CN114891826B - 改良玉米果穗形态的方法 - Google Patents
改良玉米果穗形态的方法 Download PDFInfo
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- CN114891826B CN114891826B CN202210337659.0A CN202210337659A CN114891826B CN 114891826 B CN114891826 B CN 114891826B CN 202210337659 A CN202210337659 A CN 202210337659A CN 114891826 B CN114891826 B CN 114891826B
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Abstract
本发明涉及玉米基因ZmTPK1在改良玉米果穗形态中的应用，属于分子遗传学领域。本发明公开了玉米果穗形态基因ZmTPK1，其特征在于所述基因的核苷酸序列如SEQ ID NO.2‑SEQ ID NO.3任意一个所示；所编码的氨基酸序列如SEQ ID NO.1所示。进一步地，本发明提供了改良玉米果穗形态的方法，该方法可用于提高鲜食玉米品质。
Description
技术领域
本发明涉及玉米基因ZmTPK1在改良玉米果穗形态中的应用，属于分子遗传学领域。
背景技术
鲜食玉米兼具粮、果、蔬三类食物特性，受到民众喜爱。甜玉米更是世界范围内第三大蔬菜作物。我国鲜食玉米年种植面积已超过2000多万亩，并有继续扩大的趋势。在我国很多地区，由于种植成本相对较高，消费群体大，市场销售旺盛，且销售价格看好，鲜食玉米消费的主要形式是鲜穗鲜食，加工比例很低。这就要求果穗品质优良、外观漂亮、排列紧密、果穗大小适中。因此，对玉米果穗形态性状的改良在鲜食玉米品种选育和产品开发中具有重要的应用前景。
为解决上述问题，本发明筛选到一个控制玉米果穗形态的基因ZmTPK1，敲除该基因后玉米穗长和穗宽均降低，果穗整体变小，适合作为可以鲜食的鲜穗，能够用于改良鲜食玉米的果穗形态，提高鲜食玉米品质。
发明内容
本发明的目的之一在于提供一个影响玉米果穗形态的基因ZmTPK1的核酸序列及其编码的氨基酸序列。
本发明的目的之二在于公开一种改良玉米果穗形态的方法。
本发明的目的之三在于公开一种玉米小果穗突变基因。
为实现上述目的，本发明采用如下技术方案：
本发明提供一种基因在调控玉米果穗形态中的应用，其特征在于：所述基因序列如SEQ ID NO.2-SEQ ID NO.3任意一个所示。
本发明还提供一种改良玉米果穗形态的方法，其特征在于：抑制玉米中SEQ IDNO.1所示基因编码蛋白的表达和/或活性，选择玉米果穗变小的植株。
在一些实施方案中，上述抑制蛋白表达和/或活性的方法包括基因编辑、RNA干扰、T-DNA插入中任一种。
在一些实施方案中，上述基因编辑采用CRISPR/Cas9方法。
在一些实施方案中，上述CRISPR/Cas9方法在玉米中的基因组靶标区域的DNA序列如SEQ ID NO.4或SEQ ID NO.5所示。
本发明还提供一种改良玉米果穗形态的试剂盒，其特征在于：包括如下任一种：
(1)能够识别上述靶标序列的RNA分子；该RNA分子可以是包含gRNA、crRNAs和tracrRNA结构的sgRNA分子，也可以是gRNA、crRNAs和tracrRNA单独形成的复合体，或包括gRNA、crRNAs的复合体；在一些实施方案中，所述的RNA分子的序列如SEQ ID NO.6或SEQ IDNO.7所示；
(2)编码(1)所述RNA的DNA分子；
(3)表达(1)所述RNA的载体。
本发明还提供一种玉米突变基因，其特征在于：所述突变基因序列为SEQ ID NO.8或SEQ ID NO.9所示。
本发明还提供上述的玉米突变基因在改良玉米果穗形态中的应用。
与现有的技术相比，本发明的有益效果是，本发明提供的ZmTPK1基因及其编码的蛋白具有调控玉米果穗形态的功能，该基因的功能是之前的公开资料中未报道的。利用CRISPR/Cas9方法敲除该基因能够改良玉米果穗形态。本发明鉴定了基因敲除后的突变基因序列，利用这些突变基因可以进一步改良其他玉米品种尤其是鲜食玉米的果穗形态，从而提高鲜食玉米品质。
附图说明
图1基因编辑载体图。主要元件标注在图上。
图2基因编辑的靶标位置、靶标序列(灰色)和编辑后的序列展示。WT：未编辑的玉米；KO：编辑的玉米；“-”表示序列缺失。
图3基因编辑玉米的果穗表现。WT：未编辑的玉米；KO：编辑的玉米。
具体实施方式
提供以下定义和方法用以更好地界定本申请以及在本申请实践中指导本领域普通技术人员。除非另作说明，术语按照相关领域普通技术人员的常规用法理解。本文所引用的所有专利文献、学术论文、行业标准及其他公开出版物等，其中的全部内容整体并入本文作为参考。
本发明在本实验室构建的高通量CRISPR/Cas9突变体库中筛选到一个短穗突变体材料，通过基因型鉴定确认所编辑基因为Zm00001d040798。为进一步确认该基因的功能，对Zm00001d040798基因重新设计靶点进行基因编辑敲除操作并获得了该基因的敲除系，确定敲除Zm00001d040798基因后玉米穗长变短，可用于改良鲜食玉米果穗形态，提高鲜食玉米品质。
以下实施例用于说明本发明，但不用来限制本发明的范围。在不背离本发明精神和实质的情况下，对本发明方法、步骤或条件所作的修改或替换，均属于本申请的范围。若无特别指明，实施例按照常规实验条件，如Sambrook等人的分子克隆实验手册(SambrookJ&Russell D W,Molecular cloning:a laboratory manual,2001)，或按照制造厂商说明书建议的条件。若未特别指明，实施例中所用的化学试剂均为常规市售试剂，实施例中所用的技术手段为本领域技术人员所熟知的常规手段。
实施例1玉米穗长基因的功能验证
本发明在本实验室构建的高通量CRISPR/Cas9突变体库(构建方法详见：Liu H J,Jian L,Xu J,et al.High-Throughput CRISPR/Cas9 Mutagenesis Streamlines TraitGene Identification in Maize[J].The Plant Cell,2020,32(5))中筛选到一个短穗突变体材料，经过鉴定，发现该突变体的突变基因是Zm00001d040798，该基因编码硫胺素焦磷酸激酶1，将其命名为ZmTPK1。根据玉米参考基因组上的注释信息，ZmTPK1编码蛋白的氨基酸序列如SEQ ID NO.1所示，基因组序列如SEQ ID NO.2所示，cDNA序列如SEQ ID NO.3所示。并无公开资料显示该基因控制玉米穗长性状。为了验证其功能，将该基因重新设计靶点进行基因编辑敲除操作。具体步骤如下：
利用在线软件CRISPR-P 2.0(http://crispr.hzau.edu.cn/CRISPR2/)在基因CDS区域上各选取两个靶标编辑位点设计gRNA(序列见表1)。直接合成约2000bp的“U6-promoter1-gRNA1-sgRNA-U6 promoter2-gRNA2-sgRNA”融合单元序列，利用双酶切体系将其从中间载体PUC57上切下，并通过同源重组的方法将酶切片段连接到骨架载体CPB-ZmUbi-hspCas9上(图1)。将重组载体转化至大肠杆菌感受态细胞DH5α，检测目标序列，序列检测无误后转化玉米自交系KN5585。对获得的T0代转化植株，每个基因的两个靶标位点进行检测。检测通过PCR扩增后测序，根据测序序列分析编辑类型。
表1基因编辑的靶标序列和检测引物信息
方框标识PAM序列
获得的ZmTPK1基因编辑敲除材料的果穗整体偏小，穗长、行粒数、穗宽和穗行数等果穗形态相关性状均显著低于野生型对照(表2)，可以用于改良一些果穗偏大的鲜食玉米品种。
表2 ZmTPK1对玉米果穗形态相关性状的影响
KO1，KO2：2个敲除材料；NT：受体对照；N：材料数。表型值以平均值±标准差表示，显著性用单因素方差分析计算获得。
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gcctcagtgt ttcctgcgcc gtttgcggta gcctgtgatg tcgcacgtga gctctctcca 300
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gaagcgtgac tccagtcctt gccaatcgat tttagtgcct tccagggtta cggctgctta 420
gtcaaatcga tctcaccggg attgcgtagt tccaggcata tggtgtcgac tttgggaggg 480
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taggctggga agttctcctt ttgaggagat tttggcatat aatcaatgac tataaaatag 600
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gccacaaagt atctagtctc agttcattgt tttttaaaga caatcatgtc tcacttaaat 3780
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agttagccaa aataagattt tatgttcaag taagtcagaa gagtatcttt caatgaagaa 4020
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aaaagtatca tgtcagtttt attgacgatt tcagcaagtt cacctcgttg aatttcagaa 4200
catggttgag caccacttta acacgaagat cattgctatc caaactaatt ggggggatga 4260
atatcagcgt ccgaactcgc ttagtgtctt gtcctcatgc tcaccacaaa gtggtcctgt 4320
tgaaagaaag agtcgccaca taatagaggt tggcctattt tacttgtaca tgcttcaatg 4380
ccacttacat tccgggatga agaatttgtt gttgttgcat accttataga tcacacaccc 4440
accatgttgc ttgattattc atcttctttt gagaaaatat ttcaacaacc tcctgattgt 4500
tcatctccca aggttttagg gtgtggctac tagcctaact tgtgtcctta caacacttag 4560
aagttgttat ttaggtatgt ttgttgtgtt tttctaggat acaacactac acaacactat 4620
gcataaaggg tctctgatca tccactggtt ttatttatat ttttcgagat gtttttgaaa 4680
aaactcggcc ggtaggggaa agaccacccc cacggcattg aattaagaag aagctcaatc 4740
ggagccccga ccgagaaagg tcacgaaagc tggtccccgt gcaacatgag ggtccgcccc 4800
tataggccag atcgtgcttt gagccctccc agcccctaca tgaggatctg ccccccatag 4860
gttagtccat ctcgtgtgca catgaccagg gaaccagcga gtgacctttt ttaacctcag 4920
cctgaaattt gctcccactg ggattcgaac tcaggaccta agaagtgcta ctcaggccac 4980
ctaaccaact tggctagagg ccctttcgtc gagatgtttt tgatgaagaa gcttttctat 5040
ttgctgagct tcaccctaat gctggtacaa acactagaca aacgcagacc cagaccccta 5100
agaaccccaa gggtgtctgt cgcagacaga tgcagacccc taagccccaa gggtgtctat 5160
cgagtcggtc ggggttcatt tagtgggacg atctcttggt cggacagaaa ataatttttg 5220
gcgttctgat ggacacatcc agttgtatcc tttttggtag gatctaacgg gtatcctttg 5280
ttgttgttcc catcaatctc ccgtctataa gtgtctacgc tttcctggag ggggtgaaca 5340
tgctgacatt gttaaggttg gtgtgaatct cgtgtccatt gtgttgtaaa cgagtagtcc 5400
gagcgagacc taggattcaa ccggtgaggg atctgtccta ttaggcagtt ccacgtggtg 5460
ggtgcctaac ccgctggggc gtctcattat ataatccttc cttactcaac cctgtctgga 5520
cttggtaagt ttgagcggca atgttgccca cgccttgagg ggatttgcag gggagtgcta 5580
ggtatgacac ttgaatggtt gatcgagatt ccccatgcat accagtgcgg ttagtcagtg 5640
cccctaccat tgagttgtca aatatcactt aagagggctc gctcataatt gccccacacc 5700
catttttgcc gattgacagg ctaacgattg tggttgtttg ggacggccta taagggcata 5760
aattcgacac taggatgtaa aagaccggtg gcttcttccc tacaacacat gggtgtaata 5820
gtaggggtat tgaacgaagt caggttttgg gagttgtact cgtaatgatt tgaagcctgg 5880
tcgcggctgt tgcgcatgcg acatagtggt cgaggcggtt ggtattcgtg agtcggcgtg 5940
cgacgtggga ctgggtggtc cactagctag acgtcaagaa tgcctttctt cacggcactc 6000
tgaccgagac gttccgctca ctcgggccta tgtgcaagct taaccgctcc ttgtatgtcc 6060
tcaagcaggc gtcacaggcc tgttacaatc gcttcgcctc ctatttggcc tctgtcggct 6120
tcattgaggc caagtcagac atgtccctgt ttatccttcg gtgcgtcaat gacactgtct 6180
acctcctgct ctatgtcgag aacatcgtgc ttatggtgtc cagccccgcc ctcctttggc 6240
gcacgatcgc ctcactacat cgtgagtttg cgatgaagga cttgggccct ctccaccact 6300
tcctggggat tgttgtggag caccaccccc aaagcctctt ccttcactag tgccagtaca 6360
ccatcagcat tctcgagcag gccggcatgt ccgactgcaa gccttgctcc acgcccgttg 6420
acacacatgc caagctctcc gatgacgaca acgacatgat gacgtaccag agttttgtcg 6480
aggctctcta gtgcctcacc ttcacccaac ccgacatcgc ctacgtcgtc cagcaggtgt 6540
tccttcacat gcacacccca cgggagccgc gcctcactat cgcaaagcgg attctttgct 6600
accttcacgg caccctcgac cacgaccttc ttctccgccc ctctctggcc tttgagctag 6660
tcatctacac tgatgctgac tggatcgatt gccctgacac ctgctggtcc acctctggct 6720
atgtcttgtt cttgggacca acctcgtctc ctggtcgttg aagtggcagc ccgtcgtctc 6780
ccattctagc gctgaggtcg agtatcgcga tgtgggcaat ggcatggccg aggcctcctg 6840
gctttgccag ctgtttcaag ggcttcacaa cgtcaagatg atctatctct ccaatccagt 6900
atagcaccag cgcacgaaac acgtggagat tgacctccac ttcattcgtg agcatgtcgc 6960
tgtcggtgat gtccgtgtcc tctgcgtcct gacaccttct agtttgccga catctacatc 7020
aagggcctcc tatccacgat cttctcggaa tttgggtcca gtctcaacgt ctataatagc 7080
tagagttgcg actgcgaggg gtattaaaca attgtgtggt tttgtctatt gtaatgggct 7140
tgtcccaacc catatactat tattatatgt atggataagc ctaccctaat cagggttagt 7200
gaaagggaaa tggactccaa tccatttcta gatttgtttt ggtgcttgat gaccatcaca 7260
atcatgtgag ctaactattt tgctaaattt atgattacaa gttcataaga tcaatggttg 7320
tgtttttatg tcaaaaaaca tctcaaatcc agccaaagag gggtaaaact tggaaaagat 7380
gaacttgtag tagtttcgca atttgtgtta gactttgtgt gtgtgtgtgt gtgtgtgtgt 7440
gtgtgtgtgt gtgtgggccg gcaccactgt tgggctgtcg gcccattagg gttagggttg 7500
tgagtctata tattgtaccc catctcctat caatacatca agcacttaca ttattctaca 7560
tggtatcaga ggttagggtt agggtttctc ccaagcactt acattattct acagtttgga 7620
taagttctaa acatccctag gaacctattt gatctatcta gagagattgg aaagctcaga 7680
tttgaagttt cacccttttg aagctaagtt gagctaaaat cagaatatgc cttagagata 7740
tcaaatcgac caaaatctgt cacttagact agttgactag tctctagata tgtttgtata 7800
agtcatagga ccaaaatcga gtgcccagag aggttgtata ctcggtttcg accggctggt 7860
gcactggacc tcctcggtgg cgcaccagac cgagtttgca gagagggtgt aaatggattt 7920
cactcgactg gcgcacggga ccctcggttc ggcgcaacgg ttggcttcag ctttcaatgg 7980
tcgaacctgc gtggcatggt ctgttgtgcc actggatcga tactgtttat tctgacctcg 8040
tcagaatggc tacacgcaac acagtccgac agagctccag tgcacattgt tcatggttcg 8100
gtgcacctgc agactgtgca atttttgccc attcacttca atggctatat gggtagttcg 8160
agactataaa tacaccccca accagcacat tgaaggtaca agagctacac caaacatcta 8220
tacattcatt gcaataactc caagccattc aaagcctcct aggtgccact ttcaagagaa 8280
ctgagctaga aaagagatag tgccttacga gtgtgatcga gttgagtttg ttgctaagtt 8340
cttgtgctct tgtgattggt tttcttttcc tcactctcat tcttgtaaaa gctagcgaga 8400
gattccgtgt tgtgtggaga tccttgcaga gactccagtc tcttgatttg agaaaggata 8460
ctcaaactcg atttagatga tcgcttgaga gggaaagggt tgaaaaagac ccgtcctaag 8520
tggactcctc aatagagacg taggtttttc tggaaaccga acttcggtaa ataaatcgcc 8580
cgtgtctttt gtgttgtgtt atttgcgatt tattcctccc ttccctctcc taagttctct 8640
ttctcgtgtt tgattctaaa ttatttgtgt tgcttccgac aaaattccgc aatcataaga 8700
gcaacactag caagaagaac atctccttca ctctgattaa tttgcctctc gttctaacac 8760
ctaaccgggg ttcaagtttt tgtatttagt gataaatttt aggtttcgcc tattcactct 8820
cctctaaacg actttcagtt agggtttcat ttcctctaac aaatagtact gcatgcgtcc 8880
agttcattta aagtaagagt cggttgggat gttgtaactg agaaatatgt gccaccggtc 8940
ctcctttgct acacgatcta tacatctgct tgccatcagc ccctgcctgt gccttccgac 9000
atccttattt tctggtaact ggttagagcc gaaaatgaac tttgattatc agtgtttatt 9060
ttatggaatc ctgtatgtta tgactctaga ttgattctag catacatgat tgtttttcat 9120
actttgatgt cattttaagt agggttcgta tactatgctt tttctagatg cttgttctct 9180
gttctcccct gagaaacagg aaaacaagat tttttaggat tcatcttagt atcgtaccta 9240
atatcattat taattgctat tatccaggta tgttccagaa ataattgaag gggacatgga 9300
ttctataaga cctgaagtaa aactgttcta ctctagtcag gttcttttat ttgcgctttg 9360
gtttctacat tgtttaccca gtcaggtttg tgtattgcaa tcagttcata ctggatgtta 9420
agttagttgt aactcccgta agaattacgc aggcagtagt accatcactt atcctaacac 9480
cttttaaagc accattgtac tcatggacag caaaaagaat taaaatcaac ttagcaccag 9540
gcaattgact attgtgcatg acatcttatt tggaaacaaa atactaatcc ttctcaagga 9600
gcccatcggt cattgtgaat gactgactct tcaatcaaat tgctatatta actgcagaaa 9660
tttgaaatcc attagaaatt tgaaatcgta tgaatggaag cttgttttat cgttatgtac 9720
accttaggat ccgccggtcc tatcttgact atagtcagag ctcaggatga atgctggctt 9780
agtgactctg gaggaaatgc caattatttt ggtttttatg tcaacaaact ttctgttttc 9840
tatttactat gttaaaacaa ataaattatt ttatacttaa ataattttac gcttctatct 9900
gttgcatacc tgaattgaaa tgagctggta aatgtctgtt tgaaataata aaatattgca 9960
ttaaccaaag catggacttt ctttgtatta ctctagtttc cacacatatg aactttcttg 10020
aagtgtttta aaaaaactcg gccggtgggg aaaagaccgt ccccacggta tttcactaag 10080
aagtgctcaa ccacagtctg gccgagaaaa gccacgaaac ctggccccct catacataca 10140
tgggcatgtc gccccctata gttggggcct tgactcatgc tgaggcctaa cccacatgac 10200
cagcccctta catgaggacc cgtccaccat aggttggttc acccatgtcg acaaagggat 10260
ccaacgggtg acttttttta acctcagcct gaaattcgct ctcacgagga gtagaaccca 10320
ggacctaagg agtgctactc ggaaggagtg ctactcagac cacctaacta actcagctag 10380
agaccctttc gctccctgaa gtgtttagtg tttaacaatc aacttaactt tttaaggtga 10440
gcattaacta gactttgatt ttactaaatg ttgaagaata ctgcacacaa gttttattca 10500
ttcattgaac tacttagctg gctactggcc agttattgac tatatacttt ggcatctaaa 10560
ttttatgatc aggtattcat atgccattcc atatagaatt tgaatctatt accagtggat 10620
taattatact gtagatgtta tcaggatgtc atgattttag taaacttctc ctattttcag 10680
ggatccaaaa tttctgataa gtcacataac caggagacaa cagatctaca caaatgtatt 10740
tctcgaatcc atcattgtac gcctgatgat gaaaaaccaa atgtaagtgg caatcttatt 10800
atttggtctt gatacttctc ttgtttcttc ttcatagttg ttggtagttt ctatcaacat 10860
tagactcaca tctgtacaaa attcattggc aatgaaactt atttttttca tcaaatactg 10920
tagctctgtg tacttgttac tggagcacta ggtggaaggt ttgatcacga ggcggcaaat 10980
atcaatgttc tgtatctgtt ttcggacatg aggattgtcc tcctatcaga cgattgcttg 11040
atccgacttc ttccaagaac acatcgccat gagctttata ttgagtcgtc tgttgaagga 11100
ccccattgtg ggcttttccc tgttggagca ccatcaacaa gcacaacaac tactggcctg 11160
aaatggaatc taagtaaatt tctttgtgct ctgttggcca taaacgtgta gaagttctgg 11220
attgaattaa tggattattg gactcatcta acaaatcaaa ttgatatata ttgcgtggaa 11280
acactgaaga caatggacga aataaaatga agtacatata ttatcaagtt tattcatgaa 11340
atattaatct agtgcaaaat gaaactcttg ctgacattta cataagcatt taattccata 11400
aggataagtt tccctctcaa ttttcatagg ggatgtacat gaacagagta ggatttatgc 11460
tttgtctcat ttattttttg gaccagtaag attagaggta gtattgattc ttttccccac 11520
ttccaggtga atcaaagatg agatttggga gcatgataag cacatccaac attgtgcagt 11580
cggagaaagt aactgtagaa tctgatgcag atctcttgtg gacaatttct ctgcgaaatc 11640
tgacataagc agttcctagt ttgttatctt ttttcttctc tttcctgtca agtggcaacc 11700
aagctcttgt acagccactt ttgtgtttat gagttgtgtg tctggtaaat aaataccatc 11760
tgatgacaaa tcgatgaggt ctcatcatta gacaggatgt ctaccacgat cttgaactgc 11820
tgatgaatta tgtgaataac acgagcaggc atgatggttg tatcctctct agttgtataa 11880
ctgtattgtt gtacttgcaa cggggtttct tgtataaaat tgttctcaac agttcacact 11940
gtatggttgg attttgtaat gagtctggca attgggtaca gtgatatatg ctaatccatc 12000
t 12001
<210> 3
<211> 1447
<212> DNA
<213> Zea mays
<400> 3
gtggtcccgt ttgacggcgc tcgtgagaac agtccatgtc cgtcgcagca catcgagcgg 60
ctacaaccaa cttcagaatt ctgaaaggga cacaccgtca taccgacagc gaagaccttt 120
gccgttttta ttcttcctcc gtctctgcct gtacagcggc ggcttggcca cctctgagcc 180
tcaagccctc aacctccctc ttcggcgcgt aacgtcagtc ctcaggtatt ggcagcataa 240
agtgttagat ttgtccttga ttgaggtacc ccttcaaagg tcaagatgtt gtggcgcgca 300
gtacgcagta tggatgtaat aatgcattct tcgagtttcc ttcttccgaa actgtatcaa 360
cctgtcaaca aaccagtcaa gaactatgct ttagttgttc taaaccagca tcttccacga 420
ttcatgcctc gactctggga tcatgaacag caaacttgag gatctgcgct gatggaggag 480
ctaatcatat ttttgatgag atgtatcaga taaccaatga tgaagacaaa aaaagcacca 540
gaaataagta tgttccagaa ataattgaag gggacatgga ttctataaga cctgaagtaa 600
aactgttcta ctctagtcag ggatccaaaa tttctgataa gtcacataac caggagacaa 660
cagatctaca caaatgtatt tctcgaatcc atcattgtac gcctgatgat gaaaaaccaa 720
atctctgtgt acttgttact ggagcactag gtggaaggtt tgatcacgag gcggcaaata 780
tcaatgttct gtatctgttt tcggacatga ggattgtcct cctatcagac gattgcttga 840
tccgacttct tccaagaaca catcgccatg agctttatat tgagtcgtct gttgaaggac 900
cccattgtgg gcttttccct gttggagcac catcaacaag cacaacaact actggcctga 960
aatggaatct aagtgaatca aagatgagat ttgggagcat gataagcaca tccaacattg 1020
tgcagtcgga gaaagtaact gtagaatctg atgcagatct cttgtggaca atttctctgc 1080
gaaatctgac ataagcagtt cctagtttgt tatctttttt cttctctttc ctgtcaagtg 1140
gcaaccaagc tcttgtacag ccacttttgt gtttatgagt tgtgtgtctg gtaaataaat 1200
accatctgat gacaaatcga tgaggtctca tcattagaca ggatgtctac cacgatcttg 1260
aactgctgat gaattatgtg aataacacga gcaggcatga tggttgtatc ctctctagtt 1320
gtataactgt attgttgtac ttgcaacggg gtttcttgta taaaattgtt ctcaacagtt 1380
cacactgtat ggttggattt tgtaatgagt ctggcaattg ggtacagtga tatatgctaa 1440
tccatct 1447
<210> 4
<211> 20
<212> DNA
<213> Zea mays
<400> 4
ttctgataag tcacataacc 20
<210> 5
<211> 20
<212> DNA
<213> Zea mays
<400> 5
gtacttgtta ctggagcact 20
<210> 6
<211> 103
<212> DNA/RNA
<213> unknown(人工合成)
<400> 6
uucugauaag ucacauaacc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc 60
cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu uuu 103
<210> 7
<211> 103
<212> DNA/RNA
<213> unknown(人工合成)
<400> 7
guacuuguua cuggagcacu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc 60
cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu uuu 103
<210> 8
<211> 11999
<212> DNA
<213> unknown(人工合成)
<400> 8
gtggtcccgt ttgacggcgc tcgtgagaac agtccatgtc cgtcgcagca catcgagcgg 60
ctacaaccaa cttcagaatt ctgaaaggga cacaccgtca taccgacagc gaagaccttt 120
gccgttttta ttcttcctcc gtctctgcct gtacagcggc ggcttggcca cctctgagcc 180
tcaagccctc aacctccctc ttcggcgcgt aacgtcagtc ctcaggtgag atctgcttgc 240
gcctcagtgt ttcctgcgcc gtttgcggta gcctgtgatg tcgcacgtga gctctctcca 300
ttcttttcct tctcggagtt ggttctgttg ccccttgtgt ttctctcggt tctgggttcc 360
gaagcgtgac tccagtcctt gccaatcgat tttagtgcct tccagggtta cggctgctta 420
gtcaaatcga tctcaccggg attgcgtagt tccaggcata tggtgtcgac tttgggaggg 480
gaggtgcaga atttcaggtt attctttgtc ttgtgttcta aaagatattc cccacatcaa 540
taggctggga agttctcctt ttgaggagat tttggcatat aatcaatgac tataaaatag 600
tatggttagt tctgatgtag atgccgtttc tgcatttggg ttctccagag tctgaatttg 660
ctactacgaa agcaaatgga gtaatcatgg catatgtcag gatagtcagc ttgtattact 720
aaactattta tgtgaatgcc attggaacat aggtcttggt gcaagcaaac ataacctcat 780
atgttttaca tgatggaata aacaaaagaa taatgctcaa gatgagtaag cactgtgcgt 840
ggggcgtcgt cgtgttttct taggccatgt atggcacgtc ttgcggagtt ggagctgctt 900
gtcccttggg tcgagctcaa catcgataac tctggatggg ttgtgggggg gggggcttcg 960
ttgtctgagc tgtttgtgca ttctgtataa gtttttgggc tcggttttcc ttaaaactag 1020
gtcaattcta tacttcttaa ttgaaaggca gagcttctgc tgttacgttc aaaaaaaagt 1080
aagcgctaca agaatccatc ctctcttcca ttctgttaca ctgaaaaatc atgtagtaat 1140
tgaatcttaa ctgggggcat aggtgtattc ttcttctttt tcttagataa tggaaagaaa 1200
tatcgatatg taactaaatg ttagtgctta aaatatgtgg ttattgttgc ctgagaatca 1260
atcaaattgt gtagtgcagt tttgttggat acttgactaa aaaactatta atacctagag 1320
tcccacatct caagacagac cttttaggga acataaaaga agaccctttt gagcagtacc 1380
gcacagaaga aaaaaatgat tgtcatataa tggcattggc atcaataata gttgtctttc 1440
gtatataggt attggcagca taaagtgtta gatttgtcct tgattgaggt accccttcaa 1500
aggtcaagat gttgtggcgc gcagtacgca gtatggatgt aataatgcat tcttcgagtt 1560
tccttcttcc gaaactgtat caacctgtca acaaaccagt caagaactat gctttagttg 1620
ttctaaacca gcatcttcca cgattcatgc ctcgactctg ggatcatggt aactagacat 1680
agctcatctg tctttcctat atttttgtag gaaagtatca ttaaaacaca tttgcatttt 1740
gatgtagaac agcaaacttg aggatctgcg ctgatggagg agctaatcat atttttgatg 1800
agatgtatca gataaccaat gatgaagaca aaaaaagcac cagaaataag tgaagttttt 1860
ttctccatta ttttatacta aagtgcttat taagttttca acattcttaa gatcatattc 1920
taataggaac tgccatacca ccatttttat tgatcttaaa aaaactaatg attgtcattt 1980
tactccaaaa tagttttctt cactagatag ttgtgcctta tgcaaagata gatagcatgg 2040
cttagtatct tatgggtttg acgatgagtt gtgattgttg gattggattt tgtttcttaa 2100
acagttgagt ttgattgggt tcgcttaagc tatgggccaa tattgtgtgg caggttaagt 2160
tatttctata tctcattcaa catttaatta gaaaaagaag gatatcatag gttcccttgt 2220
cccagtgatt tagaatggcg gaatatgtgg tagggaggcg acctgtgtgg cagttgttgc 2280
aaaatagatg aaaagaagga tatatgtgag gactcttgct cttgctcgaa gccaatctat 2340
acgctaagtg tagaagcatt attggcaatg aaacatcaat acatttttga tgtcgaatac 2400
acatacgtcc tacttgttac aatatgttat acgtaggcta cgaccgtagt tgtctacgat 2460
cctcttgggc tagattagaa atcgactcta attatatgtt agagataaaa tagaatagga 2520
ctatatgcgt cgagttcatt tagagtcgga gtgagttaga atgttgtaac taagtagaat 2580
ttctcttgta ataggcggct cttgtctgta taaatataca accccatcga acctagaggt 2640
caagggaagc attttaatat cctgatgttg tgtttctttc gtagtatcag ggccacctac 2700
ttcatctgtt agtcggcgtc ctgaccaaga gaagtcaaca tcaacctcac cagcaacaag 2760
accccatagc gtcatccggc tttgttgtac taacgtctct cctagggcaa accattgctg 2820
taaagatcaa caagaccaac ttcacatcct gaaaagcgca agttctgccc atccttagag 2880
gtgtgcaact actagtcttc ctagatggca tggctatagt gcttgatgag gagatctctg 2940
ccaagcagga tggcaagatg atcgacaacg tcaacccaga gtacatccga tggatagcac 3000
tctagtagca ggtccttagc ttcctcatga catccatgac tcttgagtca tggatcaggt 3060
taccatctac aaaacacccc attaggtgtg gtcagaacta tagaagacct atgcgtcata 3120
gtcaagagcc tggaccgtaa acacacggat tgccctggca actacaagaa aaggtaatat 3180
gacaatggtt gaatatattg ccaagatgaa atctcttgcg aatgagatgg catctgcaaa 3240
gatgattgtt gatgatgaag aactcgtctt tgtaattttt ggtggaactt gatgaagagt 3300
acaatctggt ggtctctgct ctggtggatc gtgttgagta ggtcaccata gttgaagcca 3360
cctcccaatt tctcatcttt ggcaccggga tggatctact tcatggagat catcaagcat 3420
ttgctaatgt tgctggtcat gggtgtggaa acaaccatgg cttggccatg ttcgaggcag 3480
agggcatggt gttcctccta gtgactatgg tggctaccaa ggtccatcgt catcttcgtc 3540
tgatggacat ctacaagctc tgtgcactaa caagatctgt caagtctacg acaaggtagg 3600
tcatatggcc ctcgattcct ggtaccacta cgatgagagc tacacctttg acaccaagac 3660
tgcaacagcg gccacccatg gtggttatgg tgtcaacacc aatttcttac atgtttttgt 3720
gccacaaagt atctagtctc agttcattgt tttttaaaga caatcatgtc tcacttaaat 3780
atttttcttg tcactttctg attaaggatc tagacatgca gaaggttctt ctctgtggta 3840
gacaagaaaa cagtctctac cccttatcaa gtttttggcg aggggctcta ggtgccatca 3900
agatgtcccc tcaccggata gccgccttgg tcatccgtcc ttttagacta tttagaagtt 3960
agttagccaa aataagattt tatgttcaag taagtcagaa gagtatcttt caatgaagaa 4020
ctcgtcagca ggcaaagagt catcatcttc tctatcctac atcaacaagt gtctcatcaa 4080
aaacccttga acttgtgttt tcaaatgtat ggggtactgc tttgaagtat gtaggtcgaa 4140
aaaagtatca tgtcagtttt attgacgatt tcagcaagtt cacctcgttg aatttcagaa 4200
catggttgag caccacttta acacgaagat cattgctatc caaactaatt ggggggatga 4260
atatcagcgt ccgaactcgc ttagtgtctt gtcctcatgc tcaccacaaa gtggtcctgt 4320
tgaaagaaag agtcgccaca taatagaggt tggcctattt tacttgtaca tgcttcaatg 4380
ccacttacat tccgggatga agaatttgtt gttgttgcat accttataga tcacacaccc 4440
accatgttgc ttgattattc atcttctttt gagaaaatat ttcaacaacc tcctgattgt 4500
tcatctccca aggttttagg gtgtggctac tagcctaact tgtgtcctta caacacttag 4560
aagttgttat ttaggtatgt ttgttgtgtt tttctaggat acaacactac acaacactat 4620
gcataaaggg tctctgatca tccactggtt ttatttatat ttttcgagat gtttttgaaa 4680
aaactcggcc ggtaggggaa agaccacccc cacggcattg aattaagaag aagctcaatc 4740
ggagccccga ccgagaaagg tcacgaaagc tggtccccgt gcaacatgag ggtccgcccc 4800
tataggccag atcgtgcttt gagccctccc agcccctaca tgaggatctg ccccccatag 4860
gttagtccat ctcgtgtgca catgaccagg gaaccagcga gtgacctttt ttaacctcag 4920
cctgaaattt gctcccactg ggattcgaac tcaggaccta agaagtgcta ctcaggccac 4980
ctaaccaact tggctagagg ccctttcgtc gagatgtttt tgatgaagaa gcttttctat 5040
ttgctgagct tcaccctaat gctggtacaa acactagaca aacgcagacc cagaccccta 5100
agaaccccaa gggtgtctgt cgcagacaga tgcagacccc taagccccaa gggtgtctat 5160
cgagtcggtc ggggttcatt tagtgggacg atctcttggt cggacagaaa ataatttttg 5220
gcgttctgat ggacacatcc agttgtatcc tttttggtag gatctaacgg gtatcctttg 5280
ttgttgttcc catcaatctc ccgtctataa gtgtctacgc tttcctggag ggggtgaaca 5340
tgctgacatt gttaaggttg gtgtgaatct cgtgtccatt gtgttgtaaa cgagtagtcc 5400
gagcgagacc taggattcaa ccggtgaggg atctgtccta ttaggcagtt ccacgtggtg 5460
ggtgcctaac ccgctggggc gtctcattat ataatccttc cttactcaac cctgtctgga 5520
cttggtaagt ttgagcggca atgttgccca cgccttgagg ggatttgcag gggagtgcta 5580
ggtatgacac ttgaatggtt gatcgagatt ccccatgcat accagtgcgg ttagtcagtg 5640
cccctaccat tgagttgtca aatatcactt aagagggctc gctcataatt gccccacacc 5700
catttttgcc gattgacagg ctaacgattg tggttgtttg ggacggccta taagggcata 5760
aattcgacac taggatgtaa aagaccggtg gcttcttccc tacaacacat gggtgtaata 5820
gtaggggtat tgaacgaagt caggttttgg gagttgtact cgtaatgatt tgaagcctgg 5880
tcgcggctgt tgcgcatgcg acatagtggt cgaggcggtt ggtattcgtg agtcggcgtg 5940
cgacgtggga ctgggtggtc cactagctag acgtcaagaa tgcctttctt cacggcactc 6000
tgaccgagac gttccgctca ctcgggccta tgtgcaagct taaccgctcc ttgtatgtcc 6060
tcaagcaggc gtcacaggcc tgttacaatc gcttcgcctc ctatttggcc tctgtcggct 6120
tcattgaggc caagtcagac atgtccctgt ttatccttcg gtgcgtcaat gacactgtct 6180
acctcctgct ctatgtcgag aacatcgtgc ttatggtgtc cagccccgcc ctcctttggc 6240
gcacgatcgc ctcactacat cgtgagtttg cgatgaagga cttgggccct ctccaccact 6300
tcctggggat tgttgtggag caccaccccc aaagcctctt ccttcactag tgccagtaca 6360
ccatcagcat tctcgagcag gccggcatgt ccgactgcaa gccttgctcc acgcccgttg 6420
acacacatgc caagctctcc gatgacgaca acgacatgat gacgtaccag agttttgtcg 6480
aggctctcta gtgcctcacc ttcacccaac ccgacatcgc ctacgtcgtc cagcaggtgt 6540
tccttcacat gcacacccca cgggagccgc gcctcactat cgcaaagcgg attctttgct 6600
accttcacgg caccctcgac cacgaccttc ttctccgccc ctctctggcc tttgagctag 6660
tcatctacac tgatgctgac tggatcgatt gccctgacac ctgctggtcc acctctggct 6720
atgtcttgtt cttgggacca acctcgtctc ctggtcgttg aagtggcagc ccgtcgtctc 6780
ccattctagc gctgaggtcg agtatcgcga tgtgggcaat ggcatggccg aggcctcctg 6840
gctttgccag ctgtttcaag ggcttcacaa cgtcaagatg atctatctct ccaatccagt 6900
atagcaccag cgcacgaaac acgtggagat tgacctccac ttcattcgtg agcatgtcgc 6960
tgtcggtgat gtccgtgtcc tctgcgtcct gacaccttct agtttgccga catctacatc 7020
aagggcctcc tatccacgat cttctcggaa tttgggtcca gtctcaacgt ctataatagc 7080
tagagttgcg actgcgaggg gtattaaaca attgtgtggt tttgtctatt gtaatgggct 7140
tgtcccaacc catatactat tattatatgt atggataagc ctaccctaat cagggttagt 7200
gaaagggaaa tggactccaa tccatttcta gatttgtttt ggtgcttgat gaccatcaca 7260
atcatgtgag ctaactattt tgctaaattt atgattacaa gttcataaga tcaatggttg 7320
tgtttttatg tcaaaaaaca tctcaaatcc agccaaagag gggtaaaact tggaaaagat 7380
gaacttgtag tagtttcgca atttgtgtta gactttgtgt gtgtgtgtgt gtgtgtgtgt 7440
gtgtgtgtgt gtgtgggccg gcaccactgt tgggctgtcg gcccattagg gttagggttg 7500
tgagtctata tattgtaccc catctcctat caatacatca agcacttaca ttattctaca 7560
tggtatcaga ggttagggtt agggtttctc ccaagcactt acattattct acagtttgga 7620
taagttctaa acatccctag gaacctattt gatctatcta gagagattgg aaagctcaga 7680
tttgaagttt cacccttttg aagctaagtt gagctaaaat cagaatatgc cttagagata 7740
tcaaatcgac caaaatctgt cacttagact agttgactag tctctagata tgtttgtata 7800
agtcatagga ccaaaatcga gtgcccagag aggttgtata ctcggtttcg accggctggt 7860
gcactggacc tcctcggtgg cgcaccagac cgagtttgca gagagggtgt aaatggattt 7920
cactcgactg gcgcacggga ccctcggttc ggcgcaacgg ttggcttcag ctttcaatgg 7980
tcgaacctgc gtggcatggt ctgttgtgcc actggatcga tactgtttat tctgacctcg 8040
tcagaatggc tacacgcaac acagtccgac agagctccag tgcacattgt tcatggttcg 8100
gtgcacctgc agactgtgca atttttgccc attcacttca atggctatat gggtagttcg 8160
agactataaa tacaccccca accagcacat tgaaggtaca agagctacac caaacatcta 8220
tacattcatt gcaataactc caagccattc aaagcctcct aggtgccact ttcaagagaa 8280
ctgagctaga aaagagatag tgccttacga gtgtgatcga gttgagtttg ttgctaagtt 8340
cttgtgctct tgtgattggt tttcttttcc tcactctcat tcttgtaaaa gctagcgaga 8400
gattccgtgt tgtgtggaga tccttgcaga gactccagtc tcttgatttg agaaaggata 8460
ctcaaactcg atttagatga tcgcttgaga gggaaagggt tgaaaaagac ccgtcctaag 8520
tggactcctc aatagagacg taggtttttc tggaaaccga acttcggtaa ataaatcgcc 8580
cgtgtctttt gtgttgtgtt atttgcgatt tattcctccc ttccctctcc taagttctct 8640
ttctcgtgtt tgattctaaa ttatttgtgt tgcttccgac aaaattccgc aatcataaga 8700
gcaacactag caagaagaac atctccttca ctctgattaa tttgcctctc gttctaacac 8760
ctaaccgggg ttcaagtttt tgtatttagt gataaatttt aggtttcgcc tattcactct 8820
cctctaaacg actttcagtt agggtttcat ttcctctaac aaatagtact gcatgcgtcc 8880
agttcattta aagtaagagt cggttgggat gttgtaactg agaaatatgt gccaccggtc 8940
ctcctttgct acacgatcta tacatctgct tgccatcagc ccctgcctgt gccttccgac 9000
atccttattt tctggtaact ggttagagcc gaaaatgaac tttgattatc agtgtttatt 9060
ttatggaatc ctgtatgtta tgactctaga ttgattctag catacatgat tgtttttcat 9120
actttgatgt cattttaagt agggttcgta tactatgctt tttctagatg cttgttctct 9180
gttctcccct gagaaacagg aaaacaagat tttttaggat tcatcttagt atcgtaccta 9240
atatcattat taattgctat tatccaggta tgttccagaa ataattgaag gggacatgga 9300
ttctataaga cctgaagtaa aactgttcta ctctagtcag gttcttttat ttgcgctttg 9360
gtttctacat tgtttaccca gtcaggtttg tgtattgcaa tcagttcata ctggatgtta 9420
agttagttgt aactcccgta agaattacgc aggcagtagt accatcactt atcctaacac 9480
cttttaaagc accattgtac tcatggacag caaaaagaat taaaatcaac ttagcaccag 9540
gcaattgact attgtgcatg acatcttatt tggaaacaaa atactaatcc ttctcaagga 9600
gcccatcggt cattgtgaat gactgactct tcaatcaaat tgctatatta actgcagaaa 9660
tttgaaatcc attagaaatt tgaaatcgta tgaatggaag cttgttttat cgttatgtac 9720
accttaggat ccgccggtcc tatcttgact atagtcagag ctcaggatga atgctggctt 9780
agtgactctg gaggaaatgc caattatttt ggtttttatg tcaacaaact ttctgttttc 9840
tatttactat gttaaaacaa ataaattatt ttatacttaa ataattttac gcttctatct 9900
gttgcatacc tgaattgaaa tgagctggta aatgtctgtt tgaaataata aaatattgca 9960
ttaaccaaag catggacttt ctttgtatta ctctagtttc cacacatatg aactttcttg 10020
aagtgtttta aaaaaactcg gccggtgggg aaaagaccgt ccccacggta tttcactaag 10080
aagtgctcaa ccacagtctg gccgagaaaa gccacgaaac ctggccccct catacataca 10140
tgggcatgtc gccccctata gttggggcct tgactcatgc tgaggcctaa cccacatgac 10200
cagcccctta catgaggacc cgtccaccat aggttggttc acccatgtcg acaaagggat 10260
ccaacgggtg acttttttta acctcagcct gaaattcgct ctcacgagga gtagaaccca 10320
ggacctaagg agtgctactc ggaaggagtg ctactcagac cacctaacta actcagctag 10380
agaccctttc gctccctgaa gtgtttagtg tttaacaatc aacttaactt tttaaggtga 10440
gcattaacta gactttgatt ttactaaatg ttgaagaata ctgcacacaa gttttattca 10500
ttcattgaac tacttagctg gctactggcc agttattgac tatatacttt ggcatctaaa 10560
ttttatgatc aggtattcat atgccattcc atatagaatt tgaatctatt accagtggat 10620
taattatact gtagatgtta tcaggatgtc atgattttag taaacttctc ctattttcag 10680
ggatccaaaa tttctgataa gtcacataac caggagacaa cagatctaca caaatgtatt 10740
tctcgaatcc atcattgtac gcctgatgat gaaaaaccaa atgtaagtgg caatcttatt 10800
atttggtctt gatacttctc ttgtttcttc ttcatagttg ttggtagttt ctatcaacat 10860
tagactcaca tctgtacaaa attcattggc aatgaaactt atttttttca tcaaatactg 10920
tagctctgtg tacttgttac tggagctagg tggaaggttt gatcacgagg cggcaaatat 10980
caatgttctg tatctgtttt cggacatgag gattgtcctc ctatcagacg attgcttgat 11040
ccgacttctt ccaagaacac atcgccatga gctttatatt gagtcgtctg ttgaaggacc 11100
ccattgtggg cttttccctg ttggagcacc atcaacaagc acaacaacta ctggcctgaa 11160
atggaatcta agtaaatttc tttgtgctct gttggccata aacgtgtaga agttctggat 11220
tgaattaatg gattattgga ctcatctaac aaatcaaatt gatatatatt gcgtggaaac 11280
actgaagaca atggacgaaa taaaatgaag tacatatatt atcaagttta ttcatgaaat 11340
attaatctag tgcaaaatga aactcttgct gacatttaca taagcattta attccataag 11400
gataagtttc cctctcaatt ttcatagggg atgtacatga acagagtagg atttatgctt 11460
tgtctcattt attttttgga ccagtaagat tagaggtagt attgattctt ttccccactt 11520
ccaggtgaat caaagatgag atttgggagc atgataagca catccaacat tgtgcagtcg 11580
gagaaagtaa ctgtagaatc tgatgcagat ctcttgtgga caatttctct gcgaaatctg 11640
acataagcag ttcctagttt gttatctttt ttcttctctt tcctgtcaag tggcaaccaa 11700
gctcttgtac agccactttt gtgtttatga gttgtgtgtc tggtaaataa ataccatctg 11760
atgacaaatc gatgaggtct catcattaga caggatgtct accacgatct tgaactgctg 11820
atgaattatg tgaataacac gagcaggcat gatggttgta tcctctctag ttgtataact 11880
gtattgttgt acttgcaacg gggtttcttg tataaaattg ttctcaacag ttcacactgt 11940
atggttggat tttgtaatga gtctggcaat tgggtacagt gatatatgct aatccatct 11999
<210> 9
<211> 12001
<212> DNA
<213> unknown(人工合成)
<400> 9
gtggtcccgt ttgacggcgc tcgtgagaac agtccatgtc cgtcgcagca catcgagcgg 60
ctacaaccaa cttcagaatt ctgaaaggga cacaccgtca taccgacagc gaagaccttt 120
gccgttttta ttcttcctcc gtctctgcct gtacagcggc ggcttggcca cctctgagcc 180
tcaagccctc aacctccctc ttcggcgcgt aacgtcagtc ctcaggtgag atctgcttgc 240
gcctcagtgt ttcctgcgcc gtttgcggta gcctgtgatg tcgcacgtga gctctctcca 300
ttcttttcct tctcggagtt ggttctgttg ccccttgtgt ttctctcggt tctgggttcc 360
gaagcgtgac tccagtcctt gccaatcgat tttagtgcct tccagggtta cggctgctta 420
gtcaaatcga tctcaccggg attgcgtagt tccaggcata tggtgtcgac tttgggaggg 480
gaggtgcaga atttcaggtt attctttgtc ttgtgttcta aaagatattc cccacatcaa 540
taggctggga agttctcctt ttgaggagat tttggcatat aatcaatgac tataaaatag 600
tatggttagt tctgatgtag atgccgtttc tgcatttggg ttctccagag tctgaatttg 660
ctactacgaa agcaaatgga gtaatcatgg catatgtcag gatagtcagc ttgtattact 720
aaactattta tgtgaatgcc attggaacat aggtcttggt gcaagcaaac ataacctcat 780
atgttttaca tgatggaata aacaaaagaa taatgctcaa gatgagtaag cactgtgcgt 840
ggggcgtcgt cgtgttttct taggccatgt atggcacgtc ttgcggagtt ggagctgctt 900
gtcccttggg tcgagctcaa catcgataac tctggatggg ttgtgggggg gggggcttcg 960
ttgtctgagc tgtttgtgca ttctgtataa gtttttgggc tcggttttcc ttaaaactag 1020
gtcaattcta tacttcttaa ttgaaaggca gagcttctgc tgttacgttc aaaaaaaagt 1080
aagcgctaca agaatccatc ctctcttcca ttctgttaca ctgaaaaatc atgtagtaat 1140
tgaatcttaa ctgggggcat aggtgtattc ttcttctttt tcttagataa tggaaagaaa 1200
tatcgatatg taactaaatg ttagtgctta aaatatgtgg ttattgttgc ctgagaatca 1260
atcaaattgt gtagtgcagt tttgttggat acttgactaa aaaactatta atacctagag 1320
tcccacatct caagacagac cttttaggga acataaaaga agaccctttt gagcagtacc 1380
gcacagaaga aaaaaatgat tgtcatataa tggcattggc atcaataata gttgtctttc 1440
gtatataggt attggcagca taaagtgtta gatttgtcct tgattgaggt accccttcaa 1500
aggtcaagat gttgtggcgc gcagtacgca gtatggatgt aataatgcat tcttcgagtt 1560
tccttcttcc gaaactgtat caacctgtca acaaaccagt caagaactat gctttagttg 1620
ttctaaacca gcatcttcca cgattcatgc ctcgactctg ggatcatggt aactagacat 1680
agctcatctg tctttcctat atttttgtag gaaagtatca ttaaaacaca tttgcatttt 1740
gatgtagaac agcaaacttg aggatctgcg ctgatggagg agctaatcat atttttgatg 1800
agatgtatca gataaccaat gatgaagaca aaaaaagcac cagaaataag tgaagttttt 1860
ttctccatta ttttatacta aagtgcttat taagttttca acattcttaa gatcatattc 1920
taataggaac tgccatacca ccatttttat tgatcttaaa aaaactaatg attgtcattt 1980
tactccaaaa tagttttctt cactagatag ttgtgcctta tgcaaagata gatagcatgg 2040
cttagtatct tatgggtttg acgatgagtt gtgattgttg gattggattt tgtttcttaa 2100
acagttgagt ttgattgggt tcgcttaagc tatgggccaa tattgtgtgg caggttaagt 2160
tatttctata tctcattcaa catttaatta gaaaaagaag gatatcatag gttcccttgt 2220
cccagtgatt tagaatggcg gaatatgtgg tagggaggcg acctgtgtgg cagttgttgc 2280
aaaatagatg aaaagaagga tatatgtgag gactcttgct cttgctcgaa gccaatctat 2340
acgctaagtg tagaagcatt attggcaatg aaacatcaat acatttttga tgtcgaatac 2400
acatacgtcc tacttgttac aatatgttat acgtaggcta cgaccgtagt tgtctacgat 2460
cctcttgggc tagattagaa atcgactcta attatatgtt agagataaaa tagaatagga 2520
ctatatgcgt cgagttcatt tagagtcgga gtgagttaga atgttgtaac taagtagaat 2580
ttctcttgta ataggcggct cttgtctgta taaatataca accccatcga acctagaggt 2640
caagggaagc attttaatat cctgatgttg tgtttctttc gtagtatcag ggccacctac 2700
ttcatctgtt agtcggcgtc ctgaccaaga gaagtcaaca tcaacctcac cagcaacaag 2760
accccatagc gtcatccggc tttgttgtac taacgtctct cctagggcaa accattgctg 2820
taaagatcaa caagaccaac ttcacatcct gaaaagcgca agttctgccc atccttagag 2880
gtgtgcaact actagtcttc ctagatggca tggctatagt gcttgatgag gagatctctg 2940
ccaagcagga tggcaagatg atcgacaacg tcaacccaga gtacatccga tggatagcac 3000
tctagtagca ggtccttagc ttcctcatga catccatgac tcttgagtca tggatcaggt 3060
taccatctac aaaacacccc attaggtgtg gtcagaacta tagaagacct atgcgtcata 3120
gtcaagagcc tggaccgtaa acacacggat tgccctggca actacaagaa aaggtaatat 3180
gacaatggtt gaatatattg ccaagatgaa atctcttgcg aatgagatgg catctgcaaa 3240
gatgattgtt gatgatgaag aactcgtctt tgtaattttt ggtggaactt gatgaagagt 3300
acaatctggt ggtctctgct ctggtggatc gtgttgagta ggtcaccata gttgaagcca 3360
cctcccaatt tctcatcttt ggcaccggga tggatctact tcatggagat catcaagcat 3420
ttgctaatgt tgctggtcat gggtgtggaa acaaccatgg cttggccatg ttcgaggcag 3480
agggcatggt gttcctccta gtgactatgg tggctaccaa ggtccatcgt catcttcgtc 3540
tgatggacat ctacaagctc tgtgcactaa caagatctgt caagtctacg acaaggtagg 3600
tcatatggcc ctcgattcct ggtaccacta cgatgagagc tacacctttg acaccaagac 3660
tgcaacagcg gccacccatg gtggttatgg tgtcaacacc aatttcttac atgtttttgt 3720
gccacaaagt atctagtctc agttcattgt tttttaaaga caatcatgtc tcacttaaat 3780
atttttcttg tcactttctg attaaggatc tagacatgca gaaggttctt ctctgtggta 3840
gacaagaaaa cagtctctac cccttatcaa gtttttggcg aggggctcta ggtgccatca 3900
agatgtcccc tcaccggata gccgccttgg tcatccgtcc ttttagacta tttagaagtt 3960
agttagccaa aataagattt tatgttcaag taagtcagaa gagtatcttt caatgaagaa 4020
ctcgtcagca ggcaaagagt catcatcttc tctatcctac atcaacaagt gtctcatcaa 4080
aaacccttga acttgtgttt tcaaatgtat ggggtactgc tttgaagtat gtaggtcgaa 4140
aaaagtatca tgtcagtttt attgacgatt tcagcaagtt cacctcgttg aatttcagaa 4200
catggttgag caccacttta acacgaagat cattgctatc caaactaatt ggggggatga 4260
atatcagcgt ccgaactcgc ttagtgtctt gtcctcatgc tcaccacaaa gtggtcctgt 4320
tgaaagaaag agtcgccaca taatagaggt tggcctattt tacttgtaca tgcttcaatg 4380
ccacttacat tccgggatga agaatttgtt gttgttgcat accttataga tcacacaccc 4440
accatgttgc ttgattattc atcttctttt gagaaaatat ttcaacaacc tcctgattgt 4500
tcatctccca aggttttagg gtgtggctac tagcctaact tgtgtcctta caacacttag 4560
aagttgttat ttaggtatgt ttgttgtgtt tttctaggat acaacactac acaacactat 4620
gcataaaggg tctctgatca tccactggtt ttatttatat ttttcgagat gtttttgaaa 4680
aaactcggcc ggtaggggaa agaccacccc cacggcattg aattaagaag aagctcaatc 4740
ggagccccga ccgagaaagg tcacgaaagc tggtccccgt gcaacatgag ggtccgcccc 4800
tataggccag atcgtgcttt gagccctccc agcccctaca tgaggatctg ccccccatag 4860
gttagtccat ctcgtgtgca catgaccagg gaaccagcga gtgacctttt ttaacctcag 4920
cctgaaattt gctcccactg ggattcgaac tcaggaccta agaagtgcta ctcaggccac 4980
ctaaccaact tggctagagg ccctttcgtc gagatgtttt tgatgaagaa gcttttctat 5040
ttgctgagct tcaccctaat gctggtacaa acactagaca aacgcagacc cagaccccta 5100
agaaccccaa gggtgtctgt cgcagacaga tgcagacccc taagccccaa gggtgtctat 5160
cgagtcggtc ggggttcatt tagtgggacg atctcttggt cggacagaaa ataatttttg 5220
gcgttctgat ggacacatcc agttgtatcc tttttggtag gatctaacgg gtatcctttg 5280
ttgttgttcc catcaatctc ccgtctataa gtgtctacgc tttcctggag ggggtgaaca 5340
tgctgacatt gttaaggttg gtgtgaatct cgtgtccatt gtgttgtaaa cgagtagtcc 5400
gagcgagacc taggattcaa ccggtgaggg atctgtccta ttaggcagtt ccacgtggtg 5460
ggtgcctaac ccgctggggc gtctcattat ataatccttc cttactcaac cctgtctgga 5520
cttggtaagt ttgagcggca atgttgccca cgccttgagg ggatttgcag gggagtgcta 5580
ggtatgacac ttgaatggtt gatcgagatt ccccatgcat accagtgcgg ttagtcagtg 5640
cccctaccat tgagttgtca aatatcactt aagagggctc gctcataatt gccccacacc 5700
catttttgcc gattgacagg ctaacgattg tggttgtttg ggacggccta taagggcata 5760
aattcgacac taggatgtaa aagaccggtg gcttcttccc tacaacacat gggtgtaata 5820
gtaggggtat tgaacgaagt caggttttgg gagttgtact cgtaatgatt tgaagcctgg 5880
tcgcggctgt tgcgcatgcg acatagtggt cgaggcggtt ggtattcgtg agtcggcgtg 5940
cgacgtggga ctgggtggtc cactagctag acgtcaagaa tgcctttctt cacggcactc 6000
tgaccgagac gttccgctca ctcgggccta tgtgcaagct taaccgctcc ttgtatgtcc 6060
tcaagcaggc gtcacaggcc tgttacaatc gcttcgcctc ctatttggcc tctgtcggct 6120
tcattgaggc caagtcagac atgtccctgt ttatccttcg gtgcgtcaat gacactgtct 6180
acctcctgct ctatgtcgag aacatcgtgc ttatggtgtc cagccccgcc ctcctttggc 6240
gcacgatcgc ctcactacat cgtgagtttg cgatgaagga cttgggccct ctccaccact 6300
tcctggggat tgttgtggag caccaccccc aaagcctctt ccttcactag tgccagtaca 6360
ccatcagcat tctcgagcag gccggcatgt ccgactgcaa gccttgctcc acgcccgttg 6420
acacacatgc caagctctcc gatgacgaca acgacatgat gacgtaccag agttttgtcg 6480
aggctctcta gtgcctcacc ttcacccaac ccgacatcgc ctacgtcgtc cagcaggtgt 6540
tccttcacat gcacacccca cgggagccgc gcctcactat cgcaaagcgg attctttgct 6600
accttcacgg caccctcgac cacgaccttc ttctccgccc ctctctggcc tttgagctag 6660
tcatctacac tgatgctgac tggatcgatt gccctgacac ctgctggtcc acctctggct 6720
atgtcttgtt cttgggacca acctcgtctc ctggtcgttg aagtggcagc ccgtcgtctc 6780
ccattctagc gctgaggtcg agtatcgcga tgtgggcaat ggcatggccg aggcctcctg 6840
gctttgccag ctgtttcaag ggcttcacaa cgtcaagatg atctatctct ccaatccagt 6900
atagcaccag cgcacgaaac acgtggagat tgacctccac ttcattcgtg agcatgtcgc 6960
tgtcggtgat gtccgtgtcc tctgcgtcct gacaccttct agtttgccga catctacatc 7020
aagggcctcc tatccacgat cttctcggaa tttgggtcca gtctcaacgt ctataatagc 7080
tagagttgcg actgcgaggg gtattaaaca attgtgtggt tttgtctatt gtaatgggct 7140
tgtcccaacc catatactat tattatatgt atggataagc ctaccctaat cagggttagt 7200
gaaagggaaa tggactccaa tccatttcta gatttgtttt ggtgcttgat gaccatcaca 7260
atcatgtgag ctaactattt tgctaaattt atgattacaa gttcataaga tcaatggttg 7320
tgtttttatg tcaaaaaaca tctcaaatcc agccaaagag gggtaaaact tggaaaagat 7380
gaacttgtag tagtttcgca atttgtgtta gactttgtgt gtgtgtgtgt gtgtgtgtgt 7440
gtgtgtgtgt gtgtgggccg gcaccactgt tgggctgtcg gcccattagg gttagggttg 7500
tgagtctata tattgtaccc catctcctat caatacatca agcacttaca ttattctaca 7560
tggtatcaga ggttagggtt agggtttctc ccaagcactt acattattct acagtttgga 7620
taagttctaa acatccctag gaacctattt gatctatcta gagagattgg aaagctcaga 7680
tttgaagttt cacccttttg aagctaagtt gagctaaaat cagaatatgc cttagagata 7740
tcaaatcgac caaaatctgt cacttagact agttgactag tctctagata tgtttgtata 7800
agtcatagga ccaaaatcga gtgcccagag aggttgtata ctcggtttcg accggctggt 7860
gcactggacc tcctcggtgg cgcaccagac cgagtttgca gagagggtgt aaatggattt 7920
cactcgactg gcgcacggga ccctcggttc ggcgcaacgg ttggcttcag ctttcaatgg 7980
tcgaacctgc gtggcatggt ctgttgtgcc actggatcga tactgtttat tctgacctcg 8040
tcagaatggc tacacgcaac acagtccgac agagctccag tgcacattgt tcatggttcg 8100
gtgcacctgc agactgtgca atttttgccc attcacttca atggctatat gggtagttcg 8160
agactataaa tacaccccca accagcacat tgaaggtaca agagctacac caaacatcta 8220
tacattcatt gcaataactc caagccattc aaagcctcct aggtgccact ttcaagagaa 8280
ctgagctaga aaagagatag tgccttacga gtgtgatcga gttgagtttg ttgctaagtt 8340
cttgtgctct tgtgattggt tttcttttcc tcactctcat tcttgtaaaa gctagcgaga 8400
gattccgtgt tgtgtggaga tccttgcaga gactccagtc tcttgatttg agaaaggata 8460
ctcaaactcg atttagatga tcgcttgaga gggaaagggt tgaaaaagac ccgtcctaag 8520
tggactcctc aatagagacg taggtttttc tggaaaccga acttcggtaa ataaatcgcc 8580
cgtgtctttt gtgttgtgtt atttgcgatt tattcctccc ttccctctcc taagttctct 8640
ttctcgtgtt tgattctaaa ttatttgtgt tgcttccgac aaaattccgc aatcataaga 8700
gcaacactag caagaagaac atctccttca ctctgattaa tttgcctctc gttctaacac 8760
ctaaccgggg ttcaagtttt tgtatttagt gataaatttt aggtttcgcc tattcactct 8820
cctctaaacg actttcagtt agggtttcat ttcctctaac aaatagtact gcatgcgtcc 8880
agttcattta aagtaagagt cggttgggat gttgtaactg agaaatatgt gccaccggtc 8940
ctcctttgct acacgatcta tacatctgct tgccatcagc ccctgcctgt gccttccgac 9000
atccttattt tctggtaact ggttagagcc gaaaatgaac tttgattatc agtgtttatt 9060
ttatggaatc ctgtatgtta tgactctaga ttgattctag catacatgat tgtttttcat 9120
actttgatgt cattttaagt agggttcgta tactatgctt tttctagatg cttgttctct 9180
gttctcccct gagaaacagg aaaacaagat tttttaggat tcatcttagt atcgtaccta 9240
atatcattat taattgctat tatccaggta tgttccagaa ataattgaag gggacatgga 9300
ttctataaga cctgaagtaa aactgttcta ctctagtcag gttcttttat ttgcgctttg 9360
gtttctacat tgtttaccca gtcaggtttg tgtattgcaa tcagttcata ctggatgtta 9420
agttagttgt aactcccgta agaattacgc aggcagtagt accatcactt atcctaacac 9480
cttttaaagc accattgtac tcatggacag caaaaagaat taaaatcaac ttagcaccag 9540
gcaattgact attgtgcatg acatcttatt tggaaacaaa atactaatcc ttctcaagga 9600
gcccatcggt cattgtgaat gactgactct tcaatcaaat tgctatatta actgcagaaa 9660
tttgaaatcc attagaaatt tgaaatcgta tgaatggaag cttgttttat cgttatgtac 9720
accttaggat ccgccggtcc tatcttgact atagtcagag ctcaggatga atgctggctt 9780
agtgactctg gaggaaatgc caattatttt ggtttttatg tcaacaaact ttctgttttc 9840
tatttactat gttaaaacaa ataaattatt ttatacttaa ataattttac gcttctatct 9900
gttgcatacc tgaattgaaa tgagctggta aatgtctgtt tgaaataata aaatattgca 9960
ttaaccaaag catggacttt ctttgtatta ctctagtttc cacacatatg aactttcttg 10020
aagtgtttta aaaaaactcg gccggtgggg aaaagaccgt ccccacggta tttcactaag 10080
aagtgctcaa ccacagtctg gccgagaaaa gccacgaaac ctggccccct catacataca 10140
tgggcatgtc gccccctata gttggggcct tgactcatgc tgaggcctaa cccacatgac 10200
cagcccctta catgaggacc cgtccaccat aggttggttc acccatgtcg acaaagggat 10260
ccaacgggtg acttttttta acctcagcct gaaattcgct ctcacgagga gtagaaccca 10320
ggacctaagg agtgctactc ggaaggagtg ctactcagac cacctaacta actcagctag 10380
agaccctttc gctccctgaa gtgtttagtg tttaacaatc aacttaactt tttaaggtga 10440
gcattaacta gactttgatt ttactaaatg ttgaagaata ctgcacacaa gttttattca 10500
ttcattgaac tacttagctg gctactggcc agttattgac tatatacttt ggcatctaaa 10560
ttttatgatc aggtattcat atgccattcc atatagaatt tgaatctatt accagtggat 10620
taattatact gtagatgtta tcaggatgtc atgattttag taaacttctc ctattttcag 10680
ggatccaaaa tttctgataa gtcacatacc aggagacaac agatctacac aaatgtattt 10740
ctcgaatcca tcattgtacg cctgatgatg aaaaaccaaa tgtaagtggc aatcttatta 10800
tttggtcttg atacttctct tgtttcttct tcatagttgt tggtagtttc tatcaacatt 10860
agactcacat ctgtacaaaa ttcattggca atgaaactta tttttttcat caaatactgt 10920
agctctgtgt acttgttact ggagctacta ggtggaaggt ttgatcacga ggcggcaaat 10980
atcaatgttc tgtatctgtt ttcggacatg aggattgtcc tcctatcaga cgattgcttg 11040
atccgacttc ttccaagaac acatcgccat gagctttata ttgagtcgtc tgttgaagga 11100
ccccattgtg ggcttttccc tgttggagca ccatcaacaa gcacaacaac tactggcctg 11160
aaatggaatc taagtaaatt tctttgtgct ctgttggcca taaacgtgta gaagttctgg 11220
attgaattaa tggattattg gactcatcta acaaatcaaa ttgatatata ttgcgtggaa 11280
acactgaaga caatggacga aataaaatga agtacatata ttatcaagtt tattcatgaa 11340
atattaatct agtgcaaaat gaaactcttg ctgacattta cataagcatt taattccata 11400
aggataagtt tccctctcaa ttttcatagg ggatgtacat gaacagagta ggatttatgc 11460
tttgtctcat ttattttttg gaccagtaag attagaggta gtattgattc ttttccccac 11520
ttccaggtga atcaaagatg agatttggga gcatgataag cacatccaac attgtgcagt 11580
cggagaaagt aactgtagaa tctgatgcag atctcttgtg gacaatttct ctgcgaaatc 11640
tgacataagc agttcctagt ttgttatctt ttttcttctc tttcctgtca agtggcaacc 11700
aagctcttgt acagccactt ttgtgtttat gagttgtgtg tctggtaaat aaataccatc 11760
tgatgacaaa tcgatgaggt ctcatcatta gacaggatgt ctaccacgat cttgaactgc 11820
tgatgaatta tgtgaataac acgagcaggc atgatggttg tatcctctct agttgtataa 11880
ctgtattgtt gtacttgcaa cggggtttct tgtataaaat tgttctcaac agttcacact 11940
gtatggttgg attttgtaat gagtctggca attgggtaca gtgatatatg ctaatccatc 12000
t 12001
Claims (8)
1.一种基因在负调控玉米果穗形态中的应用，其特征在于：所述基因序列如SEQ IDNO.2-SEQ ID NO.3任意一个所示。
2.一种改良玉米果穗形态的方法，其特征在于：抑制玉米中SEQ ID NO.1所示蛋白的表达和/或活性，选择玉米果穗变小的植株。
3.根据权利要求2所述改良玉米果穗形态的方法，其特征在于：所述抑制蛋白表达和/或活性的方法包括基因编辑、RNA干扰、T-DNA插入中任一种。
4.根据权利要求3所述改良玉米果穗形态的方法，其特征在于：所述基因编辑采用CRISPR/Cas9方法。
5.根据权利要求4所述改良玉米果穗形态的方法，其特征在于：所述CRISPR/Cas9方法在玉米中的基因组靶标区域的DNA序列如SEQ ID NO.4或SEQ ID NO.5所示。
6.一种改良玉米果穗形态的试剂盒，其特征在于：包括如下任一种：
(1)能够识别权利要求5中所述靶标序列的RNA分子；所述RNA分子的序列如SEQ IDNO.6或SEQ ID NO.7所示；
(2)编码(1)所述RNA的DNA分子；
(3)表达(1)所述RNA的载体。
7.一种玉米突变基因，其特征在于：所述突变基因序列为SEQ ID NO.8或SEQ ID NO.9所示。
8.权利要求7所述的玉米突变基因在改良玉米果穗形态中的应用。
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