CN113005105B - 一组抗除草剂蛋白及其在植物育种中的应用 - Google Patents

一组抗除草剂蛋白及其在植物育种中的应用 Download PDF

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CN113005105B
CN113005105B CN201911330008.3A CN201911330008A CN113005105B CN 113005105 B CN113005105 B CN 113005105B CN 201911330008 A CN201911330008 A CN 201911330008A CN 113005105 B CN113005105 B CN 113005105B
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李雪峰
罗祖勇
王明
李莹莹
李相敢
黎跃进
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Keji Dalong Beijing Biotechnology Co ltd
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Abstract

本发明公开了一组抗除草剂蛋白及其在植物育种中的应用。提供了使植物具有除草剂抗性的ALS蛋白质，与野生型相比，带有在位点168从P到L的氨基酸突变和/或在位点624从S到N的氨基酸突变。另外，本发明还提供了相应的获得具有除草剂抗性的植物的方法和应用、以及鉴定本发明的抗除草剂植物或抗除草剂植物的方法。

Description

一组抗除草剂蛋白及其在植物育种中的应用
技术领域
本发明属于植物蛋白质领域，具体而言，本发明涉及能赋予高粱或者其他植物除草剂抗性(耐受性)的蛋白及其在植物育种中的应用。
背景技术
除草剂又称杀草剂。能杀死杂草或有害植物，而不影响农作物正常生长的化学药剂。可用于防治农田杂草或杀灭非农耕地的杂草或灌木。按其作用特点可分为选择性除草剂和灭生性除草剂。选择性除草剂指在使用一定剂量和浓度的范围内，可有选择性地杀死某些种类的植物，而对其他种类植物无害。常见的除草剂，包括有磺酰脲类、咪唑啉酮类、嘧啶并三唑类、水杨酸嘧啶类等除草剂，目前已经大面积商业推广使用。然而，由于这些除草剂通常也能杀死农作物本身，因此对一般不具有除草剂抗性(耐受性)的农作物而言，除草剂的使用在时间和空间上受到了极大限制，如需要在农作物播种前一段时间使用除草剂，以达到除草的目的。传统的清除杂草方式是人工操作，所需要消耗的人力资源和成本非常高，不利于农业大规模化的生产。培育对除草剂有抗性的农作物有极大的经济价值和商业用途，这样可以在种植期间使用除草剂，对抗性农作物使用对应的除草剂，在不影响农作物生长的情况下，杀死杂草，不但拓宽除草剂的使用范围，也可以节约农业的人力资源和成本，有利于农业的大规模化的生产。
以ALS为靶点来研制开发的除草剂目前已有五大类，即磺酰脲类(SU)、咪唑啉酮类(IMI)、磺酰氨基-羰基三唑啉酮(SCT)、嘧啶水杨酸类(PTB)和三唑并嘧啶类(TP)。因此，在本文中，除草剂抗性指的是对包括磺酰脲类、咪唑啉酮类、三唑并嘧啶类、嘧啶水杨酸类在内的除草剂具有耐受性。
ALS为靶点来研制开发的除草剂的作用机理是通过抑制乙酰乳酸合酶(acetolactate synthase,ALS)的活性,破坏蛋白质合成，干扰DNA合成及细胞分裂与生长，最终造成植株死亡达到杀死植物的目的。而植物对这类除草剂的抗性则来自ALS基因一个或多个氨基酸突变而导致酶的结构变化。
目前，已经发现ALS突变蛋白在不同植物(包括玉米、小麦、水稻、油菜和向日葵等)的同源蛋白的第122、155、197、205、574、653、654等位点会产生突变，从而使得相应植物对咪唑啉酮类、磺酰脲类、三唑并嘧啶类类和嘧啶水杨酸类等除草剂产生抗性(Tan S，EvznsR R,DahmerM L,et al.1midazoIinone-tolerant crops:History,current status andFuture.Pest Management Science.2005,61:246-257；Tan S,Evzns R R,Dahmer M L,etal.1midazolinone-tolerant crops:History,current status and Future[J].PestManagement Science.2005,61:246-257)。然而，没有报道表明，在高粱的ALS基因的168位点发生从P到L和/或在624位点发生从S到N的氨基酸突变，会使得高粱植物具有除草剂抗性。
本发明人正是经过长期而艰苦的研究实践，通过化学诱变的方法在高粱突变植株中发现了一系列(突变)蛋白，其能够在除草剂存在的条件下基本保持原有蛋白的正常植物生理功能，从而使得植物具有除草剂抗性(耐受性)。本发明人还开发了这些蛋白及其编码基因在转基因，基因编辑或者非转基因，非基因编辑植物育种中的应用，可用于培育具有除草剂抗性的植物，尤其是农作物，如其他农业植物等。
发明内容
本发明的目的在于提供一个或者多个新的使植物具有除草剂抗性的氨基酸序列，本领域人员可通过已知的生物技术，例如转基因，基因编辑或者突变技术，使得植物能够表达上述的蛋白质，该植物既可具有除草剂的抗性。
具体而言，在所述目的方面，本发明提供了使植物具有除草剂抗性的ALS氨基酸序列。ALS就是乙酰乳酸合成酶(acetolactatesynthase)的简称,其酶分类编号为EC4,1.3.18。使植物具有除草剂抗性的ALS氨基酸序列与野生型相比，其在168位点上带有从P到L的氨基酸突变。野生型高粱的ALS的氨基酸序列如SEQ ID NO：2所述，突变后的ALS的氨基酸序列如SEQ ID NO：4。
具体而言，在所述目的方面，本发明提供了使植物具有除草剂抗性的ALS氨基酸序列。ALS就是乙酰乳酸合成酶(acetolactatesynthase)的简称,其酶分类编号为EC4,1.3.18。使植物具有除草剂抗性的ALS氨基酸序列与野生型相比，其在624位点上带有从S到N的氨基酸突变。野生型高粱的ALS的氨基酸基因序列如SEQ ID NO：2所述，突变后的ALS的氨基酸基因序列如SEQ ID NO：6。具体而言，在所述目的方面，本发明提供了使植物具有除草剂抗性的ALS氨基酸序列。ALS就是乙酰乳酸合成酶(acetolactatesynthase)的简称,其酶分类编号为EC4,1.3.18。使植物具有除草剂抗性的ALS氨基酸序列与野生型相比，其在168位点上带有从P到L的氨基酸突变和在624位点上带有从S到N的氨基酸突变。野生型高粱的ALS的氨基酸序列如SEQ ID NO：2所述，突变后的ALS的核酸序列如SEQ ID NO：7，突变后的ALS的氨基酸序列如SEQ ID NO：8。本发明的另一个目的在于提供具有除草剂抗性的x1)植物；x2)单子叶植物或双子叶植物；x3)高粱。示例性的植物包括水稻、玉米、小麦、黑麦、燕麦、大麦、大豆、马铃薯、油菜、甜菜、甘蔗、高粱、西红柿、南瓜、辣椒、生菜、向日葵、咖啡、茶、棉花、苜蓿、烟草或拟南芥等。
本发明的另一个目的在于提供获得具有除草剂抗性的x1)植物；x2)单子叶植物或双子叶植物；x3)高粱的方法。示例性的植物包括水稻、玉米、小麦、黑麦、燕麦、大麦、大豆、马铃薯、油菜、甜菜、甘蔗、高粱、西红柿、南瓜、辣椒、生菜、向日葵、咖啡、茶、棉花、苜蓿、烟草或拟南芥等。上述除草剂抗性植物的获得，可通过生物技术，例如转基因，基因编辑，或者突变技术，例如化学诱变等，或者通过传统育种，例如将已有除草剂抗性植物与其他植物品种杂交产生的植物后代、以及回交或自交产生的植物后代等等的方法，将上述氨基酸突变在植物中表达，从而使植物获得除草剂的抗性。
本发明的另一个目的在于提供上述基因在x1)或x2)或x3)中获得除草剂抗性的应用:x1)植物；x2)单子叶植物或双子叶植物；x3)高粱。示例性的植物包括水稻、玉米、小麦、黑麦、燕麦、大麦、大豆、马铃薯、油菜、甜菜、甘蔗、高粱、西红柿、南瓜、辣椒、生菜、向日葵、咖啡、茶、棉花、苜蓿、烟草或拟南芥等。该应用包括，通过生物技术，突变技术或者传统育种技术获得具有除草剂抗性的植物过程中的应用，例如转基因植物中的应用，基因编辑植物中的应用或者非转基因植物，非基因编辑植物中的应用等等。
本发明的另一个目的在于提供一个鉴定方法，本领域人员可通过已知的生物技术，通过检测是否在植物中是否含有上述的突变后的氨基酸序列，来判断植物是否采用本发明方法获得的植物的鉴定方法。测定的步骤可以通过常规的核酸检测和/或蛋白质检测方法来进行，只需要能够检测出蛋白质带有P168L和/或S624N突变或其相应核酸突变的方法都可以。
下面将通过具体的附图和实施例对本发明进行详细地描述，使本发明更容易理解。需要特别指出的是，这些描述仅仅是示例性的描述，并不构成对本发明范围的限制。而在阅读本说明书之后，在不脱离本发明的目的和/或所附的权利要求书的范围的前提下，本领域技术人员可以提出本发明的其它实施方式，修改和等同物。依据本说明书的论述，本发明的许多变化、改变对所属领域技术人员来说都是显而易见的了。
附图说明
图1：含有P168L突变体植株与野生型植株在喷洒除草剂后的14天在大田里的对比照片；
图2：含有P168L突变体植株与野生型植株在喷洒除草剂后的12天在温室里的对比照片；
图3：含有S624N突变体植株与野生型植株在喷洒除草剂后的14天在大田里的对比照片；
图4：含有S624N突变体植株与野生型植株在喷洒除草剂后的12天在温室里的对比照片。
具体实施方式
下述实施例子中所用方法如无特殊说明，均为本领域人员常用的方法。
实施例1：从高粱EMS诱变库M2群体中筛选抗性植株
为了对高粱突变体库进行大规模筛选，我们通过化学诱变的方法构建高粱突变体库。具体步骤如下：称取大约1kg的高粱种子，装入80目的网袋中，共计10份，自来水冲洗2次。然后用0.3％(w/w)甲磺酸乙酯(EMS)浸泡高粱种子12h，期间每1h摇动一次种子。12h后取出种子，置于自来水中浸洗5次，EMS废液及浸洗废液收集起来专门容器保存。将浸洗后的种子摊开风干，之后即刻安排播种。同时，光照培养箱测试处理后的种子芽率，控制发芽率在65％左右。待植株成熟后分地块收获，每个地块面积1亩。地块内分大中小穗3种类型，分别混收，晾干至含水量13％。将收获的种子按1万粒/m2的密度播种，待幼苗长至3～4叶期时，用咪唑啉酮类除草剂“甲氧咪草烟”(Imazamox，CAS号114311-32-9)喷雾处理，4％水剂，喷施剂量160ml/亩、兑水30L。2周后调查幼苗枯死情况，发现其中有绿色存活小苗，保留并繁殖这些绿色小苗。
实施例2：筛选出的抗除草剂高粱的分子检测
对筛选得到的2个疑似抗除草剂高粱的单株进行了分子检测。具体步骤如下：取可能带有除草剂抗性的植株叶片10-20mg，放入离心管中，加入钢珠后，置于液氮速冷，用组织研磨机将叶片充分震荡研磨至粉末状，采用SDS法提取样品DNA，然后用特异性引物进行PCR测序。测序公司：英潍捷基(上海)贸易有限公司。经过测序确认：高粱中编码了ALS基因中的两个突变位点，一个突变位点是第503位C-T的碱基突变，对应的是第168位P-L的氨基酸的突变；另一个突变位点是第1871位G-A的碱基突变，对应的是第624位的S-N的氨基酸的突变。野生型植株的核酸序列如SEQ ID NO：1所示，野生型植株的氨基酸序列如SEQ ID NO：2所示；第503位C-T的碱基突变后的核酸序列如SEQ ID NO：3所示，第503位C-T的碱基突变后的氨基酸序列如SEQ ID NO：4所示；第1871位G-A的碱基突变后的核酸序列如SEQ ID NO：5所示，第1871位G-A的碱基突变后的氨基酸序列如SEQ ID NO：6所示；
实施例3：筛选出的抗除草剂植株后代的田间抗咪唑啉酮类除草剂的测试
将鉴定为突变体的株系后代单株收种，在田间按50×20cm行株距、5m行长小区种植，野生型作为对照。待高粱长至四叶期时，按照48g ai/ha的剂量喷施甲氧咪草烟。喷施14、21天后调查植株枯死情况，发现野生型对照在喷施甲氧咪草烟之后几乎停止生长，叶片枯黄或枯死，而筛选到的抗除草剂植株则呈现对甲氧咪草烟完全耐受的表型，其正常的生长发育没有受到影响，最终正常拔节抽穗。这表明我们筛选到的抗除草剂突变体能够稳定地遗传其对咪唑啉酮类除草剂的抗性(如图1和图3)。
实施例4：筛选出的抗除草剂高粱的后代的除草剂耐受性的温室实验
将实施例3中的抗除草剂突变体留种进行后代的除草剂耐受的实验。温室除草剂测试步骤分为以下几个部分：
(1)将营养土铺在穴盘上，穴盘至于盛水的托盘上，让营养土充分吸水。
(2)实施例3收获的高粱种子播种于穴盘的营养土里，再覆一层约1cm的营养土。
(3)人工气候室培养，设置温度为22-26℃，湿度为70％，光照与暗培养分别为14小时和10小时。
(4)出苗后适当间苗，间掉不正常苗，每穴保留4株。
(5)待幼苗长至3-4叶期时，喷施除草剂甲氧咪草烟。除草剂剂量为48g ai/ha，使用扇形喷头、喷施柜喷施。
(6)除草剂喷施后14天调查和记录成苗率、植株鲜重、药害症状(如图2和图4)。
序列表
<110> 科稷达隆生物技术有限公司
<120> 一组抗除草剂蛋白及其在植物育种中的应用
<130> reference
<160> 8
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1926
<212> DNA
<213> Sorghum
<400> 1
atggccacca ccgccgccgc cgctgccgcc gcgctagccg gcgccactac cgctgcgccc 60
aaggcgaggc gccgggcgca cctcctggcc gcacggcgcg ccctcgccgc gcccatcagg 120
tgctcagcgg cgccacccgc cacgctgacg gtgacggctc ccccggccac cccgctccgg 180
ccgtggggcc ccaccgatcc ccgcaagggc gccgacatcc tcgtcgaggc tcttgagcgc 240
tgcggcgtcc gcgacgtctt cgcctacccc ggcggcgcgt ccatggagat ccaccaggca 300
ctcacccgtt cccccgtcat cgccaaccac ctcttccgcc acgagcaagg ggaggccttc 360
gccgcctctg gcttcgcgcg ctcctcgggc cgcgtcggcg tctgcgtcgc cacctccggc 420
cccggcgcca ccaacctagt ctccgcgctc gccgacgcgc tgctcgactc cgtccccatg 480
gtcgccatca cgggacaggt tccgcggcgc atgattggca ccgacgcctt ccaggagacg 540
cccatcgtcg aggtcacccg ctccatcacc aaacataact acctggtcct cgacgtcgac 600
gacatccccc gcgtcgtgca ggaggctttc ttcctcgcct cctccggtcg cccgggaccg 660
gtgcttgtcg acatccccaa ggacatccag cagcagatgg ccgtgccggt ctgggacacg 720
cccatgagtc tgcctgggta cattgcgcgc cttcccaagc ctcctgcgac tgaattgctt 780
gagcaggtgc tgcgtcttgt tggtgaatca aggcgccctg ttctttatgt tggtggtggc 840
tgcgcagcat ctggcgagga gttgcgccgc tttgtggaga tgactggaat cccagtcaca 900
actactctta tgggccttgg caatttccct ggcgacgacc cactgtctct gcgcatgctt 960
ggtatgcatg gcacggtgta tgcaaattat gcagtggata aggcggatct gttgcttgca 1020
tttggtgtgc ggtttgatga tcgtgtgaca gggaagattg aggcttttgc aagcagggct 1080
aagattgtgc acattgatat tgatcccgct gagattggca agaacaagca gccacatgtg 1140
tccatctgtg cagacgttaa gcttgctttg cagggcatga atgctcttct ggaaggaagc 1200
acatcaaaga agagctttga ctttggctca tggcaagctg agttggatca gcagaagaga 1260
gagttccccc ttgggtataa aacttttgat gacgagatcc agccacaata tgctattcag 1320
gttcttgatg agctgacaaa aggggaggcc atcattgcca caggtgttgg gcagcaccag 1380
atgtgggcgg cacagtacta cacttacaag cggccaaggc agtggttgtc ttcagctggt 1440
cttggggcta tgggatttgg tttgccggct gctgctggcg ctgctgtggc caacccaggt 1500
atcactgttg ttgacatcga cggagatggt agcttcctca tgaacattca ggagctagct 1560
atgatccgaa ttgagaacct cccagtgaag gtctttgtgc taaacaacca gcacctgggg 1620
atggtggtgc agtgggagga caggttctat aaggccaata gagcacacac atacttggga 1680
aacccagaga atgaaagtga gatatatcca gatttcgtga caattgccaa agggttcaac 1740
attccagcag tccgtgtgac aaagaagagc gaagtccatg cagcaatcaa gaagatgctt 1800
gagactccag ggccatacct cttggatata atcgtcccgc accaggagca tgtgttgcct 1860
atgatcccta gtggtggggc tttcaaggat atgatcctgg atggtgatgg caggactgtg 1920
tattga 1926
<210> 2
<211> 641
<212> PRT
<213> Sorghum
<400> 2
Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala Leu Ala Gly Ala Thr
1 5 10 15
Thr Ala Ala Pro Lys Ala Arg Arg Arg Ala His Leu Leu Ala Ala Arg
20 25 30
Arg Ala Leu Ala Ala Pro Ile Arg Cys Ser Ala Ala Pro Pro Ala Thr
35 40 45
Leu Thr Val Thr Ala Pro Pro Ala Thr Pro Leu Arg Pro Trp Gly Pro
50 55 60
Thr Asp Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala Leu Glu Arg
65 70 75 80
Cys Gly Val Arg Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu
85 90 95
Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Ala Asn His Leu Phe
100 105 110
Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Phe Ala Arg Ser
115 120 125
Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr
130 135 140
Asn Leu Val Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Val Pro Met
145 150 155 160
Val Ala Ile Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr Asp Ala
165 170 175
Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His
180 185 190
Asn Tyr Leu Val Leu Asp Val Asp Asp Ile Pro Arg Val Val Gln Glu
195 200 205
Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp
210 215 220
Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr
225 230 235 240
Pro Met Ser Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ala
245 250 255
Thr Glu Leu Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg
260 265 270
Pro Val Leu Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu
275 280 285
Arg Arg Phe Val Glu Met Thr Gly Ile Pro Val Thr Thr Thr Leu Met
290 295 300
Gly Leu Gly Asn Phe Pro Gly Asp Asp Pro Leu Ser Leu Arg Met Leu
305 310 315 320
Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp
325 330 335
Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
340 345 350
Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile Asp Ile Asp
355 360 365
Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala
370 375 380
Asp Val Lys Leu Ala Leu Gln Gly Met Asn Ala Leu Leu Glu Gly Ser
385 390 395 400
Thr Ser Lys Lys Ser Phe Asp Phe Gly Ser Trp Gln Ala Glu Leu Asp
405 410 415
Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe Asp Asp Glu
420 425 430
Ile Gln Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly
435 440 445
Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala
450 455 460
Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ala Gly
465 470 475 480
Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val
485 490 495
Ala Asn Pro Gly Ile Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe
500 505 510
Leu Met Asn Ile Gln Glu Leu Ala Met Ile Arg Ile Glu Asn Leu Pro
515 520 525
Val Lys Val Phe Val Leu Asn Asn Gln His Leu Gly Met Val Val Gln
530 535 540
Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly
545 550 555 560
Asn Pro Glu Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala
565 570 575
Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
580 585 590
His Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu
595 600 605
Asp Ile Ile Val Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser
610 615 620
Gly Gly Ala Phe Lys Asp Met Ile Leu Asp Gly Asp Gly Arg Thr Val
625 630 635 640
Tyr
<210> 3
<211> 1926
<212> DNA
<213> Sorghum
<400> 3
atggccacca ccgccgccgc cgctgccgcc gcgctagccg gcgccactac cgctgcgccc 60
aaggcgaggc gccgggcgca cctcctggcc gcacggcgcg ccctcgccgc gcccatcagg 120
tgctcagcgg cgccacccgc cacgctgacg gtgacggctc ccccggccac cccgctccgg 180
ccgtggggcc ccaccgatcc ccgcaagggc gccgacatcc tcgtcgaggc tcttgagcgc 240
tgcggcgtcc gcgacgtctt cgcctacccc ggcggcgcgt ccatggagat ccaccaggca 300
ctcacccgtt cccccgtcat cgccaaccac ctcttccgcc acgagcaagg ggaggccttc 360
gccgcctctg gcttcgcgcg ctcctcgggc cgcgtcggcg tctgcgtcgc cacctccggc 420
cccggcgcca ccaacctagt ctccgcgctc gccgacgcgc tgctcgactc cgtccccatg 480
gtcgccatca cgggacaggt tctgcggcgc atgattggca ccgacgcctt ccaggagacg 540
cccatcgtcg aggtcacccg ctccatcacc aaacataact acctggtcct cgacgtcgac 600
gacatccccc gcgtcgtgca ggaggctttc ttcctcgcct cctccggtcg cccgggaccg 660
gtgcttgtcg acatccccaa ggacatccag cagcagatgg ccgtgccggt ctgggacacg 720
cccatgagtc tgcctgggta cattgcgcgc cttcccaagc ctcctgcgac tgaattgctt 780
gagcaggtgc tgcgtcttgt tggtgaatca aggcgccctg ttctttatgt tggtggtggc 840
tgcgcagcat ctggcgagga gttgcgccgc tttgtggaga tgactggaat cccagtcaca 900
actactctta tgggccttgg caatttccct ggcgacgacc cactgtctct gcgcatgctt 960
ggtatgcatg gcacggtgta tgcaaattat gcagtggata aggcggatct gttgcttgca 1020
tttggtgtgc ggtttgatga tcgtgtgaca gggaagattg aggcttttgc aagcagggct 1080
aagattgtgc acattgatat tgatcccgct gagattggca agaacaagca gccacatgtg 1140
tccatctgtg cagacgttaa gcttgctttg cagggcatga atgctcttct ggaaggaagc 1200
acatcaaaga agagctttga ctttggctca tggcaagctg agttggatca gcagaagaga 1260
gagttccccc ttgggtataa aacttttgat gacgagatcc agccacaata tgctattcag 1320
gttcttgatg agctgacaaa aggggaggcc atcattgcca caggtgttgg gcagcaccag 1380
atgtgggcgg cacagtacta cacttacaag cggccaaggc agtggttgtc ttcagctggt 1440
cttggggcta tgggatttgg tttgccggct gctgctggcg ctgctgtggc caacccaggt 1500
atcactgttg ttgacatcga cggagatggt agcttcctca tgaacattca ggagctagct 1560
atgatccgaa ttgagaacct cccagtgaag gtctttgtgc taaacaacca gcacctgggg 1620
atggtggtgc agtgggagga caggttctat aaggccaata gagcacacac atacttggga 1680
aacccagaga atgaaagtga gatatatcca gatttcgtga caattgccaa agggttcaac 1740
attccagcag tccgtgtgac aaagaagagc gaagtccatg cagcaatcaa gaagatgctt 1800
gagactccag ggccatacct cttggatata atcgtcccgc accaggagca tgtgttgcct 1860
atgatcccta gtggtggggc tttcaaggat atgatcctgg atggtgatgg caggactgtg 1920
tattga 1926
<210> 4
<211> 641
<212> PRT
<213> Sorghum
<400> 4
Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala Leu Ala Gly Ala Thr
1 5 10 15
Thr Ala Ala Pro Lys Ala Arg Arg Arg Ala His Leu Leu Ala Ala Arg
20 25 30
Arg Ala Leu Ala Ala Pro Ile Arg Cys Ser Ala Ala Pro Pro Ala Thr
35 40 45
Leu Thr Val Thr Ala Pro Pro Ala Thr Pro Leu Arg Pro Trp Gly Pro
50 55 60
Thr Asp Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala Leu Glu Arg
65 70 75 80
Cys Gly Val Arg Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu
85 90 95
Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Ala Asn His Leu Phe
100 105 110
Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Phe Ala Arg Ser
115 120 125
Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr
130 135 140
Asn Leu Val Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Val Pro Met
145 150 155 160
Val Ala Ile Thr Gly Gln Val Leu Arg Arg Met Ile Gly Thr Asp Ala
165 170 175
Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His
180 185 190
Asn Tyr Leu Val Leu Asp Val Asp Asp Ile Pro Arg Val Val Gln Glu
195 200 205
Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp
210 215 220
Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr
225 230 235 240
Pro Met Ser Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ala
245 250 255
Thr Glu Leu Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg
260 265 270
Pro Val Leu Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu
275 280 285
Arg Arg Phe Val Glu Met Thr Gly Ile Pro Val Thr Thr Thr Leu Met
290 295 300
Gly Leu Gly Asn Phe Pro Gly Asp Asp Pro Leu Ser Leu Arg Met Leu
305 310 315 320
Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp
325 330 335
Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
340 345 350
Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile Asp Ile Asp
355 360 365
Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala
370 375 380
Asp Val Lys Leu Ala Leu Gln Gly Met Asn Ala Leu Leu Glu Gly Ser
385 390 395 400
Thr Ser Lys Lys Ser Phe Asp Phe Gly Ser Trp Gln Ala Glu Leu Asp
405 410 415
Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe Asp Asp Glu
420 425 430
Ile Gln Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly
435 440 445
Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala
450 455 460
Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ala Gly
465 470 475 480
Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val
485 490 495
Ala Asn Pro Gly Ile Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe
500 505 510
Leu Met Asn Ile Gln Glu Leu Ala Met Ile Arg Ile Glu Asn Leu Pro
515 520 525
Val Lys Val Phe Val Leu Asn Asn Gln His Leu Gly Met Val Val Gln
530 535 540
Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly
545 550 555 560
Asn Pro Glu Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala
565 570 575
Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
580 585 590
His Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu
595 600 605
Asp Ile Ile Val Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser
610 615 620
Gly Gly Ala Phe Lys Asp Met Ile Leu Asp Gly Asp Gly Arg Thr Val
625 630 635 640
Tyr
<210> 5
<211> 1926
<212> DNA
<213> Sorghum
<400> 5
atggccacca ccgccgccgc cgctgccgcc gcgctagccg gcgccactac cgctgcgccc 60
aaggcgaggc gccgggcgca cctcctggcc gcacggcgcg ccctcgccgc gcccatcagg 120
tgctcagcgg cgccacccgc cacgctgacg gtgacggctc ccccggccac cccgctccgg 180
ccgtggggcc ccaccgatcc ccgcaagggc gccgacatcc tcgtcgaggc tcttgagcgc 240
tgcggcgtcc gcgacgtctt cgcctacccc ggcggcgcgt ccatggagat ccaccaggca 300
ctcacccgtt cccccgtcat cgccaaccac ctcttccgcc acgagcaagg ggaggccttc 360
gccgcctctg gcttcgcgcg ctcctcgggc cgcgtcggcg tctgcgtcgc cacctccggc 420
cccggcgcca ccaacctagt ctccgcgctc gccgacgcgc tgctcgactc cgtccccatg 480
gtcgccatca cgggacaggt tccgcggcgc atgattggca ccgacgcctt ccaggagacg 540
cccatcgtcg aggtcacccg ctccatcacc aaacataact acctggtcct cgacgtcgac 600
gacatccccc gcgtcgtgca ggaggctttc ttcctcgcct cctccggtcg cccgggaccg 660
gtgcttgtcg acatccccaa ggacatccag cagcagatgg ccgtgccggt ctgggacacg 720
cccatgagtc tgcctgggta cattgcgcgc cttcccaagc ctcctgcgac tgaattgctt 780
gagcaggtgc tgcgtcttgt tggtgaatca aggcgccctg ttctttatgt tggtggtggc 840
tgcgcagcat ctggcgagga gttgcgccgc tttgtggaga tgactggaat cccagtcaca 900
actactctta tgggccttgg caatttccct ggcgacgacc cactgtctct gcgcatgctt 960
ggtatgcatg gcacggtgta tgcaaattat gcagtggata aggcggatct gttgcttgca 1020
tttggtgtgc ggtttgatga tcgtgtgaca gggaagattg aggcttttgc aagcagggct 1080
aagattgtgc acattgatat tgatcccgct gagattggca agaacaagca gccacatgtg 1140
tccatctgtg cagacgttaa gcttgctttg cagggcatga atgctcttct ggaaggaagc 1200
acatcaaaga agagctttga ctttggctca tggcaagctg agttggatca gcagaagaga 1260
gagttccccc ttgggtataa aacttttgat gacgagatcc agccacaata tgctattcag 1320
gttcttgatg agctgacaaa aggggaggcc atcattgcca caggtgttgg gcagcaccag 1380
atgtgggcgg cacagtacta cacttacaag cggccaaggc agtggttgtc ttcagctggt 1440
cttggggcta tgggatttgg tttgccggct gctgctggcg ctgctgtggc caacccaggt 1500
atcactgttg ttgacatcga cggagatggt agcttcctca tgaacattca ggagctagct 1560
atgatccgaa ttgagaacct cccagtgaag gtctttgtgc taaacaacca gcacctgggg 1620
atggtggtgc agtgggagga caggttctat aaggccaata gagcacacac atacttggga 1680
aacccagaga atgaaagtga gatatatcca gatttcgtga caattgccaa agggttcaac 1740
attccagcag tccgtgtgac aaagaagagc gaagtccatg cagcaatcaa gaagatgctt 1800
gagactccag ggccatacct cttggatata atcgtcccgc accaggagca tgtgttgcct 1860
atgatcccta atggtggggc tttcaaggat atgatcctgg atggtgatgg caggactgtg 1920
tattga 1926
<210> 6
<211> 641
<212> PRT
<213> Sorghum
<400> 6
Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala Leu Ala Gly Ala Thr
1 5 10 15
Thr Ala Ala Pro Lys Ala Arg Arg Arg Ala His Leu Leu Ala Ala Arg
20 25 30
Arg Ala Leu Ala Ala Pro Ile Arg Cys Ser Ala Ala Pro Pro Ala Thr
35 40 45
Leu Thr Val Thr Ala Pro Pro Ala Thr Pro Leu Arg Pro Trp Gly Pro
50 55 60
Thr Asp Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala Leu Glu Arg
65 70 75 80
Cys Gly Val Arg Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu
85 90 95
Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Ala Asn His Leu Phe
100 105 110
Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Phe Ala Arg Ser
115 120 125
Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr
130 135 140
Asn Leu Val Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Val Pro Met
145 150 155 160
Val Ala Ile Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr Asp Ala
165 170 175
Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His
180 185 190
Asn Tyr Leu Val Leu Asp Val Asp Asp Ile Pro Arg Val Val Gln Glu
195 200 205
Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp
210 215 220
Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr
225 230 235 240
Pro Met Ser Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ala
245 250 255
Thr Glu Leu Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg
260 265 270
Pro Val Leu Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu
275 280 285
Arg Arg Phe Val Glu Met Thr Gly Ile Pro Val Thr Thr Thr Leu Met
290 295 300
Gly Leu Gly Asn Phe Pro Gly Asp Asp Pro Leu Ser Leu Arg Met Leu
305 310 315 320
Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp
325 330 335
Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
340 345 350
Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile Asp Ile Asp
355 360 365
Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala
370 375 380
Asp Val Lys Leu Ala Leu Gln Gly Met Asn Ala Leu Leu Glu Gly Ser
385 390 395 400
Thr Ser Lys Lys Ser Phe Asp Phe Gly Ser Trp Gln Ala Glu Leu Asp
405 410 415
Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe Asp Asp Glu
420 425 430
Ile Gln Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly
435 440 445
Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala
450 455 460
Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ala Gly
465 470 475 480
Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val
485 490 495
Ala Asn Pro Gly Ile Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe
500 505 510
Leu Met Asn Ile Gln Glu Leu Ala Met Ile Arg Ile Glu Asn Leu Pro
515 520 525
Val Lys Val Phe Val Leu Asn Asn Gln His Leu Gly Met Val Val Gln
530 535 540
Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly
545 550 555 560
Asn Pro Glu Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala
565 570 575
Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
580 585 590
His Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu
595 600 605
Asp Ile Ile Val Pro His Gln Glu His Val Leu Pro Met Ile Pro Asn
610 615 620
Gly Gly Ala Phe Lys Asp Met Ile Leu Asp Gly Asp Gly Arg Thr Val
625 630 635 640
Tyr
<210> 7
<211> 1926
<212> DNA
<213> Sorghum
<400> 7
atggccacca ccgccgccgc cgctgccgcc gcgctagccg gcgccactac cgctgcgccc 60
aaggcgaggc gccgggcgca cctcctggcc gcacggcgcg ccctcgccgc gcccatcagg 120
tgctcagcgg cgccacccgc cacgctgacg gtgacggctc ccccggccac cccgctccgg 180
ccgtggggcc ccaccgatcc ccgcaagggc gccgacatcc tcgtcgaggc tcttgagcgc 240
tgcggcgtcc gcgacgtctt cgcctacccc ggcggcgcgt ccatggagat ccaccaggca 300
ctcacccgtt cccccgtcat cgccaaccac ctcttccgcc acgagcaagg ggaggccttc 360
gccgcctctg gcttcgcgcg ctcctcgggc cgcgtcggcg tctgcgtcgc cacctccggc 420
cccggcgcca ccaacctagt ctccgcgctc gccgacgcgc tgctcgactc cgtccccatg 480
gtcgccatca cgggacaggt tctgcggcgc atgattggca ccgacgcctt ccaggagacg 540
cccatcgtcg aggtcacccg ctccatcacc aaacataact acctggtcct cgacgtcgac 600
gacatccccc gcgtcgtgca ggaggctttc ttcctcgcct cctccggtcg cccgggaccg 660
gtgcttgtcg acatccccaa ggacatccag cagcagatgg ccgtgccggt ctgggacacg 720
cccatgagtc tgcctgggta cattgcgcgc cttcccaagc ctcctgcgac tgaattgctt 780
gagcaggtgc tgcgtcttgt tggtgaatca aggcgccctg ttctttatgt tggtggtggc 840
tgcgcagcat ctggcgagga gttgcgccgc tttgtggaga tgactggaat cccagtcaca 900
actactctta tgggccttgg caatttccct ggcgacgacc cactgtctct gcgcatgctt 960
ggtatgcatg gcacggtgta tgcaaattat gcagtggata aggcggatct gttgcttgca 1020
tttggtgtgc ggtttgatga tcgtgtgaca gggaagattg aggcttttgc aagcagggct 1080
aagattgtgc acattgatat tgatcccgct gagattggca agaacaagca gccacatgtg 1140
tccatctgtg cagacgttaa gcttgctttg cagggcatga atgctcttct ggaaggaagc 1200
acatcaaaga agagctttga ctttggctca tggcaagctg agttggatca gcagaagaga 1260
gagttccccc ttgggtataa aacttttgat gacgagatcc agccacaata tgctattcag 1320
gttcttgatg agctgacaaa aggggaggcc atcattgcca caggtgttgg gcagcaccag 1380
atgtgggcgg cacagtacta cacttacaag cggccaaggc agtggttgtc ttcagctggt 1440
cttggggcta tgggatttgg tttgccggct gctgctggcg ctgctgtggc caacccaggt 1500
atcactgttg ttgacatcga cggagatggt agcttcctca tgaacattca ggagctagct 1560
atgatccgaa ttgagaacct cccagtgaag gtctttgtgc taaacaacca gcacctgggg 1620
atggtggtgc agtgggagga caggttctat aaggccaata gagcacacac atacttggga 1680
aacccagaga atgaaagtga gatatatcca gatttcgtga caattgccaa agggttcaac 1740
attccagcag tccgtgtgac aaagaagagc gaagtccatg cagcaatcaa gaagatgctt 1800
gagactccag ggccatacct cttggatata atcgtcccgc accaggagca tgtgttgcct 1860
atgatcccta atggtggggc tttcaaggat atgatcctgg atggtgatgg caggactgtg 1920
tattga 1926
<210> 8
<211> 641
<212> PRT
<213> Sorghum
<400> 8
Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala Leu Ala Gly Ala Thr
1 5 10 15
Thr Ala Ala Pro Lys Ala Arg Arg Arg Ala His Leu Leu Ala Ala Arg
20 25 30
Arg Ala Leu Ala Ala Pro Ile Arg Cys Ser Ala Ala Pro Pro Ala Thr
35 40 45
Leu Thr Val Thr Ala Pro Pro Ala Thr Pro Leu Arg Pro Trp Gly Pro
50 55 60
Thr Asp Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala Leu Glu Arg
65 70 75 80
Cys Gly Val Arg Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu
85 90 95
Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Ala Asn His Leu Phe
100 105 110
Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Phe Ala Arg Ser
115 120 125
Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr
130 135 140
Asn Leu Val Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Val Pro Met
145 150 155 160
Val Ala Ile Thr Gly Gln Val Leu Arg Arg Met Ile Gly Thr Asp Ala
165 170 175
Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His
180 185 190
Asn Tyr Leu Val Leu Asp Val Asp Asp Ile Pro Arg Val Val Gln Glu
195 200 205
Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp
210 215 220
Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr
225 230 235 240
Pro Met Ser Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ala
245 250 255
Thr Glu Leu Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg
260 265 270
Pro Val Leu Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu
275 280 285
Arg Arg Phe Val Glu Met Thr Gly Ile Pro Val Thr Thr Thr Leu Met
290 295 300
Gly Leu Gly Asn Phe Pro Gly Asp Asp Pro Leu Ser Leu Arg Met Leu
305 310 315 320
Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp
325 330 335
Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
340 345 350
Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile Asp Ile Asp
355 360 365
Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala
370 375 380
Asp Val Lys Leu Ala Leu Gln Gly Met Asn Ala Leu Leu Glu Gly Ser
385 390 395 400
Thr Ser Lys Lys Ser Phe Asp Phe Gly Ser Trp Gln Ala Glu Leu Asp
405 410 415
Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe Asp Asp Glu
420 425 430
Ile Gln Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly
435 440 445
Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala
450 455 460
Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ala Gly
465 470 475 480
Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val
485 490 495
Ala Asn Pro Gly Ile Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe
500 505 510
Leu Met Asn Ile Gln Glu Leu Ala Met Ile Arg Ile Glu Asn Leu Pro
515 520 525
Val Lys Val Phe Val Leu Asn Asn Gln His Leu Gly Met Val Val Gln
530 535 540
Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly
545 550 555 560
Asn Pro Glu Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala
565 570 575
Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
580 585 590
His Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu
595 600 605
Asp Ile Ile Val Pro His Gln Glu His Val Leu Pro Met Ile Pro Asn
610 615 620
Gly Gly Ala Phe Lys Asp Met Ile Leu Asp Gly Asp Gly Arg Thr Val
625 630 635 640
Tyr

Claims (5)

1.一种抗除草剂高粱的ALS的突变体，其特征在于，与其野生型相比，带有S624N的突变，其氨基酸序列如SEQ ID NO： 6所示。
2.根据权利要求1所述的ALS的突变体，其特征在于，其核酸序列如SEQ ID NO： 5所示。
3.获得具有除草剂抗性的高粱的方法，其特征在于，包括如下步骤 ：
（1）使高粱包含权利要求2所述的核酸 ；
或，（2）使高粱表达具有权利要求 1所述的氨基酸序列的蛋白质。
4.鉴定具有除草剂抗性的高粱的方法，其特征在于，包括如下步骤 ：
（1）测试高粱是否包含权利要求2所述的核酸 ；
或，（2）测试高粱是否表达具有权利要求 1所述的氨基酸序列的蛋白质。
5.权利要求1或2所述的突变体在获得抗除草剂的高粱中的应用，其特征在于，所述除草剂是咪唑啉酮类和磺酰脲类除草剂。
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IL83348A (en) * 1986-08-26 1995-12-08 Du Pont Nucleic acid fragment encoding herbicide resistant plant acetolactate synthase
US5013659A (en) * 1987-07-27 1991-05-07 E. I. Du Pont De Nemours And Company Nucleic acid fragment encoding herbicide resistant plant acetolactate synthase
US9365862B1 (en) * 2011-05-26 2016-06-14 Nutech Ventures Herbicide resistant sorghum mutants
CN102559646B (zh) * 2011-11-24 2013-01-23 未名兴旺系统作物设计前沿实验室（北京）有限公司 小麦抗除草剂蛋白及其在植物育种中的应用
CN103215243B (zh) * 2013-04-07 2014-10-08 深圳兴旺生物种业有限公司 甘蓝型油菜抗除草剂蛋白及其在植物育种中的应用
CN106636028A (zh) * 2017-02-08 2017-05-10 上海市农业科学院 具有抗咪唑啉酮类除草剂活性的水稻蛋白质、其编码基因及应用
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