CN110643588A - TaCDPK1蛋白质及其编码基因在调控植物耐逆性中的应用 - Google Patents
TaCDPK1蛋白质及其编码基因在调控植物耐逆性中的应用 Download PDFInfo
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Abstract
本发明公开了TaCDPK1蛋白质及其编码基因在调控植物耐逆性中的应用。本发明的TaCDPK1蛋白质的氨基酸序列是序列2所示的蛋白质,其编码基因的核苷酸序列是序列1第280‑1878位所示的DNA分子。通过实验证明:本发明的TaCDPK1基因在干旱、GA或BR的诱导下表达;将TaCDPK1基因导入拟南芥中得到的转基因拟南芥,其对干旱逆境的抗性高于野生型拟南芥并且参与GA信号转导途径。本发明提供的蛋白和基因为人为控制抗逆和耐逆相关基因的表达提供了基础,将在培育抗逆性和耐逆性增强的植物中发挥重要的作用。
Description
技术领域
本发明涉及生物技术领域,具体涉及TaCDPK1蛋白质及其编码基因在调控植物耐逆性中的应用。
背景技术
干旱、高盐及低温等逆境胁迫是影响小麦生长、发育的障碍因子。因此,了解小麦对逆境条件的应答与信号传导机制,提高小麦品种的抗逆性,成为小麦遗传研究及小麦品种改良的重要任务之一。
在逆境胁迫下植物体内会产生一系列应答反应,伴随着许多生理生化及发育上的变化。明确植物对逆境的反应机制,将为抗逆基因工程研究和应用提供科学论据。目前,植物抗逆性研究已逐渐深入到细胞、分子水平,并与遗传学和遗传工程研究相结合,探索用生物技术来改进植物生长特性,其目的是提高植物对逆境的适应能力。
在干旱、高盐和低温等环境胁迫的逆境条件下,植物能够在分子、细胞和整体水平上做出相应的调整,以最大程度上减少环境所造成的伤害并得以生存。许多基因受胁迫诱导表达,这些基因的产物不仅能够直接参与植物的胁迫应答,而且能够调节其它相关基因的表达或参与信号传导途径,从而使植物避免或减少伤害,增强对胁迫环境的抗性。与胁迫相关的基因产物可以分为两大类:第一类基因编码的产物包括离子通道蛋白、水通道蛋白、渗透调节因子(蔗糖、脯氨酸和甜菜碱等)合成酶等直接参与植物胁迫应答的基因产物;第二类基因编码的产物包括参与胁迫相关的信号传递和基因表达调节的蛋白因子,如蛋白激酶、转录因子等。其中,蛋白激酶在植物胁迫信号的感知、传递的调控中起着重要作用。
到目前为止,已发现的蛋白激酶约有300种左右。在细胞信号传导、细胞周期调控等系统中,蛋白激酶形成了纵横交错的网络。这类酶催化从ATP转移出磷酸并共价结合到特定蛋白质分子中某些丝氨酸、苏氨酸或酪氨酸残基的羟基上,从而改变蛋白质、酶的构象和活性。根据蛋白激酶的磷酸化氨基酸残基的种类,蛋白激酶可以分为Ser/Thr蛋白激酶,Tyr蛋白激酶以及双特异性Ser/Thr/Tyr蛋白激酶。Ser/Thr蛋白激酶主要包括MACDPK1,CDCDPK1,JNK等。这种类型的蛋白激酶目前研究的较多,并且在信号传导途径中起重要作用。而酪氨酸蛋白激酶分为两类,一类是非受体型酪氨酸蛋白激酶,以src基因产物为代表,此外还有Yes、Fyn、Lck、Fgr、Lyn、Fps/Fes及Ab1等;另一类是受体型酪氨酸蛋白激酶,根据它们的结构不同,受体型酪氨酸激酶可以分为9种类型。在动物细胞中,酪氨酸蛋白激酶往往参与程序性细胞死亡、癌变等重要细胞行为。
蛋白激酶在植物响应逆境胁迫的信号传导途径中占有重要的地位。蛋白激酶通过自身的磷酸化提高或者降低自身的激酶活性,通过磷酸化底物提高或者降低底物蛋白的酶活性,然后底物蛋白又调控下游基因的酶活,形成一个级联调控途径。如RAS信号调控途径,在细胞受体酪氨酸蛋白激酶感受到配体的信号后,形成二聚体并发生自身磷酸化,激活RAS,然后由活化的RAS激活引起蛋白激酶的级联反应。这种级联调控是一种很精密很复杂的调控网络,蛋白激酶可能在上游作为一种开关在行使其功能。
发明内容
本发明所要解决的技术问题是如何调控植物耐逆性。
为解决上述技术问题,本发明首先提供了来源于麦类小麦属(Triticum aestivumL.)的TaCDPK1蛋白质的新用途。
本发明提供了TaCDPK1蛋白质在如下任一中的应用:
1)调控植物耐逆性;
2)培育耐逆性提高的转基因植物;
3)参与GA信号转导途径;
4)植物育种;
所述TaCDPK1蛋白质是如下a)或b)或c)或d)的蛋白质:
a)氨基酸序列是序列2所示的蛋白质;
b)在序列2所示的蛋白质的N端和/或C端连接标签得到的融合蛋白质;
c)将序列2所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的具有相同功能的蛋白质;
d)与序列2所示的氨基酸序列具有75%或75%以上的同源性且具有相同功能的蛋白质。
其中,序列2由532个氨基酸残基组成。
为了使a)中的蛋白质便于纯化,可在序列表中序列2所示的蛋白质的氨基末端或羧基末端连接上如表1所示的标签。
表1、标签的序列
标签 | 残基 | 序列 |
Poly-Arg | 5-6(通常为5个) | RRRRR |
Poly-His | 2-10(通常为6个) | HHHHHH |
FLAG | 8 | DYKDDDDK |
Strep-tag II | 8 | WSHPQFEK |
c-myc | 10 | EQKLISEEDL |
上述c)中的蛋白质TaCDPK1,所述一个或几个氨基酸残基的取代和/或缺失和/或添加为不超过10个氨基酸残基的取代和/或缺失和/或添加。
上述c)中的蛋白质TaCDPK1可人工合成,也可先合成其编码基因,再进行生物表达得到。
上述c)中的蛋白质TaCDPK1的编码基因可通过将序列1所示的DNA序列中缺失一个或几个氨基酸残基的密码子,和/或进行一个或几个碱基对的错义突变,和/或在其5′端和/或3′端连上表1所示的标签的编码序列得到。
为解决上述技术问题,本发明又提供了与TaCDPK1蛋白质相关的生物材料的新用途。
本发明提供了与TaCDPK1蛋白质相关的生物材料在如下任一中的应用:
1)调控植物耐逆性;
2)培育耐逆性提高的转基因植物;
3)参与GA信号转导途径;
4)植物育种;
所述生物材料为下述A1)至A12)中的任一种:
A1)编码TaCDPK1蛋白质的核酸分子;
A2)含有A1)所述核酸分子的表达盒;
A3)含有A1)所述核酸分子的重组载体;
A4)含有A2)所述表达盒的重组载体;
A5)含有A1)所述核酸分子的重组微生物;
A6)含有A2)所述表达盒的重组微生物;
A7)含有A3)所述重组载体的重组微生物;
A8)含有A4)所述重组载体的重组微生物;
A9)含有A1)所述核酸分子的转基因植物细胞系;
A10)含有A2)所述表达盒的转基因植物细胞系;
A11)含有A3)所述重组载体的转基因植物细胞系;
A12)含有A4)所述重组载体的转基因植物细胞系。
上述应用中,A1)所述核酸分子为如下1)或2)或3)所示的基因:
1)其编码序列是序列1第280-1878位所示的cDNA分子或基因组DNA分子;
2)与1)限定的核苷酸序列具有75%或75%以上同一性,且编码TaCDPK1蛋白质的cDNA分子或基因组DNA分子;
3)在严格条件下与1)或2)限定的核苷酸序列杂交,且编码TaCDPK1蛋白质的cDNA分子或基因组DNA分子。
其中,所述核酸分子可以是DNA,如cDNA、基因组DNA或重组DNA;所述核酸分子也可以是RNA,如mRNA或hnRNA等。
本领域普通技术人员可以很容易地采用已知的方法,例如定向进化和点突变的方法,对本发明的编码TaCDPK1的核苷酸序列进行突变。那些经过人工修饰的,具有与本发明分离得到的TaCDPK1的核苷酸序列75%或者更高同一性的核苷酸,只要编码TaCDPK1且具有相同功能,均是衍生于本发明的核苷酸序列并且等同于本发明的序列。
这里使用的术语“同一性”指与天然核酸序列的序列相似性。“同一性”包括与本发明的编码序列2所示的氨基酸序列组成的蛋白质的核苷酸序列具有75%或更高,或85%或更高,或90%或更高,或95%或更高同一性的核苷酸序列。同一性可以用肉眼或计算机软件进行评价。使用计算机软件,两个或多个序列之间的同一性可以用百分比(%)表示,其可以用来评价相关序列之间的同一性。
上述75%或75%以上同一性,可为80%、85%、90%或95%以上的同一性。
上述应用中,A2)所述的含有编码TaCDPK1的核酸分子的表达盒(TaCDPK1基因表达盒),是指能够在宿主细胞中表达TaCDPK1的DNA,该DNA不但可包括启动TaCDPK1转录的启动子,还可包括终止TaCDPK1转录的终止子。进一步的,所述表达盒还可包括增强子序列。可用于本发明的启动子包括但不限于:组成型启动子;组织、器官和发育特异的启动子及诱导型启动子。合适的转录终止子包括但不限于:农杆菌胭脂碱合成酶终止子(NOS终止子)、花椰菜花叶病毒CaMV 35S终止子、tml终止子和豌豆rbcS E9终止子。
可用现有的表达载体构建含有所述TaCDPK1基因表达盒的重组载体。所述植物表达载体包括双元农杆菌载体和可用于植物微弹轰击的载体等。如pAHC25、pBin438、pCAMBIA1302、pCAMBIA2300、pCAMBIA2301、pCAMBIA1301、pCAMBIA1300、pBI121、pCAMBIA1391-Xa或pCAMBIA1391-Xb(CAMBIA公司)等。所述植物表达载体还可包含外源基因的3′端非翻译区域,即包含聚腺苷酸信号和任何其它参与mRNA加工或基因表达的DNA片段。所述聚腺苷酸信号可引导聚腺苷酸加入到mRNA前体的3′端,如农杆菌冠瘿瘤诱导(Ti)质粒基因(如胭脂碱合成酶基因Nos)、植物基因(如大豆贮存蛋白基因)3′端转录的非翻译区均具有类似功能。使用本发明的基因构建植物表达载体时,还可使用增强子,包括翻译增强子或转录增强子,这些增强子区域可以是ATG起始密码子或邻接区域起始密码子等,但必需与编码序列的阅读框相同,以保证整个序列的正确翻译。所述翻译控制信号和起始密码子的来源是广泛的,可以是天然的,也可以是合成的。翻译起始区域可以来自转录起始区域或结构基因。为了便于对转基因植物细胞或植物进行鉴定及筛选,可对所用植物表达载体进行加工,如加入可在植物中表达的编码可产生颜色变化的酶或发光化合物的基因(GUS基因、萤光素酶基因等)、抗生素的标记基因(如赋予对卡那霉素和相关抗生素抗性的nptII基因,赋予对除草剂膦丝菌素抗性的bar基因,赋予对抗生素潮霉素抗性的hph基因,和赋予对氨甲喋呤抗性的dhfr基因,赋予对草甘磷抗性的EPSPS基因)或是抗化学试剂标记基因等(如抗除莠剂基因)、提供代谢甘露糖能力的甘露糖-6-磷酸异构酶基因。从转基因植物的安全性考虑,可不加任何选择性标记基因,直接以逆境筛选转化植株。
上述应用中,所述载体可为质粒、黏粒、噬菌体或病毒载体。在本发明中,所述重组表达载体为将上述TaCDPK1基因插入pBI121载体中得到的重组载体,具体为将序列表的序列1自5'端第280-1878位核苷酸所示的DNA片段插入pBI121载体的SmaI和SpeI酶切识别位点间得到的重组载体。
上述应用中,所述微生物可为酵母、细菌、藻或真菌,如农杆菌。
上述应用中,所述转基因植物细胞系均不包括繁殖材料。
上述应用中,所述耐逆性为耐旱性;所述提高植物耐旱性或参与GA信号转导途径具体体现在如下(1)-(6)中任一种:(1)在干旱或GA处理下,提高植物发芽率;(2)在干旱或GA处理下,提高植物总根长;(3)在干旱或GA处理下,提高植物存活率;(4)在干旱或GA处理下,提高植物下胚轴长度;(5)在干旱或GA处理下,提高植物干旱胁迫相关基因的表达量;所述干旱胁迫相关基因具体为RAB18和/或COR47和/或COR15A和/或KIN1和/或KIN2和/或DREB2A;(6)在干旱或GA处理下,降低植物GA途径相关基因的表达量;所述GA途径相关基因具体为KAO1和/或KAO2和/或KS。所述干旱为PEG处理或控水处理;所述PEG处理的浓度具体为4%、6%、8%或10%。所述GA处理的浓度具体为10μM。
上述应用中,所述植物为单子叶植物或双子叶植物。所述双子叶植物具体可为拟南芥;所述拟南芥具体可为拟南芥(哥伦比亚生态型col-0)。
为解决上述技术问题,本发明最后提供了一种培育耐逆性提高的转基因植物的方法。
本发明提供的培育耐逆性提高的转基因植物的方法包括提高受体植物中TaCDPK1蛋白质的表达量和/或活性,得到转基因植物的步骤;所述转基因植物的耐逆性高于所述受体植物。
上述方法中,所述提高受体植物中TaCDPK1蛋白质的表达量和/或活性的方法为在受体植物中过表达TaCDPK1蛋白质。
上述方法中,所述过表达的方法为将TaCDPK1蛋白质的编码基因导入受体植物;所述TaCDPK1蛋白质的编码基因的核苷酸序列是序列1第280-1878位所示的DNA分子。
在本发明的一个实施方式中,TaCDPK1蛋白质的编码基因(即序列1第280-1878位所示的核苷酸)通过含有TaCDPK1蛋白质的编码基因的表达盒的重组载体pBI121-TaCDPK1导入受体植物中。所述重组载体pBI121-TaCDPK1为将pBI121载体的SmaI和SpeI酶切位点之间的DNA片段替换为序列表中序列1第280-1878位所示的TaCDPK1基因,且保持pBI121载体的其他序列不变后得到的载体。
上述方法中,所述耐逆性为耐旱性。所述转基因植物的耐逆性高于所述受体植物体现在如下(1)-(6)中任一种:(1)转基因植物的发芽率高于受体植物;(2)转基因植物的总根长长于受体植物;(3)转基因植物的存活率高于受体植物;(4)转基因植物的下胚轴长度长于受体植物;(5)转基因植物的干旱胁迫相关基因表达量高于受体植物;所述干旱胁迫相关基因具体为RAB18和/或COR47和/或COR15A和/或KIN1和/或KIN2和/或DREB2A;(6)转基因植物的GA途径相关基因表达量低于受体植物;所述GA途径相关基因具体为KAO1和/或KAO2和/或KS。
上述方法中,所述转基因植物理解为不仅包含将所述TaCDPK1基因转化受体植物得到的第一代转基因植物,也包括其子代。对于转基因植物,可以在该物种中繁殖该基因,也可用常规育种技术将该基因转移进入相同物种的其它品种,特别包括商业品种中。所述转基因植物包括种子、愈伤组织、完整植株和细胞。
上述方法中,所述植物为单子叶植物或双子叶植物。所述双子叶植物具体可为拟南芥;所述拟南芥具体可为拟南芥(哥伦比亚生态型col-0)。
通过实验证明:本发明发现的TaCDPK1基因在干旱、GA或BR的诱导下表达,且将TaCDPK1基因导入拟南芥中得到的转基因拟南芥,其对干旱逆境的抗性高于野生型拟南芥并且参与GA信号转导途径。本发明提供的蛋白和基因为人为控制抗逆和耐逆相关基因的表达提供了基础,将在培育抗逆性和耐逆性增强的植物中发挥重要的作用。
附图说明
图1为TaCDPK1的表达模式。
图2为转TaCDPK1拟南芥PEG处理下的萌发实验和根长实验。
图3为转TaCDPK1拟南芥干旱处理下的存活率。
图4为转TaCDPK1拟南芥干旱处理下的干旱相关基因的表达量。
图5为转TaCDPK1拟南芥参与GA信号通路。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明,均为常规方法。
下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
下述实施例中的定量试验,均设置三次重复实验,结果取平均值。
下述实施例中的%,如无特殊说明,均为质量百分含量。
下述实施例中的普通小麦(Triticum aestivum L.)品种小白麦记载于文献“孙海桃等,小麦TaDREB6转录因子互作蛋白的筛选,中国农业科学,2011,44(22):4740-4747.”中,公众可从中国农业科学院作物科学研究所获得,该生物材料只为重复本发明的相关实验所用,不可作为其它用途使用;公众也可从国家种质资源库获得(编号为ZM242)。
实施例1、TaCDPK1的克隆
一、植物材料的处理
将水培生长10天左右的小白麦(Triticum aestivum cv.Xiaobaimai)三叶期整株幼苗用浓度为15%的PEG6000溶液浸泡处理2小时,并将处理后的幼苗用液氮速冻,-80℃保存备用。
二、总RNA的提取
采用Trizol法(TianGen)提取步骤一获得的处理后的小白麦幼苗叶片的总RNA。
三、cDNA的获得
第一链cDNA合成用反转录酶XL(AMV)。采用SMART法合成ds cDNA,PCR产物进行1.0%琼脂糖凝胶电泳检测。通过5’RACE和3’RACE的方法获得序列表中的序列1。上述序列1也可以通过人工合成的方式获得。
将序列表中序列1所示的基因命名为TaCDPK1基因,其开放阅读框架为自序列表的序列1的5′端第280-1878位核苷酸,将该基因编码的蛋白命名为TaCDPK1蛋白,该蛋白的氨基酸序列为序列表中的序列2,由532个氨基酸残基组成。
实施例2、TaCDPK1的表达模式
一、植物材料的处理
将水培生长10天左右的小白麦(Triticum aestivum cv.Xiaobaimai)三叶期整株幼苗用浓度为100μM的GA3和10μM的BR水溶液浸泡处理,分别在处理0、0.5、1、2、4、8、12、24h时间点取样(小白麦叶片),并将处理后的样品用液氮速冻,-80℃保存备用。另外,将同样水培生长10天左右的小白麦取出,放在滤纸上自然干旱,分别在处理0、0.5、1、2、4、8、12、24h时间点取样(小白麦叶片),并将处理后的样品用液氮速冻,-80℃保存备用。
二、总RNA的提取
采用Trizol法(TianGen)提取步骤一获得的处理后的小白麦叶片的总RNA。
三、cDNA的获得
样品采用Trizol法(TianGen)提取叶片总RNA,然后用RT/RIEnzyme Mix(全式金)合成第一链cDNA。
四、TaCDPK1的表达模式
以拟南芥的Actin作为内参,在7500型号的仪器(Bio-Rad)上进行实时定量PCR(qRT-PCR),然后在excel表中计算出每个基因的相对表达量。结果如图1所示。结果表明:TaCDPK1基因在干旱、GA或BR的诱导下表达。
实施例3、转TaCDPK1拟南芥的获得及其耐逆性分析
一、转TaCDPK1拟南芥的获得
1、重组表达载体的构建
(1)TaCDPK1基因的克隆
提取小白麦叶片总RNA,反转录得到cDNA,以cDNA模板,采用引物对TaCDPK1-121F和TaCDPK1-121R进行扩增,得到PCR产物。引物序列如下(引物末端分别引入Sma I和Spe I酶切识别位点,如下划线序列所示):
TaCDPK1-121F:5'-TTACCCGGGATGGGCCAGTGTTGC-3'(序列3);
TaCDPK1-121R:5'-AGAACTAGTCTTGAGCCTTATTGG-3(序列4)。
将PCR产物进行1.2%琼脂糖凝胶电泳,其大小为1.6kb。并将该PCR产物进行测序,结果表明该PCR产物具有序列表中序列1,即为TaCDPK1基因。
采用Agarose Gel DNA Purification Kit Ver.2.0(TaKaRa公司,Code No.:DV807A)回收纯化大小为1.6kb的PCR产物。
(2)用限制性内切酶Sma I和Spe I酶切步骤(1)回收纯化的PCR产物,回收酶切产物;用限制性内切酶Sma I和Spe I酶切pBI121载体(Clontech公司),回收载体骨架;将酶切产物和载体骨架连接,得到连接产物。
(3)将步骤(2)获得的连接产物热击转化TOP10菌株(Tiangen,CB104-03),37℃过夜培养,挑取阳性克隆提取质粒进行测序。
测序结果表明,该质粒为将pBI121载体的SmaI和SpeI酶切位点之间的DNA片段替换为序列表中序列1第280-1878位所示的TaCDPK1基因,且保持pBI121载体的其他序列不变后得到的载体,并将其命名为pBI121-TaCDPK1。
2、重组菌的构建
(1)将步骤1获得的重组质粒pBI121-TaCDPK1转化农杆菌C58(购自北京拜尔迪生物技术公司),得到重组农杆菌。
(2)提取重组农杆菌的质粒送去测序,结果表明该质粒为pBI121-TaCDPK1,证明该重组菌为阳性重组农杆菌,并将其命名为C58/pBI121-TaCDPK1。
3、转TaCDPK1拟南芥的获得
(1)将重组农杆菌C58/pBI121-TaCDPK1接种于YEP液体培养基中,28℃、3000rpm培养约30小时。
(2)将步骤(1)获得的菌液转至YEP液体培养基(含50μg/ml利福平)中,28℃、300rpm培养约14小时(菌液OD600达到1.5-3.0)。
(3)收集菌体,4℃、4000g离心10min,用10%蔗糖(含0.02%silwet)稀释至OD600约为1.0。
(4)将拟南芥哥伦比亚生态型Col-0(美国拟南芥信息资源网,以下简称为野生型拟南芥,购自SALK公司)整株与花盆一起倒扣在盛有步骤(3)获得的菌液的容器中,使花浸泡50s左右,浸泡完毕后,取出花盆,侧放于托盘中,盖上黑色塑料布,24hr后揭开塑料布,直立放置花盆,进行正常的光照培养,混收T0代转TaCDPK1拟南芥种子。将T0代转TaCDPK1拟南芥种子播种到含有50mg/ml卡那霉素的MS培养基上培养,得到7株T1代转TaCDPK1拟南芥幼苗。
采用同样的方法将空载体pBI121转入野生型拟南芥,得到T1代转空载体拟南芥。
分别将T1代转TaCDPK1拟南芥和T1代转空载体拟南芥播种、自交,直到得到T3代转TaCDPK1拟南芥和T3代转空载体拟南芥。
提取T3代转TaCDPK1拟南芥植株的DNA,以TaCDPK1-121F和TaCDPK1-121R为引物进行PCR检测,得到条带为1.6kb的阳性T3代转TaCDPK1拟南芥植株。选取T3代转TaCDPK1拟南芥纯合株系OE-1、OE-2和OE-3用于下一步的耐逆性分析。
二、转TaCDPK1拟南芥的耐逆性分析
1、PEG处理的种子萌发实验
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种在含有不同浓度PEG(0%、4%、6%和10%)的1/2MS培养基上,经过3天的春化处理后在正常条件下进行培养,连续观察培养12、24、36、48、60、72小时后的各植株的萌发情况并统计发芽率。每个株系30颗,实验重复三次,结果取平均值。
结果如图2A和图2B所示,从图中可以看出,编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子和野生型拟南芥种子(WT)在4%、6%和10%PEG处理下,转TaCDPK1拟南芥株系均表现一定的优势(图2A),且在PEG处理后的36小时内,转TaCDPK1拟南芥的发芽率也高于野生型拟南芥(图2B)。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
2、PEG处理的根长实验
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种在1/2MS培养基上,经过3天的春化处理后在正常条件下进行培养,正常生长6天后移至含有不同浓度PEG(0%、6%和8%)的1/2MS培养基上生长7天,观察生长7天后各植株的根部生长情况并统计总根长。每个株系30颗,实验重复三次,结果取平均值。
结果如图2C和图2D所示,从图中可以看出,编号为OE-1、OE-2和OE-3的T3代转TaCDPK1拟南芥在6%和8%PEG的处理下总根长均长于野生型拟南芥(WT)。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
3、干旱处理下的存活率实验
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种在1/2MS培养基上,经过3天的春化处理后在正常条件下进行培养,培养一周后移至培养盆(蛭石:营养土为1:1)中,在16/8h的光照/黑暗培养室(22℃)培养,2周后开始控水(自然干旱),控水两周后复水三天。干旱处理后观察各植株的生长状态并统计复水后存活率。每个株系30颗,实验重复三次,结果取平均值。
结果如图3所示,从图中可以看出,编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)的抗旱性明显优于野生型拟南芥(图3A),且存活率也明显高于野生型拟南芥(图3B)。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
4、干旱处理下的干旱胁迫相关基因的表达量
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种在1/2MS培养基上,经过3天的春化处理后在正常的光照培养条件下生长两周,然后取出各个株系,放在滤纸上干旱处理一个小时后取样。提取各株系样品的RNA,反转录获得cDNA,检测干旱胁迫相关基因RAB18、COR47、COR15A、KIN1、KIN2和DREB2A的表达量。检测引物如下:
qAtRAB18+F1:AGCTCGGAGGATGATGGACA;
qAtRAB18+R1:AGCCACCAGCATCATATCCG;
qAtCOR47+F1:AGGATTCACCAGCTGTCACG;
qAtCOR47+R1:CCTCTTCAGTGGTCTTGGCA;
qAtCOR15A+F1:GCGATGTCTTTCTCAGGAGC;
qAtCOR15A+R1:CGAACTGAGTTTTCTGGCCG;
qAtKIN2+F1:CAAGAATGCCTTCCAAGCC;
qAtKIN2+R1:TTAACACCTCCCACTGCCG;
qAtKIN1+F1:CTGGAGCTGGAGCACAACAG;
qAtKIN1+R1:TTGTTCAGGCCGGTCTTGTC;
qAtDREB2A+F1:GGTTGGCCCAATGATGTGGA;
qAtDREB2A+R1:CCATCCTTTCCCTCGAGCTG;
qAtActin2+F1:AAGTCTTGTTCCAGCCCTCG;
qAtActin2+R1:TCTGATCAATTTTTACCTGCTGGA。
结果如图4所示,从图中可以看出,编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)的干旱胁迫相关基因的表达量均明显高于野生型拟南芥。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
三、转TaCDPK1拟南芥的GA途径分析
1、GA处理下的萌发率实验
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种于如下培养基1-3中,经过3天的春化处理后在正常条件下进行培养,连续观察培养12、24、36、48、60、72和84小时后的萌发情况。
培养基1:1/2MS培养基;
培养基2:含有PAC(多效挫)的1/2MS培养基,PAC在培养基2中的浓度为2μM;
培养基3:含有PAC(多效挫)和GA3的1/2MS培养基,PAC在培养基3中的浓度为0.5μM,GA3在培养基2中的浓度为10μM。
结果如图5A所示,从图中可以看出:在2μM PAC处理下,T3代转TaCDPK1拟南芥株系的发芽率明显高于野生型;在0.5μM PAC+10μM GA3处理下,T3代转TaCDPK1拟南芥株系的萌发早于野生型,在培养36小时后就几乎全部萌发,但是最后的萌发率没有差异。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
2、GA处理下的下胚轴实验
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种于上述步骤1中的培养基3中,经过3天的春化处理后在黑暗条件下进行培养,培养7天后分别检测下胚轴长度。
结果如图5B所示,从图中可以看出:在0.5μM PAC+10μM GA3处理下,T3代转TaCDPK1拟南芥株系的下胚轴的长度显著长于野生型(OE-2的下胚轴也长于野生型,但未达到显著水平)。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
3、GA处理下的GA途径相关基因表达量检测
分别将编号为OE-1、OE-2和OE-3的阳性T3代转TaCDPK1拟南芥(TaCDPK1)种子、T3代转空载体拟南芥和野生型拟南芥种子(WT)播种于1/2MS培养基中,经过3天的春化处理后在正常的光照培养条件下生长两周,然后取出各个株系,用GA3处理一个小时后取样。提取各株系样品的RNA,反转录获得cDNA,检测GA途径相关基因KAO1、KAO2、GA20OX1、GA30OX1和KS的表达量。引物序列如下:
qAtActin2+F1:AAGTCTTGTTCCAGCCCTCG;
qAtActin2+R1:TCTGATCAATTTTTACCTGCTGGA;
qAtKAO1+F:CTGACTCCTTCACTCGC;
qAtKAO1+R:CCTGAGACGCTTGTGTT;
qAtKAO2+F:TCCATTTGGACCCTGAAATC;
qAtKAO2+R:TGTGAGGCAAGAACATCACC;
qAtGA20OX1+F:CAGACCAAGTAAACTCCTCGTA;
qAtGA20OX1+R:CACTATCCACCATGTCCTCTTA;
qAtGA30OX1+F:CCATTCACCTCCCACACTCT;
qAtGA30OX1+R:GCCAGTGATGGTGAAACCTT;
qAtKS+F:AACGGAGATTGGACTCAGAA;
qAtKS+R:CATGGTTATCAAGTCCCCAAG。
结果如图5C所示,从图中可以看出:与野生型相比,在正常条件下拟南芥中过表达TaCDPK1基因会抑制GA途径相关基因(KAO1、KAO2和KS)表达,但是在经过GA处理后,其抑制程度更加明显(如KS基因)。在正常条件和GA处理条件下,野生型和过表达株系中GA20OX1和GA3OX1基因的表达量没有差异。
T3代转空载体拟南芥和野生型拟南芥的结果无显著差异。
序列表
<110>中国农业科学院作物科学研究所
<120>TaCDPK1蛋白质及其编码基因在调控植物耐逆性中的应用
<160>4
<170>PatentIn version 3.5
<210>1
<211>1960
<212>DNA
<213>小麦(Triticum aestivum L.)
<400>1
gtgtgtatta gtcgctctca cgattggggg cgacgctaat cgcggtgcca ccacttccaa 60
tccactctcg cggtcgccgt gccgagctgt tattgctctg tcttcaacta ctccaagatt 120
tgttctttag acgcctgcag cttcggtgcg cagggacggc ggagagctcg ggacttggtt 180
cagagcttag gatctttgtc gattcaggtt tcgattggtt tcgtctgctc ggctgactgg 240
gttgtaaatt tggaactgaa aggaagaatt tgagcacaga tgggccagtg ttgcagcaga 300
gctacgtccc cggattctgt ccaaggagga gccaatggct atggctattc tcaccagcca 360
aaacaagcac aagcacctcc tagttacaac aatgctcagc caccaccaca agctgaggtg 420
aggtacacac cgccagcgat gaaccctcct gtagtcccac ctgtggttgc cccctcaaag 480
cccatgccgg acacgattct cggcaagcag tacgaggatg tgcgctctgt ctactccctt 540
ggcaaggaac ttggtagggg gcagttcggg gtgacgtacc tttgcactga gattagcact 600
ggcaggcagt atgcttgcaa gtccatatcc aagcgcaagc tcgtgagtaa ggctgacaag 660
gaggatatcc gcagggagat ccagatcatg cagcacctat ctgggcagcc aaacatagtg 720
gagttctgtg gagcatatga ggacaagggc agcgtgcatg tcgtgatgga gctctgtgca 780
ggcggggagc tgtttgatcg gattattgcc aaggggcact actcagagcg tgcagctgct 840
acaatctgca gaggagttgt gaatgttgtc aatgtttgcc atttcatggg agtgatgcac 900
cgcgatctga agccggagaa cttcttgctt gcgaccaagg atgagaatgc agtgctcaag 960
gccactgatt tcgggctttc ggtcttcatt gaagaaggaa aaatgtatag ggacatcgtt 1020
ggaagtgctt attatgttgc tcctgaagtc cttaggagaa actatggtaa agagatagat 1080
gtttggagtg caggtgttat tctgtacatt cttctcagcg gtgttcctcc attctgggct 1140
gaaactgaga agggaatatt tgatgctatt cttcaagggg agattgactt tgaaagtcag 1200
ccatggcctt caatttctga gagcgctaaa gaccttgtga ggaagatgtt ggcacaggat 1260
ccaaagaaaa gaattagttc agcccaagtt cttcaacatc catggctcag ggaaggagaa 1320
gcatcagata aacctattga cagtgctgtt ctttctagga tgaagcaatt cagagcaatg 1380
aataaactga aaaagatggc tctaaaggtt atagcttcaa atcttaatga ggaagagatc 1440
aagggcctga agcaaatgtt tatgaacatg gacacagaca acagtgggac aatcacatat 1500
gaggaactca aagcaggact agccaaactt ggatcgaagc tctcagaagc tgaagtaaag 1560
cagttgatgg atgcggctga tgtggatggc aatggatcaa ttgactacgt tgagttcata 1620
acagcaacca tgcatagaca caagctcgaa agagacgagc atttgttcaa ggcgttccag 1680
tactttgaca aagacaacag tggcttcatc acaagagatg aactggagac tgctttgatt 1740
gagcatgaaa tgggcgatgc ggataccata aaggacatca tatcagaagt cgacacagat 1800
aatgatggga ggattaacta cgaggaattc tgcgctatga tgagaggagg gatgcagcag 1860
ccaataaggc tcaagtagtc tttccttaga aatggtgtac cattagcaac tgttcttggt 1920
tccttttttt tcgctgctgc acggatgaga agttctcttt 1960
<210>2
<211>532
<212>PRT
<213>小麦(Triticum aestivum L.)
<400>2
Met Gly Gln Cys Cys Ser Arg Ala Thr Ser Pro Asp Ser Val Gln Gly
1 5 10 15
Gly Ala Asn Gly Tyr Gly Tyr Ser His Gln Pro Lys Gln Ala Gln Ala
20 25 30
Pro Pro Ser Tyr Asn Asn Ala Gln Pro Pro Pro Gln Ala Glu Val Arg
35 40 45
Tyr Thr Pro Pro Ala Met Asn Pro Pro Val Val Pro Pro Val Val Ala
50 55 60
Pro Ser Lys Pro Met Pro Asp Thr Ile Leu Gly Lys Gln Tyr Glu Asp
65 70 75 80
Val Arg Ser Val Tyr Ser Leu Gly Lys Glu Leu Gly Arg Gly Gln Phe
85 90 95
Gly Val Thr Tyr Leu Cys Thr Glu Ile Ser Thr Gly Arg Gln Tyr Ala
100 105 110
Cys Lys Ser Ile Ser Lys Arg Lys Leu Val Ser Lys Ala Asp Lys Glu
115 120 125
Asp Ile Arg Arg Glu Ile Gln Ile Met Gln His Leu Ser Gly Gln Pro
130 135 140
Asn Ile Val Glu Phe Cys Gly Ala Tyr Glu Asp Lys Gly Ser Val His
145 150 155 160
Val Val Met Glu Leu Cys Ala Gly Gly Glu Leu Phe Asp Arg Ile Ile
165 170 175
Ala Lys Gly His Tyr Ser Glu Arg Ala Ala Ala Thr Ile Cys Arg Gly
180 185 190
Val Val Asn Val Val Asn Val Cys His Phe Met Gly Val Met His Arg
195 200 205
Asp Leu Lys Pro Glu Asn Phe Leu Leu Ala Thr Lys Asp Glu Asn Ala
210 215 220
Val Leu Lys Ala Thr Asp Phe Gly Leu Ser Val Phe Ile Glu Glu Gly
225 230 235 240
Lys Met Tyr Arg Asp Ile Val Gly Ser Ala Tyr Tyr Val Ala Pro Glu
245 250 255
Val Leu Arg Arg Asn Tyr Gly Lys Glu Ile Asp Val Trp Ser Ala Gly
260 265 270
Val Ile Leu Tyr Ile Leu Leu Ser Gly Val Pro Pro Phe Trp Ala Glu
275 280 285
Thr Glu Lys Gly Ile Phe Asp Ala Ile Leu Gln Gly Glu Ile Asp Phe
290 295 300
Glu Ser Gln Pro Trp Pro Ser Ile Ser Glu Ser Ala Lys Asp Leu Val
305 310 315 320
Arg Lys Met Leu Ala Gln Asp Pro Lys Lys Arg Ile Ser Ser Ala Gln
325 330 335
Val Leu Gln His Pro Trp Leu Arg Glu Gly Glu Ala Ser Asp Lys Pro
340 345 350
Ile Asp Ser Ala Val Leu Ser Arg Met Lys Gln Phe Arg Ala Met Asn
355 360 365
Lys Leu Lys Lys Met Ala Leu Lys Val Ile Ala Ser Asn Leu Asn Glu
370 375 380
Glu Glu Ile Lys Gly Leu Lys Gln Met Phe Met Asn Met Asp Thr Asp
385 390 395 400
Asn Ser Gly Thr Ile Thr Tyr Glu Glu Leu Lys Ala Gly Leu Ala Lys
405 410 415
Leu Gly Ser Lys Leu Ser Glu Ala Glu Val Lys Gln Leu Met Asp Ala
420 425 430
Ala Asp Val Asp Gly Asn Gly Ser Ile Asp Tyr Val Glu Phe Ile Thr
435 440 445
Ala Thr Met His Arg His Lys Leu Glu Arg Asp Glu His Leu Phe Lys
450 455 460
Ala Phe Gln Tyr Phe Asp Lys Asp Asn Ser Gly Phe Ile Thr Arg Asp
465 470 475 480
Glu Leu Glu Thr Ala Leu Ile Glu His Glu Met Gly Asp Ala Asp Thr
485 490 495
Ile Lys Asp Ile Ile Ser Glu Val Asp Thr Asp Asn Asp Gly Arg Ile
500 505 510
Asn Tyr Glu Glu Phe Cys Ala Met Met Arg Gly Gly Met Gln Gln Pro
515 520 525
Ile Arg Leu Lys
530
<210>3
<211>24
<212>DNA
<213>人工序列(Artificial Sequence)
<400>3
ttacccggga tgggccagtg ttgc 24
<210>4
<211>24
<212>DNA
<213>人工序列(Artificial Sequence)
<400>4
agaactagtc ttgagcctta ttgg 24
Claims (10)
1.TaCDPK1蛋白质在如下任一中的应用:
1)调控植物耐逆性;
2)培育耐逆性提高的转基因植物;
3)参与GA信号转导途径;
4)植物育种;
所述TaCDPK1蛋白质是如下a)或b)或c)或d)的蛋白质:
a)氨基酸序列是序列2所示的蛋白质;
b)在序列2所示的蛋白质的N端和/或C端连接标签得到的融合蛋白质;
c)将序列2所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的具有相同功能的蛋白质;
d)与序列2所示的氨基酸序列具有75%或75%以上的同源性且具有相同功能的蛋白质。
2.与权利要求1中所述的TaCDPK1蛋白质相关的生物材料在如下任一中的应用:
1)调控植物耐逆性;
2)培育耐逆性提高的转基因植物;
3)参与GA信号转导途径;
4)植物育种;
所述生物材料为下述A1)至A12)中的任一种:
A1)编码TaCDPK1蛋白质的核酸分子;
A2)含有A1)所述核酸分子的表达盒;
A3)含有A1)所述核酸分子的重组载体;
A4)含有A2)所述表达盒的重组载体;
A5)含有A1)所述核酸分子的重组微生物;
A6)含有A2)所述表达盒的重组微生物;
A7)含有A3)所述重组载体的重组微生物;
A8)含有A4)所述重组载体的重组微生物;
A9)含有A1)所述核酸分子的转基因植物细胞系;
A10)含有A2)所述表达盒的转基因植物细胞系;
A11)含有A3)所述重组载体的转基因植物细胞系;
A12)含有A4)所述重组载体的转基因植物细胞系。
3.根据权利要求2所述的应用,其特征在于:A1)所述核酸分子为如下1)或2)或3)所示的基因:
1)其编码序列是序列1第280-1878位所示的cDNA分子或基因组DNA分子;
2)与1)限定的核苷酸序列具有75%或75%以上同一性,且编码权利要求1中所述的TaCDPK1蛋白质的cDNA分子或基因组DNA分子;
3)在严格条件下与1)或2)限定的核苷酸序列杂交,且编码权利要求1中所述的TaCDPK1蛋白质的cDNA分子或基因组DNA分子。
4.根据权利要求1-3任一所述的应用,其特征在于:所述耐逆性为耐旱性。
5.一种培育耐逆性提高的转基因植物的方法,包括提高受体植物中权利要求1中所述的TaCDPK1蛋白质的表达量和/或活性,得到转基因植物的步骤;所述转基因植物的耐逆性高于所述受体植物。
6.根据权利要求5所述的方法,其特征在于:所述耐逆性为耐旱性。
7.根据权利要求5或6所述的方法,其特征在于:所述转基因植物的耐逆性高于所述受体植物体现在如下(1)-(6)中任一种:
(1)转基因植物的发芽率高于受体植物;
(2)转基因植物的总根长长于受体植物;
(3)转基因植物的存活率高于受体植物;
(4)转基因植物的下胚轴长度长于受体植物;
(5)转基因植物的干旱胁迫相关基因表达量高于受体植物;
(6)转基因植物的GA途径相关基因表达量低于受体植物。
8.根据权利要求5-7任一所述的方法,其特征在于:所述提高受体植物中权利要求1中所述的TaCDPK1蛋白质的表达量和/或活性的方法为在受体植物中过表达权利要求1中所述的TaCDPK1蛋白质;
或,所述过表达的方法为将权利要求1中所述的TaCDPK1蛋白质的编码基因导入受体植物。
9.根据权利要求5-8任一所述的方法,其特征在于:所述TaCDPK1蛋白质的编码基因的核苷酸序列是序列1第280-1878位所示的DNA分子。
10.根据权利要求1-4任一所述的应用或权利要求5-9任一所述的方法,其特征在于:所述植物为单子叶植物或双子叶植物。
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103509766A (zh) * | 2012-06-15 | 2014-01-15 | 中国农业科学院作物科学研究所 | 植物耐逆性相关蛋白TaPK及其编码基因和应用 |
CN105821016A (zh) * | 2016-04-21 | 2016-08-03 | 南京农业大学 | 一种大豆cdpk类激酶及其编码基因与应用 |
-
2018
- 2018-06-27 CN CN201810677416.5A patent/CN110643588A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103509766A (zh) * | 2012-06-15 | 2014-01-15 | 中国农业科学院作物科学研究所 | 植物耐逆性相关蛋白TaPK及其编码基因和应用 |
CN105821016A (zh) * | 2016-04-21 | 2016-08-03 | 南京农业大学 | 一种大豆cdpk类激酶及其编码基因与应用 |
Non-Patent Citations (3)
Title |
---|
KUI HUANG ET AL.: "Arabidopsis calcium-dependent protein kinase AtCPK1 plays a positive role in salt/drought-stress response", 《BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS》 * |
SHIN-LON HO ET AL.: "Sugar starvation-and GA-inducible calcium-dependent protein kinase 1 feedback regulates GA biosynthesis and activates a 14-3-3 protein to confer drought tolerance in rice seedlings", 《PLANT MOL BIOL》 * |
XU ZS ET AL.: "Accession No:DQ333374.1", 《GENBANK》 * |
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