CN104946666B - 水稻OsACBP5基因及其在提高水稻抗病性中的应用 - Google Patents
水稻OsACBP5基因及其在提高水稻抗病性中的应用 Download PDFInfo
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Abstract
本发明公开了一种水稻OsACBP5基因及其在提高水稻抗病性中的应用。本发明根据同源性分析,在水稻中克隆拟南芥AtACBP3的同源基因OsACBP5。该基因的全长5579 bp,CDS长1710 bp,编码570 aa的蛋白。用CDD、PFAM、SMART、Interproscan程序分析预测,该蛋白有跨膜域和ACBP结构域。通过比较野生型和过表达转基因植物对细菌性病菌的耐受性,结果表明OsACBP5基因可以提高植物的抗病能力,可以广泛应用于基因工程领域有着巨大的经济价值和应用前景。
Description
技术领域
本发明涉及植物抗病技术领域,具体地,涉及水稻OsACBP5基因及其在提高水稻抗病性中的应用。
背景技术
水稻是我国的重要粮食作物,全世界一半以上的人口以水稻为主食,病虫害浸染直接影响水稻的生长和产量。例如,由水稻黄单胞菌(Xanthomonas oryzae pv.oryzae, Xoo)引起的白叶枯病是水稻生产中的最严重的细菌性病害,在我国南北各稻区包括广东地区普遍存在,造成的损失巨大。据估计白叶枯病发生时,会导致水稻产量减产20~30%,严重的甚至导致绝收。因此,改善和提高水稻对病原菌胁迫的抗性、培育具有高抗病性的新种质,在促进水稻生产稳步发展、保障我国的粮食安全上具有重要的作用。
开发水稻抗病基因是提高水稻对病原菌胁迫的抗性、培育具有高抗病性的新种质的有效途径。目前,水稻中已定位至少30个稻瘟病抗性位点和25个白叶枯病抗性位点。其中,抗稻瘟病的Pib和Pi-ta和抗白叶枯病的Xa21和Xa1基因已经被克隆鉴定。但是,目前水稻的抗病基因资源还不够丰富,而且已报道的这些抗病基因大多都是针对某一特定病害起作用。
发明内容
本发明的目的是为了克服现有技术的不足,提供一种水稻抗病基因OsACBP5。
本发明的另一个目的是提供一种抗病基因OsACBP5在提高水稻抗病性中的应用,尤其是提高水稻抗黄单胞菌引起的白叶枯病。
为了实现上述目的,本发明是通过以下方案予以实现的:
一种水稻抗病基因OsACBP5,该基因的全长5,579 bp(如SEQ ID NO:1所示),CDS长1,710 bp(如SEQ ID NO:2所示),编码570 aa的蛋白(如SEQ ID NO:3所示)。
发明人通过研究发现,该基因能够提高水稻抗黄单胞菌引起的白叶枯病。
经序列比对发现,本发明水稻抗病基因OsACBP5编码的蛋白与拟南芥AtACBP3蛋白是同源蛋白,同AtACBP3一样,OsACBP5是唯一一个其ACB结构域存在于C末端的ACBP蛋白,其N末端也同样检测到了与AtACBP3相似的信号肽/转膜结构域;氨基酸序列比对分析结果显示,AtACBP3和OsACBP5蛋白中氨基酸序列的相同率为56%。
水稻OsACBP5基因在提高水稻抗病中的应用,所述基因的核苷酸序列如SEQ IDNO:1或2所示。
本发明通过比较野生型和过表达转基因(OsACBP5基因)植物对细菌性病菌的耐受性,结果表明OsACBP5基因可以提高植物的抗病能力,尤其是抗稻瘟病和白叶枯病,可以广泛应用于基因工程领域有着巨大的经济价值和应用前景。实施例中虽然中做了对日本晴品种的抗病情况,但是,本发明在保护基因的用途时,对于水稻的品种并不局限于日本晴这种品种。
水稻OsACBP5基因在提高水稻抗黄单胞菌引起的白叶枯病中的应用,所述基因的核苷酸序列如SEQ ID NO:1或2所示。
一种抗黄单胞菌引起的白叶枯病的水稻的构建方法,包括如下步骤:
S1.将SEQ ID NO:1或2所述的OsACBP5基因克隆到PCXSN-MYC载体;
S2.用农杆菌转化法侵染水稻,得到稳定表达的转基因植株。
与现有技术相比,本发明具有如下有益效果:
本发明在水稻中首次发现了OsACBP5基因,该基因的全长5,579 bp,CDS长1,710bp,编码570 aa的蛋白。用CDD、PFAM、SMART、Interproscan程序分析预测,该蛋白有跨膜域和ACBP结构域。通过比较突变体、野生型和过表达转基因水稻的对细菌性病菌的耐受性,发现OsACBP5基因可以提高水稻的抗病能力,可以广泛应用于基因工程领域有着巨大的经济价值和应用前景。
附图说明
图1为SMART程序预测OsACBP5蛋白的结构域,含有跨膜结构域和ACBP结构域。
图2为黄单胞菌(Xoo)处理16天后野生型、过表达转基因水稻植株的生长情况,CK为野生型水稻。
具体实施方式
下面结合说明书附图和具体实施例对本发明作出进一步地详细阐述,所述实施例只用于解释本发明,并非用于限定本发明的范围。下述实施例中所使用的试验方法如无特殊说明,均为常规方法;所使用的材料、试剂等,如无特殊说明,为可从商业途径得到的试剂和材料。
实施例1水稻OsACBP5基因的克隆
根据拟南芥中的AtACBP3的同源蛋白的同源性,在水稻基因组数据中利用Blastp程序进行搜索,得到水稻OsACBP5蛋白,其是唯一一个其ACB结构域存在于C末端的水稻ACBP蛋白,其N末端也同样检测到了与AtACBP3相似的信号肽/转膜结构域。设计正反向引物
XS258:CATGGAGCTGTTCTACGAGCTGCTCCTC;
XS259:TCATTCAGCAGGGATGTCAGAACTCTGT;
提取水稻叶片RNA,反转录成cDNA后,以上述引物进行PCR扩增,将PCR扩增产物进行克隆测序。水稻OsACBP5基因的全长5579 bp(如SEQ ID NO:1所示),CDS长1710 bp(如SEQID NO:2所示),编码570 aa的蛋白(如SEQ ID NO:3所示)。用CDD、PFAM、SMART、Interproscan程序分析预测,水稻OsACBP5蛋白有跨膜域和ACBP结构域,见图1。
实施例2水稻OsACBP5基因的功能研究
将实施例1所述的OsACBP5基因连接到pCXSN-MYC载体上,具体步骤如下:将通过XS258和XS259引物克隆后的序列(即SEQ ID NO:2所示)序列),通过TaKaRa的加“A”试剂盒,与用XcmⅠ进行酶切的pCXSN-MYC载体相连接,在含卡那平板上进行筛选后,通过PCR鉴定并送测序,正确后转化农杆菌。
将阳性载体利用农杆菌转化法转化到日本晴水稻中,得到稳定表达的转基因植株OsACBP5-OE,具体步骤如下:
将成熟水稻种子消毒后,经盾片诱导培养基诱导产生胚性愈伤,用含有鉴定正确的农杆菌侵染,侵染20 min后将愈伤组织转至加有一张滤纸的共培养基上,然后转入筛选培养基上进行筛选培养。生长旺盛的水稻愈伤在预分化光照培养2-3周,转移到分化培养基上培养至分化出苗,再转至壮苗生根培养基中,最后移栽到人工气候室种植。
黄单胞菌处理野生型和转基因水稻:分别将野生型(日本晴水稻,Oryza sativajaponica. cv. Nipponbare,后续简称为CK)和转基因水稻OsACBP5-OE的成熟种子,去颖壳,用75%酒精浸泡30~45秒,无菌水洗3次,再用20%的次氯酸钠浸泡30分钟,不时摇晃,水洗5次以上;消毒完成后,将三种类型的种子分别置于含有1/2 MS液体培养液的无菌烧杯内培养。 然后再种植在1/2 MS固体培养基上,并移入28℃(16-h光照/8-h黑暗)的人工气候室。
待以上三种水稻生长到20天大小的植株,分别接种黄单胞菌,具体步骤如下:采用剪叶法进行接种,利用剪出2 - 3cm的缺口,将菌龄为3 d,菌液浓度3×108 cfu,,接种后于湿度大于85%的环境中培育。
选取己完全展开的水稻野生型和转基因植株叶片,用手术剪蘸取菌液,剪切叶片2-3 cm的缺口,接种后90%的环境中培育。
接种Xoo(Pxo99)处理后16天,发现转基因水稻OsACBP5-OE的叶片比对照野生型水稻的叶片病斑长度与叶片总长比值低(见图2),这说明OsACBP5基因可以提高植物对黄单胞菌的抗性。
SEQUENCE LISTING
<110> 中山大学
<120> 水稻OsACBP5基因及其在提高水稻抗病性中的应用
<130>
<160> 5
<170> PatentIn version 3.3
<210> 1
<211> 5580
<212> DNA
<213> OsACBP5基因DNA序列全长
<400> 1
ccggttgctc gatcgtctcg ctctcctctc ctcttctctt cgtctctcct cctgaccctt 60
tttccgcttt cccttttccc ccctcctcga tccgcggcgc aaccgaacac gaaagcgaga 120
gagagaggag tgcagccgct gccctgtccc ctcaccgcgc cgcgccgaca tggagctgtt 180
ctacgagctg ctcctcacgg cggctgcctc cctcctcgtc gccttcctgc tggccaggct 240
gctggcctcc gccgccaccg ccagtgatcc ccgccgccgc gcgcccgatc acgccgccgt 300
gatcgcggag gaggaggcgg tggtggtgga ggaggagagg atcatcgagg tcgatgaggt 360
cgaggtgaag agcgcgcgcg cgagggagtg cgtggtttcg gaggggtggg tcgaggtggg 420
gagggcctcg tcggcggagg ggaagctcga gtgcttgccg gaggaagagg aggctcccgc 480
gaaggccgct cgggagcttg ttctcgatgc cgttttggag gaacgcgagg aggaaggaca 540
agttggcgaa gagcggtgcg atttggccgc tgcggtggcg gaggtcgtgg gagtgaagcc 600
gcatgagttg ggggttgaag ctgctcccgg ggaggtatct gacgtgacgc tggaggaagg 660
gaaggtgcag gatgttgggg tggagcagca tgatttggtc gccgaggctg ctccaaggga 720
agctcttgac acagggttgg agaaacaggg tgttcccatc attgaagcgg ttgaaatcaa 780
gcggcaggat gatctgggtg ctgaggtagc tccgagtgac gttcctgagg tggaatttga 840
gcaacaggga gttcgcatta ttgaagctat tgatgtgaat caacatcatc gggttgctct 900
ggctgctcct gcggaagttg ttgatgcggg attggaggag agggtccaag ctattgaagc 960
agggtcatct ggattgactt ccgagacagt tcctgaagag gttcttgacg agttatctga 1020
gaagcaagaa gagcaagtta ttgaagagaa ggaacatcaa ttggctgcag cgactgctcc 1080
agtagcaatt cctggtgtgg cattggcaga gacggaggaa cttaaagaag aacaatcctc 1140
tgaaaaagct gtcaatgttc atgaagaagt tcagagtaag gacgaagcta aatgcaagct 1200
ccatctggtt gatcaacaag aaggttcggc ttctaaggtg gagctggtgg ggaggaatac 1260
cgacaatgtg gaaattagcc atggaagcag ttctggtgac aaaatgattg ctgagttgac 1320
cgaggaggaa ttgacattgc aaggtgtgcc cgcagatgag actcagacag acatggaatt 1380
tggggagtgg gaagggattg aaagaactga gatagaaaag aggtttggtg tggcagcagc 1440
gtttgcatct agtgacgccg ggatggctgc cctgtcaaag cttgatagtg atgtgcagct 1500
gcagctgcag ggactcctta aggttgccat tgatggtcca tgctatgact ctacacagcc 1560
acttaccttg aggccttcat ctcgtgcaaa atggtaattg tttttctcat tgatatccta 1620
gctccaactt tttttttggt atgctgcaca actgcggtag gttatatagt tcaaggacaa 1680
caaaaccaat atactaccca aatgccttac tttgcactgg ctagaatgta atcggcaatg 1740
gacttgtgat ttataggcag atcaggtata ctggtctgaa atagcttaaa ctagtccttt 1800
ttttggcaaa agaccaaagt acagaacttt gaactctgaa ttctgggtag ttggtagata 1860
gactgtaatg acatattaga aaactgaatt gtttgatctg ctggtctgtt tgatctagtt 1920
gggacttctt atggctacta tgacacttat gtatgaaatt tggtcctagc tatttggcta 1980
aacctaaaaa agagccaaag aactggttca aaattttgaa tcccatttta ctgcggaaca 2040
tagaacttct ctgtggaaca ataaacgact catataatgc aaaagttttt aatcttttgc 2100
ataattgatt gttaaaatta gaaaaatatt ttcctacctt atgttgtgac ttgagacaat 2160
gagaatctgt cctctaattt gttaagggga aaaaaagctt ctgtcaactt ttctgggcta 2220
tctaaacaaa gacattaaat gtgccatttt tgttttatat tgtttaataa acttgttatg 2280
ctaaaagaag ataattatag ttgtccatca tcagtgaaga taaagataag tgaattataa 2340
ttgcccatca tcaatgggat aatgcactta aaccagtgat atagtgtgta atagtgtaag 2400
gatgtcactg atacgctgat cttctgtcat gcataatggt gtgtggttca catgagaatc 2460
tatcaagttt ttgggataag ttgcaagtga aggcaaagca aagcatatga aaagcatagt 2520
atttttagag gtatcaatcc agtgttactt ggccgcactg caaggtagtt attcatataa 2580
acaggccaaa acacttcatc tatgtatgcc caacatgata taactgtgac atgagtgcag 2640
gacacacata ttgcaatatt ccatgcttaa tgcagtcaga tttgtaattt ggagtgcttc 2700
tccgagatag tttgtaacta atcatgtagc tactatttgg aatccaaaaa tggacaaaat 2760
ccaatgtcaa atatatgatg taagcagtta ttgagatgct gatacgctag gtatttctct 2820
tgttggacat aattatatat gagtatatca ttgaactata ggtattttgt caaagctacc 2880
taaaacgatg catttgagtt gattcaataa ttatatctta actgaatacc tttatatatg 2940
acaaaaccta aatggtgggc gacccccttt tcatctccca tatttggcat gtgggaccca 3000
cctgttagtt gcatacttgc atgtgataga caattatagt actcctgtca cttttgaaca 3060
tttattcttt ttactatgag tttttccaca aatacttgtc acttcacagt ctcaatcaat 3120
caatttttgt ttgcatagag cgcccctgtt agttctatat gtgccaccat gtaattcaat 3180
atcatatact tggagtgcag aggtagcata tacttacaaa atgggaccta tcaaaaaaat 3240
tactagttga atttttttca ctggtattat gatatgccat cgaatgactg attccagcca 3300
gatttcctat gataaatgag agtaatgatc acaaattcaa agaattttca tatgttttgc 3360
atgtgcgcag caccttatct ctcatttgtt ggttgtgtag cctttttata ttgattcttt 3420
aaatgggttg ctgccaaaag cggagcgggg cctgtgtcta accataacaa ttaccctttg 3480
gttgagtgag aacctagatg ttttagaata tggaatgctg gtcacataat atacatgatg 3540
tgacagcatc acagcactga catcttgcac ctatctctct atgttaaggc tttctattta 3600
gtataatggt agttggttga caatctatag tcatattcat atatgcctgt aagtaattct 3660
ggagttatac ttgcagtaac tatatataac ttggctactt atatttttta tttccaaaaa 3720
ttatagggaa tgtatgttgg actatatgaa aaattataag ttgtaaatag acataaggaa 3780
agacacacaa gatggtgggc tatttgtttg taggagttgt acaattctgt taagctgcat 3840
acgagtcaca gtctgtaatg ataacccata actatcgtgt attattctct cagggctgct 3900
tggcaaaagc tagggaacat gtatccggag acagctatgg aaagatacat gaatcttttg 3960
tcagaggcta ttccaggatg gatgggtgac aatatctcgg taagtttttc tatgggtgcc 4020
aaagtgttac ttctgtttat tgcattgttt ttttacacca gactggacaa tagttctaat 4080
catttcgaaa tactgtactc tagggcacaa aggaacatga agctggtgat gatgctgtag 4140
ggtctgtctt aacaatgact tcaaatacaa tcaaccaaca tgacagtcaa gggtaattta 4200
ctcatttact gtagtgtgaa acctggaatg tagcaaagtg ttgacagggc tgaccatacc 4260
gatgatatac ctgccatact aactagtagt atttgttagt gctggtataa gaatatctta 4320
aatgaaattc ctggctcttt agctgtacca gttctttttt acaatggcaa tatctacgag 4380
tcccaactgg tataactcaa tctggtacgg taaccttgat ttagaagtgt ttctaatgtt 4440
tattaccatg gcttgctcct gctgatggat ttcttggcag gaatgaagac aatactggca 4500
tgtatgaagg tcacttgaca agttccccta acccagagaa aggtacgtac ttgtcatatt 4560
ttcggcagtg aaccaggatt ttttttatct ggctcctaaa agtaaaatat tgcaattcat 4620
tagtgaccag tgttgtgtat ttatgcatga cgactggttc aggtgcattt gtgaaattaa 4680
cttctgatag ctatctatca tttttttgcc tgcaccctct gtgatctcca ctttgttaac 4740
cacattttgt aaggaaaact aaattacaag gagtatctcc tgcgtgtttg agaaaaaaaa 4800
acttatggag tatattgcat ttcattagtg gtagttctcc tgccatgcat gtgttgtgct 4860
tgctaaccat ttaattccaa gttaatgttt ttctacacca taattaaaca aactactccc 4920
tcattctttt tttacttgct atttcaggca taggtgcaga tcacaatgca tgcatttgac 4980
cgttacatta ctgaacttat ttagaaaatg atatctaaag taattttgtg aaaaaaaaat 5040
ctgctaatac catttcagtg tgaaaatagt ataaacattt ttagtcaaag aatggaagtg 5100
taccgttcct ctatttcagt atgtttctgt ttgattactt cgtcaatcaa tttcatgaat 5160
tcaacccgcc actaaatttt caggacagag ttctgacatc cctgctgaat gaacataaca 5220
ctcatggaag atttcttgca tcagtgattt cagcccattg ctgagaccgc taccgatgcc 5280
ttcctcataa gattagattt gtacagtgct cctgtggtgc taatattagt gttgtcatgg 5340
tgttggtttg tgtatatata tgactgtata tatcagtaac tgccggtcct actgcgtgta 5400
ttccgccagc agcagactta tcatctggta ttctggtgta gtctgtagaa caatacatct 5460
ggtgagtcgt aattatatgt cgatacgatg cagtttggaa ggagaaattg aagtggttgg 5520
aaaaccagtt gaaaggactg aaaaataatc tcatcagaag gaacataaat tctttatccc 5580
<210> 2
<211> 1710
<212> DNA
<213> OsACBP5基因cDNA序列全长
<400> 2
atggagctgt tctacgagct gctcctcacg gcggctgcct ccctcctcgt cgccttcctg 60
ctggccaggc tgctggcctc cgccgccacc gccagtgatc cccgccgccg cgcgcccgat 120
cacgccgccg tgatcgcgga ggaggaggcg gtggtggtgg aggaggagag gatcatcgag 180
gtcgatgagg tcgaggtgaa gagcgcgcgc gcgagggagt gcgtggtttc ggaggggtgg 240
gtcgaggtgg ggagggcctc gtcggcggag gggaagctcg agtgcttgcc ggaggaagag 300
gaggctcccg cgaaggccgc tcgggagctt gttctcgatg ccgttttgga ggaacgcgag 360
gaggaaggac aagttggcga agagcggtgc gatttggccg ctgcggtggc ggaggtcgtg 420
ggagtgaagc cgcatgagtt gggggttgaa gctgctcccg gggaggtatc tgacgtgacg 480
ctggaggaag ggaaggtgca ggatgttggg gtggagcagc atgatttggt cgccgaggct 540
gctccaaggg aagctcttga cacagggttg gagaaacagg gtgttcccat cattgaagcg 600
gttgaaatca agcggcagga tgatctgggt gctgaggtag ctccgagtga cgttcctgag 660
gtggaatttg agcaacaggg agttcgcatt attgaagcta ttgatgtgaa tcaacatcat 720
cgggttgctc tggctgctcc tgcggaagtt gttgatgcgg gattggagga gagggtccaa 780
gctattgaag cagggtcatc tggattgact tccgagacag ttcctgaaga ggttcttgac 840
gagttatctg agaagcaaga agagcaagtt attgaagaga aggaacatca attggctgca 900
gcgactgctc cagtagcaat tcctggtgtg gcattggcag agacggagga acttaaagaa 960
gaacaatcct ctgaaaaagc tgtcaatgtt catgaagaag ttcagagtaa ggacgaagct 1020
aaatgcaagc tccatctggt tgatcaacaa gaaggttcgg cttctaaggt ggagctggtg 1080
gggaggaata ccgacaatgt ggaaattagc catggaagca gttctggtga caaaatgatt 1140
gctgagttga ccgaggagga attgacattg caaggtgtgc ccgcagatga gactcagaca 1200
gacatggaat ttggggagtg ggaagggatt gaaagaactg agatagaaaa gaggtttggt 1260
gtggcagcag cgtttgcatc tagtgacgcc gggatggctg ccctgtcaaa gcttgatagt 1320
gatgtgcagc tgcagctgca gggactcctt aaggttgcca ttgatggtcc atgctatgac 1380
tctacacagc cacttacctt gaggccttca tctcgtgcaa aatgggctgc ttggcaaaag 1440
ctagggaaca tgtatccgga gacagctatg gaaagataca tgaatctttt gtcagaggct 1500
attccaggat ggatgggtga caatatctcg ggcacaaagg aacatgaagc tggtgatgat 1560
gctgtagggt ctgtcttaac aatgacttca aatacaatca accaacatga cagtcaaggg 1620
aatgaagaca atactggcat gtatgaaggt cacttgacaa gttcccctaa cccagagaaa 1680
ggacagagtt ctgacatccc tgctgaatga 1710
<210> 3
<211> 569
<212> PRT
<213> OsACBP5蛋白序列
<400> 3
Met Glu Leu Phe Tyr Glu Leu Leu Leu Thr Ala Ala Ala Ser Leu Leu
1 5 10 15
Val Ala Phe Leu Leu Ala Arg Leu Leu Ala Ser Ala Ala Thr Ala Ser
20 25 30
Asp Pro Arg Arg Arg Ala Pro Asp His Ala Ala Val Ile Ala Glu Glu
35 40 45
Glu Ala Val Val Val Glu Glu Glu Arg Ile Ile Glu Val Asp Glu Val
50 55 60
Glu Val Lys Ser Ala Arg Ala Arg Glu Cys Val Val Ser Glu Gly Trp
65 70 75 80
Val Glu Val Gly Arg Ala Ser Ser Ala Glu Gly Lys Leu Glu Cys Leu
85 90 95
Pro Glu Glu Glu Glu Ala Pro Ala Lys Ala Ala Arg Glu Leu Val Leu
100 105 110
Asp Ala Val Leu Glu Glu Arg Glu Glu Glu Gly Gln Val Gly Glu Glu
115 120 125
Arg Cys Asp Leu Ala Ala Ala Val Ala Glu Val Val Gly Val Lys Pro
130 135 140
His Glu Leu Gly Val Glu Ala Ala Pro Gly Glu Val Ser Asp Val Thr
145 150 155 160
Leu Glu Glu Gly Lys Val Gln Asp Val Gly Val Glu Gln His Asp Leu
165 170 175
Val Ala Glu Ala Ala Pro Arg Glu Ala Leu Asp Thr Gly Leu Glu Lys
180 185 190
Gln Gly Val Pro Ile Ile Glu Ala Val Glu Ile Lys Arg Gln Asp Asp
195 200 205
Leu Gly Ala Glu Val Ala Pro Ser Asp Val Pro Glu Val Glu Phe Glu
210 215 220
Gln Gln Gly Val Arg Ile Ile Glu Ala Ile Asp Val Asn Gln His His
225 230 235 240
Arg Val Ala Leu Ala Ala Pro Ala Glu Val Val Asp Ala Gly Leu Glu
245 250 255
Glu Arg Val Gln Ala Ile Glu Ala Gly Ser Ser Gly Leu Thr Ser Glu
260 265 270
Thr Val Pro Glu Glu Val Leu Asp Glu Leu Ser Glu Lys Gln Glu Glu
275 280 285
Gln Val Ile Glu Glu Lys Glu His Gln Leu Ala Ala Ala Thr Ala Pro
290 295 300
Val Ala Ile Pro Gly Val Ala Leu Ala Glu Thr Glu Glu Leu Lys Glu
305 310 315 320
Glu Gln Ser Ser Glu Lys Ala Val Asn Val His Glu Glu Val Gln Ser
325 330 335
Lys Asp Glu Ala Lys Cys Lys Leu His Leu Val Asp Gln Gln Glu Gly
340 345 350
Ser Ala Ser Lys Val Glu Leu Val Gly Arg Asn Thr Asp Asn Val Glu
355 360 365
Ile Ser His Gly Ser Ser Ser Gly Asp Lys Met Ile Ala Glu Leu Thr
370 375 380
Glu Glu Glu Leu Thr Leu Gln Gly Val Pro Ala Asp Glu Thr Gln Thr
385 390 395 400
Asp Met Glu Phe Gly Glu Trp Glu Gly Ile Glu Arg Thr Glu Ile Glu
405 410 415
Lys Arg Phe Gly Val Ala Ala Ala Phe Ala Ser Ser Asp Ala Gly Met
420 425 430
Ala Ala Leu Ser Lys Leu Asp Ser Asp Val Gln Leu Gln Leu Gln Gly
435 440 445
Leu Leu Lys Val Ala Ile Asp Gly Pro Cys Tyr Asp Ser Thr Gln Pro
450 455 460
Leu Thr Leu Arg Pro Ser Ser Arg Ala Lys Trp Ala Ala Trp Gln Lys
465 470 475 480
Leu Gly Asn Met Tyr Pro Glu Thr Ala Met Glu Arg Tyr Met Asn Leu
485 490 495
Leu Ser Glu Ala Ile Pro Gly Trp Met Gly Asp Asn Ile Ser Gly Thr
500 505 510
Lys Glu His Glu Ala Gly Asp Asp Ala Val Gly Ser Val Leu Thr Met
515 520 525
Thr Ser Asn Thr Ile Asn Gln His Asp Ser Gln Gly Asn Glu Asp Asn
530 535 540
Thr Gly Met Tyr Glu Gly His Leu Thr Ser Ser Pro Asn Pro Glu Lys
545 550 555 560
Gly Gln Ser Ser Asp Ile Pro Ala Glu
565
<210> 4
<211> 28
<212> DNA
<213> XS258
<400> 4
catggagctg ttctacgagc tgctcctc 28
<210> 5
<211> 28
<212> DNA
<213> XS259
<400> 5
tcattcagca gggatgtcag aactctgt 28
Claims (3)
1.水稻OsACBP5基因在提高水稻抗黄单胞菌引起的白叶枯病中的应用,所述基因的核苷酸序列如SEQ ID NO:1或2所示。
2.根据权利要求1所述的应用,其特征在于,所述的水稻品种为日本晴。
3.一种抗黄单胞菌引起的白叶枯病的水稻的构建方法,其特征在于,包括如下步骤:
S1.将权利要求1所述的OsACBP5基因克隆到PCXSN-MYC载体;
S2.用农杆菌转化法侵染水稻,得到稳定表达的转基因植株。
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CN1241630A (zh) * | 1999-08-18 | 2000-01-19 | 中国农业科学院作物育种栽培研究所 | 普通野生稻抗白叶枯病近等基因系及其培育方法 |
CN101367001A (zh) * | 2007-04-11 | 2009-02-18 | 香港大学 | 在植物修复中使用表达源自植物酰基辅酶a结合蛋白的转化植物的方法 |
CN103740723A (zh) * | 2014-01-15 | 2014-04-23 | 海南大学 | 受水稻白叶枯病菌与细菌性条斑病菌诱导的启动子及应用 |
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CN101367001A (zh) * | 2007-04-11 | 2009-02-18 | 香港大学 | 在植物修复中使用表达源自植物酰基辅酶a结合蛋白的转化植物的方法 |
CN103740723A (zh) * | 2014-01-15 | 2014-04-23 | 海南大学 | 受水稻白叶枯病菌与细菌性条斑病菌诱导的启动子及应用 |
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