
aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880
aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940
gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000
caatctgttc ctcgtgctgg acaagtgttg tttccagatt ctgcaaccaa gactgcgcag 6060
gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120
ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180
gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240
cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagtgctat 6300
gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360
aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420
gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480
atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540
gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600
tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660
cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720
ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780
gttgcttgga tggataaccc tgaaaactac aaggattata ttcaatatct taaggctcaa 6840
agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900
cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960
gaaatcagga aggtccttgg ttga 6984
<210> 2
<211> 2327
<212> PRT
<213> 水稻禾广丝苗ACCase-2010突变氨基酸序列(Oryza sativa)
<400> 2
Met Thr Ser Thr His Val Ala Thr Leu Gly Val Gly Ala Gln Ala Pro
1 5 10 15
Pro Arg His Gln Lys Lys Ser Ala Gly Thr Ala Phe Val Ser Ser Gly
20 25 30
Ser Ser Arg Pro Ser Tyr Arg Lys Asn Gly Gln Arg Thr Arg Ser Leu
35 40 45
Arg Glu Glu Ser Asn Gly Gly Val Ser Asp Ser Lys Lys Leu Asn His
50 55 60
Ser Ile Arg Gln Gly Leu Ala Gly Ile Ile Asp Leu Pro Asn Asp Ala
65 70 75 80
Ala Ser Glu Val Asp Ile Ser His Gly Ser Glu Asp Pro Arg Gly Pro
85 90 95
Thr Val Pro Gly Ser Tyr Gln Met Asn Gly Ile Ile Asn Glu Thr His
100 105 110
Asn Gly Arg His Ala Ser Val Ser Lys Val Val Glu Phe Cys Thr Ala
115 120 125
Leu Gly Gly Lys Thr Pro Ile His Ser Val Leu Val Ala Asn Asn Gly
130 135 140
Met Ala Ala Ala Lys Phe Met Arg Ser Val Arg Thr Trp Ala Asn Asp
145 150 155 160
Thr Phe Gly Ser Glu Lys Ala Ile Gln Leu Ile Ala Met Ala Thr Pro
165 170 175
Glu Asp Leu Arg Ile Asn Ala Glu His Ile Arg Ile Ala Asp Gln Phe
180 185 190
Val Glu Val Pro Gly Gly Thr Asn Asn Asn Asn Tyr Ala Asn Val Gln
195 200 205
Leu Ile Val Glu Ile Ala Glu Arg Thr Gly Val Ser Ala Val Trp Pro
210 215 220
Gly Trp Gly His Ala Ser Glu Asn Pro Glu Leu Pro Asp Ala Leu Thr
225 230 235 240
Ala Lys Gly Ile Val Phe Leu Gly Pro Pro Ala Ser Ser Met His Ala
245 250 255
Leu Gly Asp Lys Val Gly Ser Ala Leu Ile Ala Gln Ala Ala Gly Val
260 265 270
Pro Thr Leu Ala Trp Ser Gly Ser His Val Glu Val Pro Leu Glu Cys
275 280 285
Cys Leu Asp Ser Ile Pro Asp Glu Met Tyr Arg Lys Ala Cys Val Thr
290 295 300
Thr Thr Glu Glu Ala Val Ala Ser Cys Gln Val Val Gly Tyr Pro Ala
305 310 315 320
Met Ile Lys Ala Ser Trp Gly Gly Gly Gly Lys Gly Ile Arg Lys Val
325 330 335
His Asn Asp Asp Glu Val Arg Thr Leu Phe Lys Gln Val Gln Gly Glu
340 345 350
Val Pro Gly Ser Pro Ile Phe Ile Met Arg Leu Ala Ala Gln Ser Arg
355 360 365
His Leu Glu Val Gln Leu Leu Cys Asp Gln Tyr Gly Asn Val Ala Ala
370 375 380
Leu His Ser Arg Asp Cys Ser Val Gln Arg Arg His Gln Lys Ile Ile
385 390 395 400
Glu Glu Gly Pro Val Thr Val Ala Pro Arg Glu Thr Val Lys Glu Leu
405 410 415
Glu Gln Ala Ala Arg Arg Leu Ala Lys Ala Val Gly Tyr Val Gly Ala
420 425 430
Ala Thr Val Glu Tyr Leu Tyr Ser Met Glu Thr Gly Glu Tyr Tyr Phe
435 440 445
Leu Glu Leu Asn Pro Arg Leu Gln Val Glu His Pro Val Thr Glu Trp
450 455 460
Ile Ala Glu Val Asn Leu Pro Ala Ala Gln Val Ala Val Gly Met Gly
465 470 475 480
Ile Pro Leu Trp Gln Ile Pro Glu Ile Arg Arg Phe Tyr Gly Met Asn
485 490 495
His Gly Gly Gly Tyr Asp Leu Trp Arg Lys Thr Ala Ala Leu Ala Thr
500 505 510
Pro Phe Asn Phe Asp Glu Val Asp Ser Lys Trp Pro Lys Gly His Cys
515 520 525
Val Ala Val Arg Ile Thr Ser Glu Asp Pro Asp Asp Gly Phe Lys Pro
530 535 540
Thr Gly Gly Lys Val Lys Glu Ile Ser Phe Lys Ser Lys Pro Asn Val
545 550 555 560
Trp Ala Tyr Phe Ser Val Lys Ser Gly Gly Gly Ile His Glu Phe Ala
565 570 575
Asp Ser Gln Phe Gly His Val Phe Ala Tyr Gly Thr Thr Arg Ser Ala
580 585 590
Ala Ile Thr Thr Met Ala Leu Ala Leu Lys Glu Val Gln Ile Arg Gly
595 600 605
Glu Ile His Ser Asn Val Asp Tyr Thr Val Asp Leu Leu Asn Ala Ser
610 615 620
Asp Phe Arg Glu Asn Lys Ile His Thr Gly Trp Leu Asp Thr Arg Ile
625 630 635 640
Ala Met Arg Val Gln Ala Glu Arg Pro Pro Trp Tyr Ile Ser Val Val
645 650 655
Gly Gly Ala Leu Tyr Lys Thr Val Thr Ala Asn Thr Ala Thr Val Ser
660 665 670
Asp Tyr Val Gly Tyr Leu Thr Lys Gly Gln Ile Pro Pro Lys His Ile
675 680 685
Ser Leu Val Tyr Thr Thr Val Ala Leu Asn Ile Asp Gly Lys Lys Tyr
690 695 700
Thr Ile Asp Thr Val Arg Ser Gly His Gly Ser Tyr Arg Leu Arg Met
705 710 715 720
Asn Gly Ser Thr Val Asp Ala Asn Val Gln Ile Leu Cys Asp Gly Gly
725 730 735
Leu Leu Met Gln Leu Asp Gly Asn Ser His Val Ile Tyr Ala Glu Glu
740 745 750
Glu Ala Ser Gly Thr Arg Leu Leu Ile Asp Gly Lys Thr Cys Met Leu
755 760 765
Gln Asn Asp His Asp Pro Ser Lys Leu Leu Ala Glu Thr Pro Cys Lys
770 775 780
Leu Leu Arg Phe Leu Val Ala Asp Gly Ala His Val Asp Ala Asp Val
785 790 795 800
Pro Tyr Ala Glu Val Glu Val Met Lys Met Cys Met Pro Leu Leu Ser
805 810 815
Pro Ala Ser Gly Val Ile His Val Val Met Ser Glu Gly Gln Ala Met
820 825 830
Gln Ala Gly Asp Leu Ile Ala Arg Leu Asp Leu Asp Asp Pro Ser Ala
835 840 845
Val Lys Arg Ala Glu Pro Phe Glu Asp Thr Phe Pro Gln Met Gly Leu
850 855 860
Pro Ile Ala Ala Ser Gly Gln Val His Lys Leu Cys Ala Ala Ser Leu
865 870 875 880
Asn Ala Cys Arg Met Ile Leu Ala Gly Tyr Glu His Asp Ile Asp Lys
885 890 895
Val Val Pro Glu Leu Val Tyr Cys Leu Asp Thr Pro Glu Leu Pro Phe
900 905 910
Leu Gln Trp Glu Glu Leu Met Ser Val Leu Ala Thr Arg Leu Pro Arg
915 920 925
Asn Leu Lys Ser Glu Leu Glu Gly Lys Tyr Glu Glu Tyr Lys Val Lys
930 935 940
Phe Asp Ser Gly Ile Ile Asn Asp Phe Pro Ala Asn Met Leu Arg Val
945 950 955 960
Ile Ile Glu Glu Asn Leu Ala Cys Gly Ser Glu Lys Glu Lys Ala Thr
965 970 975
Asn Glu Arg Leu Val Glu Pro Leu Met Ser Leu Leu Lys Ser Tyr Glu
980 985 990
Gly Gly Arg Glu Ser His Ala His Phe Val Val Lys Ser Leu Phe Glu
995 1000 1005
Glu Tyr Leu Tyr Val Glu Glu Leu Phe Ser Asp Gly Ile Gln Ser Asp
1010 1015 1020
Val Ile Glu Arg Leu Arg Leu Gln His Ser Lys Asp Leu Gln Lys Val
1025 1030 1035 1040
Val Asp Ile Val Leu Ser His Gln Ser Val Arg Asn Lys Thr Lys Leu
1045 1050 1055
Ile Leu Lys Leu Met Glu Ser Leu Val Tyr Pro Asn Pro Ala Ala Tyr
1060 1065 1070
Arg Asp Gln Leu Ile Arg Phe Ser Ser Leu Asn His Lys Ala Tyr Tyr
1075 1080 1085
Lys Leu Ala Leu Lys Ala Ser Glu Leu Leu Glu Gln Thr Lys Leu Ser
1090 1095 1100
Glu Leu Arg Ala Arg Ile Ala Arg Ser Leu Ser Glu Leu Glu Met Phe
1105 1110 1115 1120
Thr Glu Glu Ser Lys Gly Leu Ser Met His Lys Arg Glu Ile Ala Ile
1125 1130 1135
Lys Glu Ser Met Glu Asp Leu Val Thr Ala Pro Leu Pro Val Glu Asp
1140 1145 1150
Ala Leu Ile Ser Leu Phe Asp Cys Ser Asp Thr Thr Val Gln Gln Arg
1155 1160 1165
Val Ile Glu Thr Tyr Ile Ala Arg Leu Tyr Gln Pro His Leu Val Lys
1170 1175 1180
Asp Ser Ile Lys Met Lys Trp Ile Glu Ser Gly Val Ile Ala Leu Trp
1185 1190 1195 1200
Glu Phe Pro Glu Gly His Phe Asp Ala Arg Asn Gly Gly Ala Val Leu
1205 1210 1215
Gly Asp Lys Arg Trp Gly Ala Met Val Ile Val Lys Ser Leu Glu Ser
1220 1225 1230
Leu Ser Met Ala Ile Arg Phe Ala Leu Lys Glu Thr Ser His Tyr Thr
1235 1240 1245
Ser Ser Glu Gly Asn Met Met His Ile Ala Leu Leu Gly Ala Asp Asn
1250 1255 1260
Lys Met His Ile Ile Gln Glu Ser Gly Asp Asp Ala Asp Arg Ile Ala
1265 1270 1275 1280
Lys Leu Pro Leu Ile Leu Lys Asp Asn Val Thr Asp Leu His Ala Ser
1285 1290 1295
Gly Val Lys Thr Ile Ser Phe Ile Val Gln Arg Asp Glu Ala Arg Met
1300 1305 1310
Thr Met Arg Arg Thr Phe Leu Trp Ser Asp Glu Lys Leu Ser Tyr Glu
1315 1320 1325
Glu Glu Pro Ile Leu Arg His Val Glu Pro Pro Leu Ser Ala Leu Leu
1330 1335 1340
Glu Leu Asp Lys Leu Lys Val Lys Gly Tyr Asn Glu Met Lys Tyr Thr
1345 1350 1355 1360
Pro Ser Arg Asp Arg Gln Trp His Ile Tyr Thr Leu Arg Asn Thr Glu
1365 1370 1375
Asn Pro Lys Met Leu His Arg Val Phe Phe Arg Thr Leu Val Arg Gln
1380 1385 1390
Pro Ser Val Ser Asn Lys Phe Ser Ser Gly Gln Ile Gly Asp Met Glu
1395 1400 1405
Val Gly Ser Ala Glu Glu Pro Leu Ser Phe Thr Ser Thr Ser Ile Leu
1410 1415 1420
Arg Ser Leu Met Thr Ala Ile Glu Glu Leu Glu Leu His Ala Ile Arg
1425 1430 1435 1440
Thr Gly His Ser His Met Tyr Leu His Val Leu Lys Glu Gln Lys Leu
1445 1450 1455
Leu Asp Leu Val Pro Val Ser Gly Asn Thr Val Leu Asp Val Gly Gln
1460 1465 1470
Asp Glu Ala Thr Ala Tyr Ser Leu Leu Lys Glu Met Ala Met Lys Ile
1475 1480 1485
His Glu Leu Val Gly Ala Arg Met His His Leu Ser Val Cys Gln Trp
1490 1495 1500
Glu Val Lys Leu Lys Leu Asp Cys Asp Gly Pro Ala Ser Gly Thr Trp
1505 1510 1515 1520
Arg Ile Val Thr Thr Asn Val Thr Ser His Thr Cys Thr Val Asp Ile
1525 1530 1535
Tyr Arg Glu Met Glu Asp Lys Glu Ser Arg Lys Leu Val Tyr His Pro
1540 1545 1550
Ala Thr Pro Ala Ala Gly Pro Leu His Gly Val Ala Leu Asn Asn Pro
1555 1560 1565
Tyr Gln Pro Leu Ser Val Ile Asp Leu Lys Arg Cys Ser Ala Arg Asn
1570 1575 1580
Asn Arg Thr Thr Tyr Cys Tyr Asp Phe Pro Leu Ala Phe Glu Thr Ala
1585 1590 1595 1600
Val Arg Lys Ser Trp Ser Ser Ser Thr Ser Gly Ala Ser Lys Gly Val
1605 1610 1615
Glu Asn Ala Gln Cys Tyr Val Lys Ala Thr Glu Leu Val Phe Ala Asp
1620 1625 1630
Lys His Gly Ser Trp Gly Thr Pro Leu Val Gln Met Asp Arg Pro Ala
1635 1640 1645
Gly Leu Asn Asp Ile Gly Met Val Ala Trp Thr Leu Lys Met Ser Thr
1650 1655 1660
Pro Glu Phe Pro Ser Gly Arg Glu Ile Ile Val Val Ala Asn Asp Ile
1665 1670 1675 1680
Thr Phe Arg Ala Gly Ser Phe Gly Pro Arg Glu Asp Ala Phe Phe Glu
1685 1690 1695
Ala Val Thr Asn Leu Ala Cys Glu Lys Lys Leu Pro Leu Ile Tyr Leu
1700 1705 1710
Ala Ala Asn Ser Gly Ala Arg Ile Gly Ile Ala Asp Glu Val Lys Ser
1715 1720 1725
Cys Phe Arg Val Gly Trp Ser Asp Asp Gly Ser Pro Glu Arg Gly Phe
1730 1735 1740
Gln Tyr Ile Tyr Leu Ser Glu Glu Asp Tyr Ala Arg Ile Gly Thr Ser
1745 1750 1755 1760
Val Ile Ala His Lys Met Gln Leu Asp Ser Gly Glu Ile Arg Trp Val
1765 1770 1775
Ile Asp Ser Val Val Gly Lys Glu Asp Gly Leu Gly Val Glu Asn Ile
1780 1785 1790
His Gly Ser Ala Ala Ile Ala Ser Ala Tyr Ser Arg Ala Tyr Lys Glu
1795 1800 1805
Thr Phe Thr Leu Thr Phe Val Thr Gly Arg Thr Val Gly Ile Gly Ala
1810 1815 1820
Tyr Leu Ala Arg Leu Gly Ile Arg Cys Ile Gln Arg Leu Asp Gln Pro
1825 1830 1835 1840
Ile Ile Leu Thr Gly Tyr Ser Ala Leu Asn Lys Leu Leu Gly Arg Glu
1845 1850 1855
Val Tyr Ser Ser His Met Gln Leu Gly Gly Pro Lys Ile Met Ala Thr
1860 1865 1870
Asn Gly Val Val His Leu Thr Val Ser Asp Asp Leu Glu Gly Val Ser
1875 1880 1885
Asn Ile Leu Arg Trp Leu Ser Tyr Val Pro Ala Tyr Ile Gly Gly Pro
1890 1895 1900
Leu Pro Val Thr Thr Pro Leu Asp Pro Pro Asp Arg Pro Val Ala Tyr
1905 1910 1915 1920
Ile Pro Glu Asn Ser Cys Asp Pro Arg Ala Ala Ile Arg Gly Val Asp
1925 1930 1935
Asp Ser Gln Gly Lys Trp Leu Gly Gly Met Phe Asp Lys Asp Ser Phe
1940 1945 1950
Val Glu Thr Phe Glu Gly Trp Ala Lys Thr Val Val Thr Gly Arg Ala
1955 1960 1965
Lys Leu Gly Gly Ile Pro Val Gly Val Ile Ala Val Glu Thr Gln Thr
1970 1975 1980
Met Met Gln Thr Ile Pro Ala Asp Pro Gly Gln Leu Asp Ser Arg Glu
1985 1990 1995 2000
Gln Ser Val Pro Arg Ala Gly Gln Val Leu Phe Pro Asp Ser Ala Thr
2005 2010 2015
Lys Thr Ala Gln Ala Leu Leu Asp Phe Asn Arg Glu Gly Leu Pro Leu
2020 2025 2030
Phe Ile Leu Ala Asn Trp Arg Gly Phe Ser Gly Gly Gln Arg Asp Leu
2035 2040 2045
Phe Glu Gly Ile Leu Gln Ala Gly Ser Thr Ile Val Glu Asn Leu Arg
2050 2055 2060
Thr Tyr Asn Gln Pro Ala Phe Val Tyr Ile Pro Met Ala Ala Glu Leu
2065 2070 2075 2080
Arg Gly Gly Ala Trp Val Val Val Asp Ser Lys Ile Asn Pro Asp Arg
2085 2090 2095
Ile Glu Cys Tyr Ala Glu Arg Thr Ala Lys Gly Asn Val Leu Glu Pro
2100 2105 2110
Gln Gly Leu Ile Glu Ile Lys Phe Arg Ser Glu Glu Leu Gln Asp Cys
2115 2120 2125
Met Ser Arg Leu Asp Pro Thr Leu Ile Asp Leu Lys Ala Lys Leu Glu
2130 2135 2140
Val Ala Asn Lys Asn Gly Ser Ala Asp Thr Lys Ser Leu Gln Glu Asn
2145 2150 2155 2160
Ile Glu Ala Arg Thr Lys Gln Leu Met Pro Leu Tyr Thr Gln Ile Ala
2165 2170 2175
Ile Arg Phe Ala Glu Leu His Asp Thr Ser Leu Arg Met Ala Ala Lys
2180 2185 2190
Gly Val Ile Lys Lys Val Val Asp Trp Glu Glu Ser Arg Ser Phe Phe
2195 2200 2205
Tyr Lys Arg Leu Arg Arg Arg Ile Ser Glu Asp Val Leu Ala Lys Glu
2210 2215 2220
Ile Arg Ala Val Ala Gly Glu Gln Phe Ser His Gln Pro Ala Ile Glu
2225 2230 2235 2240
Leu Ile Lys Lys Trp Tyr Ser Ala Ser His Ala Ala Glu Trp Asp Asp
2245 2250 2255
Asp Asp Ala Phe Val Ala Trp Met Asp Asn Pro Glu Asn Tyr Lys Asp
2260 2265 2270
Tyr Ile Gln Tyr Leu Lys Ala Gln Arg Val Ser Gln Ser Leu Ser Ser
2275 2280 2285
Leu Ser Asp Ser Ser Ser Asp Leu Gln Ala Leu Pro Gln Gly Leu Ser
2290 2295 2300
Met Leu Leu Asp Lys Met Asp Pro Ser Arg Arg Ala Gln Leu Val Glu
2305 2310 2315 2320
Glu Ile Arg Lys Val Leu Gly
2325
<210> 3
<211> 6984
<212> DNA
<213> 水稻禾广丝苗ACCase-6029、6295突变核苷酸序列 (Oryza sativa)
<400> 3
atgacatcca cacatgtggc gacattggga gttggtgccc aggcacctcc tcgtcaccag 60
aaaaagtcag ctggcactgc atttgtatca tctgggtcat caagaccctc ataccgaaag 120
aatggtcagc gtactcggtc acttagggaa gaaagcaatg gaggagtgtc tgattccaaa 180
aagcttaacc actctattcg ccaaggtctt gctggcatca ttgacctccc aaatgacgca 240
gcttcagaag ttgatatttc acatggttcc gaagatccca gggggcctac ggtcccaggt 300
tcctaccaaa tgaatgggat tatcaatgaa acacataatg ggaggcatgc ttcagtctcc 360
aaggttgttg agttttgtac ggcacttggt ggcaaaacac caattcacag tgtattagtg 420
gccaacaatg gaatggcagc agctaagttc atgcggagtg tccgaacatg ggctaatgat 480
acttttggat cagagaaggc aattcagctg atagctatgg caactccgga ggatctgagg 540
ataaatgcag agcacatcag aattgccgat caatttgtag aggtacctgg tggaacaaac 600
aacaacaact atgcaaatgt ccaactcata gtggagatag cagagagaac aggtgtttct 660
gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720
gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780
gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840
catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900
gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960
atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020
gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080
atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140
aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200
gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260
cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320
atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380
gtcactgagt ggatagctga agtaaatttg cctgcggctc aagttgctgt tggaatgggt 1440
ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500
tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560
tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620
gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680
tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740
ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800
ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860
ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920
gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980
tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040
ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100
gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160
aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220
ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280
attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340
acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400
ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460
gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520
ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580
caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640
aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700
ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760
gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820
tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880
ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940
gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000
tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060
attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120
gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180
atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240
tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300
acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360
actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420
gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480
agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540
catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600
gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660
tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720
ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780
ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840
aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900
ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960
tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020
tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080
ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140
ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200
tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260
accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320
actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380
ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440
ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500
gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560
aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620
gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680
catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740
tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800
gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860
tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920
ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980
aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040
acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100
ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160
ggcatagcag atgaagtgaa atcttgcttc cgtgttgggt ggtctgatga tggcagccct 5220
gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280
gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340
gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400
gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460
ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520
attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580
cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640
tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700
attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760
attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820
aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880
aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940
gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000
caatctgttc ctcgtgctgg acaagtgttg tttccagatt ctgcaaccaa gactgcgcag 6060
gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120
ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180
gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240
cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagcgctat 6300
gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360
aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420
gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480
atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540
gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600
tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660
cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720
ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780
gttgcttgga tggataaccc tgaaaactac aaggattata ttcaatatct taaggctcaa 6840
agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900
cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960
gaaatcagga aggtccttgg ttga 6984
<210> 4
<211> 2327
<212> PRT
<213> 水稻禾广丝苗ACCase-2010、2099氨基酸突变(Oryza sativa)
<400> 4
Met Thr Ser Thr His Val Ala Thr Leu Gly Val Gly Ala Gln Ala Pro
1 5 10 15
Pro Arg His Gln Lys Lys Ser Ala Gly Thr Ala Phe Val Ser Ser Gly
20 25 30
Ser Ser Arg Pro Ser Tyr Arg Lys Asn Gly Gln Arg Thr Arg Ser Leu
35 40 45
Arg Glu Glu Ser Asn Gly Gly Val Ser Asp Ser Lys Lys Leu Asn His
50 55 60
Ser Ile Arg Gln Gly Leu Ala Gly Ile Ile Asp Leu Pro Asn Asp Ala
65 70 75 80
Ala Ser Glu Val Asp Ile Ser His Gly Ser Glu Asp Pro Arg Gly Pro
85 90 95
Thr Val Pro Gly Ser Tyr Gln Met Asn Gly Ile Ile Asn Glu Thr His
100 105 110
Asn Gly Arg His Ala Ser Val Ser Lys Val Val Glu Phe Cys Thr Ala
115 120 125
Leu Gly Gly Lys Thr Pro Ile His Ser Val Leu Val Ala Asn Asn Gly
130 135 140
Met Ala Ala Ala Lys Phe Met Arg Ser Val Arg Thr Trp Ala Asn Asp
145 150 155 160
Thr Phe Gly Ser Glu Lys Ala Ile Gln Leu Ile Ala Met Ala Thr Pro
165 170 175
Glu Asp Leu Arg Ile Asn Ala Glu His Ile Arg Ile Ala Asp Gln Phe
180 185 190
Val Glu Val Pro Gly Gly Thr Asn Asn Asn Asn Tyr Ala Asn Val Gln
195 200 205
Leu Ile Val Glu Ile Ala Glu Arg Thr Gly Val Ser Ala Val Trp Pro
210 215 220
Gly Trp Gly His Ala Ser Glu Asn Pro Glu Leu Pro Asp Ala Leu Thr
225 230 235 240
Ala Lys Gly Ile Val Phe Leu Gly Pro Pro Ala Ser Ser Met His Ala
245 250 255
Leu Gly Asp Lys Val Gly Ser Ala Leu Ile Ala Gln Ala Ala Gly Val
260 265 270
Pro Thr Leu Ala Trp Ser Gly Ser His Val Glu Val Pro Leu Glu Cys
275 280 285
Cys Leu Asp Ser Ile Pro Asp Glu Met Tyr Arg Lys Ala Cys Val Thr
290 295 300
Thr Thr Glu Glu Ala Val Ala Ser Cys Gln Val Val Gly Tyr Pro Ala
305 310 315 320
Met Ile Lys Ala Ser Trp Gly Gly Gly Gly Lys Gly Ile Arg Lys Val
325 330 335
His Asn Asp Asp Glu Val Arg Thr Leu Phe Lys Gln Val Gln Gly Glu
340 345 350
Val Pro Gly Ser Pro Ile Phe Ile Met Arg Leu Ala Ala Gln Ser Arg
355 360 365
His Leu Glu Val Gln Leu Leu Cys Asp Gln Tyr Gly Asn Val Ala Ala
370 375 380
Leu His Ser Arg Asp Cys Ser Val Gln Arg Arg His Gln Lys Ile Ile
385 390 395 400
Glu Glu Gly Pro Val Thr Val Ala Pro Arg Glu Thr Val Lys Glu Leu
405 410 415
Glu Gln Ala Ala Arg Arg Leu Ala Lys Ala Val Gly Tyr Val Gly Ala
420 425 430
Ala Thr Val Glu Tyr Leu Tyr Ser Met Glu Thr Gly Glu Tyr Tyr Phe
435 440 445
Leu Glu Leu Asn Pro Arg Leu Gln Val Glu His Pro Val Thr Glu Trp
450 455 460
Ile Ala Glu Val Asn Leu Pro Ala Ala Gln Val Ala Val Gly Met Gly
465 470 475 480
Ile Pro Leu Trp Gln Ile Pro Glu Ile Arg Arg Phe Tyr Gly Met Asn
485 490 495
His Gly Gly Gly Tyr Asp Leu Trp Arg Lys Thr Ala Ala Leu Ala Thr
500 505 510
Pro Phe Asn Phe Asp Glu Val Asp Ser Lys Trp Pro Lys Gly His Cys
515 520 525
Val Ala Val Arg Ile Thr Ser Glu Asp Pro Asp Asp Gly Phe Lys Pro
530 535 540
Thr Gly Gly Lys Val Lys Glu Ile Ser Phe Lys Ser Lys Pro Asn Val
545 550 555 560
Trp Ala Tyr Phe Ser Val Lys Ser Gly Gly Gly Ile His Glu Phe Ala
565 570 575
Asp Ser Gln Phe Gly His Val Phe Ala Tyr Gly Thr Thr Arg Ser Ala
580 585 590
Ala Ile Thr Thr Met Ala Leu Ala Leu Lys Glu Val Gln Ile Arg Gly
595 600 605
Glu Ile His Ser Asn Val Asp Tyr Thr Val Asp Leu Leu Asn Ala Ser
610 615 620
Asp Phe Arg Glu Asn Lys Ile His Thr Gly Trp Leu Asp Thr Arg Ile
625 630 635 640
Ala Met Arg Val Gln Ala Glu Arg Pro Pro Trp Tyr Ile Ser Val Val
645 650 655
Gly Gly Ala Leu Tyr Lys Thr Val Thr Ala Asn Thr Ala Thr Val Ser
660 665 670
Asp Tyr Val Gly Tyr Leu Thr Lys Gly Gln Ile Pro Pro Lys His Ile
675 680 685
Ser Leu Val Tyr Thr Thr Val Ala Leu Asn Ile Asp Gly Lys Lys Tyr
690 695 700
Thr Ile Asp Thr Val Arg Ser Gly His Gly Ser Tyr Arg Leu Arg Met
705 710 715 720
Asn Gly Ser Thr Val Asp Ala Asn Val Gln Ile Leu Cys Asp Gly Gly
725 730 735
Leu Leu Met Gln Leu Asp Gly Asn Ser His Val Ile Tyr Ala Glu Glu
740 745 750
Glu Ala Ser Gly Thr Arg Leu Leu Ile Asp Gly Lys Thr Cys Met Leu
755 760 765
Gln Asn Asp His Asp Pro Ser Lys Leu Leu Ala Glu Thr Pro Cys Lys
770 775 780
Leu Leu Arg Phe Leu Val Ala Asp Gly Ala His Val Asp Ala Asp Val
785 790 795 800
Pro Tyr Ala Glu Val Glu Val Met Lys Met Cys Met Pro Leu Leu Ser
805 810 815
Pro Ala Ser Gly Val Ile His Val Val Met Ser Glu Gly Gln Ala Met
820 825 830
Gln Ala Gly Asp Leu Ile Ala Arg Leu Asp Leu Asp Asp Pro Ser Ala
835 840 845
Val Lys Arg Ala Glu Pro Phe Glu Asp Thr Phe Pro Gln Met Gly Leu
850 855 860
Pro Ile Ala Ala Ser Gly Gln Val His Lys Leu Cys Ala Ala Ser Leu
865 870 875 880
Asn Ala Cys Arg Met Ile Leu Ala Gly Tyr Glu His Asp Ile Asp Lys
885 890 895
Val Val Pro Glu Leu Val Tyr Cys Leu Asp Thr Pro Glu Leu Pro Phe
900 905 910
Leu Gln Trp Glu Glu Leu Met Ser Val Leu Ala Thr Arg Leu Pro Arg
915 920 925
Asn Leu Lys Ser Glu Leu Glu Gly Lys Tyr Glu Glu Tyr Lys Val Lys
930 935 940
Phe Asp Ser Gly Ile Ile Asn Asp Phe Pro Ala Asn Met Leu Arg Val
945 950 955 960
Ile Ile Glu Glu Asn Leu Ala Cys Gly Ser Glu Lys Glu Lys Ala Thr
965 970 975
Asn Glu Arg Leu Val Glu Pro Leu Met Ser Leu Leu Lys Ser Tyr Glu
980 985 990
Gly Gly Arg Glu Ser His Ala His Phe Val Val Lys Ser Leu Phe Glu
995 1000 1005
Glu Tyr Leu Tyr Val Glu Glu Leu Phe Ser Asp Gly Ile Gln Ser Asp
1010 1015 1020
Val Ile Glu Arg Leu Arg Leu Gln His Ser Lys Asp Leu Gln Lys Val
1025 1030 1035 1040
Val Asp Ile Val Leu Ser His Gln Ser Val Arg Asn Lys Thr Lys Leu
1045 1050 1055
Ile Leu Lys Leu Met Glu Ser Leu Val Tyr Pro Asn Pro Ala Ala Tyr
1060 1065 1070
Arg Asp Gln Leu Ile Arg Phe Ser Ser Leu Asn His Lys Ala Tyr Tyr
1075 1080 1085
Lys Leu Ala Leu Lys Ala Ser Glu Leu Leu Glu Gln Thr Lys Leu Ser
1090 1095 1100
Glu Leu Arg Ala Arg Ile Ala Arg Ser Leu Ser Glu Leu Glu Met Phe
1105 1110 1115 1120
Thr Glu Glu Ser Lys Gly Leu Ser Met His Lys Arg Glu Ile Ala Ile
1125 1130 1135
Lys Glu Ser Met Glu Asp Leu Val Thr Ala Pro Leu Pro Val Glu Asp
1140 1145 1150
Ala Leu Ile Ser Leu Phe Asp Cys Ser Asp Thr Thr Val Gln Gln Arg
1155 1160 1165
Val Ile Glu Thr Tyr Ile Ala Arg Leu Tyr Gln Pro His Leu Val Lys
1170 1175 1180
Asp Ser Ile Lys Met Lys Trp Ile Glu Ser Gly Val Ile Ala Leu Trp
1185 1190 1195 1200
Glu Phe Pro Glu Gly His Phe Asp Ala Arg Asn Gly Gly Ala Val Leu
1205 1210 1215
Gly Asp Lys Arg Trp Gly Ala Met Val Ile Val Lys Ser Leu Glu Ser
1220 1225 1230
Leu Ser Met Ala Ile Arg Phe Ala Leu Lys Glu Thr Ser His Tyr Thr
1235 1240 1245
Ser Ser Glu Gly Asn Met Met His Ile Ala Leu Leu Gly Ala Asp Asn
1250 1255 1260
Lys Met His Ile Ile Gln Glu Ser Gly Asp Asp Ala Asp Arg Ile Ala
1265 1270 1275 1280
Lys Leu Pro Leu Ile Leu Lys Asp Asn Val Thr Asp Leu His Ala Ser
1285 1290 1295
Gly Val Lys Thr Ile Ser Phe Ile Val Gln Arg Asp Glu Ala Arg Met
1300 1305 1310
Thr Met Arg Arg Thr Phe Leu Trp Ser Asp Glu Lys Leu Ser Tyr Glu
1315 1320 1325
Glu Glu Pro Ile Leu Arg His Val Glu Pro Pro Leu Ser Ala Leu Leu
1330 1335 1340
Glu Leu Asp Lys Leu Lys Val Lys Gly Tyr Asn Glu Met Lys Tyr Thr
1345 1350 1355 1360
Pro Ser Arg Asp Arg Gln Trp His Ile Tyr Thr Leu Arg Asn Thr Glu
1365 1370 1375
Asn Pro Lys Met Leu His Arg Val Phe Phe Arg Thr Leu Val Arg Gln
1380 1385 1390
Pro Ser Val Ser Asn Lys Phe Ser Ser Gly Gln Ile Gly Asp Met Glu
1395 1400 1405
Val Gly Ser Ala Glu Glu Pro Leu Ser Phe Thr Ser Thr Ser Ile Leu
1410 1415 1420
Arg Ser Leu Met Thr Ala Ile Glu Glu Leu Glu Leu His Ala Ile Arg
1425 1430 1435 1440
Thr Gly His Ser His Met Tyr Leu His Val Leu Lys Glu Gln Lys Leu
1445 1450 1455
Leu Asp Leu Val Pro Val Ser Gly Asn Thr Val Leu Asp Val Gly Gln
1460 1465 1470
Asp Glu Ala Thr Ala Tyr Ser Leu Leu Lys Glu Met Ala Met Lys Ile
1475 1480 1485
His Glu Leu Val Gly Ala Arg Met His His Leu Ser Val Cys Gln Trp
1490 1495 1500
Glu Val Lys Leu Lys Leu Asp Cys Asp Gly Pro Ala Ser Gly Thr Trp
1505 1510 1515 1520
Arg Ile Val Thr Thr Asn Val Thr Ser His Thr Cys Thr Val Asp Ile
1525 1530 1535
Tyr Arg Glu Met Glu Asp Lys Glu Ser Arg Lys Leu Val Tyr His Pro
1540 1545 1550
Ala Thr Pro Ala Ala Gly Pro Leu His Gly Val Ala Leu Asn Asn Pro
1555 1560 1565
Tyr Gln Pro Leu Ser Val Ile Asp Leu Lys Arg Cys Ser Ala Arg Asn
1570 1575 1580
Asn Arg Thr Thr Tyr Cys Tyr Asp Phe Pro Leu Ala Phe Glu Thr Ala
1585 1590 1595 1600
Val Arg Lys Ser Trp Ser Ser Ser Thr Ser Gly Ala Ser Lys Gly Val
1605 1610 1615
Glu Asn Ala Gln Cys Tyr Val Lys Ala Thr Glu Leu Val Phe Ala Asp
1620 1625 1630
Lys His Gly Ser Trp Gly Thr Pro Leu Val Gln Met Asp Arg Pro Ala
1635 1640 1645
Gly Leu Asn Asp Ile Gly Met Val Ala Trp Thr Leu Lys Met Ser Thr
1650 1655 1660
Pro Glu Phe Pro Ser Gly Arg Glu Ile Ile Val Val Ala Asn Asp Ile
1665 1670 1675 1680
Thr Phe Arg Ala Gly Ser Phe Gly Pro Arg Glu Asp Ala Phe Phe Glu
1685 1690 1695
Ala Val Thr Asn Leu Ala Cys Glu Lys Lys Leu Pro Leu Ile Tyr Leu
1700 1705 1710
Ala Ala Asn Ser Gly Ala Arg Ile Gly Ile Ala Asp Glu Val Lys Ser
1715 1720 1725
Cys Phe Arg Val Gly Trp Ser Asp Asp Gly Ser Pro Glu Arg Gly Phe
1730 1735 1740
Gln Tyr Ile Tyr Leu Ser Glu Glu Asp Tyr Ala Arg Ile Gly Thr Ser
1745 1750 1755 1760
Val Ile Ala His Lys Met Gln Leu Asp Ser Gly Glu Ile Arg Trp Val
1765 1770 1775
Ile Asp Ser Val Val Gly Lys Glu Asp Gly Leu Gly Val Glu Asn Ile
1780 1785 1790
His Gly Ser Ala Ala Ile Ala Ser Ala Tyr Ser Arg Ala Tyr Lys Glu
1795 1800 1805
Thr Phe Thr Leu Thr Phe Val Thr Gly Arg Thr Val Gly Ile Gly Ala
1810 1815 1820
Tyr Leu Ala Arg Leu Gly Ile Arg Cys Ile Gln Arg Leu Asp Gln Pro
1825 1830 1835 1840
Ile Ile Leu Thr Gly Tyr Ser Ala Leu Asn Lys Leu Leu Gly Arg Glu
1845 1850 1855
Val Tyr Ser Ser His Met Gln Leu Gly Gly Pro Lys Ile Met Ala Thr
1860 1865 1870
Asn Gly Val Val His Leu Thr Val Ser Asp Asp Leu Glu Gly Val Ser
1875 1880 1885
Asn Ile Leu Arg Trp Leu Ser Tyr Val Pro Ala Tyr Ile Gly Gly Pro
1890 1895 1900
Leu Pro Val Thr Thr Pro Leu Asp Pro Pro Asp Arg Pro Val Ala Tyr
1905 1910 1915 1920
Ile Pro Glu Asn Ser Cys Asp Pro Arg Ala Ala Ile Arg Gly Val Asp
1925 1930 1935
Asp Ser Gln Gly Lys Trp Leu Gly Gly Met Phe Asp Lys Asp Ser Phe
1940 1945 1950
Val Glu Thr Phe Glu Gly Trp Ala Lys Thr Val Val Thr Gly Arg Ala
1955 1960 1965
Lys Leu Gly Gly Ile Pro Val Gly Val Ile Ala Val Glu Thr Gln Thr
1970 1975 1980
Met Met Gln Thr Ile Pro Ala Asp Pro Gly Gln Leu Asp Ser Arg Glu
1985 1990 1995 2000
Gln Ser Val Pro Arg Ala Gly Gln Val Leu Phe Pro Asp Ser Ala Thr
2005 2010 2015
Lys Thr Ala Gln Ala Leu Leu Asp Phe Asn Arg Glu Gly Leu Pro Leu
2020 2025 2030
Phe Ile Leu Ala Asn Trp Arg Gly Phe Ser Gly Gly Gln Arg Asp Leu
2035 2040 2045
Phe Glu Gly Ile Leu Gln Ala Gly Ser Thr Ile Val Glu Asn Leu Arg
2050 2055 2060
Thr Tyr Asn Gln Pro Ala Phe Val Tyr Ile Pro Met Ala Ala Glu Leu
2065 2070 2075 2080
Arg Gly Gly Ala Trp Val Val Val Asp Ser Lys Ile Asn Pro Asp Arg
2085 2090 2095
Ile Glu Arg Tyr Ala Glu Arg Thr Ala Lys Gly Asn Val Leu Glu Pro
2100 2105 2110
Gln Gly Leu Ile Glu Ile Lys Phe Arg Ser Glu Glu Leu Gln Asp Cys
2115 2120 2125
Met Ser Arg Leu Asp Pro Thr Leu Ile Asp Leu Lys Ala Lys Leu Glu
2130 2135 2140
Val Ala Asn Lys Asn Gly Ser Ala Asp Thr Lys Ser Leu Gln Glu Asn
2145 2150 2155 2160
Ile Glu Ala Arg Thr Lys Gln Leu Met Pro Leu Tyr Thr Gln Ile Ala
2165 2170 2175
Ile Arg Phe Ala Glu Leu His Asp Thr Ser Leu Arg Met Ala Ala Lys
2180 2185 2190
Gly Val Ile Lys Lys Val Val Asp Trp Glu Glu Ser Arg Ser Phe Phe
2195 2200 2205
Tyr Lys Arg Leu Arg Arg Arg Ile Ser Glu Asp Val Leu Ala Lys Glu
2210 2215 2220
Ile Arg Ala Val Ala Gly Glu Gln Phe Ser His Gln Pro Ala Ile Glu
2225 2230 2235 2240
Leu Ile Lys Lys Trp Tyr Ser Ala Ser His Ala Ala Glu Trp Asp Asp
2245 2250 2255
Asp Asp Ala Phe Val Ala Trp Met Asp Asn Pro Glu Asn Tyr Lys Asp
2260 2265 2270
Tyr Ile Gln Tyr Leu Lys Ala Gln Arg Val Ser Gln Ser Leu Ser Ser
2275 2280 2285
Leu Ser Asp Ser Ser Ser Asp Leu Gln Ala Leu Pro Gln Gly Leu Ser
2290 2295 2300
Met Leu Leu Asp Lys Met Asp Pro Ser Arg Arg Ala Gln Leu Val Glu
2305 2310 2315 2320
Glu Ile Arg Lys Val Leu Gly
2325
<210> 5
<211> 25
<212> DNA
<213> 特异引物(Artificial Sequence)
<400> 5
atgacatcca cacatgtggc gacat 25
<210> 6
<211> 25
<212> DNA
<213> 特异引物(Artificial Sequence)
<400> 6
tcaaccaagg accttcctga tttct 25
<210> 7
<211> 20
<212> DNA
<213> 正向引物35SF (Artificial Sequence)
<400> 7
atggttagag aggcttacgc 20
<210> 8
<211> 23
<212> DNA
<213> 反向引物5R(Artificial Sequence)
<400> 8
agcaacaggt cagccttatc cac 23
<210> 9
<211> 24
<212> DNA
<213> 正向引物(Artificial Sequence)
<400> 9
gtcagatttc acacatctgg ggat 24
<210> 10
<211> 24
<212> DNA
<213> 反向引物(Artificial Sequence)
<400> 10
acttgcactt tcatctggca gaac 24
<210> 11
<211> 6984
<212> DNA
<213> 禾广丝苗野生型ACCase序列全长基因(Oryza sativa)
<400> 11
atgacatcca cacatgtggc gacattggga gttggtgccc aggcacctcc tcgtcaccag 60
aaaaagtcag ctggcactgc atttgtatca tctgggtcat caagaccctc ataccgaaag 120
aatggtcagc gtactcggtc acttagggaa gaaagcaatg gaggagtgtc tgattccaaa 180
aagcttaacc actctattcg ccaaggtctt gctggcatca ttgacctccc aaatgacgca 240
gcttcagaag ttgatatttc acatggttcc gaagatccca gggggcctac ggtcccaggt 300
tcctaccaaa tgaatgggat tatcaatgaa acacataatg ggaggcatgc ttcagtctcc 360
aaggttgttg agttttgtac ggcacttggt ggcaaaacac caattcacag tgtattagtg 420
gccaacaatg gaatggcagc agctaagttc atgcggagtg tccgaacatg ggctaatgat 480
acttttggat cagagaaggc aattcagctg atagctatgg caactccgga ggatctgagg 540
ataaatgcag agcacatcag aattgccgat caatttgtag aggtacctgg tggaacaaac 600
aacaacaact atgcaaatgt ccaactcata gtggagatag cagagagaac aggtgtttct 660
gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720
gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780
gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840
catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900
gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960
atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020
gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080
atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140
aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200
gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260
cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320
atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380
gtcactgagt ggatagctga agtaaatttg cctgcggctc aagttgctgt tggaatgggt 1440
ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500
tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560
tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620
gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680
tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740
ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800
ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860
ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920
gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980
tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040
ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100
gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160
aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220
ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280
attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340
acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400
ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460
gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520
ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580
caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640
aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700
ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760
gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820
tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880
ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940
gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000
tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060
attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120
gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180
atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240
tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300
acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360
actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420
gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480
agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540
catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600
gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660
tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720
ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780
ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840
aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900
ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960
tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020
tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080
ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140
ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200
tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260
accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320
actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380
ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440
ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500
gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560
aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620
gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680
catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740
tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800
gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860
tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920
ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980
aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040
acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100
ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160
ggcatagcag atgaagtgaa atcttgcttc cgtgttgggt ggtctgatga tggcagccct 5220
gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280
gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340
gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400
gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460
ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520
attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580
cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640
tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700
attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760
attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820
aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880
aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940
gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000
caatctgttc ctcgtgctgg acaagtgtgg tttccagatt ctgcaaccaa gactgcgcag 6060
gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120
ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180
gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240
cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagtgctat 6300
gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360
aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420
gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480
atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540
gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600
tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660
cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720
ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780
gttgcttgga tggataaccc tgaaaactac aaggattata ttcaatatct taaggctcaa 6840
agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900
cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960
gaaatcagga aggtccttgg ttga 6984

Claims (12)

1.一种水稻ACCase突变型蛋白，其特征在于，所述突变型蛋白的氨基酸序列对应于野生型水稻ACCase的氨基酸序列的第2010位和/或2099发生突变。
2.根据权利要求1所述的ACCase突变型蛋白，其特征在于，所述蛋白的氨基酸序列的第2010位由色氨酸突变为亮氨酸和/或2099位由半胱氨酸突变为精氨酸。
3.根据权利要求2所述的ACCase突变型蛋白，其特征在于，其氨基酸序列如SEQ ID NO.2或SEQ ID NO. 4所示。
4.一种ACCase突变型基因或核酸，其特征在于，其编码权利要求1~3中任一项所述的突变型蛋白。
5.根据权利要求4所述的ACCase突变型基因或核酸，其特征在于，其核苷酸序列SEQ IDNO:1 或SEQ ID NO:3所示。
6.表达盒、重组载体或细胞，其含有权利要求4或5所述的突变型基因或核酸。
7.权利要求1~3任一项所述的突变型蛋白，权利要求4或5所述的突变型基因或核酸，权利要求6所述的表达盒、重组载体或细胞在植物抗除草剂方面的应用。
8.根据权利要求7所述的应用，其特征在于，所述除草剂为盖草能或精喹禾灵。
9.获得具有除草剂抗性的植物的方法，其特征在于，包括如下步骤：
1）使植物包含权利要求4或5所述的突变型基因或核酸；或
2）使植物表达权利要求1~3之任一所述的水稻ACCase突变型蛋白。
10.鉴定植物的方法，其中植物是包含权利要求4或5所述的突变型基因或核酸的植物、表达权利要求1~3之任一所述的ACCase突变型蛋白的植物或权利要求9所述的方法获得的植物，其特征在于，包括以下步骤：
1) 测定所述植物是否包含权利要求4或5所述的突变型基因或核酸；或，
2) 测定所述植物是否表达权利要求1~3之任一所述的ACCase突变型蛋白。
11.一种控制杂草的方法，其特征在于，包括：对种植作物的大田施用有效剂量的除草剂，所述植物包含权利要求4或5所述的核酸或基因或权利要求6所述的表达盒、重组载体或细胞，所述除草剂为精喹禾灵和盖草能中的一种或两种。
12.一种用于保护植物免受由除草剂引起的损伤的方法，其特征在于，包括：对种植作物的大田施用有效剂量的除草剂，所述植物包含权利要求4或5所述的核酸或基因或将权利要求6所述的表达盒、重组载体导入植物，导入后的植物产生除草剂抗性蛋白，所述除草剂为精喹禾灵和盖草能中的一种或两种。
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