CN1908171A - 水稻胚乳直链淀粉含量控制基因du1及其应用 - Google Patents
水稻胚乳直链淀粉含量控制基因du1及其应用 Download PDFInfo
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- CN1908171A CN1908171A CN 200510088978 CN200510088978A CN1908171A CN 1908171 A CN1908171 A CN 1908171A CN 200510088978 CN200510088978 CN 200510088978 CN 200510088978 A CN200510088978 A CN 200510088978A CN 1908171 A CN1908171 A CN 1908171A
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Abstract
本发明是从水稻粳稻品种秀水11中克隆并鉴定了控制水稻胚乳直链淀粉含量的基因DU1,该基因的核苷酸序列选自:(1)编码SEQ ID NO:2所示的氨基酸序列的核苷酸序列;(2)与(1)中的核苷酸序列能够在严谨条件下杂交,并同时编码具有控制植物器官淀粉组成和水稻胚乳直链淀粉含量功能的核苷酸序列。本发明还提供了一种控制水稻胚乳直链淀粉含量的蛋白质、含有本发明的基因的植物表达载体和一种培育植物的方法,该方法可使植物淀粉的主要储藏器官的淀粉构成发生变化,包括用含有本发明的基因的表达载体转化植物细胞;和将转化的植物细胞培育成植株的步骤。
Description
技术领域
本发明涉及植物基因工程领域,更具体地,本发明涉及水稻胚乳直链淀粉含量控制基因DU1,该基因编码的蛋白质及其功能类似物,编码其的核苷酸序列,含有该核苷酸的载体和含有该载体的宿主细胞;另外,本发明还涉及控制植物胚乳直链淀粉含量的方法和改良植物胚乳直链淀粉含量育种的方法。
背景技术
水稻胚乳淀粉通常由直链淀粉和支链淀粉组成,含量高达90%左右[1]。直链淀粉是指葡萄糖单元之间以α-1,4糖苷键连接形成的、长约1000个葡萄糖残基的线性长链分子,而支链淀粉则是在此基础上以α-1,6糖苷键衍生出小的链状分子,支链平均长度约20个葡萄糖残基[2]。直链淀粉和支链淀粉的比例和特性决定淀粉的属性,也决定稻米食味品质和加工品质的最重要因素。
Wx是影响稻米品质的重要基因,水稻Wx基因首先由Okagaki克隆[3],随后Wang等克隆了Wx基因的DNA全序列[4]。研究表明,水稻Wx基因的转录能力与GBSS蛋白含量及直链淀粉含量关系密切。颗粒结合淀粉合酶(GBSS)紧密地结合在淀粉粒上,催化水稻胚乳直链淀粉的合成,其大小约为60kD[3,4]。
1981年,Satoh等[5]发现一突变体,种子在完全干燥后才表现云雾状特征,称为暗胚乳(dull endosperm,简称du),其胚乳直链淀粉含量普遍下降。研究表明,该性状受一隐性基因控制,突变材料胚乳直链淀粉含量通常在6%左右,米饭外观油润而富有光泽,食味独特,冷不回生,尤其适合开发各种方便米饭等,也是改良稻米直链淀粉含量和食味品质的优良材料[6,7]。
Isshiki等[8]通过对du突变体的研究认为DU基因编码的蛋白可能通过对调节Wx从而影响水稻胚乳淀粉的生物合成。但至今未见克隆该基因的报道。
发明内容
针对上述研究背景,本发明的一个目的是提供一种控制水稻胚乳直链淀粉含量的基因。
本发明的另一个目的是提供一种由本发明的控制水稻胚乳直链淀粉含量的基因所编码的蛋白质。
本发明的再一个目的是提供一种含有本发明的控制水稻胚乳直链淀粉含量的基因的植物表达载体。
本发明的又一个目的是提供一种培育植物的方法,该方法可使植物淀粉的主要储藏器官的淀粉构成发生变化。
本发明提供了一种控制水稻胚乳直链淀粉含量的基因,该基因的核苷酸序列选自:
(1)编码SEQ ID NO:2所示的氨基酸序列的核苷酸序列;
(2)与(1)中的核苷酸序列能够在严谨条件下杂交,并同时编码具有控制水稻胚乳直链淀粉含量的功能的核苷酸序列。
严谨杂交条件是指,将杂交膜置于预杂交液(0.25mol/L磷酸钠缓冲液,pH 7.2,7%SDS)中,65℃预杂交30min;弃预杂交液,加入杂交液(0.25mol/L磷酸钠缓冲液,pH 7.2,7%SDS,同位素标记的核苷酸片段),65℃杂交12小时;弃杂交液,加入洗膜液I(20mmol/L磷酸钠缓冲液,pH 7.2,5%SDS),65℃洗膜2次,每次30min;加入洗膜液II(20mmol/L磷酸钠缓冲液,pH 7.2,1%SDS),65℃洗膜30min。
本发明的控制水稻胚乳直链淀粉含量的基因优选具有如图7和SEQID NO:1所示的DNA序列。
本发明还提供了一种由上述核苷酸序列编码的蛋白质。该蛋白质优选具有如图8和SEQ ID NO:2所示的氨基酸序列。本发明中的SEQ ID NO:2和图8所示的蛋白质属于PRP蛋白家族,与人类的Prp1/Zer1蛋白和Prp6蛋白的同源性为51%,参与mRNA前体的剪接而调控支链淀粉的生物合成。
本发明还提供了一种含有本发明的控制水稻胚乳直链淀粉含量的基因的植物表达载体。这种表达载体可以是如图5所示的pCAMDU1,该载体可以表达由上述核酸序列编码的多肽。
本发明还提供了一种培育植物的方法,该方法可使植物淀粉的主要储藏器官的淀粉构成发生变化,包括用本发明的表达载体转化植物细胞;和将转化的植物细胞培育成植株的步骤。
附图说明
下面结合附图对理解本发明作进一步的详细描述,但并非对本发明作限定。
附图1.水稻F2代植株所结种子的表型鉴定(A,普通水稻籽粒表现为阳性;B低直链淀粉水稻胚乳表现为阴性)
附图2.DU1在水稻第10条染色体上的初步定位图
附图3.DU1基因的精细定位及物理定位
附图4.DU1基因转化du1突变体恢复正常表型(A.du1突变体胚乳表现低直链淀粉特征;B.转DU1基因后表型恢复正常)
附图5.载体pCAMDU1质粒图谱
附图6.载体pCAMDU1质粒表达的包含DU1的8.3kb基因组片段
附图7.DU1的核苷酸序列
附图8.DU1所编码的氨基酸序列
具体实施方式
下面结合附图对理解本发明作进一步的详细描述,但并非对本发明作限定。
实施例1 水稻胚乳直链淀粉含量控制基因DU1的克隆
1.水稻材料
水稻(Oryza sativa ssp.)胚乳低直链淀粉突变体du1(中国水稻研究所使用EMS(甲基磺酸乙酯)诱变获得)[6,7]和常规水稻品种明恢63(购自中国水稻研究所)。
2.分析和定位群体
纯合的籼稻品种明恢63和粳稻突变体du1进行杂交,F1代自交,共得到7,150个F2个体,并从中选出1,618个个体作为定位群体。在苗期每株取2克左右的嫩叶,用来提取DNA。
3、通过SSR(Simple Sequence Repeat),STS(Sequence-tagged Sites),和CAPS(Cleaved amplified polymorphic sequence)标记定位DU1基因
采用改进的CTAB(Cetyltrimethyl Ammonium Bromide)方法[9]从水稻叶片中提取用于基因定位的基因组DNA。取大约100mg水稻叶片,经液氮冷冻,在直径5cm的小研钵中磨成粉状,转移到1.5ml离心管里提取DNA,获得的DNA沉淀溶解于100μl超纯水中。每一个SSR、STS或CAPS反应用1μl DNA样品。
根据水稻经典遗传学的研究,DU1基因被定位于第10染色体的长臂上。我们选取了两对SSR引物(序列见表1)。我们发现DU1基因与这两个标记连锁并位于二者之间。在此基础上,我们设计的PCR引物分别以两个亲本的基因组DNA为模板进行PCR扩增(94℃预变性5min,94℃1min,55℃1min,72℃1min,35个循环,72℃延伸10mins),将DU1基因初步定位在SSR标记M2和M3之间。
为了将DU1定位在一个PAC克隆上,我们用已公布的水稻品种Nipponbare的PAC文库(http://rgp.dna.affrc.go.jp)序列构建了DU1位点附近的重叠群,设计的PCR引物分别以两个亲本(籼稻品种明恢63和du1突变体)的基因组DNA为模板进行PCR扩增(94℃预变性5min,94℃1min,55℃1min,72℃1min,35个循环,72℃延伸10mins)。以此发展了STS和SSR标记(序列见表1),并用于群体精细定位,最终将其定位在一个PAC克隆AC068923上。
4.DU1基因的精细定位
根据公布的PAC克隆AC068923的序列(http://rgp.dna.affrc.go.jp),2个新标记(序列见表1),最后通过连锁分析将DU1定位在66kb的范围之内。
表1 克隆DU1基因所用引物序列
名称 | 正向序列 | 反向序列 |
DU1 | atgtataccttggtatgcgtgc | tatgctttcccactcctgcg |
DU2 | attcacccaaaggtcccaaacc | ctagggatctgcagcatttgg |
DU3 | aaaggaagctgttggaggaagg | tacgatggcatctcctggaactg |
M1 | tcagatctacaattccatcc | tcggtgagacctagagagcc |
M2 | tcgataacacagtattcagccagg | acaaggacaaatgctatgggactc |
M3 | atgtgcaatacagtgccatgtgg | tgctattgccattgtactgctgc |
M4 | tgcactttcacctagcagtatgcc | ttccttgtgcctcacagtccatc |
a | attagccggtaaatggatgagttc | aagcaatactaatccctccaaacc |
b | aggtcttgggtcgtaccaccctgc | tcgttcgctccctggcttctcc |
c | tgttccttgtgcggttgtgc | aacacccacctccgaacacacc |
d | tgtggtgccttttattccctcc | tttcctgcacggcatacagtg |
e | aacgcgaggacacgtacttac | acgagatacgtacgcctttg |
f | aatccaacgcatcaaggctggc | acaatgccaaacaccaggaactcg |
g | tgagctttacctcccctcctaacc | tccacctttctctctcatcccac |
5.DU1部分cDNA基因的获得与功能的预测
首先对66kb的全长基因组序列用GENSCAN软件(
http://genes.mit.edu/GENSCAN.html)预测可能的编码区(ORF),发现这个区间共有12个ORF。其中一个ORF具有典型的PRP蛋白的开放阅读框;扩大序列范围预测,并将基因产物用blastX软件预测(
http://www.ncbi.nlm.nih.gov/BLAST)。用DNAStar软件(Lasergene)MegAlign程序中的ClustalW方法进行蛋白质序列比较和进化树分析。同时又设计一对引物DUF(5’tgtgaagctgtggttgcagg 3’)和DUR(5’ttcatccagaccctctcagtgc 3’)以籼稻明恢63总RNA为模板进行RT-PCR反应(70℃10mins,42℃60mins,99℃5mins,4℃5mins)并用ABI3730DNAanalyzers型测序仪测序,得到了DU1基因的cDNA序列。在上述研究的基础上推测该基因可能编码一种剪接因子,与人类的Prp1/Zer1蛋白和Prp6蛋白极其相似。
6.不同品种间DU1基因序列的比较
通过水稻不同品种间,包括野生型秀水11、亲本明恢63等多个品种(由中国水稻研究所提供)的DU1基因所在位点的DNA序列的比较,并与表型结果相对应,发现了引起表型差异的基因改变位置,表明碱基替换是造成水稻胚乳直链含量改变的遗传基础。
实施例2 水稻胚乳直链淀粉含量控制基因SU1的功能互补及转基因研究
根据籼稻明恢63 DU1基因的序列设计引物,用引物DU1F,DU1R,DU2F,DU2R,DU3F和DU3R(序列见表1)分3段高保真PCR(序列见表1,94℃预变性5min,94℃1min,60℃1min,72℃1min,35个循环,72℃延伸10mins并用ABI3730 DNA analyzers型测序仪测序(ABI公司,美国),挑选序列完全正确的克隆利用共有的Sal I和Xba I位点将它们连接成一个8.3kb片段,包含起始密码子ATG上游的3,055个碱基和终止密码子TAG后的2,115个碱基的全长序列,克隆到双元载体pCAMBIA1300(购自CAMIA公司,澳大利亚)中,获得了用于转化的质粒pCAMBIDU1(图5)。质粒通过电击的方法转入农杆菌(AgroBacterium tumefaciens)株系EHA105(购自CAMIA公司,澳大利亚)中转化水稻。将du1突变体的幼胚脱壳灭菌,接种到诱导愈伤组织的MS培养基[10]中。在28℃的培养室中暗培养3周,从盾片处生长出愈伤组织,挑选生长旺盛,颜色浅黄,比较松散的胚性愈伤组织,用作转化的受体。用含有双元质粒载体的农杆菌EHA105菌株侵染水稻愈伤组织,在黑暗处25℃培养3天后,在含有50mg/L潮霉素的选择培养基上筛选抗性愈伤组织和转基因植株。将潮霉素抗性植株在阴凉处炼苗,几天后移栽到水田,结实后收种进行表型鉴定。共收获8个株系的T0代种子,其中胚乳直链淀粉含量表现为阳性的有5个株系,证明DU1基因已经整合进受体基因组内并能够正确表达(见附图4)。
参考文献
1.闵绍楷等主编,《水稻育种学》,中国农业科学院,1996年出版,ISBN7-109-04338-X/S.2689,P:324
2.French D(1984).Organization of starch granules.In:Whistler RL BeMillerJN.Paschall E(eds).Starch:Chemistry and Technology.Orlando:Academic,183-247
3.Okagaki,R.J.,和Wessler,S.R.Genetics.1988,120:1137-1143.
4.Wang,Z.Y.和Wu,Z.L.,.Nucleic Acids Res.1990,18,5898.
5.Satoh和Omura.,Japan.J.Breed..1981,316-326.
6.钱前、朱旭东、曾大力等.浙江农业科学,1996,(4):155-156
7.钱前、曾大力、滕胜等.中国水稻科学,2000,14(3):173-176
8.Isshiki,M.和Nakajima,M.Plant J.2000,23,451-460.
9.Xueyong Li,Qian Qian and Jiayang Li.,Nature,2003,422(6932):618-21
10.Linsmaier,E.M.and Skoog,F.,Organic growth factor requirements oftobacco tissue cultures.Physiol.Plant.1965,18,100-127
序列表
<110>中国科学院遗传与发育生物学研究所
<120>水稻胚乳直链淀粉含量控制基因DU1及其应用
<130>IB053806
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<170>PatentIn version 3.1
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atggtgttcg tccgcgcgcc ggacgggagg acccaccacg tcgacctcga cccctccacc 60
gccacgctcg ccgacctcac ggcctccgcc tcccgcgtct gcggcggcgt cccgccggag 120
cagctgcggc tctacctcgc ccaccgccgc ctcctcccgg ccgagccgtc cccgctgctg 180
tcctccctcc gggtctcggc ctcctcctcc ctgctactcc acctccccct gctcggaggg 240
atgaccggcc cgacgacgac ccccgcggca cccccgcccc cgccgccgcc gtcggcgcag 300
ccgcccgccc gccccgcgcg ctacgacttc ctcaactcca agccgccccc gaactacgtg 360
gccggtctgg ggcgtggcgc caccgggttc accacccgtt cggatatcgg gccggcccgc 420
gcggcgcccg atctgcctga ccggtccgcc gccgccgccg ccgcccccgc cgtcgggcgc 480
ggccgtggga agccacccgg ggacgacgac ggcgacgacg atggcggcga cgaggagaag 540
gggtacgacg agaaccagaa gttcgacgag ttcgagggca acgacgccgg gctgttctcc 600
aacgccgact acgacgacga cgaccgcgag gcggatgcgg tctgggagag catcgaccag 660
aggatggact ctcgccggaa ggatcggcgg gaggcgcggc tgaagcagga gatcgagaag 720
taccgtgctt ccaaccctaa gatcaccgag caattcgctg atttgaagcg taagttggtc 780
gatttgtcgg cgcaggagtg ggaaagcata cctgaaattg gggactactc gctgcgcaac 840
aagaagaagc gatttgagag cttcgttccc gtgccggaca ccctgctcga gaaggctcgg 900
caggagcagg agcatgtcac ggcactggat cccaagagcc gtgcagctgg tggcaccgag 960
acgccatggg cgcagactcc ggttaccgat ctgacggctg tgggcgaagg tcgtggcacc 1020
gtgctctcct tgaagctgga caggttgtcg gattcggtat ctggtcttac tgttgttgat 1080
ccaaagggtt acttgacgga cctgaaaagt atgaagatta ctagtgatgc tgagatttct 1140
gacattaaaa aggcgcgatt gttgcttaag tcagtgacac agacaaaccc gaagcatcca 1200
ccaggatgga ttgctgctgc taggcttgaa gaggttgctg gcaagcttca ggttgctcgg 1260
cagcttatcc agcgtggctg tgaggagtgc cccacaaatg aggatgtttg ggtcgaggca 1320
tgccggctgg ccagcccaga cgaggcaaag gcagtgattg ctaggggcgt gaaggcaatt 1380
cccaattctg tgaagctgtg gttgcaggca gcaaagttgg aaactagtga tttgaataag 1440
agcagggttt tgagaaaagg gttggaacac attcctgatt cagtcagact gtggaaagca 1500
gtagtagagc ttgcaaatga ggaggatgca agactgttgc ttcacagggc tgtggagtgc 1560
tgcccactcc atgtggaact gtggcttgcc ctagcaaggc tggagacata tgaccaagca 1620
aagaaggtac ttaacaaggc aagagaaaag cttcctaagg aacctgccat ctggattaca 1680
gctgcaaagc tggaggaagc taatggaaac acccagtcag taatcaaggt gattgagaga 1740
agtataaaaa ctttacagag agaaggattg gatattgaca gggaggcatg gctaaaggaa 1800
gcagaagctg ctgagcgtgc tggatctgta ttgacttgcc aggctattgt taagagcact 1860
attggcattg gtgttgatga ggaagacaga aaacgcacat gggttgccga tgctgaggaa 1920
tgcaagaagc gtggttcaat tgagacagcc cgtgccatct atgcgcatgc actcagtgtc 1980
ttcgtttcca agaagagtat ttggctgaaa gcggctcagc ttgagaagag ccatggaacc 2040
aaggagtctc tttataatct cctcagaaag gctgttacct acaatccacg tgcagaagtt 2100
ttatggctta tgagtgcaaa ggagaaatgg ctggctggag atgtcccggc tgcccgagcc 2160
attcttcagg aagcttatgc ttctctcccc aattcagagg agatctggct agctgccttc 2220
aagcttgagt ttgagaacaa tgaaccagag agagcaagaa ttcttttgtc aaaggccagg 2280
gaaagaggag gcactgagag ggtctggatg aaatctgcga ttgttgaaag ggagttaggg 2340
aatgtagacg aagaaaggaa gctgttggag gaaggtctga agttattccc ctcattcttc 2400
aagctgtggt taatgcttgg acaaatggaa gaccggcttg gccatggatc caaggcaaag 2460
gaggtttacg agaatgcact gaagcactgc ccgagttgca tccctctttg gctctctcta 2520
gctaatctag aggagaagat aaatggcttg agcaagtcac gtgctgtcct caccatggca 2580
agaaagaaga acccagctac acctgaactc tggcttgcag cagttagggc tgaattgaga 2640
catgggaaca agaaggaagc tgatgctcta ctagccaagg cattacagga atgcccgaca 2700
agtggtattt tgtgggctgc agctatagag atggtgccac gtccccagcg taaagcaaag 2760
agctcagatg ctataaaacg atgtgaccat gatccccatg tcattgcagc tgtggccaaa 2820
cttttctggc atgataggaa ggttgataaa gctagaagtt ggttgaatag agctgttact 2880
cttgctccag acattggaga tttttgggcc ttgtactaca aatttgaact gcaacatgga 2940
aatgctgata cacaaaagga tgtcctacaa agatgtgttg cagcagaacc aaagcatgga 3000
gagagatggc aagcaataac aaaggctgtt gagaactcac atctgtcaat tgaggccctt 3060
ctgaagaaag ctgtgttggc tcttggccag gaagaaaatc caaatgctgc agatccctag 3120
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Val Pro Val Pro Asp Thr Leu Leu Glu Lys Ala Arg Gln Glu Gln Glu
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His Val Thr Ala Leu Asp Pro Lys Ser Arg Ala Ala Gly Gly Thr Glu
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Val Ser Gly Leu Thr Val Val Asp Pro Lys Gly Tyr Leu Thr Asp Leu
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Lys Ser Met Lys Ile Thr Ser Asp Ala Glu Ile Ser Asp Ile Lys Lys
370 375 380
Ala Arg Leu Leu Leu Lys Ser Val Thr Gln Thr Asn Pro Lys His Pro
385 390 395 400
Pro Gly Trp Ile Ala Ala Ala Arg Leu Glu Glu Val Ala Gly Lys Leu
405 410 415
Gln Val Ala Arg Gln Leu Ile Gln Arg Gly Cys Glu Glu Cys Pro Thr
420 425 430
Asn Glu Asp Val Trp Val Glu Ala Cys Arg Leu Ala Ser Pro Asp Glu
435 440 445
Ala Lys Ala Val Ile Ala Arg Gly Val Lys Ala Ile Pro Asn Ser Val
450 455 460
Lys Leu Trp Leu Gln Ala Ala Lys Leu Glu Thr Ser Asp Leu Asn Lys
465 470 475 480
Ser Arg Val Leu Arg Lys Gly Leu Glu His Ile Pro Asp Ser Val Arg
485 490 495
Leu Trp Lys Ala Val Val Glu Leu Ala Asn Glu Glu Asp Ala Arg Leu
500 505 510
Leu Leu His Arg Ala Val Glu Cys Cys Pro Leu His Val Glu Leu Trp
515 520 525
Leu Ala Leu Ala Arg Leu Glu Thr Tyr Asp Gln Ala Lys Lys Val Leu
530 535 540
Asn Lys Ala Arg Glu Lys Leu Pro Lys Glu Pro Ala Ile Trp Ile Thr
545 550 555 560
Ala Ala Lys Leu Glu Glu Ala Asn Gly Asn Thr Gln Ser Val Ile Lys
565 570 575
Val Ile Glu Arg Ser Ile Lys Thr Leu Gln Arg Glu Gly Leu Asp Ile
580 585 590
Asp Arg Glu Ala Trp Leu Lys Glu Ala Glu Ala Ala Glu Arg Ala Gly
595 600 605
Ser Val Leu Thr Cys Gln Ala Ile Val Lys Ser Thr Ile Gly Ile Gly
610 615 620
Val Asp Glu Glu Asp Arg Lys Arg Thr Trp Val Ala Asp Ala Glu Glu
625 630 635 640
Cys Lys Lys Arg Gly Ser Ile Glu Thr Ala Arg Ala Ile Tyr Ala His
645 650 655
Ala Leu Ser Val Phe Val Ser Lys Lys Ser Ile Trp Leu Lys Ala Ala
660 665 670
Gln Leu Glu Lys Ser His Gly Thr Lys Glu Ser Leu Tyr Asn Leu Leu
675 680 685
Arg Lys Ala Val Thr Tyr Asn Pro Arg Ala Glu Val Leu Trp Leu Met
690 695 700
Ser Ala Lys Glu Lys Trp Leu Ala Gly Asp Val Pro Ala Ala Arg Ala
705 710 715 720
Ile Leu Gln Glu Ala Tyr Ala Ser Leu Pro Asn Ser Glu Glu Ile Trp
725 730 735
Leu Ala Ala Phe Lys Leu Glu Phe Glu Asn Asn Glu Pro Glu Arg Ala
740 745 750
Arg Ile Leu Leu Ser Lys Ala Arg Glu Arg Gly Gly Thr Glu Arg Val
755 760 765
Trp Met Lys Ser Ala Ile Val Glu Arg Glu Leu Gly Asn Val Asp Glu
770 775 780
Glu Arg Lys Leu Leu Glu Glu Gly Leu Lys Leu Phe Pro Ser Phe Phe
785 790 795 800
Lys Leu Trp Leu Met Leu Gly Gln Met Glu Asp Arg Leu Gly His Gly
805 810 815
Ser Lys Ala Lys Glu Val Tyr Glu Asn Ala Leu Lys His Cys Pro Ser
820 825 830
Cys Ile Pro Leu Trp Leu Ser Leu Ala Asn Leu Glu Glu Lys Ile Asn
835 840 845
Gly Leu Ser Lys Ser Arg Ala Val Leu Thr Met Ala Arg Lys Lys Asn
850 855 860
Pro Ala Thr Pro Glu Leu Trp Leu Ala Ala Val Arg Ala Glu Leu Arg
865 870 875 880
His Gly Asn Lys Lys Glu Ala Asp Ala Leu Leu Ala Lys Ala Leu Gln
885 890 895
Glu Cys Pro Thr Ser Gly Ile Leu Trp Ala Ala Ala Ile Glu Met Val
900 905 910
Pro Arg Pro Gln Arg Lys Ala Lys Ser Ser Asp Ala Ile Lys Arg Cys
915 920 925
Asp His Asp Pro His Val Ile Ala Ala Val Ala Lys Leu Phe Trp His
930 935 940
Asp Arg Lys Val Asp Lys Ala Arg Ser Trp Leu Asn Arg Ala Val Thr
945 950 955 960
Leu Ala Pro Asp Ile Gly Asp Phe Trp Ala Leu Tyr Tyr Lys Phe Glu
965 970 975
Leu Gln His Gly Asn Ala Asp Thr Gln Lys Asp Val Leu Gln Arg Cys
980 985 990
Val Ala Ala Glu Pro Lys His Gly Glu Arg Trp Gln Ala Ile Thr Lys
995 1000 1005
Ala Val Glu Asn Ser His Leu Ser Ile Glu Ala Leu Leu Lys Lys
1010 1015 1020
Ala Val Leu Ala Leu Gly Gln Glu Glu Asn Pro Asn Ala Ala Asp
1025 1030 1035
Pro
Claims (7)
1.一种控制水稻胚乳直链淀粉含量的基因,该基因的核苷酸序列选自:
(1)编码SEQ ID NO:2所示的氨基酸序列的核苷酸序列;
(2)与(1)中的核苷酸序列能够在严谨条件下杂交,并同时编码具有控制植物器官淀粉组成和水稻胚乳直链淀粉含量功能的核苷酸序列。
2.按照权利要求1所述的基因,它具有SEQ ID NO:1所示的DNA序列。
3.一种由权利要求1或2所述的核苷酸序列编码的蛋白质。
4.按照权利要求3所述的蛋白质,它具有SEQ ID NO:2所示的氨基酸序列。
5.一种含有权利要求1或2所述的控制水稻胚乳直链淀粉含量的基因的植物表达载体。
6.按照权利要求5所述的表达载体,该表达载体是pCAMDU1。
7.一种培育植物方法,该方法可使植物淀粉的主要储藏器官的淀粉构成发生变化,包括用权利要求5或6所述的表达载体转化植物细胞;和将转化的植物细胞培育成植株的步骤。
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106591435A (zh) * | 2016-11-22 | 2017-04-26 | 南京农业大学 | 粳稻品种越光暗胚乳突变体w54低直链淀粉含量基因位点的分子标记方法 |
CN113637688A (zh) * | 2021-09-23 | 2021-11-12 | 上海师范大学 | 水稻稻米直链淀粉含量调控基因OsACF1及其应用 |
CN114262710A (zh) * | 2021-12-31 | 2022-04-01 | 西南大学 | 水稻胞间连丝基因及其突变基因、编码的蛋白和应用 |
CN114438101A (zh) * | 2022-03-10 | 2022-05-06 | 江苏省农业科学院 | 一种稻米外观透明的低直链淀粉含量的等位基因及其应用 |
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106591435A (zh) * | 2016-11-22 | 2017-04-26 | 南京农业大学 | 粳稻品种越光暗胚乳突变体w54低直链淀粉含量基因位点的分子标记方法 |
CN113637688A (zh) * | 2021-09-23 | 2021-11-12 | 上海师范大学 | 水稻稻米直链淀粉含量调控基因OsACF1及其应用 |
CN113637688B (zh) * | 2021-09-23 | 2023-10-13 | 上海师范大学 | 水稻稻米直链淀粉含量调控基因OsACF1及其应用 |
CN114262710A (zh) * | 2021-12-31 | 2022-04-01 | 西南大学 | 水稻胞间连丝基因及其突变基因、编码的蛋白和应用 |
CN114262710B (zh) * | 2021-12-31 | 2023-10-31 | 西南大学 | 水稻胞间连丝基因及其突变基因、编码的蛋白和应用 |
CN114438101A (zh) * | 2022-03-10 | 2022-05-06 | 江苏省农业科学院 | 一种稻米外观透明的低直链淀粉含量的等位基因及其应用 |
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