CN102643831A - 一种火把梨耐干旱基因Pp14-3-3及其应用 - Google Patents
一种火把梨耐干旱基因Pp14-3-3及其应用 Download PDFInfo
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Abstract
本发明公开了一种火把梨耐干旱基因Pp14-3-3及其应用,Pp14-3-3基因具有SEQIDNO:1所示核苷酸序列,编码14-3-3蛋白。本发明采用功能基因组学相关技术证实火把梨Pp14-3-3基因具有提高植物耐干旱胁迫的功能。将本发明耐旱基因Pp14-3-3构建到植物表达载体上并转入植物中过量表达,转基因植物表现出很强的耐干旱胁迫的能力。
Description
技术领域
本发明涉及一种火把梨耐干旱基因Pp14-3-3及其应用,属于分子生物学以及基工程领域。
背景技术
随着全球性的气候异常和生态平衡的破坏,土地日趋沙漠化和盐碱化,水资源短缺成为全人类面临的严重生态问题。我国是一个人口大国,粮食问题始终是关系国家安全、社会稳定的重大战略问题,与其它自然灾害相比,旱灾是影响我国粮食生产的主要因素,因为旱灾造成的粮食损失占全部自然灾害粮食损失的一半以上。据统计分析,我国受旱面积50年代为1.7亿多亩,90年代年均3.64亿亩,因干旱损失粮食50年代年均43.5亿公斤,90年代为195.7亿公斤。干旱始终困扰着我国经济、社会的发展,特别是农业的发展。
干旱对植物造成伤害的机理比较复杂,最直接的伤害是使植物细胞脱水、破坏细胞结构、造成机械伤害,从而导致细胞死亡(Mahajan S, Tuteja N. Cold,
salinity and drought stresses: An overview. Arch Biochem Biophys, 2005, 444:
139~158)。干旱胁迫除了造成直接伤害,还能引起一系列间接伤害,如抑制光合作用系统Ⅱ的活性,光合作用系统Ⅱ活性的下降使电子的产生和利用失衡,产生的活性氧会破坏细胞的膜脂结构、DNA和其他生物大分子,这也是干旱对植物造成间接破坏的主要原因(Pehzer D, Dreyer E, Polle
A. Differential temperature dependencies of antioxidative enzymes in two
contrasting species: Fagus sylvatica and Coleus blumei. Plant
Physiol Biochem, 2002, 40: 14l~150)。植物在长期的自然选择中形成了渗透调节、减少活性氧伤害、增强蛋白质的稳定性等一些列有效的抵御干旱胁迫的机制(邵艳军, 山仑. 植物耐旱机制研究进展. 中国农业生态学报, 2006, 14(4): 16~20)。研究表明,转录因子、激酶等胁迫相关蛋白参与逆境胁迫应答的信号通路。14-3-3蛋白做为一种信号调控因子,已被证实广泛参与植物对干旱等非生物胁迫的应答(Roberts MR, Salinas J,
Collinge DB. 14-3-3 proteins and the response to abiotic and biotic stress.
Plant Mol Biol, 2002, 50:1031~1039)。
14-3-3蛋白是一类在真核生物中起重要作用且结构高度保守的蛋白,广泛分布于真核生物中,与多种蛋白结合并相互作用,参与调控细胞内的生命代谢过程。植物中的14-3-3可分为ε和非ε两大类,拟南芥(Arabidopsis thaliana)
μ、π、ι、ο、ε类14-3-3蛋白属于ε大类,κ、λ、ψ、ν、υ、ω、φ、χ属于非ε大类(Ferl RJ, Manak MS, Reyes
MF. The 14-3-3s. Genome Biol, 2002, 3(7): reviews 3010.1~3010.7)。14-3-3为酸性可溶性蛋白,单体分子量在27-32kDa之间,主要以同源或异源二聚体形式存在 (Ferl RJ, Manak MS, Reyes
MF. The 14-3-3s. Genome Biol, 2002, 3(7): reviews 3010.1~3010.7)。14-3-3蛋白通过蛋白相互作用以及结合靶蛋白的磷酸丝氨酸/磷酸苏氨酸来实现其功能,目前已发现14-3-3蛋白参与细胞生长分化、细胞周期和凋亡的调控、信号转导和迁移等多种生理过程(van Hemert MJ, Steensma
HY, van Heusden GP. 14-3-3 proteins: key regulators of cell division,
signalling and apoptosis. Bioessays, 2001, 23(10): 936~946)。
近年来多个植物14-3-3基因被分离出来。研究表明植物14-3-3与能量代谢的关键酶结合,从而调控能量代谢过程,如14-3-3蛋白通过调节硝酸还原酶 (Weiner H, Kaiser WM.
14-3-3 proteins control proteolysis of nitrate reductase in spinach leaves.
FEBS Lett, 1999, 455: 75~78)、蔗糖磷酸合酶、海藻糖-6磷酸合酶 (Toroser D, Athwal GS, Huber SC. Site-specific regulatory
interaction between spinach leaf sucrose-phosphate synthase and 14-3-3
proteins. FEBS Lett, 1998, 435(1): 110~114)、甘油三磷酸脱氢酶、谷氨酰胺合成酶 (Moorhead G, Douglas P,
Cotelle V, et al. Phosphorylation-dependent interactions between enzymes of
plant metabolism and 14-3-3 proteins. Plant J, 1999, 18(1): 1~12)等碳、氮代谢关键酶的活性从而参与植物碳、氮代谢途径。14-3-3蛋白还能结合质膜H+-ATPase来控制物质的跨膜运输过程 (Olsson A, Svennelid F,
Ek B, et al. A phosphothreonine residue at the C-terminal end of the plasma
membrane H+-ATPase is protected by fusicoccin-induced 14-3-3
binding. Plant Physiol, 1998, 118(2): 551~555)。研究发现,14-3-3在大麦(Hordeum distichum.)种子吸涨后被诱导表达,暗示14-3-3参与大麦的种子萌发 (Testerink C, van der
Meulen RM, Oppedijk BJ, et al. Differences in spatial expression between 14-3-3
isoforms in germinating barley embryos. Plant Physiol, 1999, 121(1): 81~87)。14-3-3还能通过与转录因子结合来调控细胞的信号转导过程,14-3-3能结合玉米(Zea mays) (de
Vetten NC, Lu G, Feri RJ. A maize protein associated with the G-box binding
complex has homology to brain regulatory proteins. Plant Cell, 1992, 4(10):
1295~1307)和拟南芥 (Lu G, DeLisle AJ, de
Vetten NC, et al. Brain proteins in plants: an Arabidopsis homolog to
neurotransmitter pathway activators is part of a DNA binding complex. Proc Natl
Acad Sci U S A, 1992, 89(23): 11490~11494) 的G-box复合体,此外,TFIIB、TBP2、VPl、EmBPl、RSG (repression of shoot growth) 等多种转录因子均为14-3-3蛋白的靶蛋白 (Schultz TF, Medina J,
Hill A, et al. 14-3-3 proteins are part of an abscisic acid-VIVIPAROUS1 (VP1)
response complex in the Em promoter and interact with VP1 and EmBP1. Plant
Cell, 1998, 10(5): 837~847)。
14-3-3广泛参与植物对逆境胁迫应答的信号转导过程。拟南芥的2个14-3-3基因RCI1/RCI1A和RCI2/RCI1B的表达能被低温胁迫所调控 (arillo JA, Capel J,
Leyva A, et al.Two related low-temperature-inducible genes of Arabidopsis
encode proteins showing high homology to 14-3-3 proteins, a family of putative
kinase regulators. Plant Mol Biol, 1994, 25(4): 693-704);拟南芥14-3-3同工蛋白能结合并激活胁迫反应的重要蛋白-钙依赖蛋白激酶(CPK-1)(Camoni L, Harper JF,
Palmgren MG. 14-3-3 proteins activate a plant calcium-dependent protein kinase
(CDPK). FEBS Lett, 1998, 430(3): 381~384)。盐胁迫下,烟草(Nicotiana tabacum)和玉米14-3-3基因表达量上调,表明14-3-3受盐胁迫诱导,将番茄 (Lycopersicon
esculentum) 14-3-3基因(TFT7)转入拟南芥中表达,盐胁迫下转基因拟南芥相对于非转基因拟南芥型有较高的种子萌发率、干重和较长的根长,暗示14-3-3过表达能提高拟南芥对盐胁迫的抵抗能力 (Xu WF, Shi WM.
Mechanisms of salt tolerance in transgenic Arabidopsis thaliana
constitutively overexpressing the tomato 14-3-3 protein TFT7. Plant Soil, 2007,
301:17~28)。14-3-3同样能被干旱胁迫诱导表达,经1%聚乙二醇(PEG)处理过的棉花(Gossypium hirsutum
L.),其根部和叶中6种14-3-3基因家族成员的表达量均上调,其中根部最高上调了10.8倍,在叶中最高上调了3.8倍 (Sun G, Xie F, Zhang B.
Transcriptome-wide identification and stress properties of the 14-3-3 gene
family in cotton (Gossypium hirsutum L.). Funct Integr Genom, 2011,
11(4): 627~636)。转入拟南芥14-3-3基因GF14 λ的棉花较野生型对干旱胁迫具有更高的抗性,并表现“常绿”的表型 (Yan J, He C, Wang J, Mao Z, et al.
Overexpression of the Arabidopsis 14-3-3 protein GF14 lambda in cotton leads to
a "stay-green" phenotype and improves stress tolerance under moderate
drought conditions. Plant Cell Physiol, 2004, 45(8): 1007~1014)。通过干旱处理检测转入玉米14-3-3基因(ZmGF14-6)的水稻 (Oryza sativa)和非转基因水稻的耐干旱能力,在停止灌溉18天后转ZmGF14-6水稻表现出良好的生长态势,胁迫停止后,经过7天的正常灌溉,所有转ZmGF14-6水稻能恢复正常生长,而非转基因水稻只有小部分能恢复生长 (Campo S, Peris-Peris C,
Montesinos L, Peñas G, et al). Expression of the maize ZmGF14-6 gene in
rice confers tolerance to drought stress while enhancing susceptibility to
pathogen infection. J Exp Bot, 2012, 63(2): 983~999)。上述研究结果表明14-3-3蛋白在植物抵御干旱胁迫中起重要作用。
本发明的耐干旱基因Pp14-3-3是从云南本地的砂梨品种火把梨中克隆得到,云南火把梨不仅具有稳定遗传的红色果皮表型,而且对生物胁迫和非生物胁迫的抗性均较强,抗黑星病和腐烂病,抗晚霜、耐低温,对梨木虱也有较强的抗性。此外,火把梨对土壤适应性也很强。
发明内容
本发明的目的是提供一种火把梨耐干旱基因Pp14-3-3,具体是从云南火把梨(Pyrus pyrifolia)中获得一个新的耐干旱基因Pp14-3-3,该基因具有如SEQ ID NO:1所示的核苷酸序列,编码14-3-3蛋白。
本发明中Pp14-3-3基因的编码区是SEQ ID NO:1中所述第85-870位所示的核苷酸序列或编码如SEQ ID NO:2所示氨基酸序列的蛋白质。
本发明另一目的是将火把梨耐干旱基因Pp14-3-3核苷酸序列应用在提高植物耐干旱胁迫中,将基因Pp14-3-3构建到植物表达载体上并导入烟草中表达,对获得的Pp14-3-3转基因烟草进行干旱胁迫处理,确认Pp14-3-3具有耐干旱胁迫的活性,为后期利用该基因改良烟草以及其他植物耐干旱能力奠定基础。
本发明分离克隆云南火把梨中携带的一个抗逆境胁迫相关基因的完整cDNA片段,利用农杆菌介导使目的基因转入受体植物中并过量表达,进而通过实验验证该基因在植物抗逆过程中所起的作用,为后期利用该基因改良烟草以及其他植物抗逆能力奠定基础,增强烟草和其他植物对干旱的抗性。本发明将这个基因命名为Pp14-3-3。
具有该基因片段的植物在一定程度上具有耐干旱胁迫的表型。其中所述DNA片段如序列表SEQ ID所示。对该基因进行序列分析,表明Pp14-3-3全长cDNA为1107 bp,具有786bp的开放阅读框(ORF)、84bp的5’非翻译区和237bp的3’非翻译区,编码含有261个氨基酸的蛋白质。Pp14-3-3编码蛋白具有14-3-3家族的保守结构域,与来自苹果(Malus domestica)、龙眼(Dimocarpus longan)和棉花以及其他物种的14-3-3蛋白高度相似,结构域分析显示该序列包含2个14-3-3典型结构域,分别位于第48–58和220-239氨基酸残基处。生物信息学分析表明本发明克隆基因编码的蛋白质属于火把梨中的14-3-3蛋白。超量表达序列表SEQ ID所示序列可以增强烟草对干旱胁迫的抗性。
本发明通过如下具体技术方案实现本发明目的:
(1)基因的获得:以火把梨红色果皮为材料,提取果皮的总RNA,利用RT-PCR(reverse
transcription-polymerase chain reaction)扩增到Pp14-3-3的基因序列,然后将其连接到pMD-18T载体上,经测序获得具有目的基因的克隆。
(2)植物表达载体构建与遗传转化:用限制性内切酶BamHⅠ和EcoRⅠ酶切pMD-18T-Pp14-3-3载体,通过胶回收获得目的基因片段。用同样的内切酶消化植物表达载体pCAMBIA2300s,胶回收获得载体大片段。将目的基因片段与pCAMBIA2300s载体片段连接,构建超表达载体pCAMBIA2300s-Pp14-3-3。通过液氮冻融法将pCAMBIA2300s-Pp14-3-3植物表达载体导入农杆菌菌株LBA4404中。利用农杆菌介导的遗传转化法,将Pp14-3-3编码序列导入烟草等植物中表达。通过抗生素筛选、基因组DNA PCR和RT-PCR筛选阳性转基因植株。
(3)转基因植株抗性分析:将生长健壮的转基因植株和野生型植株(非转基因对照),分别种植于花盆中。待生长稳定之后,停止灌溉15天,观察干旱胁迫下转基因植株和野生型植株的表型,从而验证Pp14-3-3转基因植株对干旱胁迫的抗性。
本发明为增强植物对干旱的抗性提供了一种方法,通过基因工程手段培育抗干旱植物能克服传统育种的不足,不仅育种周期短,而且操作简单。将来自火把梨的Pp14-3-3基因导入植物中表达,能增强转基因植物对干旱的抗性,将该基因导入烟草中,可以产生耐干旱的新型烟草种质和品种。利用基因工程技术增加植物抗逆性具有明显的优势和不可取代的重要性。它可以为作物、花卉等大规模生产提供方便,大量减少因自然条件恶劣而造成的减产,为农业生产节约成本且提高管理水平,因此本发明具有广阔的市场应用前景。
附图说明
图1是本发明部分Pp14-3-3转基因烟草基因组DNA的PCR检测结果。其中Marker:DL2000 DNA Marker (大连宝生物),由2,000bp、1,000bp、750bp、500bp、250bp以及100bp六条DNA片段组成。正对照:以质粒pMD-18T-Pp14-3-3为模板的PCR产物;WT:以非转基因烟草(即野生型烟草)总DNA为模板的PCR产物;空白对照:PCR体系中不包含模板DNA的反应产物。
图2是本发明部分阳性转基因烟草中Pp14-3-3转录水平的表达分析结果。其中Marker:DL2000 DNA Marker (大连宝生物);1-16分别为不同的阳性转基因烟草单株;WT:非转基因烟草总RNA逆转录cDNA为模板的PCR产物;空白对照:PCR体系中不包含模板DNA的反应产物;正对照:质粒pMD-18T-Pp14-3-3为模板的PCR产物。
图3是Pp14-3-3转基因烟草耐干旱特性分析示意图。其中A为WT,即野生型烟草;B为阴性对照植株,即转入pCAMBIA2300S空载体的烟草植株;C为编号为2的Pp14-3-3转基因烟草;D为编号为7的Pp14-3-3转基因烟草。
具体实施方式
下面通过附图和实施例对本发明作进一步详细说明,但本发明保护范围不局限于所述内容。
实施例1:Pp14-3-3全长基因的克隆以及序列分析
用液氮将火把梨的红色果皮研磨成粉末以后,转入离心管中,采用异硫氰酸胍法提取总RNA。采用逆转录酶M-MLV (天根)以总RNA为模板合成cDNA 第一链,反应体系和操作过程为:取5 μg Total RNA,依次加入50 ng oligo(dT) 15、2 μL dNTP (2.5mM each)、ddH2O
(RNase-free)至反应体积为13.5 μL;混匀后,70℃加热变性5min后迅速在冰上冷却5min,然后依次加入4 μL 5×First-stand buffer、0.5 μL RNasin (200U)、1 μL M-MLV (200U)、1 μL DTT (0.1M),混匀并短时离心,42℃温浴1.5h,取出后95℃加热5min,终止反应。cDNA第一链合成后置于-20℃保存备用。
以合成的第一链cDNA为模板,扩增目的基因Pp14-3-3,所用引物序列分别为5’
GACCAGAGAAGGTTTCAGATTTAGG3’以及5’ TGCCATCTATTACAAAGGTCCCAAG3’。采用大连宝生物的高保真DNA聚合酶ex taq扩增出目的基因。PCR反应条件:94℃ 3min;94℃ 30s,60℃ 30s,72℃ 1min,28cycles;72℃ 5 min。反应体系(20 μL)为1 μL cDNA、2 μL 10×Buffer、0.4 μL dNTP (10mM each)、0.1 μL 正向引物(20μM)、0.1 μL 反向引物(20μM)、0.2 μL ex Taq DNA polymerase
(5U/μL)、16.2 μL ddH2O。PCR结束后,取5 μL用于琼脂糖凝胶电泳,检测扩增产物的特异性以及大小。
由于PCR产物只有一条DNA带,故直接对PCR产物进行TA克隆,使用的试剂盒为pMD-18T vector kit (大连宝生物),反应体系和操作过程为:取1.5 μL PCR产物,依次加入1 μL pMD18-T vector (50ng/μL) 和2.5 μL 2×Ligation solution I,混匀置于16℃过夜反应。采用热激转化法将连接产物转入大肠杆菌DH5α中。在涂于含有X-Gal、IPTG、氨苄青霉素(ampicillin,Amp)的LB固体培养基筛选阳性克隆。挑选若干个白色菌落,摇菌后用扩增Pp14-3-3的特异引物鉴定出多克隆位点插入Pp14-3-3的克隆。将鉴定的克隆进行测序,最终获得的Pp14-3-3全长cDNA为1107 bp,通过NCBI ORF finder
(http://www.ncbi.nlm.nih.gov/gorf/ gorf.html)分析发现其包含一个786bp的开放读码框(见序列表)。Pp14-3-3编码一个含261个氨基酸的蛋白质,Pp14-3-3的分子量为29.6KDa,等电为4.71。SignalP 3.0分析结果显示Pp14-3-3没有信号肽的存在,而且未发现线粒体、过氧化酶体、溶酶体和细胞核等亚细胞定位序列,可见Pp14-3-3属于非分泌蛋白。
实施例2:植物表达载体的构建
采用碱裂解法提取插入Pp14-3-3的大肠杆菌质粒pMD-18T-Pp14-3-3以及植物表达载体pCAMBIA2300S的质粒,取1 μL用于琼脂糖凝胶电泳以检测所提取质粒的完整性和浓度高低。用EcoRI (Fermentas)和BamHI (Fermentas)分别对质粒pMD-18T-Pp14-3-3和pCAMBIA2300S进行双酶切(100 μL体系),反应体系和操作过程为:取10 μL pMD-18T-Pp14-3-3或pCAMBIA2300S质粒、依次加入10 μL 10×Tango buffer、2 μL EcoRI、2 μL BamHI、76 μL ddH2O,混匀后短时离心,置于37℃过夜反应。将所有酶切产物点于琼脂糖凝胶中进行电泳,然后对Pp14-3-3片段和pCAMBIA2300S大片段分别进行胶回收,整个过程使用UNIQ-10柱式DNA胶回收试剂盒 (上海生工)。具体操作流程为:切下含有目的DNA条带的琼脂糖凝胶块置于1.5 mL离心管中,按400 μL/100 mg琼脂糖凝胶的比例加入Binding Buffer Ⅱ,置于60℃水浴10min,使胶彻底融化;将融化的凝胶溶液转移至2 mL收集管内的UNIQ-10柱中,室温放置2min后离心1min (6,000g/min);取下UNIQ-10柱,倒掉收集管中的废液,再将UNIQ-10柱放入收集管中,加入500 μL Wash Solution,室温离心1min (8,000g/min),再重复此步骤一次;取下UNIQ-10柱,倒掉收集管中的废液,再将UNIQ-10柱放入收集管中,室温离心2min (12,000g/min);取下UNIQ-10柱放入新的1.5 mL离心管中,在膜中央加入预热的20 μL Elution Buffer,室温放置2min,室温离心1min (12,000g/min),离心管中收集液体即为回收的DNA片段。取1 μL回收产物通过琼脂糖凝胶电泳检测回收片段的大小以及浓度,置于-20℃保存备用。
利用T4 DNA Ligase (大连宝生物),将回收的Pp14-3-3 DNA片段和pCAMBIA2300S载体片段连接起来,反应体系(20 μL)和操作过程为:取10 μL Pp14-3-3 DNA片段依次加入2 μL pCAMBIA2300S载体DNA、2 μL 10×T4 DNA Ligase Buffer、1 μL T4 DNA Ligase、5 μL ddH2O,混匀后短时离心,置于16℃金属浴过夜反应。接着采用热激转化法将连接产物转入大肠杆菌DH5α中,用含有50mg/L卡那霉素(kanamycin,Km)的固体培养基筛选阳性克隆。挑选单菌落摇菌,以菌液为模板用扩增Pp14-3-3的特异引物进行PCR,挑选出Pp14-3-3与pCAMBIA2300S成功连接的克隆,所检测的菌株若为阳性,加入适量甘油并置于-80℃保存备用。
用碱裂解法提取并纯化上述大肠杆菌中的pCAMBIA2300S-Pp14-3-3质粒。并制备农杆菌LBA4404菌株的感受态细胞,操作过程为:将实验室保存的LBA4404菌株在LB固体培养基(含利福平20mg/L)上划线,28℃培养至长出单菌落后,挑取一个单克隆于含20mg/L利福平的2 mL LB液体培养基,28℃振荡培养至混浊;取5 mL菌液转接于含20mg/L利福平的100 mL LB液体培养基中,28℃振荡培养至OD600为0.5;4℃离心5min (5,000g/min)收集菌体,弃上清,加入10 mL预冷的0.1M CaCl2,轻轻充分悬浮菌体,冰浴20min;接着4℃离心5min (5,000g/min)收集菌体,弃上清,加入4 mL预冷的含15%甘油的0.1M CaCl2溶液,充分悬浮后分装于1.5 mL离心管中,每管200 μL,液氮速冻后置于-80℃保存备用。
采用液氮冻融法将上述构建的植物表达载体pCAMBIA2300S-Pp14-3-3转入所制备的农杆菌LBA4404感受态细胞中。操作步骤为:取2 μg pCAMBIA2300S-Pp14-3-3质粒加入含有200 μL感受态细胞的离心管中,轻轻混匀后冰浴5min,接着转入液氮中冷冻1min,然后迅速置于37℃水浴5min,之后立即冰浴2min,加入800 μL LB液体培养基,28℃振荡培养4h。将活化后的农杆菌涂于含有50mg/L Km的LB固体培养基上,28℃静止培养。挑选单菌落摇菌,用扩增Pp14-3-3的特异引物进行PCR,检测pCAMBIA2300S-Pp14-3-3是否转入农杆菌中。对于阳性克隆,加入甘油后置于-80℃保存备用。
实施例3:农杆菌介导的植物遗传转化以及转基因植物筛选
本实验的转基因受体是烟草。将烟草种子用75%的酒精浸泡30s,用无菌水洗涤后用0.1%的HgCl2浸泡8min,然后再用无菌水洗涤若干次,播种于1/2 MS培养基上,28℃暗培养5-8d,发芽后转至光照培养箱(25℃,16h/d光照),以后每月用MS培养基继代一次。
从-80℃冰箱中取出保存的含有pCAMBIA2300S-Pp14-3-3质粒的农杆菌LBA4404菌种,接种于5 mL含有50mg/L Km和20mg/L利福平的LB液体培养基中,28℃培养至浑浊。吸取1 mL浑浊的菌液至含有50mg/L Km的LB固体培养基上,28℃培养48h。将LB固体培养基上的农杆菌刮下适量接种于MGL液体培养基中,附加一定量的乙酰丁香酮,28℃振荡培养2-3h以活化农杆菌。
取烟草无菌苗叶子切成1cm2左右的叶盘,完全浸泡于上述含有活化农杆菌的MGL液体培养基中,浸染15min。用无菌滤纸吸干叶片表面的菌液,将叶盘置于共培养基上进行室温培养。共培养基为MS+0.02 mg/L 6-BA+2 mg/L
NAA+30g/L蔗糖,培养时间为2d。
将共培养后的叶盘转到加有抗生素的MS筛选培养基中分化成苗,同时筛选转基因植株。筛选培养基为MS+0.5 mg/L 6-BA+0.1 mg/L
NAA+30g/L蔗糖+50 mg/L Km+200 mg/L 头孢霉素(cefotaxime sodium salt,Cef);筛选培养时将培养瓶转移至光照培养箱培养(25℃,16h/d光照,8h/d黑暗)。出芽后用含有50 mg/L Km和200 mg/L Cef的MS培养基继代培养。因烟草愈伤分化率较高,故需要对再生植株进行进一步筛选。将烟草再生苗移至含有50mg/L Km和200mg/L Cef的MS培养基上使其生根,最后选用生根较好的再生苗做进一步的分子水平检测。
采用CTAB法提取转基因烟草植株叶片的基因组DNA,取1 μL基因组DNA通过琼脂糖凝胶电泳检测其完整性和浓度。以转基因植株的基因组DNA为模板用扩增Pp14-3-3的特异引物进行PCR。PCR结束后,取8 μL产物用于琼脂糖凝胶电泳以检测阳性转基因植株。部分转基因烟草植株的扩增结果如图1所示。Pp14-3-3转化烟草共筛选到41株阳性转基因植株,分别编号为1~41。
实施例4:Pp14-3-3表达分析以及转基因植株抗干旱能力分析
取阳性转基因单株以及非转基因烟草(野生型)的嫩叶提取总RNA,逆转录生成第一链cDNA,并以此为模板扩增Pp14-3-3基因所用引物序列与实施例1中相同。PCR反应条件:94℃ 3min;94℃ 30s,60℃ 30s,72℃ 1min,26个循环;72℃ 5min。根据PCR结果分析各转基因单株中Pp14-3-3转录水平的表达。总RNA提取以及RT-PCR的方法与步骤1)相同。PCR结束后,取5 μL用于琼脂糖凝胶电泳,部分单株的检测结果如图2所示。共检测到24株转基因单株中Pp14-3-3在转录水平有表达,这些植株分别编号为1、2、3、5、6、7、8、11、12、13、14、15、16、19、21、22、23、27、29、30、33、34、36、39。
对检测出的阳性转基因烟草进行组培快繁,并挑选生长健壮的Pp14-3-3转基因烟草、野生型烟草和阴性对照烟草植株(转入pCAMBIA2300S空载体烟草植株)组培苗分别种于花盆中,待生长稳定后,停止灌溉15天,观察干旱胁迫下各植株的生长情况。结果如图3所示,干旱处理15天后,野生型和阴性对照出现明显脱水枯萎现象,而Pp14-3-3转基因烟草(编号分别为2和7)则未出现明显的枯萎和脱水症状。在干旱胁迫下, Pp14-3-3转基因烟草相对于野生型烟草表现出更好的生长态势。上述实验结果表明,转入Pp14-3-3基因可大大增强烟草对干旱胁迫的抵抗能力。
序列表
(SEQ
ID)
<110> 昆明理工大学
<120> 一种火把梨耐干旱基因Pp14-3-3及其应用
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 1107
<212> DNA
<213> Pyrus pyrifolia Nakai
<220>
<221> mRNA
<222> (1)..(1107)
<220>
<221> 5'UTR
<222> (1)..(84)
<220>
<221> CDS
<222> (85)..(870)
<220>
<221> 3'UTR
<222> (871)..(1107)
<400> 1
acgcggggag ttttgcttcg agcttagatc ttcttcttcc tcgtggacca
gagaaggttt 60
cagatttagg cttacatagc aaag atg ccg cca act gat tct tca cgc
gag 111
Met
Pro Pro Thr Asp Ser Ser Arg
Glu
1
5
gag aat gtg tac atg gcc aag ttg gcc gaa cag gct gag aga tat
gag 159
Glu Asn Val Tyr Met Ala Lys Leu Ala Glu Gln Ala Glu Arg Tyr
Glu
10
15
20
25
gag atg gtc gag ttt atg gag aaa gtt gcc aag act gtc gat gtt
gag 207
Glu Met Val Glu Phe Met Glu Lys Val Ala Lys Thr Val Asp Val
Glu
30
35
40
gag cta aca gtg gag gaa aga aat ctc ctc tct gtg gct tat aag
aat 255
Glu Leu Thr Val Glu Glu Arg Asn Leu Leu Ser Val Ala Tyr Lys
Asn
45
50
55
gtg att gga gct agg aga gct tcg tgg agg atc ata tct tcc att
gag 303
Val Ile Gly Ala Arg Arg Ala Ser Trp Arg Ile Ile Ser Ser Ile
Glu
60
65
70
cag aag gag gag agc aga ggg aat gag gat cat gtt gcg att atc
aag 351
Gln Lys Glu Glu Ser Arg Gly Asn Glu Asp His Val Ala Ile Ile
Lys
75
80
85
gag tac agg agc aat att gag act gag ctc agc aag atc ttt gat
ggg 399
Glu Tyr Arg Ser Asn Ile Glu Thr Glu Leu Ser Lys Ile Phe Asp
Gly
90
95
100
105
atc ttg aac ctt ctt gag tct cac ctc att cca tct gcc tca tcc
gcc 447
Ile Leu Asn Leu Leu Glu Ser His Leu Ile Pro Ser Ala Ser Ser
Ala
110
115
120
gag tcc aag gtg ttt tac ctc aag atg aaa ggt gat tac cac agg
tac 495
Glu Ser Lys Val Phe Tyr Leu Lys Met Lys Gly Asp Tyr His Arg
Tyr
125
130
135
ctt gct gag ttt aag act gga ggc gaa aga aag gaa gca gct gag
agc 543
Leu Ala Glu Phe Lys Thr Gly Gly Glu Arg Lys Glu Ala Ala Glu
Ser
140
145
150
act ttg ttg gct tac aag tct gct cag gat att gcc ctg gct gag
ctg 591
Thr Leu Leu Ala Tyr Lys Ser Ala Gln Asp Ile Ala Leu Ala Glu
Leu
155
160
165
gct cca acc cac ccg ata agg ctt gga ctt gca ctc aac ttc tca
gtc 639
Ala Pro Thr His Pro Ile Arg Leu Gly Leu Ala Leu Asn Phe Ser Val
170
175
180
185
ttt tac tat gaa atc ctt aat tca ccc gat cgt gct tgc aat ctg
gca 687
Phe Tyr Tyr Glu Ile Leu Asn Ser Pro Asp Arg Ala Cys Asn Leu
Ala
190
195
200
aaa cag gct ttt gat gag gct att tct gag ctt gac aca ttg ggt
gag 735
Lys Gln Ala Phe Asp Glu Ala Ile Ser Glu Leu Asp Thr Leu Gly
Glu
205
210
215
gaa tcc tac aag gat agc act ttg atc atg cag ctt ttc cga gac
aat 783
Glu Ser Tyr Lys Asp Ser Thr Leu Ile Met Gln Leu Phe Arg Asp
Asn
220
225
230
ctg aca ctc tgg act tct gat atc acg gat gat gcc gga gac gaa
atc 831
Leu Thr Leu Trp Thr Ser Asp Ile Thr Asp Asp Ala Gly Asp Glu
Ile
235
240
245
aag gaa gca tca aag cgc gaa tca agt gaa gca cag taa
caggtttttt 880
Lys Glu Ala Ser Lys Arg Glu Ser Ser Glu Ala
Gln
250
255
260
ggttcataga atttgtaccc tgtacccctt ttatcttggg acctttgtaa
tagatggcat 940
gtgggttgta gaatgatttg tctatgaact tgacgatctt gagcgcggca
gcgcttaggg 1000
ttgaaatgtt ccaacagatt gcaaaattgc gttttataat ttggtgagat
tttaatgctg 1060
tgctatatga tttagtgtct aaaaaaaaaa aaaaaaaaaa
aaaaaaa
1107
<210> 2
<211> 261
<212> PRT
<213> Pyrus pyrifolia Nakai
<400> 2
Met Pro Pro Thr Asp Ser Ser Arg Glu Glu Asn Val Tyr Met Ala Lys
1
5
10
15
Leu Ala Glu Gln Ala Glu Arg Tyr Glu Glu Met Val Glu Phe Met Glu
20
25
30
Lys Val Ala Lys Thr Val Asp Val Glu Glu Leu Thr Val Glu Glu Arg
35
40
45
Asn Leu Leu Ser Val Ala Tyr Lys Asn Val Ile Gly Ala Arg Arg Ala
50
55
60
Ser Trp Arg Ile Ile Ser Ser Ile Glu Gln Lys Glu Glu Ser Arg Gly
65
70
75
80
Asn Glu Asp His Val Ala Ile Ile Lys Glu Tyr Arg Ser Asn Ile Glu
85
90
95
Thr Glu Leu Ser Lys Ile Phe Asp Gly Ile Leu Asn Leu Leu Glu Ser
100
105
110
His Leu Ile Pro Ser Ala Ser Ser Ala Glu Ser Lys Val Phe Tyr Leu
115
120
125
Lys Met Lys Gly Asp Tyr His Arg Tyr Leu Ala Glu Phe Lys Thr Gly
130
135
140
Gly Glu Arg Lys Glu Ala Ala Glu Ser Thr Leu Leu Ala Tyr Lys Ser
145
150
155
160
Ala Gln Asp Ile Ala Leu Ala Glu Leu Ala Pro Thr His Pro Ile Arg
165
170
175
Leu Gly Leu Ala Leu Asn Phe Ser Val Phe Tyr Tyr Glu Ile Leu Asn
180
185
190
Ser Pro Asp Arg Ala Cys Asn Leu Ala Lys Gln Ala Phe Asp Glu Ala
195
200
205
Ile Ser Glu Leu Asp Thr Leu Gly Glu Glu Ser Tyr Lys Asp Ser Thr
210
215
220
Leu Ile Met Gln Leu Phe Arg Asp Asn Leu Thr Leu Trp Thr Ser Asp
225
230
235
240
Ile Thr Asp Asp Ala Gly Asp Glu Ile Lys Glu Ala Ser Lys Arg Glu
245
250
255
Ser Ser Glu Ala Gln
260
Claims (3)
1.一种火把梨耐干旱基因Pp14-3-3,其特征在于:其具有如SEQ ID NO:1所示核苷酸序列,编码14-3-3蛋白。
2.根据权利要求1所述火把梨耐干旱基因Pp14-3-3,其特征在于:Pp14-3-3基因的编码区是SEQ ID NO:1中所述第85-870位所示的核苷酸序列或编码如SEQ
ID NO:2所示氨基酸序列的蛋白质。
3.权利要求1所述火把梨耐干旱基因Pp14-3-3核苷酸序列在提高植物耐干旱胁迫中的应用。
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103060360A (zh) * | 2013-01-04 | 2013-04-24 | 昆明理工大学 | 黑大豆14-3-3j蛋白基因的原核表达载体及其应用 |
CN109912705A (zh) * | 2019-04-09 | 2019-06-21 | 贵州大学 | 一种高粱14-3-3蛋白GF14c基因及其重组载体和表达方法 |
CN110656125A (zh) * | 2019-09-23 | 2020-01-07 | 四川育良生物科技有限公司 | 一种耐干旱玉米的遗传转化方法 |
CN114644702A (zh) * | 2020-12-21 | 2022-06-21 | 中国农业大学 | Tango蛋白、其相关生物材料以及植物育种方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102391369A (zh) * | 2011-12-07 | 2012-03-28 | 中国科学院南京土壤研究所 | 耐逆性相关的14-3-3蛋白grf9及其应用 |
-
2012
- 2012-04-13 CN CN2012101070498A patent/CN102643831A/zh active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102391369A (zh) * | 2011-12-07 | 2012-03-28 | 中国科学院南京土壤研究所 | 耐逆性相关的14-3-3蛋白grf9及其应用 |
Non-Patent Citations (6)
Title |
---|
《Gene Bank》 20110101 Liu D. et al Accession No:JF810599.1 全文 1-3 , * |
《Plant Cell Physiol. 》 20041231 Juqiang Yan et al. Overexpression of the Arabidopsis 14-3-3 Protein GF14lambda in Cotton Leads to a "Stay-Green" Phenotype and Improves Stress Tolerance under Moderate Drought Conditions 摘要 1 第45卷, 第8期 * |
《生命中的化学》 20021231 陈明 等 14-3-3蛋白 第447页右边第5段 1 第22卷, 第5期 * |
JUQIANG YAN ET AL.: "Overexpression of the Arabidopsis 14-3-3 Protein GF14λ in Cotton Leads to a "Stay-Green" Phenotype and Improves Stress Tolerance under Moderate Drought Conditions", 《PLANT CELL PHYSIOL. 》 * |
LIU D. ET AL: "Accession No:JF810599.1", 《GENE BANK》 * |
陈明 等: "14-3-3蛋白", 《生命中的化学》 * |
Cited By (5)
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CN103060360A (zh) * | 2013-01-04 | 2013-04-24 | 昆明理工大学 | 黑大豆14-3-3j蛋白基因的原核表达载体及其应用 |
CN109912705A (zh) * | 2019-04-09 | 2019-06-21 | 贵州大学 | 一种高粱14-3-3蛋白GF14c基因及其重组载体和表达方法 |
CN110656125A (zh) * | 2019-09-23 | 2020-01-07 | 四川育良生物科技有限公司 | 一种耐干旱玉米的遗传转化方法 |
CN114644702A (zh) * | 2020-12-21 | 2022-06-21 | 中国农业大学 | Tango蛋白、其相关生物材料以及植物育种方法 |
CN114644702B (zh) * | 2020-12-21 | 2023-10-03 | 中国农业大学 | Tango蛋白、其相关生物材料以及植物育种方法 |
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