CN114085854A - 一种水稻抗旱、耐盐基因OsSKL2及其应用 - Google Patents
一种水稻抗旱、耐盐基因OsSKL2及其应用 Download PDFInfo
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- CN114085854A CN114085854A CN202111540270.8A CN202111540270A CN114085854A CN 114085854 A CN114085854 A CN 114085854A CN 202111540270 A CN202111540270 A CN 202111540270A CN 114085854 A CN114085854 A CN 114085854A
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Abstract
本发明公开了一种水稻抗旱、耐盐基因OsSKL2及其应用,涉及基因工程技术领域,该基因具有如SEQ ID NO.1所示的核苷酸序列。该基因能够调控水稻在干旱、高盐下的存活率,对水稻干旱、高盐具有正调控作用;该基因的发现为莽草酸在非生物胁迫的调控作用提供了直接证据,为改良作物的非生物逆境提供了新的研究途径,对培育抗旱、耐盐的材料具有重要的理论和实践意义。
Description
技术领域
本发明涉及基因工程技术领域,具体涉及一种水稻抗旱、耐盐基因OsSKL2及其应用。
背景技术
水稻是全球主要的粮食作物,为全世界一半以上人口提供了口粮。然而,随着人口增长、环境条件恶化以及耕地面积减少,导致人口增多与粮食需求间的供需矛盾日益突出。目前,引起水稻产量下降的主要环境因素是干旱和土壤盐碱化。据统计依靠传统淹水灌溉方式的水稻种植面积和产量约占世界水稻总面积和总产量的55%和75%。近年来,干旱和土壤盐碱化造成的粮食减产呈现加重趋势,已威胁到粮食安全生产。因此挖掘抗旱耐盐基因、解析其作用机制是生物技术育种的一个重要目标,对保障水稻高产稳产和粮食安全生产具有重要意义。
为了适应外界环境的变化,植物在其长期演化过程中已进化出多种途径和调控机制来应对环境刺激从而保障正常的生长发育。研究表明,位于植物细胞表面(如细胞壁、质膜)和细胞器(如细胞质、细胞核)的胁迫感受器捕捉到外界胁迫信号后,迅速将刺激信号转化传递到细胞内影响第二信使(如Ca2+、ROS、NO、磷脂等)的水平,进而改变调控基因和转录因子的表达。转录因子通过结合、调控下游功能基因的表达诱导植物保护机制如解毒、修复应激反应来保证植物的正常生长。玉米转录因子ZmbHLH124直接结合并激活ZmDREB2A的表达,进而提高玉米的耐旱性。水稻中,OsMYB3R-2通过调控OsCycB1;1和OsKNOLLE2等细胞周期蛋白基因的表达,降低ABA的敏感性,增强水稻对冷、高盐和干旱的抵抗能力。虽然对于植物干旱耐盐的基因已有不少报道,但是初生代谢与次生代谢途径直接参与到干旱高盐胁迫中的功能主效基因知之甚少。
莽草酸循环广泛存在于植物和微生物中,是连接碳水化合物代谢和次生代谢的重要枢纽,并为其他次生代谢提供底物。莽草酸途径是由多种代谢酶催化的生化反应,其产物是苯丙氨酸、酪氨酸和色氨酸。其中莽草酸激酶(SK)是催化莽草酸途径的第五步反应,水稻中SK的编码基因为SK-1,SK-2,SK-3和SKL1,SKL2。研究发现SK1和SK2和叶绿体的形成与发育密切相关。虽然已有研究表示莽草酸含量在干旱、高盐条件下有显著变化,但是具体如何调控干旱高盐胁迫反应没有直接证据,其功能基因也不清楚。因此,提出一种水稻抗旱、耐盐基因OsSKL2及其应用。
发明内容
本发明的目的在于克服现有技术的不足,提供了一种水稻抗旱、耐盐基因OsSKL2及其应用。
本发明通过以下技术方案来实现上述目的:
本发明提供了一种水稻抗旱、耐盐基因OsSKL2,该基因具有如SEQID NO.1所示的核苷酸序列,全长1128bp。
本发明还提供了一种上述水稻抗旱、耐盐基因OsSKL2在提高水稻抗旱性、耐盐性中的应用。
进一步改进在于,所述基因OsSKL2对水稻的抗旱性、耐盐性具有正调控作用,过量表达转基因水稻植株可提高水稻在干旱、高盐下的存活率,而RNAi沉默转基因水稻植株干旱、高盐下的存活率显著降低
本发明还提供了一种上述水稻抗旱、耐盐基因OsSKL2的编码蛋白,该编码蛋白具有如SEQID NO.2所示的氨基酸序列。
进一步改进在于,所述蛋白定位于叶绿体中,受干旱、高盐诱导表达,其编码的莽草酸激酶是催化莽草酸途径的关键代谢酶。
本发明还提供了一种质粒载体,所述质粒载体为通过在pEASY-T1载体上***上述水稻抗旱、耐盐基因OsSKL2获得。
本发明还提供了一种遗传工程化的宿主细胞,所述宿主细胞为上述质粒载体的大肠杆菌感受态Trans5α细胞。
本发明还提供了一种水稻抗旱、耐盐基因OsSKL2的获得方法,以OsSKL2基因的核苷酸序列为模板设计特异性扩增引物,以水稻日本晴品种为材料,提取RNA并反转录成cDNA,通过PCR扩增技术获得基因OsSKL2。
进一步改进在于,所述PCR扩增的特异性扩增引物为:
SKL2–F-1:ATGAGGATAGGGGCAGGGGCGAACA;
SKL2–R-1:TCAGATATTGGTCGGTGGGGCGTCG。
本发明还提供了一种过量表达载体,所述过量表达载体为多克隆位点区域依次连接有35S启动子、上述基因OsSKL2和NOS终止子的pCAMBIA1301植物表达载体。
本发明还提供了一种亚细胞定位载体,所述亚细胞融合载体为将上述基因OsSKL2和pCAMBIA1305载体同时用SpeI和BamHI切割后连接获得,具体为pCAMBIA1305-SKL2,连接时用上下游引物如下:
OsSKL2-F:ATGTTGGCCTCCACTTGCTTCTCCG;
OsSKL2-R:TATGTTGGTGGGTGGTGCGTCGGA。
本发明提具有如下有益效果:
本发明相比现有技术具有以下优点:本发明提供了一种水稻抗旱、耐盐基因OsSKL2及其应用,该基因能够调控水稻在干旱、高盐下的存活率,对水稻干旱、高盐具有正调控作用;该基因的发现为莽草酸在非生物胁迫的调控作用提供了直接证据,为改良作物的非生物逆境提供了新的研究途径,对培育抗旱、耐盐的材料具有重要的理论和实践意义。
附图说明
图1为RNAi沉默载体pEASY-T1图谱;
图2为过表达pCAMBIA1301载体图谱;
图3为OsSKL2的高盐、干旱诱导表达模式图;100mM mannitol溶液、200mM NaCl溶液分别处理四周大小野生型水稻中花11,12h后取样分析;
图4为OsSKL2亚细胞定位分析图;
图5为OsSKL2过表达和RNAi沉默株系的分子鉴定图;WT为野生型,OE3、OE6为过表达转基因植株,Ri6、Ri9为沉默转基因植株;
图6为OsSKL2转基因水稻在正常与盐处理下的表型图和存活率统计图;图6a为野生型与转基因植株在高盐处理下的表型,图6b为为萌发率统计图。四周大小的野生型与转基因水稻植株分别进行120mM、140mM NaCl处理,WT为野生型植株,OE3、OE6为过表达转基因植株,Ri6、Ri9为沉默转基因植株;
图7为OsSKL2转基因水稻在正常与干旱处理下的表型图和存活率统计图;图7a为野生型与转基因植株在干旱处理下的表型,图7b为萌发率统计图。四周大小的野生型与转基因水稻植株分别进行20%PEG处理,WT为野生型植株,OE3、OE6为过表达转基因植株,Ri6、Ri9为沉默转基因植株。
具体实施方式
下面结合附图对本申请作进一步详细描述,有必要在此指出的是,以下具体实施方式只用于对本申请进行进一步的说明,不能理解为对本申请保护范围的限制,该领域的技术人员可以根据上述申请内容对本申请作出一些非本质的改进和调整。
1、材料
本实施例所用方法如无特别说明均为本领域的技术人员所知晓的常规方法,所用的试剂等材料,如无特别说明,均为市售购买产品。
2、方法
2.1OsSKL2基因的获得
选取水稻日本晴品种,提取RNA并反转录成cDNA,以水稻cDNA为模板,根据OsSKL2基因序列和水稻基因组数据库,结合植物表达载体的多克隆位点设计引物SKL2–F-1、SKL2–R-1,进行PCR扩增,获得PCR扩增产物。
引物序列为:
SKL2–F-1:ATGAGGATAGGGGCAGGGGCGAACA;
SKL2–R-1:TCAGATATTGGTCGGTGGGGCGTCG;
PCR反应程序为:98℃,预变性10min;98℃,变性20s;65℃,退火20s;72℃,延伸2min,30个循环;72℃,复性10min;10℃保存。
将PCR扩增产物用质量比为2%的琼脂糖凝胶电泳检测,回收目的片段并连接至pEASY-T1载体(购于全式金生物技术有限公司,载体图谱如图1所示),获得连接产物。将连接产物转化到大肠杆菌感受态Trans5α细胞中,提取质粒。以提取的质粒做模板,以SKL2-F-1、SKL2-R-1为引物进行PCR扩增验证,同时用Kpn I和Pst I双酶切质粒进行检测,筛选阳性克隆子,将阳性克隆子送去华大生物公司进行测序,测序结果使用Sequencher软件比对,结果与预测一致。获得的重组质粒命名为T-SKL2。
2.2OsSKL2基因过量表达载体和沉默表达载体的构建
以pCAMBIA1301(购于上海捷兰生物技术有限公司,载体图谱如图2所示)为原始载体,在其多克隆位点的EcoRI和SacI酶切位点之间连接一个35S启动子,并在SphI和HindIII酶切位点之间加上一段NOS终止子,获得改造的载体p1。用Kpn I和PstI双酶切T-SKL2得到目的基因片段,同时用KpnI和PstI双酶切p1得到载体片段,将上述两片段使用T4连接酶连接,构建获得载体p1-OsSKL2,作为转水稻的OsSKL2基因过量表达载体。以水稻cDNA为模板,用OsSKL2特异引物扩增出OsSKL2部分基因片断,产物分别用BamHI和HindIII双酶切,克隆到经相同酶切pRNAi-Ubi载体中并转化TOP10感受态细胞,经抗生素(卡那霉素)和PCR筛选鉴定,获得目的片断一次正向组装成功的中间重组质粒。用载体特异通用引物RNAi-Pst/RNAi-Mlu扩增中间重组质粒,产物用PstI和MluI双酶切后克隆到同样酶切的中间重组质粒中并转化DH10B感受态细胞,经卡那霉素LB平板筛选\PCR\酶切和测序鉴定,完成pRNAi-ubiSKL2转化载体的构建。用BamH I单酶切pRNAi-ubiSKL2,克隆到经相同酶切的pRNAi-IP(单酶切后的pRNAi-IP载体经过碱性磷酸酶处理)中并转化DH10B感受态细胞,经卡那霉素LB平板筛选、酶切检测和测序鉴定,完成pRNAi-IP-SKL2载体的构建。
2.3OsSKL2的诱导表达模式的分析
用100mM的mannitol溶液、200mM NaCl溶液分别处理四周大小野生型水稻中花11,然后选取处理12h后的水稻根、叶,提取其RNA,通过反转录分别获得其cDNA,再通过qRT-PCR来确定处理下根、叶的表达量。
设计定量引物如下:
SKL2-F:TTGGCTTCGGAAGGAGTGGATT
SKL2-R:TTCCCGAGCTCACACTTCTACG
qPCR反应程序为:94℃,预变性5min;94℃,变性20s;60℃,退火20s;72℃,延伸1min,30个循环。将结果进行作图分析。
结果如图3所示,基因OsSKL2受到干旱、高盐的显著诱导表达。
2.4OsSKL2的亚细胞定位分析
首先构建pCAMBIA1305-SKL2的载体。将SKL2和p1305载体同时用SpeI和BamHI切割,然后用凝胶电泳回收目的条带,再用T4连接酶过夜连接回收pCAMBIA1305-SKL2载体。利用PEG瞬时转化体系将上述载体转到水稻叶片原生质体中,转化体系为:100μL原生质体、100μL的pCAMBIA1305-SKL2载体质粒和200μL的45%PEG。诱导转化30分钟,用440μL W5溶液稀释离心,再加入200μL的WI溶液在暗处培养12-18小时后,通过激光共聚焦显微镜观察。
亚细胞定位中用到的引物如下:
OsSKL2-F:ATGTTGGCCTCCACTTGCTTCTCCG
OsSKL2-R:TATGTTGGTGGGTGGTGCGTCGGA
结果如图4所示,发现基因OsSKL2定位于叶绿体中。
2.5农杆菌介导的OsSKL2基因过量表达和沉默表达植株的获得与鉴定
农杆菌介导的过量表达植株和沉默表达植株的获得方法及培养过程中使用的培养基配方参见文献:Methods in Molecular Biology,vol.343:AgrobacteriumProtocols,2/e,volume 1,具体包括以下步骤:
(1)水稻成熟胚诱导愈伤的获得
选取成熟饱满的水稻种子,剥壳备用。准备1-2L的无菌水,75%(质量分数)酒精,次氯酸和吐温配制的杀菌水(具体比例:灭菌水:次氯酸=1:1,总体积:吐温=1mL:1ul)。先用75%的酒精洗5分钟左右,再用杀菌水消毒15分钟,分两段时间各7-8分钟,接着用75%的酒精消泡3次,最后用灭菌水清洗数遍,直至水变澄清,再将成熟种子放在灭菌滤纸上吸干水分后,取其于愈伤诱导培养基上,26±1℃培养;10-15d后,将诱导出的乳黄色愈伤转入继代培养基上继代培养。每两周继代培养一次,继代两次后挑选继代培养5-7d,将色泽淡黄的愈伤用于共培养。
(2)用于转化水稻的农杆菌菌液的准备
将p1-OsSKL2和pRNAi-IP-SKL2载体转化农杆菌LBA4404中,获得重组农杆菌LBA4404/p1-OsSKL2和LBA4404/pRNAi-IP-SKL2。将重组农杆菌LBA4404/p1-OsSKL2和LBA4404/pRNAi-IP-SKL2接种于YEP液体培养基(含有50μg/mL卡那霉素和50μg/mL利福平)中,28℃振荡培养至OD600=0.6-1.0;室温下5000rpm离心5min收集菌体,随后将其悬浮于液体共培养基中,调整菌体浓度至OD600=0.4,即为共培养转化水稻用的农杆菌悬浮液。
(3)农杆菌侵染水稻营养器官
挑选色泽鲜嫩、呈乳黄的愈伤,集中放入100mL无菌三角瓶中,加入上述制备好的农杆菌悬浮液;28℃,220rpm摇床上培养20-30min;倒掉菌液,将侵染后的愈伤置于含无菌滤纸的培养皿上吸去多余菌液,随即转移到固体共培养基上,22℃暗培养2-3d。
(4)选择培养
将共培的愈伤先用灭菌水清洗直至水变澄清,弃去清洗液,转入含有羧苄青霉素(100mg/ml)的溶液中振荡清洗30min以上,用无菌滤纸吸干,置于含无菌滤纸的培养皿中干燥处理;再将愈伤挑选到含有羧苄青霉素(500mg/ml)和潮霉素(50mg/L)筛选培养基上,28℃光照培养30d左右,直至长出新的抗性愈伤。
(5)抗性愈伤的分化
从筛选培养基上挑选长势较好呈乳黄色的新愈伤于含有50mg/L潮霉素的分化培养基上,先28℃暗培养3d,然后转移到30℃条件下进行全光照培养,15-30d,有绿点出现;30-40d后会分化出幼苗。
(6)生根、壮苗和移栽
待分化出的幼苗3cm高时,转移至生根培养基上,培养2-3周;选择15cm高、根系发达的幼苗,加水室温下炼苗2-3d后,用温水洗去培养基,移入含水稻培养土的水桶中栽培。待苗生长健壮后移入水田中生长。32株独立转化事件的转基因水稻苗中有21株同时扩增出目的基因OsSKL2片段和潮霉素基因,有8株RNAi沉默的植株。T0代转基因植株成熟后,收获T1代种子,T1代种子自交繁殖得到T2代种子。
2.6转基因水稻的鉴定
为了检测转基因水稻,以提取的转基因水稻总DNA为模板,用目的基因OsSKL2片段和潮霉素基因为检测对象,设计引物,进行PCR扩增,用以初步鉴定转基因植株。
检测目的基因OsSKL2片段的引物:
SKL2–F-2:ATGAGGATAGGGGCAGGGGCGAACA
SKL2–R-2:TCAGATATTGGTCGGTGGGGCGTCG
PCR反应条件为:95℃预变性5min;95℃变性30s;60℃退火30s;72℃延伸1min,35个循环;72℃总延伸5min。
检测潮霉素基因的引物:
Hyg-F:5’-TAGGAGGGCGTGGATATGGC-3’
Hyg-R:5’-TACACAGCCATCGGTCCAGA-3’
PCR反应程序为:95℃预变性5min;95℃变性30s,62℃退火30s,72℃延伸1min,34个循环;72℃总延伸5min。
检测pRNAi-IP-SKL2的引物:
SKL2-Ri-N-F:TCCAGTCTGCACAAGTGAGC
SKL2-Ri-N-R:TCCAGCACAACACATTCAGC
95℃预变性5min;95℃变性30s;60℃退火30s;72℃延伸1min,35个循环;72℃总延伸5min。
结果如图5所示,获得了过表达(OE3和OE6)和RNAi沉默(Ri6和Ri9)转基因水稻植株。
2.7OsSKL2转基因水稻在高盐、干旱处理下的表型鉴定与存活率统计
选取上述已鉴定的OsSKL2转基因过量表达和沉默表达的水稻进行研究,每组处理以野生型中花11做对照。四周大小的野生型和转基因植株分别用120mM NaCl、140mM NaCl和20%PEG处理以及干旱处理后复水1天。处理后分别进行拍照和存活率统计。
结果如图6、7所示,120mM NaCl、140mM NaCl处理后,相比较于野生型,过表达植株存活率存活率明显较高,而沉默植株的存活率明显偏低;同样干旱处理后复水实验结果也显示过表达植株存活率存活率明显较高,而沉默植株的存活率明显偏低;这些结果说明OsSKL2基因具有抗盐和抗旱性。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。
序列表
<110> 安徽农业大学
<120> 一种水稻抗旱、耐盐基因OsSKL2及其应用
<141> 2021-12-16
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1128
<212> DNA
<213> 水稻( Oryza sativa)
<400> 1
atgttggcct ccacttgctt ctccgccccg ccgccctcct cctcctcccc gtcaatcccg 60
acccacctcg ccactctctg ctgctgcttc aggcctcccg ctcgcccacc ttggcctcgc 120
tctcttcttc ttcttggtgc cttcccaccg ccgacccgcc ccctcccccg cgcctccgtc 180
tcgtcatcga ccgccccagc gaaagactac gagtttactg atggtggcgg agaggtggag 240
ctgaggctgg acataggaaa gctcggcatt gagaattcaa gagatgtatt tgttgacgtc 300
gatgatacgt ctctgttggt cagagccaag tcggatggga cactgcggac tttgatcaat 360
gttaaacaac tctttgatag gattaagtct tctgagacta tatggttcat tgatgaggat 420
caattggtag tgaatctaaa gaaagttgag caagagctga aatggcccga cattgatgaa 480
tcttgggaat cccttacttc tggaatcact cagcttttga cagggattag tgttcatatt 540
gttggtgatt ccacagatat aaacgaggca gttgctaaag aaatagctga gggaattggt 600
taccttccag tctgcacaag tgagctgcta gaaagtgcca ccgaaaagtc tattgacaaa 660
tggttggctt cggaaggagt ggattcggta gcagaagctg aatgtgttgt gctggaaagc 720
cttagcagcc atgttcgtac agtcgtagca actctggggg gaaagcaagg agcagctagc 780
agatttgata aatggcagta tcttcatgct ggatttacgg tttggttgtc ggtctccgat 840
gccagcgatg aagcttctgc caaagaagag gcccgtagaa gtgtgagctc gggaaatgtt 900
gcgtacgcga aagctgatgt agtagtgaag cttggtggat gggatccgga gtacacacga 960
gctgttgccc agggttgcct tgtggccttg aagcagctaa cattggcaga caagaagcta 1020
gcaggtaaga agagcctata catgaggctg ggatgccgag gggattggcc caacatcgag 1080
cctcccggct gggatcctga ctccgacgca ccacccacca acatatga 1128
<210> 2
<211> 375
<212> PRT
<213> 水稻( Oryza sativa)
<400> 2
Met Leu Ala Ser Thr Cys Pro Ser Ala Pro Pro Pro Ser Ser Ser Ser
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Pro Ser Ile Pro Thr His Leu Ala Thr Leu Cys Cys Cys Pro Ala Pro
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Pro Ala Ala Pro Pro Thr Pro Ala Ser Leu Leu Leu Leu Gly Ala Pro
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Pro Pro Pro Thr Ala Pro Leu Pro Ala Ala Ser Val Ser Ser Ser Thr
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Ala Pro Ala Leu Ala Thr Gly Pro Thr Ala Gly Gly Gly Gly Val Gly
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Leu Ala Leu Ala Ile Gly Leu Leu Gly Ile Gly Ala Ser Ala Ala Val
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Pro Val Ala Val Ala Ala Thr Ser Leu Leu Val Ala Ala Leu Ser Ala
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Gly Thr Leu Ala Thr Leu Ile Ala Val Leu Gly Leu Pro Ala Ala Ile
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Leu Ser Ser Gly Thr Ile Thr Pro Ile Ala Gly Ala Gly Leu Val Val
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Ala Leu Leu Leu Val Gly Gly Gly Leu Leu Thr Pro Ala Ile Ala Gly
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Ser Thr Gly Ser Leu Thr Ser Gly Ile Thr Gly Leu Leu Thr Gly Ile
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Ser Val His Ile Val Gly Ala Ser Thr Ala Ile Ala Gly Ala Val Ala
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Leu Gly Ile Ala Gly Gly Ile Gly Thr Leu Pro Val Cys Thr Ser Gly
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Leu Leu Gly Ser Ala Thr Gly Leu Ser Ile Ala Leu Thr Leu Ala Ser
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Gly Gly Val Ala Ser Val Ala Gly Ala Gly Cys Val Val Leu Gly Ser
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Leu Ser Ser His Val Ala Thr Val Val Ala Thr Leu Gly Gly Leu Gly
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Gly Ala Ala Ser Ala Pro Ala Leu Thr Gly Thr Leu His Ala Gly Pro
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Thr Val Thr Leu Ser Val Ser Ala Ala Ser Ala Gly Ala Ser Ala Leu
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Gly Gly Ala Ala Ala Ser Val Ser Ser Gly Ala Val Ala Thr Ala Leu
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Ala Ala Val Val Val Leu Leu Gly Gly Thr Ala Pro Gly Thr Thr Ala
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Ala Val Ala Gly Gly Cys Leu Val Ala Leu Leu Gly Leu Thr Leu Ala
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Ala Leu Leu Leu Ala Gly Leu Leu Ser Leu Thr Met Ala Leu Gly Cys
340 345 350
Ala Gly Ala Thr Pro Ala Ile Gly Pro Pro Gly Thr Ala Pro Ala Ser
355 360 365
Ala Ala Pro Pro Thr Ala Ile
370 375
Claims (10)
1.一种水稻抗旱、耐盐基因OsSKL2,其特征在于,该基因具有如SEQ ID NO.1所示的核苷酸序列。
2.一种如权利要求1所述的水稻抗旱、耐盐基因OsSKL2在提高水稻抗旱性、耐盐性中的应用。
3.根据权利要求2所述的应用,其特征在于,所述基因OsSKL2过表达对水稻的抗旱性、耐盐性具有正调控作用。
4.一种如权利要求1所述的水稻抗旱、耐盐基因OsSKL2的编码蛋白,其特征在于,该编码蛋白具有如SEQ ID NO.2所示的氨基酸序列。
5.一种质粒载体,其特征在于,所述质粒载体为通过在pEASY-T1载体上***如权利要求1所述的水稻抗旱、耐盐基因OsSKL2获得。
6.一种遗传工程化的宿主细胞,所述宿主细胞为含有如权利要求5所述质粒载体的大肠杆菌感受态Trans5α细胞。
7.一种水稻抗旱、耐盐基因OsSKL2的获得方法,其特征在于,以OsSKL2基因的核苷酸序列为模板设计特异性扩增引物,以水稻日本晴品种为材料,提取RNA并反转录成cDNA,通过PCR扩增技术获得基因OsSKL2。
8.根据权利要求7所述的一种水稻抗旱、耐盐基因OsSKL2的获得方法,其特征在于,所述PCR扩增的特异性扩增引物为:
SKL2–F-1:ATGAGGATAGGGGCAGGGGCGAACA;
SKL2–R-1:TCAGATATTGGTCGGTGGGGCGTCG。
9.一种过量表达载体,其特征在于,所述过量表达载体为多克隆位点区域依次连接有35S启动子、如权利要求1所述基因OsSKL2和NOS终止子的pCAMBIA1301植物表达载体。
10.一种亚细胞定位载体,其特征在于,所述亚细胞融合载体为将权利要求1所述的基因OsSKL2和pCAMBIA1305载体同时用SpeI和BamHI切割后连接获得,具体为pCAMBIA1305-SKL2,连接时用上下游引物如下:
OsSKL2-F:ATGTTGGCCTCCACTTGCTTCTCCG;
OsSKL2-R:TATGTTGGTGGGTGGTGCGTCGGA。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116179567A (zh) * | 2022-11-11 | 2023-05-30 | 中国科学院青岛生物能源与过程研究所 | 一种调控芒草耐盐碱基因MsGRAS60及其应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002061072A2 (en) * | 2001-01-05 | 2002-08-08 | Monsanto Technology Llc | Transgenic plants containing altered levels of steroid compounds |
WO2008046069A2 (en) * | 2006-10-12 | 2008-04-17 | Ceres, Inc. | Nucleotide sequences and polypetides encoded thereby useful for increasing tolerance to oxidative stress in plants |
US20090100539A1 (en) * | 2006-03-14 | 2009-04-16 | Ceres, Inc. | Nucleotide sequences and corresponding polypeptides conferring modulated growth rate and biomass in plants grown in saline conditions |
CN102942622A (zh) * | 2012-11-14 | 2013-02-27 | 中国科学院植物研究所 | MtSKL1蛋白及其编码基因在控制植物性状中的应用 |
-
2021
- 2021-12-16 CN CN202111540270.8A patent/CN114085854B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002061072A2 (en) * | 2001-01-05 | 2002-08-08 | Monsanto Technology Llc | Transgenic plants containing altered levels of steroid compounds |
US20090100539A1 (en) * | 2006-03-14 | 2009-04-16 | Ceres, Inc. | Nucleotide sequences and corresponding polypeptides conferring modulated growth rate and biomass in plants grown in saline conditions |
WO2008046069A2 (en) * | 2006-10-12 | 2008-04-17 | Ceres, Inc. | Nucleotide sequences and polypetides encoded thereby useful for increasing tolerance to oxidative stress in plants |
CN102942622A (zh) * | 2012-11-14 | 2013-02-27 | 中国科学院植物研究所 | MtSKL1蛋白及其编码基因在控制植物性状中的应用 |
Non-Patent Citations (3)
Title |
---|
KIKUCHI, S. ET AL.: "Oryza sativa Japonica Group cDNA clone:J023052F21, full insert sequence GenBank: AK100237.1", 《GENBANK》, pages 1 - 2 * |
MUHAMMAD SALMAN HAIDER ET AL.: "Insights into grapevine defense response against drought as revealed by biochemical, physiological and RNA-Seq analysis", 《SCIENTIFIC REPORTS》, vol. 7, pages 1 - 15 * |
YINGLI JIANG ET AL.: "Regulation of Drought and Salt Tolerance by OsSKL2 and OsASR1 in Rice", 《RICE》, vol. 15, pages 1 - 17, XP021307205, DOI: 10.1186/s12284-022-00592-2 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116179567A (zh) * | 2022-11-11 | 2023-05-30 | 中国科学院青岛生物能源与过程研究所 | 一种调控芒草耐盐碱基因MsGRAS60及其应用 |
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