CN114752605A - 一种水稻OsOFP22s基因及利用其提高水稻粒长、千粒重和改良直链淀粉含量的方法 - Google Patents
一种水稻OsOFP22s基因及利用其提高水稻粒长、千粒重和改良直链淀粉含量的方法 Download PDFInfo
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- CN114752605A CN114752605A CN202210594164.6A CN202210594164A CN114752605A CN 114752605 A CN114752605 A CN 114752605A CN 202210594164 A CN202210594164 A CN 202210594164A CN 114752605 A CN114752605 A CN 114752605A
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Abstract
本发明公开一种水稻OsOFP22s基因及利用其提高水稻粒长、千粒重和改良直链淀粉含量的方法，该水稻OsOFP22s基因在种子中高表达，可较为特异改良水稻籽粒相关性状，包括粒长、千粒重和直链淀粉含量，而对水稻植株其他主要农艺性状无显著影响，利用基因工程技术在水稻中过量表达OsOFP22s可显著提高稻米千粒重，并改良粒形和蒸煮食味品质；通过比较，OsOFP22s过量表达水稻在粒长和千粒重方面均优于对照，直链淀粉含量与对照相比显著降低，蒸煮食味品质也有明显改良，OsOFP22s基因在水稻的高产优质育种中具有很好的应用前景。
Description
技术领域
本发明属于植物基因工程技术领域，具体涉及一种提高水稻粒长、千粒重和改良直链淀粉含量的基因OsOFP22s及其应用，还涉及由该基因编码的多肽序列。
背景技术
水稻(Oryza sativa L.)是世界上一半以上人口的主粮，我国是全球水稻产量和消费第一大国。随着人口的增长和耕地面积的逐渐减少，为确保国家粮食安全，提高水稻单位面积产量成为提高粮食总产量的最重要途径。同时，随着人们生活水平的提高和稻米市场的开放，稻米作为商品进行广泛交易，人们对稻米外观和蒸煮食味等品质也提出了越来越高的要求。而此前我国水稻育种一直以高产作为主要目标，品质改良进展相对缓慢，导致我国优质稻米品种较少，且在国际稻米市场上竞争力不强。因此，在保证产量的基础上，进一步改良稻米品质显得尤为重要。目前，选育优质高产的水稻新品种己成为我国水稻育种的最重要目标。
水稻粒形不仅是决定水稻产量的一个重要因素，还是衡量稻米外观、加工等品质的重要指标。此外，粒形受环境影响较小，遗传效应显著。因此，改良稻米粒形是实现水稻高产优质育种的一条有效途径。为加快水稻粒形改良的育种进程，除采用常规育种方法外，更重要的是发掘更多可利用的粒形基因，完善水稻粒形调控的分子网络，构建高效的分子育种技术体系，通过分子设计育种策略培育特定粒形的水稻新品种。
水稻粒形性状的主要指标包括粒长、粒宽、粒厚和长宽比等方面。其中粒长不仅影响稻米的外观和碾磨品质，而且与水稻千粒重呈显著正相关。此前，通过图位克隆的方法分离克隆到一个特异性调控水稻粒形的基因GS9，其敲除水稻材料的籽粒细长、外观品质提高，而对株高、千粒重等主要农艺性状无明显影响。在不同水稻品种中敲除GS9基因均显著改良了粒形性状。以GS9蛋白为诱饵，在水稻种子cDNA文库中筛选与之有互作的蛋白，筛选到一个卵形蛋白家族成员OsOFP22s与GS9可直接互作，并进一步利用萤火素酶互补实验在植物体内验证了两个蛋白的互作。
发明内容
本发明针对上述问题，提供一种提高水稻粒长、千粒重和改良直链淀粉含量的基因OsOFP22s及其应用。
本发明的目的是通过以下技术方案来实现的：一种水稻OsOFP22s基因，OsOFP22s基因位于水稻第1号染色体上，基因编号为Os05g0477200(NCBI编号)，LOC_Os05g39950(MSU编号)。OsOFP22s基因编码区CDS全长1155bp，没有内含子，所述OsOFP22s基因的编码区序列如SEQ ID NO.1所示(ATGGGCCGGCGGAAGTTCAGGCTCTCCGACATGATGCCCAACGCGTGGTTCTACAAGCTCCGCGACATGCGCGCGCGGGGCGGCCGCGGTGCAACTGCGATGCAGCCGCCGTCGTCGTCGTCGTTGATGAGGGGGAGCAGGGCGGCGCAGCAGCAGGCGGGCACGTGGAGGCTGGGGACGTCGTCGTCGTCGTCGTCGTTGCTGCCGCACAGGGCGTCGTACTACTACACCACCCGGGACAGGGAGGTCCCGCCGCTGCCGCCGCCGCCACCGCCGAGGGGCGTGGATGATCAATTCCCTTCCCTCACGCTGTCGCCGCCGTTGCCGACGAGGAACAGCAGGAGGCGGCACAGGGTTGGGAGATTTGGTTCGACGGAGATGGATGGCGGCGAGCTCGTACTAGCGCCGTCCGACGACCACGACGGCTGCAGCCACCAGGAGCCGCCAGTGGCCGATGCGTCCGGGAGCTCCCGGTGCCGTCGCGACATGTTCATCGGGAGAGATGGCGGCCGGGGCGTGGAGTTCCGGCGCCGGGCGACGACGGTGGATGGTCCTGAGGAGGACGCCGCCGTCGATGTCAAGGTGATCACGTCGGACGCGGACATAATCATCGACCTCGGCGCTGACGACGACGACGACACGCCGGAGAGGGTGCTCCGGCCTGTCGTGACCAGGCCCGCGAGGAGGGAGCTCGACTGGTGCGAGCCGGCGGAGGTGAAGCACGTCGACCTCGCCGAGCTGATGACACCGAGAGCGAGCTCTGCCTCTGCCTCCTCGGAGAAGAGCATCAGCACGGGCAAGCCGAGGCGTTCGTCCGTGTCGTCTCGACGCCGCCTCAAGACGCGCACCAACAGCCCGCGCCTCGCCGCGTGCAGGAAAGGCAAGCCGACGGCGCGGGCAACGACGACGACGCCGACGCAGCCGCCGCTCGCGCACAGCTTCGCGGTGGTGAAGACGTCGTCGGACCCGAGGAGGGACTTCCTCGAGTCCATGGAGGAGATGATCGCCGAGAACGGCATCCGCGACGCCGGCGACCTGGAGGACCTCCTCGCCTGCTACCTCTCCCTCAACTCCGGCGAGTACCATGACCTCATCGTCGAGGTGTTCGAGCAGGTCTGGACCGGCCTCGCCGCTGCCTGTGGCGTCATGCCATGA)；OsOFP22s基因编码384个氨基酸，所述OsOFP22s编码的氨基酸序列如SEQ ID NO.2所示(MGRRKFRLSDMMPNAWFYKLRDMRARGGRGATAMQPPSSSSLMRGSRAAQQQAGTWRLGTSSSSSSLLPHRASYYYTTRDREVPPLPPPPPPRGVDDQFPSLTLSPPLPTRNSRRRHRVGRFGSTEMDGGELVLAPSDDHDGCSHQEPPVADASGSSRCRRDMFIGRDGGRGVEFRRRATTVDGPEEDAAVDVKVITSDADIIIDLGADDDDDTPERVLRPVVTRPARRELDWCEPAEVKHVDLAELMTPRASSASASSEKSISTGKPRRSSVSSRRRLKTRTNSPRLAACRKGKPTARATTTTPTQPPLAHSFAVVKTSSDPRRDFLESMEEMIAENGIRDAGDLEDLLACYLSLNSGEYHDLIVEVFEQVWTGLAAACGVMP)。
所述的水稻OsOFP22s基因的过表达重组载体pActin-OFP22s-3Flag，其特征在于，所述pActin-OFP22s-3Flag包含OsOFP22s基因，载体为植物双元表达载体pActin-3Flag。
经由PCR对反转录取得的cDNA第一链进行扩增出目的条带。扩增后将含有目标基因的产物和中间载体连接，将连接产物转化到大肠杆菌感受态细胞中。摇菌，遴选单菌落酶切鉴定并送测序。若测序结果正确，提取质粒，切出目标基因并连入最终表达载体pActin-OFP22s-3Flag。
进一步地，所述过表达重组载体pActin-OFP22s-3Flag的制备方法如下：
将植物双元表达载体pCAMBIA1300用Sam I和Xba I双酶切，用同源重组酶将酶切的载体与包含OsOFP22s基因全长cDNA的PCR扩增产物连接。
优选的，所述包含OsOFP22s基因全长cDNA的PCR扩增方法如下：
以野生型日本晴的RNA为模板，引物序列如下：
| 序列名称 | 序列 | 序列编号 |
| OFP22<sup>s</sup>-OE-F | 5-TAGGTAGAAGAGGTACCCGGGCTCTGGCCTGGCCCCCCA-3 | SEQ ID NO.3 |
| OFP22<sup>s</sup>-OE-R | 5-GTAATCTCCGTCGACTCTAGATGGCATGACGCCACAGGC-3 | SEQ ID NO.4 |
。
进一步地，所述的水稻OsOFP22s基因在提高水稻粒长、千粒重和改良直链淀粉含量方面的应用。
优选的，所述应用的方法如下：用所述水稻OsOFP22s基因序列构建可在水稻中过表达的重组载体，并转化农杆菌，再用农杆菌介导的水稻遗传转化方法将载体转入水稻愈伤组织，经抗性筛选和组织培养后变成水稻植株。
优选的，在水稻中过表达的重组载体为所述的过表达重组载体pActin-OFP22s-3Flag。
优选的，所述的应用方法如下：
(1)构建工程菌：将pActin-OFP22s-3Flag载体通过电击法转化到农杆菌菌株EHA105中，通过卡那霉素和利福平筛选，获得含有pActin-OFP22s-3Flag载体的农杆菌；
(2)pActin-OFP22s-3Flag载体转化水稻愈伤并获得水稻再生苗：用含有pActin-OFP22s-3Flag载体的EHA105侵染水稻愈伤组织，并于28℃培养室中共培养3天，再用液体培养基洗去农杆菌后，将水稻愈伤放置在含有合适抗生素的筛选培养基上培养；经过两轮培养后，可获得抗性愈伤，将抗性愈伤转移至分化培养基是进行分化培养获得小苗，分化出的小苗转移至生根培养基上培养，经炼苗后移栽。
转基因植株的分子检测：根据OsOFP22s基因编码区的CDS序列和pActin-OFP22s-3Flag载体序列分别创建正向和反向引物，使用这些引物鉴定过表达材料是否创建成功。引物序列如下：
| 序列名称 | 序列 | 序列编号 |
| pActin-F | 5-TGCTGCTTCGTCAGGCTTAG-3 | SEQ ID NO.5 |
| OFP22s-OE-cs-R | 5-CGTTGGGCATCATGTCGG-3 | SEQ ID NO.6 |
PCR产物为320bp。
荧光试剂定量RT-PCR分析：取提取保存的水稻叶片RNA，经反转录后获得第一链cDNA。进行试剂定量PCR分析，水稻基因Actin01为内源参照基因，检测其表达量是否提高。引物序列如下：
| 序列名称 | 序列 | 序列编号 |
| OFP22<sup>s</sup>-qRT-F | 5-GACATGATGCCCAACGC-3 | SEQ ID NO.7 |
| OFP22<sup>s</sup>-qRT-R | 5-AACCAAATCTCCCAACCCT-3 | SEQ ID NO.8 |
。
本发明具有以下有益效果：(1)本发明中的OsOFP22s蛋白氨基酸与OFP22蛋白在N端少88个氨基酸，表达分析结果显示鉴定到的短转录本OsOFP22s在水稻种子中的表达量远高于OFP22长转录本，且水稻发育种子的三代转录组测序分析结果显示OsOFP22s转录本是水稻种子中最主要的存在形式。
(2)本发明中的水稻OsOFP22s基因在水稻种子中具有高表达，其蛋白产物可与GS9直接互作，更重要的是，OsOFP22s过表达后可协同改良水稻粒形、千粒重和直链淀粉等多个性状，而对其他主要农艺性状无明显负面效应，体现出良好的育种应用价值。
(3)本发明中用水稻OsOFP22s基因序列构建可在水稻中过表达的重组载体，水稻OsOFP22s基因过表达后，其籽粒长度和千粒重都比对照提高，对水稻粒宽、千粒重及稻米理化品质具有显著影响，OsOFP22s过表达稻米的口感更加软糯，在一定程度上改良了稻米的蒸熟品质，对改良稻米产量和品质方面具有重要意义。
附图说明
图1是GS9和OsOFP22s蛋白的酵母双杂交互作验证。
图2是在烟草体内萤火素酶互补实验验证GS9和OsOFP22s的相互作用。
图3是OsOFP22s和OFP22两种转录本在种子中的表达模式比较。
图4是中花11和OsOFP22s过表达材料成熟籽粒的表型，最上面是中花11、下面三行分别为OsOFP22s过表达材料的三个不同系。
图5野生型中花11和OsOFP22s过表达材料稻谷粒型的统计比较。
图6是野生型中花11和OsOFP22s过表达材料稻谷千粒重的统计比较。
图7是野生型中花11和OsOFP22s过表达材料的株型比较。
图8是野生型中花11和OsOFP22s过表达材料稻谷直链淀粉含量(AAC)的比较。
具体实施方式
为理解本发明，以下实施例用于说明本发明，但不限制本发明的范围。
以下实施例中未注明具体条件的实验方法，均按照常规步骤进行，所用材料和实际均为市售商品。
实施例1
GS9与OsOFP22s相互作用的验证
蛋白质与蛋白质之间的相互作用是细胞生命活动的基础，也是细胞生化反应网络的重要组成部分，对调控细胞信号有重要意义。GS9是一个重要的粒型基因，为了验证GS9与OsOFP22s蛋白的相互作用，并进一步研究OsOFP22s蛋白与GS9相互作用的具体蛋白区段，我们将OsOFP22s全长以及N端和C端截短的基因片段，分别构建在pGADT7载体上，形成了pGADT7-OsOFP22s、pGADT7-OsOFP22sN、pGADT7-OsOFP22sC多个构建(图1)。引物序列分别如下：
| 序列名称 | 序列 | 序列编号 |
| OFP22<sup>s</sup>-F | 5-CGGAATTCATGGGCCGGCGGAAGTTCAG-3 | SEQ ID NO.9 |
| OFP22<sup>s</sup>-R | 5-CGGGATCCTCATGGCATGACGCCACAGG-3 | SEQ ID NO.10 |
| OFP22<sup>s</sup>-N-F | 5-CGGAATTCATGGGCCGGCGGAAGTTCAG-3 | SEQ ID NO.11 |
| OFP22<sup>s</sup>-N-R | 5-CGGGATCCTCAGCTGTGCGCGAGCGGCG-3 | SEQ ID NO.12 |
| OFP22<sup>s</sup>-C-F | 5-CGGAATTCATGTTCGCGGTGGTGAAGAC-3 | SEQ ID NO.13 |
| OFP22<sup>s</sup>-C-R | 5-CGGGATCCTCATGGCATGACGCCACAGG-3 | SEQ ID NO.14 |
| GS9C1-F | 5-GGAATTCCATATGCAGAGCAGCAGCAAGCG-3 | SEQ ID NO.15 |
| GS9C1-R | 5-CGGAATTCCTAGCCTCTGGTTCGTATG-3 | SEQ ID NO.16 |
。
把以上载体和pGBKT7-GS9C1共转化到酵母菌AH109后，在相应的营养缺陷培养基上筛选鉴定。结果显示全长OsOFP22s蛋白与GS9C1有相互作用，截短的OsOFP22s中仅OsOFP22sC与GS9C1有相互作用(图1)，说明OsOFP22s的C端是介导其与GS9互作的关键区域。
为进一步验证GS9和OsOFP22s在植物体内是否具有互作，我们开展了荧光素酶互补实验(Split Firefly Luciferase Comp lamentation Assay)验证。将全长OsOFP22s和GS9C1分别克隆到JW771(35S-NLuc)和JW772(35S-CLuc)载体上，引物序列如下所示：
将以上构建正确的载体分别转化到农杆菌GV3101中，在含有卡那霉素的LB培养基中(5mL)震荡培养过夜。设置合适的组合，注射到烟草表皮细胞，48h后利用植物活体分子影像系统(CCD imaging system)检测荧光素酶的活性，验证互作。结果如图2所示。空载体组合NLuc和CLuc，以及空载体和OsOFP22s或GS9C1的组合都没有荧光信号，只有含有OsOFP22s和GS9C1的组合才有荧光信号。该实验说明GS9和OsOFP22s在植物细胞中也具有相互作用。
实施例2
OsOFP22s基因过表达重组载体pActin-OFP22s-3Flag的创建
以野生型日本晴的RNA为模板，合成第一链cDNA，用该OsOFP22s基因编码序列5’和3’端的寡核苷酸作为PCR引物，引物序列如下：
| 序列名称 | 序列 | 序列编号 |
| OFP22<sup>s</sup>-OE-F | 5-TAGGTAGAAGAGGTACCCGGGCTCTGGCCTGGCCCCCCA-3 | SEQ ID NO.3 |
| OFP22<sup>s</sup>-OE-R | 5-GTAATCTCCGTCGACTCTAGATGGCATGACGCCACAGGC-3 | SEQ ID NO.4 |
。
将植物双元表达载体pActin-3Flag(详见www.cambia.org)用Sam I和Xba I双酶切，用同源重组酶将酶切的载体与包含OsOFP22s基因全长cDNA的PCR扩增产物连接；得到过表达重组载体pActin-OFP22s-3Flag；转化DH5α大肠杆菌感受态细胞，在LB平板培养基上通过卡那霉素筛选并经过测序验证。其中测序引物序列如下：
| 序列名称 | 序列 | 序列编号 |
| p1300-3Flag-F | 5-ATGGGGCTCTCGGATGTAGA-3 | SEQ ID NO.21 |
| p1300-3Flag-R | 5-CGATCATAGGCGTCTCGCAT-3 | SEQ ID NO.22 |
。
实施例3
水稻OsOFP22s基因在提高水稻粒长、千粒重和直链淀粉含量方面应用的方法
(1)构建工程菌：将pActin-OFP22s-3Flag载体通过电击法转化到农杆菌菌株EHA105中，通过卡那霉素和利福平筛选，获得含有pActin-OFP22s-3Flag载体的农杆菌；
(2)pActin-OFP22s-3Flag载体转化水稻愈伤并获得水稻再生苗：首先将成熟的中花11种子去壳，用2％的次氯酸钠溶液浸泡消毒1-2h，在消毒期间不停地震荡洗涤种子然后用灭菌蒸馏水清洗3-5次，在无菌超净工作台上用尖头镊子和解剖刀剥出幼胚.将胚转至诱导培养基上，长出的愈伤组织用于后续的遗传转化。挑菌落PCR验证后的农杆菌单菌落接种到4mL含50mg/L卡那霉素的LB液体培养基中，28℃，250rpm过夜培养；第二天按1％的接种量转入50mL相同培养基、相同条件下扩大培养6-8h至对数生长期；4000rpm、4℃离心5min，收集菌体后重悬在10mL含有100-400μmol/L乙酰丁香酮的AAM液体培养基中，随后侵染剥离的水稻愈伤组织20min；侵染后，倒掉菌液并用无菌滤纸吸去残留的菌液，将愈伤组织转入N6D2C培养基上，在28℃、避光条件下培养3天，3天后将愈伤组织转接到含有600mg/L头孢霉素和25mg/L潮霉素的N6D2S1培养基上进行第一轮筛选培养；两周后转移到新的含有300mg/L头孢霉素和50mg/L潮霉素的N6D2S2培养基上进行第二轮筛选培养；然后将活性强的抗性愈伤转移至分化培养基上进行分化培养，分化出的小苗转至1/2MS0生根培养基上培养，经炼苗后移栽至转基因水稻试验田。
(3)转基因植株的分子检测：提取待检测转基因水稻叶片中目的基因基因组DNA，根据OsOFP22s基因编码区的CDS序列和pActin-OFP22s-3Flag载体序列分别创建正向和反向引物，使用这些引物鉴定过表达材料是否创建成功。引物序列如下：
| 序列名称 | 序列 | 序列编号 |
| pActin-F | 5-TGCTGCTTCGTCAGGCTTAG-3 | SEQ ID NO.5 |
| OFP22s-OE-cs-R | 5-CGTTGGGCATCATGTCGG-3 | SEQ ID NO.6 |
。
通过PCR扩增和凝胶电泳检测表达载体是否已成功转入水稻。PCR产物的电泳条带大小应为320bp，若扩增出与目的片段大小一致的植株则为阳性植株，否则为阴性植株；另一方面通过离体检测水稻叶片的潮霉素抗性，进一步确认筛选阳性的转基因植株。潮霉素检测引物如下：
| 序列名称 | 序列 | 序列编号 |
| Hyg-F | 5-GCTTCTGCGGGCGATTTGTGT-3 | SEQ ID NO.23 |
| Hyg-R | 5-GGTCGCGGAGGCTATGGATGC-3 | SEQ ID NO.24 |
。
实施例4
OsOFP22基因过表达在改良水稻粒长、千粒重方面的考察
OsOFP22s基因在过表达后水稻粒长和千粒重方面具有很高的应用价值。过表达后，其籽粒长度比对照显著提高，具体见图4所示，具体数据统计比较见图5。千粒重与野生型对照相比极显著增加，具体见图6。本发明提供的基因和基因工程技术手段在不影响水稻其他农艺性状的前提下(图7)，可显著提高水稻产量，具有很大的应用价值。
实施例5
OsOFP22s基因过表达显著降低水稻直链淀粉含量
对影响稻米蒸煮食味品质(ECQ)的最重要因素直链淀粉含量(AAC)开展分析，结果表明OsOFP22s的过量表达使水稻的AAC极显著降低(图8)。该结果表明OsOFP22s过量表达稻米的口感更加软糯，在一定程度上改良了稻米的蒸煮品质。
以上所述仅为本发明的较佳实施例，然而并非用以限定本发明，本领域技术人员，在不脱离本发明技术方案范围的情况下，可利用上述揭示的技术内容对本发明做出可能的变动和修饰，或修改为等同变化的等效实施例，在未脱离本发明技术方案的内容，依据本发明技术实质对以上实施例所做的任何简单修改、等同变化及修饰，均落在本发明技术方案保护范围内。
序列表
<110> 扬州大学
<120> 一种水稻OsOFP22s基因及利用其提高水稻粒长、千粒重和改良直链淀粉含量的方法
<160> 22
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1155
<212> DNA
<213> 水稻(Oryza sativa var.)
<400> 1
atgggccggc ggaagttcag gctctccgac atgatgccca acgcgtggtt ctacaagctc 60
cgcgacatgc gcgcgcgggg cggccgcggt gcaactgcga tgcagccgcc gtcgtcgtcg 120
tcgttgatga gggggagcag ggcggcgcag cagcaggcgg gcacgtggag gctggggacg 180
tcgtcgtcgt cgtcgtcgtt gctgccgcac agggcgtcgt actactacac cacccgggac 240
agggaggtcc cgccgctgcc gccgccgcca ccgccgaggg gcgtggatga tcaattccct 300
tccctcacgc tgtcgccgcc gttgccgacg aggaacagca ggaggcggca cagggttggg 360
agatttggtt cgacggagat ggatggcggc gagctcgtac tagcgccgtc cgacgaccac 420
gacggctgca gccaccagga gccgccagtg gccgatgcgt ccgggagctc ccggtgccgt 480
cgcgacatgt tcatcgggag agatggcggc cggggcgtgg agttccggcg ccgggcgacg 540
acggtggatg gtcctgagga ggacgccgcc gtcgatgtca aggtgatcac gtcggacgcg 600
gacataatca tcgacctcgg cgctgacgac gacgacgaca cgccggagag ggtgctccgg 660
cctgtcgtga ccaggcccgc gaggagggag ctcgactggt gcgagccggc ggaggtgaag 720
cacgtcgacc tcgccgagct gatgacaccg agagcgagct ctgcctctgc ctcctcggag 780
aagagcatca gcacgggcaa gccgaggcgt tcgtccgtgt cgtctcgacg ccgcctcaag 840
acgcgcacca acagcccgcg cctcgccgcg tgcaggaaag gcaagccgac ggcgcgggca 900
acgacgacga cgccgacgca gccgccgctc gcgcacagct tcgcggtggt gaagacgtcg 960
tcggacccga ggagggactt cctcgagtcc atggaggaga tgatcgccga gaacggcatc 1020
cgcgacgccg gcgacctgga ggacctcctc gcctgctacc tctccctcaa ctccggcgag 1080
taccatgacc tcatcgtcga ggtgttcgag caggtctgga ccggcctcgc cgctgcctgt 1140
ggcgtcatgc catga 1155
<210> 2
<211> 384
<212> PRT
<213> 水稻(Oryza sativa var.)
<400> 2
Met Gly Arg Arg Lys Phe Arg Leu Ser Asp Met Met Pro Asn Ala Trp
1 5 10 15
Phe Tyr Lys Leu Arg Asp Met Arg Ala Arg Gly Gly Arg Gly Ala Thr
20 25 30
Ala Met Gln Pro Pro Ser Ser Ser Ser Leu Met Arg Gly Ser Arg Ala
35 40 45
Ala Gln Gln Gln Ala Gly Thr Trp Arg Leu Gly Thr Ser Ser Ser Ser
50 55 60
Ser Ser Leu Leu Pro His Arg Ala Ser Tyr Tyr Tyr Thr Thr Arg Asp
65 70 75 80
Arg Glu Val Pro Pro Leu Pro Pro Pro Pro Pro Pro Arg Gly Val Asp
85 90 95
Asp Gln Phe Pro Ser Leu Thr Leu Ser Pro Pro Leu Pro Thr Arg Asn
100 105 110
Ser Arg Arg Arg His Arg Val Gly Arg Phe Gly Ser Thr Glu Met Asp
115 120 125
Gly Gly Glu Leu Val Leu Ala Pro Ser Asp Asp His Asp Gly Cys Ser
130 135 140
His Gln Glu Pro Pro Val Ala Asp Ala Ser Gly Ser Ser Arg Cys Arg
145 150 155 160
Arg Asp Met Phe Ile Gly Arg Asp Gly Gly Arg Gly Val Glu Phe Arg
165 170 175
Arg Arg Ala Thr Thr Val Asp Gly Pro Glu Glu Asp Ala Ala Val Asp
180 185 190
Val Lys Val Ile Thr Ser Asp Ala Asp Ile Ile Ile Asp Leu Gly Ala
195 200 205
Asp Asp Asp Asp Asp Thr Pro Glu Arg Val Leu Arg Pro Val Val Thr
210 215 220
Arg Pro Ala Arg Arg Glu Leu Asp Trp Cys Glu Pro Ala Glu Val Lys
225 230 235 240
His Val Asp Leu Ala Glu Leu Met Thr Pro Arg Ala Ser Ser Ala Ser
245 250 255
Ala Ser Ser Glu Lys Ser Ile Ser Thr Gly Lys Pro Arg Arg Ser Ser
260 265 270
Val Ser Ser Arg Arg Arg Leu Lys Thr Arg Thr Asn Ser Pro Arg Leu
275 280 285
Ala Ala Cys Arg Lys Gly Lys Pro Thr Ala Arg Ala Thr Thr Thr Thr
290 295 300
Pro Thr Gln Pro Pro Leu Ala His Ser Phe Ala Val Val Lys Thr Ser
305 310 315 320
Ser Asp Pro Arg Arg Asp Phe Leu Glu Ser Met Glu Glu Met Ile Ala
325 330 335
Glu Asn Gly Ile Arg Asp Ala Gly Asp Leu Glu Asp Leu Leu Ala Cys
340 345 350
Tyr Leu Ser Leu Asn Ser Gly Glu Tyr His Asp Leu Ile Val Glu Val
355 360 365
Phe Glu Gln Val Trp Thr Gly Leu Ala Ala Ala Cys Gly Val Met Pro
370 375 380
<210> 3
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
taggtagaag aggtacccgg gctctggcct ggcccccca 39
<210> 4
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
gtaatctccg tcgactctag atggcatgac gccacaggc 39
<210> 5
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
tgctgcttcg tcaggcttag 20
<210> 6
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
cgttgggcat catgtcgg 18
<210> 7
<211> 17
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gacatgatgc ccaacgc 17
<210> 8
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
aaccaaatct cccaaccct 19
<210> 9
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
cggaattcat gggccggcgg aagttcag 28
<210> 10
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
cgggatcctc atggcatgac gccacagg 28
<210> 11
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
cggaattcat gggccggcgg aagttcag 28
<210> 12
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
cgggatcctc agctgtgcgc gagcggcg 28
<210> 13
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
cggaattcat gttcgcggtg gtgaagac 28
<210> 14
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
cgggatcctc atggcatgac gccacagg 28
<210> 15
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
ggaattccat atgcagagca gcagcaagcg 30
<210> 16
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
cggaattcct agcctctggt tcgtatg 27
<210> 17
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
acgggggacg agctcggtac catgggccgg cggaagttc 39
<210> 18
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
cgcgtacgag atctggtcga ctggcatgac gccacaggc 39
<210> 19
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
tacgcgtccc ggggcggtac ccagagcagc agcaagcg 38
<210> 20
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
cgaaagctct gcaggtcgac ctagcctctg gttcgtatg 39
<210> 21
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
atggggctct cggatgtaga 20
<210> 22
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
cgatcatagg cgtctcgcat 20
Claims (9)
1.一种水稻OsOFP22s基因，其特征在于，所述OsOFP22s基因的编码区序列如SEQ IDNO.1所示。
2.根据权利要求1所述一种水稻OsOFP22s基因，其特征在于，所述OsOFP22s编码的氨基酸序列如SEQ ID NO.2所示。
3.一种如权利要求1或2所述的水稻OsOFP22s基因的过表达重组载体pActin-OFP22s-3Flag，其特征在于，所述pActin-OFP22s-3Flag包含OsOFP22s基因，载体为植物双元表达载体pActin-3Flag。
4.权利要求3所述过表达重组载体pActin-OFP22s-3Flag的制备方法，其特征在于，所述的方法如下：
将植物双元表达载体pActin-3Flag用Sam I和Xba I双酶切，用同源重组酶将酶切的载体与包含OsOFP22s基因全长cDNA的PCR扩增产物连接。
5.根据权利要求4所述过表达重组载体pActin-OFP22s-3Flag的制备方法，其特征在于，所述包含OsOFP22s基因全长cDNA的PCR扩增方法如下：
以野生型日本晴的RNA为模板，引物序列如下：
。
6.一种如权利要求1或2所述的水稻OsOFP22s基因在提高水稻粒长、千粒重和改良直链淀粉含量方面的应用。
7.根据权利要求6所述的应用，其特征在于，所述应用的方法如下：用所述水稻OsOFP22s基因序列构建可在水稻中过表达的重组载体，并转化农杆菌，再用农杆菌介导的水稻遗传转化方法将载体转入水稻愈伤组织，经抗性筛选和组织培养后变成水稻植株。
8.根据权利要求7所述的应用，其特征在于，在水稻中过表达的重组载体为权利要求3所述的过表达重组载体pActin-OFP22s-3Flag。
9.根据权利要求8所述的应用，其特征在于，所述的应用方法如下：
(1)构建工程菌：将pActin-OFP22s-3Flag载体通过电击法转化到农杆菌菌株EHA105中，通过卡那霉素和利福平筛选，获得含有pActin-OFP22s-3Flag载体的农杆菌；
(2)pActin-OFP22s-3Flag载体转化水稻愈伤并获得水稻再生苗：用含有pActin-OFP22s-3Flag载体的EHA105侵染水稻愈伤组织，并于28℃培养室中共培养3天，再用液体培养基洗去农杆菌后，将水稻愈伤放置在含有合适抗生素的筛选培养基上培养；经过两轮培养后，可获得抗性愈伤，将抗性愈伤转移至分化培养基是进行分化培养获得小苗，分化出的小苗转移至生根培养基上培养，经炼苗后移栽。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116676317A (zh) * | 2023-05-18 | 2023-09-01 | 扬州大学 | 一种水稻OsEns150基因及其在改良水稻株型、品质和增强穗发芽抗性中的应用 |
| CN118581101A (zh) * | 2024-05-21 | 2024-09-03 | 扬州大学 | 一种水稻OsSLRL2基因的应用 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102786599A (zh) * | 2012-08-01 | 2012-11-21 | 中国农业科学院作物科学研究所 | 水稻转录因子Os05g39950基因的应用 |
| CN105950633A (zh) * | 2016-06-16 | 2016-09-21 | 复旦大学 | 基因OsARF4在控制水稻粒长和千粒重中的应用 |
-
2022
- 2022-05-27 CN CN202210594164.6A patent/CN114752605B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102786599A (zh) * | 2012-08-01 | 2012-11-21 | 中国农业科学院作物科学研究所 | 水稻转录因子Os05g39950基因的应用 |
| CN105950633A (zh) * | 2016-06-16 | 2016-09-21 | 复旦大学 | 基因OsARF4在控制水稻粒长和千粒重中的应用 |
Non-Patent Citations (1)
| Title |
|---|
| 无: "CM000130.1", 《GENBANK》, pages 1 - 2 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116676317A (zh) * | 2023-05-18 | 2023-09-01 | 扬州大学 | 一种水稻OsEns150基因及其在改良水稻株型、品质和增强穗发芽抗性中的应用 |
| CN118581101A (zh) * | 2024-05-21 | 2024-09-03 | 扬州大学 | 一种水稻OsSLRL2基因的应用 |
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