CN102485896B - 水稻穗粒数调控基因OsNAC2及其表达系统和应用 - Google Patents
水稻穗粒数调控基因OsNAC2及其表达系统和应用 Download PDFInfo
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- CN102485896B CN102485896B CN 201010567349 CN201010567349A CN102485896B CN 102485896 B CN102485896 B CN 102485896B CN 201010567349 CN201010567349 CN 201010567349 CN 201010567349 A CN201010567349 A CN 201010567349A CN 102485896 B CN102485896 B CN 102485896B
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Abstract
本发明公开了一种水稻穗粒数调控基因OsNAC2及其表达系统和应用。本发明水稻穗粒数调控基因OsNAC2的核苷酸序列如SEQIDNO:1所示,其编码的蛋白质的氨基酸序列如SEQIDNO:2所示。本发明水稻穗粒数调控基因OsNAC2的克隆有助于研究水稻穗发生发育的分子机制,对生产上培育具有优良株型结构的高产水稻品种,或是通过转基因的方法改良其它作物的高产株型结构具有很大的应用价值。
Description
技术领域
本发明属于植物基因工程领域,具体涉及一种水稻穗粒数调控基因OsNAC2及其表达系统和应用。
背景技术
水稻是世界上重要的粮食作物之一,其产量和品质的不断提高是育种工作的主要目标。穗粒数是水稻产量构成最重要的因素之一,国际水稻研究所把穗粒数多作为水稻理想株型的目标之一。因而,穗粒数相关基因的克隆和研究,不但对理解水稻穗型发育具有重要的意义,同时,对水稻育种具有非常重要的应用价值。
NAC 转录因子是近十几年来发现的具有多种生物功能的一类植物特有的转录因子。第一个NAC转录因子是由Souer等(Souer E, van Houwelingen A, Kloos D, Mol J, Koes R (1996). The No Apical Meristem gene of Petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primordia boundaries. Cell 85, 159–170.)从矮牵牛中克隆得到的,它影响矮牵牛顶端分生组织的形成与分化。Aida 等(Aida M, Ishida T, Fukaki H, Fujisawa H, Tasaka M (1997). Genes involved in organ separation in Arabidopsis: an analysis of cup-shaped cotyledonmutant. Plant Cell 9, 841–857.)报道,NAC 结构域在矮牵牛NAM、拟南芥ATAF1/2 和CUC2 编码蛋白的N 端都包含一段保守的氨基酸序列,取三基因首字母命名为NAC。此后,人们就将含有NAC 结构域的蛋白统称为NAC转录因子。水稻中有140多个NAC 成员,已有的研究表明, NAC 蛋白参与植物生长发育和器官的模式建成,在植物信号转导以及非生物损伤和胁迫应答过程中,NAC作为转录因子起激活或抑制应答的功能。
在侧根发育过程中,NAC基因参与调节侧根的形成和发育,拟南芥NAC1基因主要在根尖和侧根生长原基表达,受IAA的诱导,能促进侧根的形成(Xie Q, Frugis G, Colgan D, Chua NH (2000). Arabidopsis NAC1 transduces auxin signal downstream of TIR1 to promote lateral root development. Genes Dev 14, 3024–3036.)。拟南芥的AtNAC2基因也在根中特异表达,并受到IAA的上调,能显著增加侧根数目(He XJ, Mu RL, Cao WH, Zhang ZG, Zhang JS, Chen SY (2005). AtNAC2, a transcription factor downstream of ethylene and auxin signaling pathways, is involved in salt stress response and lateral root development. Plant J 44, 903–916.)。植物开花受到多种逆境的影响,例如高盐和低温等,盐胁迫诱导NTL8的表达,并延迟开花;过表达NTL8蛋白的拟南芥出现生长缓慢及开花延迟的现象(Kim SG, Kim SY, Park CM (2007). A membraneassociated NAC transcription factor regulates saltresponsive flowering via FLOWERING LOCUST in Arabidopsis. Planta 226, 647–654.)。拟南芥中近1/5(20/107)的NAC基因与叶片衰老相关(Guo Y, Cai Z, Gan S (2004). Transcriptome of Arabidopsis leaf senescence. Plant Cell Environ 27, 521–549.)。其中几个基因已被证实参与了衰老的调控过程。最近从水稻和大麦(Hordeum vulgare)中同时鉴定出一个新的铁匮乏应答元件IDE2(iron deficiencyresponsivecis-acting element 2)结合蛋白IDEF2,它是NAC蛋白家族中的一员。
目前,没有发现与水稻穗粒数有关的NAC类转录因子。
发明内容
本发明的目的在于根据现有的水稻穗粒数相关的NAC类转录因子尚未有研究,提供一种与水稻穗粒数相关的NAC转录因子OsNAC2。
本发明另一目的在于提供上述OsNAC2基因编码的蛋白质。
本发明另一目的在于提供包含上述OsNAC2基因的质粒。
本发明另一目的在于提供包含上述质粒的表达载体。
本发明另一目的在于提供包含上述表达载体的转基因植物细胞。
本发明还有一个目的在于提供上述OsNAC2基因的应用。
本发明上述目的通过以下技术方案予以实现:
一种与水稻穗粒数相关的NAC转录因子,命名为OsNAC2,具体可如下:
(1)其核苷酸序列如SEQ ID NO:1或3所示;
(2)在SEQ ID NO:1或3所示序列中添加、取代、插入或缺失一个或多个核苷酸生成突变体、等位基因或衍生物;
(3)在严格条件下可与SEQ ID NO:1或3所限定的DNA序列杂交并且编码上述与水稻的穗粒数相关蛋白的DNA分子,其同源性最好为95%以上;
(4)与SEQ ID NO:1或3所示的基因序列具有90%以上的同源性,并且编码与水稻穗粒数相关的DNA分子。
本发明水稻穗粒数调控基因OsNAC2编码的蛋白质的氨基酸序列具体可以如下所示:
(1)其氨基酸序列如SEQ ID NO:2所示;
(2)在SEQ ID NO:2所示序列中添加、取代、插入或缺失一个或者多个氨基酸所生成的衍生物。
SEQ ID NO:1由1401个碱基组成,自5’末端第105个碱基开始为编码区,编码具有序列表中序列2的氨基酸残基序列的蛋白质。
SEQ ID NO:3由1401个碱基组成,编码序列为自5’端105碱基到1032 碱基,编码具有序列表中序列2的氨基酸残基序列的蛋白质。
本发明所述的OsNAC2启动子也属于本发明的保护范围。
OsNAC2基因启动子,是如下①或②的DNA 分子:
(1)其核苷酸序列如SEQ ID NO:4所示;
(2)在严格条件下可与如SEQ ID NO:4所示的DNA 分子杂交具有启动子功能的DNA分子。
上述的严格条件可为在6×SSC,0.5%SDS的溶液中,在65℃下杂交,然后用2×SSC,0.1% SDS和1×SSC,0.1% SDS各洗杂交膜一次。
扩增上述OsNAC2基因及其启动子全长或是任一片段的引物对也属于本发明的保护范围。
含有上述与水稻的穗粒数相关蛋白编码基因及其启动子的重组载体、转基因细胞系或是重组菌也属于本发明的保护范围。
可用现在的植物表达载体构建含有OsNAC2基因及其启动子的重组表达载体。所述植物表达载体包括双元农杆菌载体和可用于将植物微蛋轰击的载体等,如pCAMBIA3301 、pCAMBIA1300、pCAMBIA2301、pBI121或其他衍生植物表达载体。携带有本发明的与水稻的穗粒数相关蛋白编码基因OsNAC2及其启动子的植物表达载体可以通过Ti质粒、Ri质粒、植物病毒载体、直接DNA转化、显微注射、电导、农杆菌介导等常规生物学方法转化到植物细胞或是组织中。被转化的宿主植物可以是水稻。
使用OsNAC2基因构建重组植物表达载体时,其转录起始核苷酸前可加上任何一种增强型、组成型、组织特异型或诱导型启动子,如花椰菜花叶病毒(CaMV)35S启动子、泛生素基因Ubiquitin启动子(Pubi)等,他们可单独使用或与其他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,还可以使用增强子,包括翻译增强子或者转录增强子,这些增强子区域可以是ATG起始密码子或是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区域。
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可以产生颜色变化的酶或是发光化合物的基因(如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(如抗除秀剂基因)等。
本发明水稻穗粒数调控基因OsNAC2可用于改变水稻穗粒数。
本发明所提供的改变水稻穗粒数的方法有两种,一种是将OsNAC2基因导入目的水稻组织或是细胞中,得到穗粒数改变的转基因水稻。另一种是沉默目的水稻中含有的OsNAC2基因,得到穗粒数改变的转基因水稻。
与现有技术相比,本发明具有如下有益效果:
本发明转OsNAC2基因实验结果表明,转OsNAC2基因植株表现为穗粒数增加,其蛋白质和编码基因对水稻穗粒数的调控具有重要的实际意义,在实际应用中可以将OsNAC2基因转入不同的水稻品种中,以培育更加理想的水稻栽培品种。OsNAC2蛋白及其编码基因在农业领域具有广阔的应用和市场前景。
附图说明
图1 经农杆菌介导得到的转化植株分化及生根;
图2 转化植株外源基因PCR检测,泳道M为分子量标记DL2000(购自TAKARA);泳道1为空白对照,泳道2为对照植株中花11,泳道3为阳性植株对照,泳道4-25为OsNAC2基因转化植株;
图3 转化植株与对照植株形态图,CK:对照植株;13-3:转化植株;
图4:转化植株与对照植株穗部特征图,CK:对照植株;13-3:转化植株。
具体实施方式
下列实施例中所用方法如无特别说明均为常规方法。所用引物合成及测序工作由英潍捷基(上海)贸易有限公司完成。
实施例 OsNAC2基因的获得
根据NCBI(http://www.ncbi.nlm.nih.gov/)提供的关于该基因的cDNA序列设计引物,引物序列如下:(3-1234bp)
NACF如SEQ ID NO:5所示;
NACR如SEQ ID NO:6所示。
以粳稻品种中花11号2周的幼苗cDNA为模版,在引物NACF和NACR的引导下,用常规方法扩增OsNAC2基因。反应结束后,对PCR扩增产物进行1%的琼脂糖凝胶电泳,回收并纯化大约1200bp左右的DNA片段,将该片段克隆到pMD18-T载体(购自TAKARA公司)上,获得pMD18-T-OsNAC2载体,送英潍捷基(上海)贸易有限公司测序,测序结果表明,该DNA片段的序列如如SEQ ID NO:1所示;编码序列表中序列2所示的蛋白。
根据序列表中序列1的核苷酸序列设计引物对NACEF和NACER,并在引物两端分别引入限制性内切酶HindIII 和SpeI识别位点和保护碱基,引物序列如下:
NACEF如SEQ ID NO:7所示: aaaaaaagcttacaacgatttcctcttgtcacc (下划线为限制性内切酶HindIII识别位点)
NACER如SEQ ID NO:8所示: aaaaaactagtgtagatgcctcgatcgcgatc (下划线为限制性内切酶SpeI识别位点)
以pMD18-T-OsNAC2载体DNA为模版,在引物NACEF和NACER的引导下,用常规方法扩增OsNAC2基因。反应结束后,对PCR扩增产物进行1%的琼脂糖凝胶电泳,回收并纯化大约1200bp左右的DNA片段,将该片段克隆到pMD18-T载体(购自TAKARA公司)上,获得pMD18-T-OsNAC2-E载体,送英潍捷基(上海)贸易有限公司测序,测序结果表明,该DNA片段的序列如SEQ ID NO:1所示,在其DNA两端引入了合适的没切位点;编码序列表中如SEQ ID NO:2所示的蛋白。
根据序列表中如SEQ ID NO:1所示的核苷酸序列设计引物对MNACF2和MNACR2,引物序列如下:
MNACF2如SEQ ID NO:9所示;
MNACR2如SEQ ID NO:10所示。
以pMD18-T-OsNAC2-E载体DNA为模版,在引物NACEF和MNACR2的引导下,用常规方法扩增OsNAC2-F基因片段;在引物NACER和MNACF2的引导下,用常规方法扩增OsNAC2-R基因片段,反应结束后,分别对PCR扩增产物进行1%的琼脂糖凝胶电泳,分别回收并纯化大约800bp和400bp左右的DNA片段,即OsNAC2-F和OsNAC2-R片段,以OsNAC2-F和OsNAC2-R片段为模版,在引物NACEF和NACER的引导下,用常规方法扩增mOsNAC2基因全长,反应结束后,对PCR扩增产物进行1%的琼脂糖凝胶电泳,回收并纯化大约1200bp左右的DNA片段,将该片段克隆到pMD18-T载体上,获得pMD18-T-mOsNAC2载体,送英潍捷基(上海)贸易有限公司测序,测序结果表明,该DNA片段的序列如序列3所示,编码序列表中序列2所示的蛋白。
将mOsNAC2基因克隆入植物表达载体pCAMBIA1380 多克隆位点的HindIII和SpeI酶切位点之间,得到含有OsNAC2基因的植物表达载体,然后利用农杆菌介导的方法转化粳稻品种中花11号的成熟胚的愈伤组织,方法如下述文献描述:周玲艳,姜大刚,吴豪等,基于TAC载体的水稻转化系统的建立. 遗传学报,2005,32:514-518。经过预分化、分化,得到21株转化植株,图1,经过PCR鉴定,全部为阳性,PCR引物序列如下:
引物1如SEQ ID NO:11所示;
引物2如SEQ ID NO:12所示。
PCR扩增条件为:94 ℃ 2 min;94℃ 30 sec,58 ℃ 30 sec,72℃ 1 min,35个循环;72 ℃ 5 min。反应结束,对扩增产物进行1%琼脂糖凝胶电泳检测,检测结果如图2 所示,阳性植株可以扩增得到约800bp的条带。
将经过PCR鉴定为阳性的转基因植株培养至成熟,观察不同时期的植株形态,结果如图 3所示,所得到的21株植株皆表现出穗粒数明显增多的表型。转化植株与对照植株相比,在以下四个产量性状有明显变化:一级枝梗数增加;二级枝梗数增加;主穗粒数增加;单株产量增加。
图4显示的是一株转OsNAC2基因和一株对照植株的穗部图片。
SEQUENCE LISTING
<110> 华南农业大学
<120> 水稻穗粒数调控基因OsNAC2及其表达系统和应用
<130>
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<170> PatentIn version 3.2
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atacaacgat ttcctcttgt caccctgaat ctacttctgc tgcaaaagca ataagcaagg 60
agcagttagc caggtaaagc tctagctagc tagcttaggc agcaatggag cagcatcagg 120
gccaggcagg catggacttg ccccctggct tccgcttcca cccgaccgac gaggagctga 180
tcacgcacta cctcgccaag aaggtcgccg acgcccgctt cgccgccctc gccgtcgccg 240
aggccgacct caacaagtgc gagccctggg acctgccatc tctggcgaag atgggggaga 300
aggagtggta cttcttctgc ctcaaggaca ggaagtaccc gacggggctg aggacgaaca 360
gggcgacgga gtccgggtac tggaaggcca cggggaagga caaggacatc ttcagacgga 420
aggccctcgt cggcatgaag aagacgctcg ttttctacac ggggcgcgct cccaaggggg 480
agaagtctgg ctgggtcatg cacgagtacc gcctccacgg caagctccac gccgccgccc 540
tcggcttcct ccacggcaag cccgcgtcgt ccaagaacga gtgggtgttg tgcagggtgt 600
tcaagaagag cctcgtggag gtgggcgcgg cgggagggaa gaaggcggcc gtggtgacga 660
tggagatggc gaggggaggg tcgacgtcgt cgtccgtggc ggacgagatc gccatgtcgt 720
ccgtcgtcct ccctccgctg atggacatgt ccggagccgg cgccggcgcc gtcgacccgg 780
cgacgacggc gcacgtgacc tgcttctcca acgcgctgga gggccagttc tttaacccga 840
cggcagtaca cgggcacggc ggcggcgact cctcgccgtt catggcgagc ttcacgcagt 900
acgggcagct gcaccacggc gtgagcctgg tgcaactcct ggagagctgc aacggctacg 960
gcggcctcgt cgacatggca gcgtccggca gccagctgca gccggcggcg tgcggcggcg 1020
agcgggagag gcttagcgcg tcgcaggaca ccggcctcac ctccgacgtg aacccggaga 1080
tctcgtcatc ctccggccaa aaattcgacc acgaggccgc gctatggggc tactaaggtt 1140
tgatatgatc agcgccgtgt acgttaatcg cgggcgatta gcgaagtagc gattactgta 1200
aataaaacca tgagatcgcg atcgaggcat ctaccaaggt tcgcttaatt tgcttgtacc 1260
tataggtgta gattatttgg tgatttggga gatgtaatta gcattgtttg tttgtaattc 1320
gtatcagaat gatcgatgca gtattgtttg gagggttagt taagcaattg gccactcctc 1380
agttgccagt taattagtta g 1401
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Met Glu Gln His Gln Gly Gln Ala Gly Met Asp Leu Pro Pro Gly Phe
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Arg Phe His Pro Thr Asp Glu Glu Leu Ile Thr His Tyr Leu Ala Lys
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Lys Val Ala Asp Ala Arg Phe Ala Ala Leu Ala Val Ala Glu Ala Asp
35 40 45
Leu Asn Lys Cys Glu Pro Trp Asp Leu Pro Ser Leu Ala Lys Met Gly
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Glu Lys Glu Trp Tyr Phe Phe Cys Leu Lys Asp Arg Lys Tyr Pro Thr
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Gly Leu Arg Thr Asn Arg Ala Thr Glu Ser Gly Tyr Trp Lys Ala Thr
85 90 95
Gly Lys Asp Lys Asp Ile Phe Arg Arg Lys Ala Leu Val Gly Met Lys
100 105 110
Lys Thr Leu Val Phe Tyr Thr Gly Arg Ala Pro Lys Gly Glu Lys Ser
115 120 125
Gly Trp Val Met His Glu Tyr Arg Leu His Gly Lys Leu His Ala Ala
130 135 140
Ala Leu Gly Phe Leu His Gly Lys Pro Ala Ser Ser Lys Asn Glu Trp
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Val Leu Cys Arg Val Phe Lys Lys Ser Leu Val Glu Val Gly Ala Ala
165 170 175
Gly Gly Lys Lys Ala Ala Val Val Thr Met Glu Met Ala Arg Gly Gly
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Ser Thr Ser Ser Ser Val Ala Asp Glu Ile Ala Met Ser Ser Val Val
195 200 205
Leu Pro Pro Leu Met Asp Met Ser Gly Ala Gly Ala Gly Ala Val Asp
210 215 220
Pro Ala Thr Thr Ala His Val Thr Cys Phe Ser Asn Ala Leu Glu Gly
225 230 235 240
Gln Phe Phe Asn Pro Thr Ala Val His Gly His Gly Gly Gly Asp Ser
245 250 255
Ser Pro Phe Met Ala Ser Phe Thr Gln Tyr Gly Gln Leu His His Gly
260 265 270
Val Ser Leu Val Gln Leu Leu Glu Ser Cys Asn Gly Tyr Gly Gly Leu
275 280 285
Val Asp Met Ala Ala Ser Gly Ser Gln Leu Gln Pro Ala Ala Cys Gly
290 295 300
Gly Glu Arg Glu Arg Leu Ser Ala Ser Gln Asp Thr Gly Leu Thr Ser
305 310 315 320
Asp Val Asn Pro Glu Ile Ser Ser Ser Ser Gly Gln Lys Phe Asp His
325 330 335
Glu Ala Ala Leu Trp Gly Tyr
340
<210> 3
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<212> DNA
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<400> 3
atacaacgat ttcctcttgt caccctgaat ctacttctgc tgcaaaagca ataagcaagg 60
agcagttagc caggtaaagc tctagctagc tagcttaggc agcaatggag cagcatcagg 120
gccaggcagg catggacttg ccccctggct tccgcttcca cccgaccgac gaggagctga 180
tcacgcacta cctcgccaag aaggtcgccg acgcccgctt cgccgccctc gccgtcgccg 240
aggccgacct caacaagtgc gagccctggg acctgccatc tctggcgaag atgggggaga 300
aggagtggta cttcttctgc ctcaaggaca ggaagtaccc gacggggctg aggacgaaca 360
gggcgacgga gtccgggtac tggaaggcca cggggaagga caaggacatc ttcagacgga 420
aggccctcgt cggcatgaag aagacgctcg ttttctacac ggggcgcgct cccaaggggg 480
agaagtctgg ctgggtcatg cacgagtacc gcctccacgg caagctccac gccgccgccc 540
tcggcttcct ccacggcaag cccgcgtcgt ccaagaacga gtgggtgttg tgcagggtgt 600
tcaagaagag cctcgtggag gtgggcgcgg cgggagggaa gaaggcggcc gtggtgacga 660
tggagatggc gaggggaggg tcgacgtcgt cgtccgtggc ggacgagatc gccatgtcgt 720
ccgtcgtcct ccctccgctg atggacatgt ccggagccgg cgccggcgcc gtcgacccgg 780
cgacgacggc ccatgtcacg tgcttctcga acgcgctgga gggccagttc tttaacccga 840
cggcagtaca cgggcacggc ggcggcgact cctcgccgtt catggcgagc ttcacgcagt 900
acgggcagct gcaccacggc gtgagcctgg tgcaactcct ggagagctgc aacggctacg 960
gcggcctcgt cgacatggca gcgtccggca gccagctgca gccggcggcg tgcggcggcg 1020
agcgggagag gcttagcgcg tcgcaggaca ccggcctcac ctccgacgtg aacccggaga 1080
tctcgtcatc ctccggccaa aaattcgacc acgaggccgc gctatggggc tactaaggtt 1140
tgatatgatc agcgccgtgt acgttaatcg cgggcgatta gcgaagtagc gattactgta 1200
aataaaacca tgagatcgcg atcgaggcat ctaccaaggt tcgcttaatt tgcttgtacc 1260
tataggtgta gattatttgg tgatttggga gatgtaatta gcattgtttg tttgtaattc 1320
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tgtagtagtg taaattagac atatgaattc tttaaccaga catttgaatt ccccatcagc 60
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ctgacagccg cggagaataa tgcacagtag catggcgttt gtcacacata tccgttctaa 180
gccccacaaa accaagtgga tcctctgtgc tgtgctgtgc attgcatgta cctgcagggc 240
tgcaaagaga acacaggtac attggctctg cagatctgca tattatctta cggcctttgc 300
agaaacggag agaataattc tcccccattg gattgcatgt atacaagcag cagaatgttg 360
tgcacatcca ctgctattac acaatagtat cagcctgtga ccaatgtgct gtcactgggc 420
tacaggatca tcgtgccact gccagtttgt caccgatttg attggcttat ctcctgggag 480
attctccttt taagggttta agtacccctg attctaatca ggttcagggc aaatgattat 540
acgtactact gtacaatccc tgcagaaaat tacgcacaaa gaccaaatat caaggtttgc 600
tatggactga atatatcttt tgtttgcgcc tggacgtcgg tagctacccc agatattaat 660
taattatttg aattagtgat tttgtaccaa atgatcgaac cagctcatgc ttagcatgaa 720
gctaataaat tctcccgttt attgcagtac tcgaatacaa atagtaggtg tactactcgc 780
catggagtct gctctaattc gttctcaaaa gcttctggag acgaaggatt ggtttggtcg 840
ctttctcggt tgctgctcgg ctcgcgtgtc gcatatgacg ctttacttaa tgctgctagt 900
agtactcata atttccatta attaagactc atttaattag taatgcacca aatccttaaa 960
tccagccaga tcagatcaaa gtcatcctag ggtaaaacac tctgatctga tggatcacta 1020
gccactaaat taatccggtg attgatcatg tgtggaagca agttttggct cgttacaagg 1080
tactattaca taacactacg ctgtcctgta tagctatctc atctatctct tcccatgcat 1140
ttttcaccta ctagctagta ctccatccgt tttaaaatgt ttgacaccgt tgacttttta 1200
gcacatgttt gaccgttcgt cttattcaaa aatttttgtg aaatatgtaa aactatatgt 1260
gtacatgaaa gtatatttaa caataaatcg aatgatatga aaagaataaa taactactta 1320
aattttttga ataagacgaa tggttaaaca cgtactaaaa agtcaacggt gtcaaacatt 1380
ttgaaatgga gggagtagta acctgtgatg agtttccttc atcacccacg tctctttcta 1440
ctaaaaagac catcatgtag ctcgcctcta gtatttaagg ggcaccacca cccctcccac 1500
tttatttcct ctatacaacg atttcctctt gtcaccctga atctacttct gctgcaaaag 1560
gtaattagca tatcctaact atttatcacc tcatccctca agtgtgttct tctactttct 1620
tagatgtact ttttaaactt ttctgtgcat aaacatctca tgtgctgatt ctttttgtgt 1680
gttctatgtg cttgcatacc atgtccgttt ggttcgattc gattttttaa gcatccgtgt 1740
ggtttgtttt tgttctagct tagctagcta gctagcaaag aggtgtcatg gatacatacc 1800
agctttaatt tgatttggtt tgctctacct ttaaaagtgt gtgaatcatc tccggtccta 1860
gtttatgcat atctgtgtgc atatgtacaa atgtttcagg acaggagttg gcataacagc 1920
taacaaatct tacatatata gcagcaataa gcaaggagca gttagccagg taaagctcta 1980
gctagctagc ttaggcagca 2000
<210> 5
<211> 22
<212> DNA
<213> 人工序列
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acaacgattt cctcttgtca cc 22
<210> 6
<211> 22
<212> DNA
<213> 人工序列
<400> 6
gtagatgcct cgatcgcgat ct 22
<210> 7
<211> 33
<212> DNA
<213> 人工序列
<400> 7
aaaaaaagct tacaacgatt tcctcttgtc acc 33
<210> 8
<211> 32
<212> DNA
<213> 人工序列
<400> 8
aaaaaactag tgtagatgcc tcgatcgcga tc 32
<210> 9
<211> 44
<212> DNA
<213> 人工序列
<400> 9
gcccatgtca cgtgcttctc gaacgcgctg gagggccagt tctt 44
<210> 10
<211> 41
<212> DNA
<213> 人工序列
<400> 10
cgagaagcac gtgacatggg ccgtcgtcgc cgggtcgacg g 41
<210> 11
<211> 18
<212> DNA
<213> 人工序列
<400> 11
cgccgatggt ttctacaa 18
<210> 12
<211> 18
<212> DNA
<213> 人工序列
<400> 12
ggcgtcggtt tccactat 18
Claims (2)
1.一种水稻OsNAC2基因的应用,其特征在于所述OsNAC2基因用于对水稻穗粒数进行改造;水稻OsNAC2基因的核苷酸序列如SEQ ID NO:1或3所示,蛋白质序列如SEQ ID NO:2所示。
2.根据权利要求1所述的水稻OsNAC2基因的应用,其特征在于将OsNAC2基因转化水稻细胞,再将转化后的水稻细胞培育成植株;或者是沉默目的水稻中含有的OsNAC2基因,得到穗粒数改变的转基因水稻。
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CN113215296B (zh) * | 2021-04-28 | 2022-08-09 | 广西大学 | 水稻芒长基因gna1的分子标记及其鉴定方法和应用 |
CN114085846B (zh) * | 2021-12-21 | 2022-10-04 | 华南农业大学 | 水稻OsNAC2基因在调控水稻叶片夹角中的应用 |
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