CN117511965A - 一种与水稻苗期耐盐性相关的基因OsGRAS34及其应用 - Google Patents
一种与水稻苗期耐盐性相关的基因OsGRAS34及其应用 Download PDFInfo
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Abstract
本发明公开了一种与水稻苗期耐盐性相关的基因OsGRAS34及其应用,该基因具有如下(1)或(2)所述的核苷酸序列:(1)如SEQ ID NO:3所示的编码区核苷酸序列;(2)与如SEQ ID NO:3所示的核苷酸序列具有90%以上的同源性,且编码相同功能蛋白质的核苷酸序列。本发明的植物耐盐基因OsGRAS34影响水稻苗期耐盐性。敲除该基因可导致苗期耐盐性增加,从而可以培育苗期高耐盐转基因水稻。OsGRAS34基因可以作为一个提高苗期耐盐性的基因应用于水稻育种工作中,具有巨大的开发价值和应用前景。
Description
技术领域
本发明属于农业生物工程技术领域,涉及一种与水稻苗期耐盐性相关的基因OsGRAS34及其应用。本发明还涉及该基因编码的多肽序列,以及通过基因编辑技术培育苗期高耐盐性水稻品种的方法。
背景技术
我国是全球第三大盐碱地分布国家,拥有各类可利用盐碱地资源5.5亿亩,开发潜力大,是我国耕地“提质、扩容、增效”的重要来源,是粮食增产的“潜在粮仓”。水稻是我国重要粮食作物之一,高产耐盐水稻种质资源缺乏,农业生产中迫切需要高产耐盐碱的水稻新品种。大量研究表明,盐碱胁迫影响水稻生长发育,导致水稻减产,其技术瓶颈为缺乏具有育种应用价值、协同调控高产与耐盐性的优异等位基因。
苗期耐盐是耐盐水稻培育的基础,同时,目前对于水稻耐盐资源筛选也主要集中在苗期,用到的有地上部的苗高、地上部鲜重、地上部干重、含水量、地上部Na+含量、地上部K+含量、地上部Na+/K+含量比和存活率等各项指标。前人利用正向遗传学QTL定位以及反向遗传学等方式克隆到SKC1(Ren et al.2005),OsWRKY53(Yu et al.,2023)以及OsCIPK9(Zhou et al.2023)等基因。其中SKC1是通过调控茎的钾离子浓度来实现耐盐,OsWRKY53通过MKK10-2和SKC1通路来调控盐胁迫;OsCIPK9通过与OsSOS3互作来调控SOS途径,进而达到耐盐效果。
尽管目前已经发现SOS途径,转运蛋白家族参与植物体内离子稳态调节,仍迫切的需要挖掘新的离子转运蛋白和调控网络来拓宽人们对植物体内钠、钾离子平衡机制的认识。同时,如何利用好种质资源,并从种质资源中发掘更多的耐盐基因也是需要面对的问题。
GRAS家族基因是一类含有GRAS domain的基因,主要参与到根发育、赤霉素信号、光信号传导等生物进程。拟南芥中GRAS蛋白SCL7参与到耐盐中(Ma et al.,2010)。水稻中的GRAS蛋白MOC1调控水稻分蘖的产生(Li et al.,2003)。OsGRAS23可以通过影响多个胁迫响应基因调控水稻耐旱性(Xu et al.,2015)。但我们对于水稻中的GRAS类基因对于苗期耐盐性的影响的认识还非常有限。
发明内容
本发明的目的在于提供一种与水稻苗期耐盐性相关的OsGRAS34基因及其编码的蛋白和在调控水稻苗期耐盐性中的应用。
本发明的目的可以通过以下技术方案实现:
一种水稻OsGRAS34基因,该基因具有如下(1)或(2)所述的核苷酸序列:
(1)如SEQ ID NO:3所示的编码区核苷酸序列;
(2)与如SEQ ID NO:3所示的核苷酸序列具有90%以上的同源性,且编码相同功能蛋白质的核苷酸序列。
本发明通过对群体的OsGRAS34基因以及启动子变异检测,发现了该基因与苗期耐盐性相关,并利用基因编辑构建转基因植株,验证了敲除OsGRAS34基因可以提高苗期耐盐性。
该基因还具有如SEQ ID NO:2所示的基因组核苷酸序列。
上述水稻OsGRAS34基因编码的蛋白也属于本发明的保护范围。该蛋白具有如下(a)或(b)所示的氨基酸残基序列:
(a)如SEQ ID NO:1所示的氨基酸序列;
(b)将SEQ ID NO:1所示的氨基酸序列经过一个或若干个氨基酸残基的取代和/或缺失和/或添加衍生得到的具有相同功能的氨基酸序列。
含有上述水稻OsGRAS34基因的表达盒、重组载体、转基因细胞系或转基因重组菌,或者用于干扰、抑制、沉默、靶向敲除或定点突变所述水稻OsGRAS34基因的生物材料均属于本发明的保护范围。
优选的,用于干扰、抑制、沉默、靶向敲除或定点突变所述水稻OsGRAS34基因的生物材料包括以下(1)~(6)中的至少一种:
(1)针对水稻OsGRAS34基因的干扰序列;
(2)用于干扰的水稻OsGRAS34基因的干扰载体;
(3)包含有水稻OsGRAS34基因干扰载体的转基因细胞系;
(4)基于基因编辑技术靶向敲除或定点突变水稻OsGRAS34基因的引物序列;
(5)基于基因编辑技术靶向敲除或定点突变水稻OsGRAS34基因的载体;
(6)基于基因编辑技术靶向敲除或定点突变水稻OsGRAS34基因的细胞系。
所述的含有水稻OsGRAS34基因的重组载体可以为重组克隆载体或重组表达载体,可用现有的植物表达载体构建所述基因的重组表达载体。
所述植物表达载体包括双元农杆菌载体和可用于植物微弹轰击的载体等。所述植物表达载体还可包含外源基因的3’端非翻译区域,即包含聚腺苷酸信号和任何其它参与mRNA加工或基因表达的DNA片段。所述聚腺苷酸信号可引导聚腺苷酸加入到mRNA前体的3’端,如农杆菌冠瘦瘤诱导(Ti)质粒基因(如胭脂合成酶N')s基因)、植物基因(如大豆贮存蛋白基因)3’端转录的非翻译区均具有类似功能。
使用所述基因构建重组植物表达载体时,在其转录起始核苷酸前可加上任何一种增强型启动子或组成型启动子,如花椰菜花叶病毒(CAMV)35S启动子、玉米的泛素启动子(Ubiquitin),它们可单独使用或与其它的植物启动子结合使用;此外,使用本发明的基因构建植物表达载体时,还可使用增强子,包括翻译增强子或转录增强子,这些增强子区域可以是ATG起始密码子或邻接区域起始密码子等,但必需与编码序列的阅读框相同,以保证整个序列的正确翻译。所述翻译控制信号和起始密码子的来源是广泛的,可以是天然的,也可以是合成的。翻译起始区域可以来自转录起始区域或结构基因。
为了便于对转基因植物细胞或植物进行鉴定及筛选,可对所用植物表达载体进行加工,如加入可在植物中表达的编码可产生颜色变化的酶或发光化合物的基因(GUS基因、荧光素酶基因等)、具有抗性的抗生素标记物(庆大霉素标记物、卡那霉素标记物等)或是抗化学试剂标记基因(如抗除草剂基因)等。从转基因植物的安全性考虑,可不加任何选择性标记基因,直接以逆境筛选转化植株。
上述的水稻OsGRAS34基因,上述的蛋白,或上述的表达盒、重组载体、转基因细胞系或转基因重组菌在调控水稻苗期耐盐性或培育高苗期耐盐性水稻中的应用。
上述的用于干扰、抑制、沉默、靶向敲除或定点突变水稻OsGRAS34基因的生物材料在提高水稻苗期耐盐性或培育高苗期耐盐性水稻中的应用。例如,水稻OsGRAS34基因的定点突变载体在提高水稻苗期耐盐性或培育高苗期耐盐性水稻中的应用。
上述的应用,具体为通过基因编辑、干扰、抑制或沉默所述水稻OsGRAS34基因的表达提高水稻苗期耐盐性,培育高苗期耐盐性的水稻。
一种培育苗期耐盐性高水稻的方法,通过基因编辑、干扰、抑制或沉默所述水稻OsGRAS34基因提高水稻苗期耐盐性,培育高苗期耐盐性的水稻。
本发明研究结果显示,向受体水稻导入基因编辑载体,使得水稻OsGRAS34基因受到编辑,得到转基因水稻;转基因水稻与受体水稻相比,苗期耐盐性增加。所述的受体水稻为粳稻品种日本晴。
本发明的有益效果:
本发明的植物耐盐基因OsGRAS34影响水稻苗期耐盐性。敲除该基因可导致苗期耐盐性增加,从而可以培育苗期高耐盐转基因水稻。所述蛋白及其编码基因可以应用于植物遗传改良。
附图说明
图1为OsGRAS34基因的启动子在不同水稻品种中的变异和耐盐相关表型分析。
图2OsGRAS34基因编辑家系测序分析,OsGRAS34-cas1和OsGRAS34-cas2为纯和敲除家系。
图3OsGRAS34-cas1和OsGRAS34-cas2家系在OsGRAS34基因编辑后蛋白翻译分析。
图4受体水稻,OsGRAS34-cas1和OsGRAS34-cas2家系的苗期耐盐性统计分析。
具体实施方式
以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为自常规生化试剂商店购买得到的。
实施例1、
通过对大规模水稻品种进行进化分析,结果表明:含有GRAS domain的蛋白编码基因OsGRAS34(Os07g0545800)在不同品种中具有不同的单倍型(图l)。序列分析的结果表明,Os07g0545800的启动子区中含有一个CCAAT motif,motif内变异将群体分成3种单倍型,如图1左所示。
本发明通过对群体的OsGRAS34启动子变异检测,发现了该基因与苗期耐盐性相关,该基因具有如SEQ ID NO:3所示的编码区序列,该基因的基因组序列如SEQ ID NO:2所示。
实施例2、不同品种的耐盐相关性状测定
利用含有OsGRAS34不同单倍型品种进行耐盐处理,将催芽的种子放在人工气候室中培养至2叶1心期,约10天,并进行140mM NaCl处理10天,统计地上部分鲜重和干重,并计算比率,不同单倍型间存在差异,HapB具有较高地上部分鲜重和干重比,说明OsGRAS34具有耐盐性。如图1中,右所示。
实施例3、OsGRAS34基因编辑转基因株系的获得及其耐盐相关表型的测定
首先通过网站CRISR-P(http://cbi.hzau.edu.cn/cgi-bin/CRISPR)找到合适的spacer序列(如SEQ ID NO:3中第333-352位核苷酸所示),并合成两条带有接头的互补引物(前引物:GGCATAATATGCAACACGCCTTAC,后引物:AAACGTAAGGCGTGTTGCATATTA)。PCR仪中缓慢退火后得到双链,将双链Oligo连接到经BsaI酶切的BGK03载体中。构建好的载体经农杆菌EHA105转化受体水稻愈伤组织,获得T0代基因编辑转基因植株,通过一代测序共鉴定到8株OsGRAS34基因被编辑的敲除植株,其中OsGRAS34-cas1和OsGRAS34-cas2为纯合突变植株(图2)。OsGRAS34-cas1的OsGRAS34基因ATG后350位点上缺失一个碱基T,导致117个氨基酸的移码突变和翻译的提前终止(比野生型截短448个氨基酸)。OsGRAS34-cas2的OsGRAS34基因在ATG后350bp处插入了1个碱基T,导致116个氨基酸的移码突变和翻译的提前终止(比野生型截短451个氨基酸)(图3)。对上述转基因敲除家系进行120mMNaCl处理7天后耐盐相关表型的测定,结果表明:敲除植株OsGRAS34-cas1和OsGRAS34-cas2在盐处理后地上和地下部分鲜重和干重均比野生型高,证明敲除材料苗期耐盐性显著高于受体水稻(图4)。
以上结果表明,OsGRAS34是一个与苗期耐盐性有关的基因。通过基因编辑可以得到苗期耐盐性高的水稻。
SEQ ID NO:1
MDLHQLLKYRLTGANVVYEIPTENNLQNSPWQANPLKYEFSDSPYTPLSSQFECDNLSALTNTPDNQSSTETISAQPISP
LEADSSYRQAGILLQENIQVGADPLYATSRHNMQHALREIETVLMAPDTDDATTSTKHEFEEIKPAQLVRQRSRTWSHES
RQPLPGVGRSQFASGGYPTASYEFRPEKRQRELREDPQIIVKQLLTRCAEALSEDRTEEFHKLVQEARGVVSINGEPIQR
LGAYLLEGLVARHGNSGTNIYRALKCREPESKELLSYMRILYNICPYFKFGYMAANGAIAEALRTENNIHIIDFQIAQGT
QWITLIQALAARPGGPPRVRITGIDDPVSEYARGEGLDIVGKMLKSMSEEFKIPLEFTPLSVYATQVTKEMLEIRPGEAL
SVNFTLQLHHTPDESVDVNNPRDGLLRMVKGLSPKVTTLVEQESHTNTTPFLMRFGETMEYYSAMFESIDANLPRDNKER
ISVEQHCLAKDIVNIIACEGKDRVERHELLGKWKSRLTMAGFRPYPLSSYVNSVIRKLLACYSDKYTLDEKDGAMLLGWR
SRKLISASAWH
SEQ ID NO:2
ATCGCAGCACACGAAAAGAGAACCAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGGAGTGGAAGTGGATCGGGGCAT
CGCTGCGGCGTCGGGGCTGTGCAGATCGGAGGAGGTGAGAGCTCTCGTCCTCGTACCATCCATCTCATCGGATTCCGTGC
TGCCGTGGTTGCTAATTTTGGCTTGGTTTGTCGAATTCTTTTTGGGCTTTTTTTGGGGTGCTTTTTTTGGTTGGTATTAC
CCGGGGTTTGCTTCTTGAGGGTGAAGGCGGCGTCTCCGAGCGGTTTCCGGTGGAAGTCTAGTGGTTTTTCGGCGAGGAGA
TGGGTTTCAGTTGGGTGAGCTAGGTTTATTGGACGAATTTGTTGGGGATTTCTGTTGGTTTGAGCCGATTCGATCGCGTC
CGAGAGCATTTTGCTGTTGGGGACGAGAGGGCGCCTCCATTTTCGGGATTACTGAGTTTATCCTGTGCAAATTTTGCAAA
AAAAATTCTACTAATTTTGGTCCGTTTCATAGGTGAATCGTCGCTTATTGAGTGGTTCGTTAGGTTAGCTCTTAACTGCG
CCGCTTGCATTTGTTGCCGGAATTTATTATTATCTACTCCACGGGTAACTCGAGGGCGTGAAGTGAAAGCTAATTATGTC
GCCCAAAGTTACTGATCCCTTTTTTTTTTTGTTGAAACTGAGCTAGCTTAATTTGCCTCTGTTGGCTTGTGGAGACATGA
TGCAGTACAACCAAGTAGCAGCAACAGTTTTTGCCCTGTGAAACACTGGAACCATGATCCCCCTTTTTTGTGTGTGTTTG
ATTAGGACATAACTTACAAAATTCTATGTTTCACCTGATTAGTCAATTGGCTTACCTCTCCTCCCGATGACAAATCCATC
AAAGAAATTTTCTTGCTGGGGGGTAAAGTGCCACTGGAGAAGAACAAATCGAGGTTGTTCTGTGACTTGGTTGTGTAGTT
TTATATAGTTTTTAGCATTGTCAACCAGGCTCCAGGATCCCAGAAATTTCACTCCTTTGCTTACTGCATAGACTATACAA
GTTGCCCTCTTGTTTGAGCGATGAAGGCGACGTTGATTCCACACGGTTGATTGGTCAAACCTCTTGTTGGCGGAGGTACC
TTCTTCAGAATTCAGAGGGGTGATAGACCAATAGTCAAACAACTAGGTTGGTCAATGATCAATCGTCTTCATCCAACTAG
AGAATCTTATAGCCTGCTTGTGAGTAGTCCTAATTGTGTATTTTGGAAGGATCAAAGAACAGTCCACGTCAATTGTCTTC
AACCAACTAGAGAATGTTTGAGTCTGCTTATCTTAATTGCAAGGGAAGTCGATGAATATTTTGATGGGGGATGTATAACT
TTTTTTCCTTATCTTTTCTCCTGTTGTTTGCTCTCCAAACCTCATTGTTAGGCTGGTTCTCCTCGGTAGTACTTAGAATA
GCTTCATTTCTCCAGAGCTTGTTGTCCTCTAGTTGTATGCTATGTGGCTGAGCTTGACATTCTGGCTAGATTAATTGGAG
TTATGTGGGAGTGATTGGGATATTTCATAGATAATGTTCTTCTTTTGGAGTTGCGAGCACACGGTATGTGATGCAATGAT
GGACTTAAGTAAGAGAATCGCTATCAGTTTGTCTTGATCTATGTTAAATAAGATTAGAATGAAGATCTAATACTCTTTAG
TCTTTAACAGAATAGTATAACATGGTAGTGTGTAAGATCATAACGTGATGGGATTCTAGTCAATTGGCATGTTGACTCTG
GAAGGCGAACTCTTGTCATGTTGGTTGTTTGCAATCTTTCTTCCTATATGATGGTACATGCTGGAAATTTTGGATCTTAA
GCTTTGACCCACTCATGAATTCATGATTGACGATCACTTCTTTTGGCCGAAATGTTCGATTTTGACTTCTGTGGTTCTCC
TTTAAGCTTGAGGAAAGAAGGATAGAGTGATGACGGGGACACCTTGACAGAGAAACATTGCGGTTTATGCATAACTCTAG
AGAGCTACTTTACAAATTGTGCTCTATTTATAGATGAATGCCAAGCAGAAAATTCACTAAATTGTTTTAAGTTTGTTTTG
ATAAAATAAAAAAAAGTGCTCGAGTCATGAATGTTTAATTTTGGTATTTAAGAAATCACGGATGCTGTTGCCGAGAATTC
TTTTTACCTTGTTAGCATTAATTTCATCATGATCTGAAGTAGCTCTAGCTTAATTTGGTATTCTTTGTTAGCTAAGACAT
GATATTTCCTCTTTTGGTGCATATGCAAATTCCTATCTTTTCCCCCTAAAATCTGCAAATGTTTAATCAGATTTTTATAT
GCCATCTCAATTAACATTTATATACTGTTATGGATTCATCTGGGTAAACTTTTTTAGTCTTATCTCAGTCTGGTTGATTC
ATCTTGTTCCACATGTGACATGTCTTGATAATGTAGTGACAGACAAGCCATTTATGCTAATTTCATACATCCTCATGATA
ACCGGGCTGCGTTTGTGTGCAATGTTGGCTGTCTGTTTGATGTGCTTTTCTTTCTTTCTATGATTTATATGTTTGTAATT
TGCACTGATAGAGCAAAATGTTCTGTTCTGCTTGTTTGTTTTTACCACCTCCCATGGAATAAATGGTTTATAGTTTCCTT
CTTTTGCTATGTACAGGTCATGAACGACTAATCTCCGTTCATTTATTATGGACTTGCACCAGTTATTAAAGTACAGATTG
ACTGGCGCTAACGTTGTGTACGAAATTCCTACAGAGAACAACTTGCAAAACTCTCCCTGGCAAGCTAACCCACTGAAGTA
CGAGTTCAGCGATTCCCCATACACCCCTCTCTCTTCCCAATTTGAGTGTGACAATTTGTCTGCTCTTACCAACACTCCAG
ATAACCAGAGCTCTACAGAAACCATTTCAGCCCAACCAATCTCCCCATTGGAAGCAGACAGCTCATATAGACAGGCGGGT
ATTCTTCTCCAGGAGAACATTCAAGTGGGAGCTGATCCGTTGTATGCTACATCAAGACATAATATGCAACACGCCTTACG
GGAGATTGAGACTGTTCTGATGGCACCTGATACAGATGATGCAACAACTAGCACCAAGCATGAGTTTGAGGAAATCAAGC
CTGCTCAGCTTGTGAGGCAGCGGTCGAGAACATGGAGTCATGAATCACGGCAGCCGTTACCTGGAGTTGGTCGGTCACAG
TTTGCATCTGGTGGATACCCCACAGCAAGCTATGAATTTCGTCCAGAGAAACGGCAAAGGGAGTTAAGGGAAGACCCTCA
GATAATTGTGAAGCAGCTATTAACCAGGTGTGCTGAGGCTCTGAGTGAGGACAGGACAGAGGAGTTCCATAAGCTTGTTC
AGGAGGCTCGTGGAGTGGTCTCAATCAACGGGGAACCAATCCAACGTCTAGGTGCTTACCTACTGGAGGGTTTGGTTGCT
AGACATGGAAACTCTGGCACAAACATCTACCGTGCTCTGAAGTGCCGTGAGCCAGAGAGCAAGGAGCTCCTGTCCTACAT
GAGAATTCTATACAATATCTGCCCTTACTTCAAGTTTGGCTATATGGCAGCCAATGGGGCGATTGCAGAAGCATTGAGAA
CTGAGAACAATATCCACATAATTGATTTTCAGATTGCTCAAGGGACTCAATGGATCACACTGATACAAGCATTAGCTGCA
AGGCCTGGTGGTCCTCCTCGTGTGCGGATCACCGGGATAGATGACCCAGTGTCAGAGTATGCTCGTGGTGAAGGTCTTGA
CATTGTGGGGAAAATGTTGAAAAGCATGTCTGAAGAATTCAAAATACCTCTGGAGTTTACGCCTCTGTCTGTCTATGCCA
CACAAGTCACGAAAGAGATGCTTGAGATCAGGCCAGGTGAAGCACTGTCTGTAAACTTCACACTCCAGCTACACCACACC
CCGGACGAGAGCGTGGATGTCAACAACCCACGCGATGGTCTGCTCCGGATGGTGAAAGGGCTGTCCCCGAAGGTGACTAC
TTTGGTAGAGCAGGAGTCACACACCAACACAACGCCTTTCTTGATGAGGTTTGGGGAGACCATGGAGTACTACTCCGCCA
TGTTCGAGTCGATCGACGCCAACCTGCCGCGGGACAACAAGGAGAGGATCAGCGTGGAGCAGCACTGCCTCGCCAAGGAC
ATCGTCAACATCATCGCCTGCGAGGGGAAGGACAGGGTGGAGAGGCATGAGCTGCTTGGCAAGTGGAAGTCGAGGCTGAC
CATGGCCGGCTTCAGGCCTTACCCGTTGAGCTCGTACGTCAACTCGGTGATAAGGAAGCTTCTCGCCTGCTACTCCGATA
AATACACATTGGATGAGAAGGACGGCGCGATGCTTCTCGGCTGGAGGAGCAGAAAGCTGATATCTGCTTCTGCGTGGCAC
TGACAGTAGTATGTTGAGAAGAAGAAATGATAGAGAGCATATATATACATACCGATTCCCTGGTCCAATTTACAGAATGA
TACCCTTTGATGTACCCTCAGTTTGATTGACAGCGGGTGTTGCCTTCATTTATAGACCAGGATCAAACATTGACATTAGA
TTTGTAGAGAAGTTGTATGATAATAATTTTCTTTGCGATTCAATGTATGATGTATCCATGGCTGCCATAGTTAATAATAT
TCTTGCATAGTCATATTGATCGGCTAATCACCCGTGGATTAGCGTTGTGTCTGTCTTGACAAT
SEQ ID NO:3
ATGGACTTGCACCAGTTATTAAAGTACAGATTGACTGGCGCTAACGTTGTGTACGAAATTCCTACAGAGAACAACTTGCA
AAACTCTCCCTGGCAAGCTAACCCACTGAAGTACGAGTTCAGCGATTCCCCATACACCCCTCTCTCTTCCCAATTTGAGT
GTGACAATTTGTCTGCTCTTACCAACACTCCAGATAACCAGAGCTCTACAGAAACCATTTCAGCCCAACCAATCTCCCCA
TTGGAAGCAGACAGCTCATATAGACAGGCGGGTATTCTTCTCCAGGAGAACATTCAAGTGGGAGCTGATCCGTTGTATGC
TACATCAAGACATAATATGCAACACGCCTTACGGGAGATTGAGACTGTTCTGATGGCACCTGATACAGATGATGCAACAA
CTAGCACCAAGCATGAGTTTGAGGAAATCAAGCCTGCTCAGCTTGTGAGGCAGCGGTCGAGAACATGGAGTCATGAATCA
CGGCAGCCGTTACCTGGAGTTGGTCGGTCACAGTTTGCATCTGGTGGATACCCCACAGCAAGCTATGAATTTCGTCCAGA
GAAACGGCAAAGGGAGTTAAGGGAAGACCCTCAGATAATTGTGAAGCAGCTATTAACCAGGTGTGCTGAGGCTCTGAGTG
AGGACAGGACAGAGGAGTTCCATAAGCTTGTTCAGGAGGCTCGTGGAGTGGTCTCAATCAACGGGGAACCAATCCAACGT
CTAGGTGCTTACCTACTGGAGGGTTTGGTTGCTAGACATGGAAACTCTGGCACAAACATCTACCGTGCTCTGAAGTGCCG
TGAGCCAGAGAGCAAGGAGCTCCTGTCCTACATGAGAATTCTATACAATATCTGCCCTTACTTCAAGTTTGGCTATATGG
CAGCCAATGGGGCGATTGCAGAAGCATTGAGAACTGAGAACAATATCCACATAATTGATTTTCAGATTGCTCAAGGGACT
CAATGGATCACACTGATACAAGCATTAGCTGCAAGGCCTGGTGGTCCTCCTCGTGTGCGGATCACCGGGATAGATGACCC
AGTGTCAGAGTATGCTCGTGGTGAAGGTCTTGACATTGTGGGGAAAATGTTGAAAAGCATGTCTGAAGAATTCAAAATAC
CTCTGGAGTTTACGCCTCTGTCTGTCTATGCCACACAAGTCACGAAAGAGATGCTTGAGATCAGGCCAGGTGAAGCACTG
TCTGTAAACTTCACACTCCAGCTACACCACACCCCGGACGAGAGCGTGGATGTCAACAACCCACGCGATGGTCTGCTCCG
GATGGTGAAAGGGCTGTCCCCGAAGGTGACTACTTTGGTAGAGCAGGAGTCACACACCAACACAACGCCTTTCTTGATGA
GGTTTGGGGAGACCATGGAGTACTACTCCGCCATGTTCGAGTCGATCGACGCCAACCTGCCGCGGGACAACAAGGAGAGG
ATCAGCGTGGAGCAGCACTGCCTCGCCAAGGACATCGTCAACATCATCGCCTGCGAGGGGAAGGACAGGGTGGAGAGGCA
TGAGCTGCTTGGCAAGTGGAAGTCGAGGCTGACCATGGCCGGCTTCAGGCCTTACCCGTTGAGCTCGTACGTCAACTCGG
TGATAAGGAAGCTTCTCGCCTGCTACTCCGATAAATACACATTGGATGAGAAGGACGGCGCGATGCTTCTCGGCTGGAGG
AGCAGAAAGCTGATATCTGCTTCTGCGTGGCACTGA。
Claims (10)
1.一种水稻OSGRAS34基因,该基因具有如下(1)或(2)所述的核苷酸序列:
(1)如SEQ ID NO:3所示的编码区核苷酸序列;
(2)与SEQ ID NO:3所示的核苷酸序列具有90%以上的同源性,且编码相同功能蛋白质的核苷酸序列。
2.根据权利要求1所述的水稻OSGRAS34基因,其特征在于,该基因具有如SEQ ID NO:2所示的基因组核苷酸序列。
3.权利要求1所述的水稻OSGRAS34基因编码的蛋白。
4.根据权利要求3所述的蛋白,其特征在于,该蛋白具有如下(a)或(b)所示的氨基酸残基序列:
(a)如SEQ ID NO:1所示的氨基酸序列;
(b)将SEQ ID NO:1所示的氨基酸序列经过一个或若干个氨基酸残基的取代和/或缺失和/或添加衍生得到的具有相同功能的氨基酸序列。
5.含有权利要求1或2所述水稻OSGRAS34基因的表达盒、重组载体、转基因细胞系或转基因重组菌。
6.用于干扰、抑制、沉默、靶向敲除或定点突变权利要求1或2所述水稻OSGRAS34基因的生物材料,其特征在于,该生物材料包括以下(1)~(6)中的至少一种:
(1)针对水稻OSGRAS34基因的干扰序列;
(2)用于干扰的水稻OSGRAS34基因的干扰载体;
(3)包含有水稻OSGRAS34基因干扰载体的转基因细胞系;
(4)基于基因编辑技术靶向敲除或定点突变水稻OSGRAS34基因的引物序列;
(5)基于基因编辑技术靶向敲除或定点突变水稻OSGRAS34基因的载体;
(6)基于基因编辑技术靶向敲除或定点突变水稻OSGRAS34基因的细胞系。
7.权利要求1或2中所述的水稻OSGRAS34基因,权利要求3或4中所述的蛋白,或权利要求5中所述的表达盒、重组载体、转基因细胞系或转基因重组菌在调控水稻苗期耐盐性或培育高耐盐性水稻中的应用。
8.权利要求6所述的用于干扰、抑制、沉默、靶向敲除或定点突变水稻OSGRAS34基因的生物材料在提高水稻苗期耐盐性或培育高耐盐性水稻中的应用。
9.根据权利要求7或8所述的应用,其特征在于,通过基因编辑、干扰、抑制或沉默权利要求1或2所述水稻OSGRAS34基因的表达提高水稻苗期耐盐性,培育高耐盐性水稻。
10.一种培育高耐盐性水稻的方法,其特征在于,通过基因编辑、干扰、抑制或沉默权利要求1或2所述水稻OSGRAS34基因提高水稻苗期耐盐性,培育高耐盐性水稻。
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