CN114934055A - 茶树CsAMT1.3基因在调控植物氮代谢中的应用 - Google Patents
茶树CsAMT1.3基因在调控植物氮代谢中的应用 Download PDFInfo
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- CN114934055A CN114934055A CN202210738575.8A CN202210738575A CN114934055A CN 114934055 A CN114934055 A CN 114934055A CN 202210738575 A CN202210738575 A CN 202210738575A CN 114934055 A CN114934055 A CN 114934055A
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Abstract
本发明公开了茶树CsAMT1.3基因在调控植物氮代谢中的应用,属于植物学技术领域。所述CsAMT1.3基因的编码序列如SEQ ID NO.1所示,所述氮代谢包括对铵态氮的吸收和同化。本发明首次从茶树‘龙井43’品种中鉴定并克隆了CsAMT1.3基因,茶树中CsAMT1.3主要在叶中表达,其生物学功能为促进铵态氮的吸收和同化,加快碳氮代谢。该基因在优质茶树品种培育方面具有潜在的应用价值。
Description
技术领域
本发明涉及植物学技术领域,具体涉及茶树CsAMT1.3基因在调控植物氮代谢中的应用。
背景技术
茶是世界三大饮料作物之一,富含丰富的代谢产物,对人体健康有重要作用。氮代谢是植物体内重要的物质和能量代谢过程,该代谢途径能够影响茶树的代谢产物。土壤基质中的氮素主要以无机态氮和有机态氮两种形式存在,一般从土壤中吸收的主要是无机态氮,即铵盐(NH4 +)和硝酸盐(NO3 -)。铵离子是植物根系吸收的最为经济有效的一种氮素形式,虽然土壤中硝态氮比铵态氮含量高,但很多植物会优先选择同化耗能较低的铵态氮。
茶树作为一种喜铵性作物,以叶片为收获目标,不仅要经历多次采摘,还需要进行定期修剪,这些都会带走大量的氮素,然而茶叶本身并不能固氮,因此,茶树栽培过程中需要大量的氮素供应。
NH4 +的吸收是逆浓度梯度需能量的主动转运过程,植物根系主要通过位于质膜上的铵转运蛋白AMT吸收铵态氮。AMT属于AMT/Rh类蛋白,广泛存在于细菌、真菌和动植物中,AMT分为两个亚类AMT1和AMT2。目前,编码铵转运蛋白的基因在很多植物中得到了克隆与鉴定,每个基因的吸收特征不同,调控机制也存在差异。
目前,关于铵转运蛋白的AMT的研究对象主要集中在拟南芥和水稻等模式作物中。拟南芥AtAMT1.3主要在根毛、表皮和皮层细胞表达(Loque,D.,Yuan,L.,Kojima,S.,etal.Additive contribution of AMT1;1and AMT1;3to high-affinity ammonium uptakeacross the plasma membrane of nitrogen-deficient Arabidopsis roots[J].PlantJ,2006,48(4):522-534.),起到从土壤中吸收铵的作用。Lima等人研究证明了拟南芥AtAMT1.3能够起到信号转导的作用,能够响应外部NH4 +,并调节侧根分枝(Lima,J.E.,Kojima,S.,Takahashi,H.,et al.Ammonium triggers lateral root branching inArabidopsis in an AMMONIUM TRANSPORTER1;3-dependent manner[J].Plant Cell,2010,22(11):3621-3633.)。水稻OsAMT1.3主要在根尖和侧根表达(Ferreira,L.M.,deSouza,V.M.,Tavares,O.C.H.,et al.OsAMT1.3expression alters rice ammoniumuptake kinetics and root morphology[J].Plant Biotechnology Reports,2015,9(4):221-229.),在缺氮条件下表达量上升(Sonoda,Y.,Ikeda,A.,Saiki,S.,et al.Distinctexpression and function of three ammonium transporter genes(OsAMT1;1-1;3)inrice[J].Plant Cell Physiol,2003,44(7):726-734.),说明其可能与铵的吸收相关。
茶树中的NH4 +转运蛋白相关基因研究尚属起步阶段,主要集中在对茶树CsAMT1.1和CsAMT1.2克隆方面,但具体功能未有详细报道。本发明首次克隆了茶树CsAMT1.3基因并对其功能进行了鉴定,其在调控茶树氮素代谢方面的功能未见报道。
发明内容
本发明的目的在于挖掘对茶树氮素代谢具有调控作用的基因,将其应用到茶树育种当中以提高茶树氮素利用效率,改良茶叶品质。
为实现上述目的,本发明采用如下技术方案:
本发明提供了茶树CsAMT1.3基因在调控植物氮代谢中的应用,所述CsAMT1.3基因的编码序列如SEQ ID NO.1所示,所述氮代谢包括对铵态氮的吸收和同化。
本发明从茶树龙井43品种幼苗新梢中鉴定并克隆了CsAMT1.3基因,该基因编码蛋白的氨基酸序列如SEQ ID NO.2所示。
CsAMT1.3基因在茶树中的表达模式分析显示,该基因主要在叶中表达。通过不同氮素处理验证了不同氮素条件下CsAMT1.3的表达差异,得出NH4 +的供应更有利于CsAMT1.3的表达。
本发明通过酵母功能互补试验发现,茶树CsAMT1.3蛋白具有吸收铵的能力。通过构建茶树CsAMT1.3过表达的拟南芥纯合子,发现在不同铵浓度下,相较于野生型植株,转基因拟南芥植株的生长势更好,侧根更发达。同时,该基因能够使碳氮代谢关键基因表达上调,加速碳氮代谢,促进铵的同化,合成叶绿素和氨基酸。
进一步的,所述应用包括:利用生物学技术手段过表达所述CsAMT1.3基因,获得促进氮代谢的转基因植株。
进一步的,所述CsAMT1.3基因过表达,促进植物对铵态氮的吸收。
进一步的,所述CsAMT1.3基因过表达,转基因植株根长增加,促进侧根分枝。
进一步的,所述转基因植株通过上调磷酸烯醇丙酮酸羧化酶编码基因(PEPC)和谷氨酸合成酶编码基因(GLT)的表达,加快碳氮代谢,促进铵态氮同化。所述铵态氮同化形成叶绿素、氨基酸。
进一步的,所述CsAMT1.3基因过表达,促进转基因植株合成叶绿素、氨基酸。
进一步的,所述植物为茶树、拟南芥。具体的,在茶树品种改良培育中,通过过表达CsAMT1.3基因,以促进茶叶对铵态氮的吸收以及同化,增加氨基酸合成,有利于茶叶品质改良。
本发明具备的有益效果:
本发明首次从茶树‘龙井43’品种中鉴定并克隆了CsAMT1.3基因,茶树中CsAMT1.3主要在叶中表达,其生物学功能主要为促进铵态氮的吸收和同化,加快碳氮代谢。该基因在优质茶树品种培育方面具有潜在的应用价值。
附图说明
图1为不同茶树组织的CsAMT1.3在不同氮源下的表达情况,其中(A)为一芽二叶,(B)为成熟叶,(C)为根。
图2为野生型(Col-0)株系与转基因株系在不同铵浓度下的生长表型。
图3为不同铵浓度下野生型植株(Col-0)和转基因植株叶片的生理指标,其中(A)为叶绿素含量,(B)为可溶性糖含量,(C)为游离氨基酸含量。
图4为正常铵浓度下野生型植株和转基因植株碳氮代谢相关基因表达热图。
图5为CsAMT1.3的酵母功能互补结果。
具体实施方式
下面结合具体实施例对本发明做进一步说明。以下实施例仅用于说明本发明,不用来限制本发明的适用范围。在不背离本发明精神和本质的情况下,对本发明方法、步骤或条件所做的修改或替换,均属于本发明的范围。
下述实施例中所使用的试验方法如无特殊说明,均为常规方法;所使用的材料、试剂等,如无特殊说明,为可从商业途径得到的试剂和材料。
实施例1茶树CsAMT1.3的发掘和鉴定
1、植物材料的样品的准备
取茶树(Camellia sinensis)龙井43品种一年生扦插苗,置于人工气候室中水培,采摘茶树幼苗的一芽二叶,液氮速冻后,-80℃冰箱冷冻备用。
2、茶树RNA的提取
利用RNAprep Pure多糖多酚植物总RNA提取试剂盒(离心柱型)目录号:DP441进行RNA提取。将获得的RNA样品,进行浓度、纯度和完整性检验。
所提取RNA OD260/OD280在1.8左右,电泳显示28S和18S条带清晰,无降解,表明提取RNA质量合格。
3、cDNA的合成
利用takera公司的PrimeScript II 1st Strand cDNA Synthesis Kit试剂盒进行cDNA的合成。
4、PCR扩增
上游引物F:5’-ATGGAGGCATCATGGGAAG-3’;
下游引物R:5’-TCATTGGTTTTGCAGCCTC-3’;
配置PCR反应体系(50μL),见表1:
表1
反应程序为:94℃预变性5min,94℃变性45s,55℃退火5s,72℃延伸2min,30个循环,72℃后延伸10min,4℃保存。反应结束后,取上述扩增产物50μL,加入3μL10×loadingbuffer混匀,上样于1.0%琼脂糖凝胶,进行电泳分析,与marker对比确定产物长度,凝胶成像仪上观察并拍照,PCR产物于-20℃保存。
5、目的片段回收
利用TaKaRa的TaKaRa MiniBEST Agarose Gel DNAExtraction Kit Ver4.0割胶回收试剂盒进行目的片段回收。
6、目的片段插入T载体
采用TaKaRa pMDTM18-T Vector Cloning Kit试剂盒进行目的片段与T载体的连接。于灭菌PCR管中,加入1.0μL pMDTM18-T Vector、加A尾后的目的片段4μL,再加入5μLSolution I,16℃反应过夜。
7、连接转化大肠杆菌
1)将上述连接液全部(10μL)加至100μL DH5α感受态细胞中,冰中放置30min。
2)42℃热激45s后,再在冰中放置2min。
3)加入890ml SOC液体培养液,37℃振荡60min。
4)4000rpm离心4min,弃去大部分上清液,用枪吹匀沉淀。
5)涂布于含有Amp的L-琼脂平板培养基上,37℃倒置培养12h,形成单菌落。
8、转化子扩增及验证
挑选平板培养基上大小适中、形状规则的白色单菌落,先涂于空白平板上,再置于菌液PCR体系中,利用Premix TaqTM(TaKaRaTaqTMVersion 2.0plus dye)进行菌液PCR。反应程序为:94℃预变性4min,94℃变性30s,55℃退火30s,72℃延伸2min,20个循环,72℃再延伸10min,4℃保存。取上述反应液,上样于1%琼脂糖凝胶,进行电泳分析,与Marker对比确定阳性克隆。
9、测序
经菌液PCR验证的阳性克隆,加入含有1ml Amp-LB营养液的离心管中,37℃振荡12h,委托上海生工公司测序。
通过该方法鉴定出茶树mRNA的序列如SEQ ID NO.1所示,其编码的蛋白序列如SEQID NO.2所示。
实施例2茶树CsAMT1.3在不同氮源下的表达
1、采用茶树‘龙井43’品种一年生扦插苗,预培养一个月后,将生长状况相似的茶苗移栽至无氮生长条件下1周后,恢复4mM不同形态氮素(NO3 -,NH4 +,NO3 -:NH4 +=1:1),取样后液氮速冻,保存于-80℃冰箱。
2、总RNA的提取
参照实施例1方法,提取茶树的总RNA,检测RNA的纯度及浓度。
3、qRT-PCR检测CsAMT1.3的表达
1)mRNA反转录合成cDNA
使用PrimeScriptTMRT reagent Kit with gDNA Eraser(Perfect Real Time)(Takara)试剂盒进行反转录。具体步骤如下,
a、去除基因组DNA。按下列组分在冰上配置反应液。反应体系(10μL)如下:5×gDNAEraser Buffer 2μL,gDNA Eraser 1μL,Total RNA 800ng,RNaseFree dH2O补充至10μL。42℃2min,4℃保存。
b、反转录反应。按下列组分在冰上配置反应液,轻柔混匀后立即进行反转录反应。反应体系(20μL)如下:10μL步骤a的反应液,1μLPrimeScript RT Enzyme Mix I,1μL RTPrimer Mix,4μL 5×PrimeScript Buffer 2,剩余用RNase Free dH2O补足。上述反应液反应条件:37℃15min,80℃5sec,4℃保存。
2)实时荧光定量PCR
采用TB Green Premix Ex Taq(TliRNaseH Plus)试剂盒(Takara)15μL体系,1.2μL cDNAs,上下游引物(20μM)各3μL,7.5μL TB Green,3μLROX Reference Dye。反应条件:94℃预变性5min,94℃变性30s,60℃退火30s,40个循环,2△△T法计算基因相对表达量。以CsGADPH为内参基因,引物序列参见表2:
表2
结果如图1所示,在一芽二叶和成熟叶中CsAMT1.3在仅供应NH4 +的处理下表达量显著高于其他三个处理,在成熟叶中,NH4 +对CsAMT1.3的影响最明显。而在根中CsAMT1.3对单一氮源(NH4 +/NO3 -)的响应更高,缺氮条件下响应最低。
实施例3过表达茶树CsAMT1.3对拟南芥碳氮代谢表达的影响
1.过表达载体的构建
设计同源重组引物:
F:5’-acgggggactcttgaccatggggATGGAGGCATCATGGGAAGC-3’;
R:5’-aactagtcagatctaccatggTCATTGGTTTTGCAGCCTC-3’。
以‘龙井43’cDNA为模板,扩增CsAMT1.3基因。按照诺唯赞公司IIOne Step Cloning Kit说明书,将纯化的目的片段和线性载体进行连接,连接的反应体系如下(20μL):线性化片段(~0.03pmol)、目的片段(~0.06pmol)、5×CE II Buffer 4μL、Exnase II 2μL,用ddH2O补足。37℃反应30min后,将连接产物转入DH5α大肠杆菌,进行重组产物转化,参照实施例1,涂布与含有卡那霉素抗性LB固体培养基,37℃培养12小时后挑取单克隆进行验证,得到过表达载体。
2.电击法转化农杆菌
1)将GV3101农杆菌感受态从-80℃冰箱取出插入冰中5分钟,待其融化,加入1μL构建好的质粒,用手拨打管底混匀,立即插入冰中,将感受态和质粒混合物快速移到预冷好的干净电击杯中,盖上杯盖,空管保留待用。
2)启动电转仪,将电击杯迅速放入电转槽(设置参数:C=25μF,PC=200ohm,V=2400V)中,电击完成插入冰中,加入600μL无抗生素的LB培养基并转移到空离心管中,28℃振荡培养2-3小时。
3)6000rpm离心5分钟收菌,留100μL左右上清轻轻吹打并重悬菌块,涂布于含利福平和卡那霉素的LB平板上,倒放于28℃培养箱培养2-3天。
4)挑取单菌落进行菌液PCR,检测阳性克隆。
3.拟南芥的转化及筛选
采用花序侵染法转化拟南芥,具体步骤如下:
1)将单克隆农杆菌接种到含有卡那霉素的LB液体培养基,28℃,220rpm摇菌至OD600到0.5-0.8左右,4000rpm离心后收集菌体,用100mL拟南芥转化缓冲液悬浮。配方如下:1/10MS,蔗糖10g/L,Silwet-L77 200μL。
2)选取生长状况良好的拟南芥,将其地上部花朵浸泡在农杆菌悬浮液中1分钟左右,取出后去除多余液体。用保鲜膜将整株拟南芥密封,保持高湿的环境。暗处理1天后,去除保鲜膜,给予正常光照和水分,直至种子成熟。
3)消毒拟南芥种子:将种子分装至1.5mL离心管,并加入1mL70%乙醇振荡10分钟。将乙醇吸收出后,用100%乙醇灭菌2次,再用灭菌水清洗3-5次,超净台中吹干待用;
4)将灭菌好的种子播种在含有潮霉素的抗性固体平板上密封好,4℃春化2天后,正常培养10天左右。固体培养基配方如下:MS 2.2g/L,MES0.5g/L,蔗糖10g/L,琼脂粉7g/L,调节pH 5.8。
5)将生长状况较好的,有较长根系和真叶的拟南芥阳性植株移栽到灭菌过的基质中。保鲜膜覆盖3天后,进行正常培养至结实;
6)按单株收获T1代种子,按上述方法继续筛选,单株获得T2代种子;
7)按单株筛选T2代种子,统计阳性与阴性植株数目,获得纯合系。
4.叶绿素的测定
称取拟南芥叶片0.05g,利用5mL无水乙醇/丙酮(1/1,v/v)混合浸提液进行浸提。浸提至无色后,利用分光光度计检测A663、A646的吸光度值,并计算叶绿素a、叶绿素b和总叶绿素的含量。
5.游离氨基酸的测定
取1mL浸提液于25mL容量瓶中,加入0.5mLpH 8.0磷酸缓缓冲液和0.5mL 2%茚三酮溶液。在沸水浴中加热15分钟后取出,冷却至室温后加水定容至25mL。放置10分钟后,在波长570nm处测定吸光值。
6.可溶性糖的测定
称取拟南芥0.05g,加入5mL蒸馏水,在沸水浴中浸提20分钟,12000rpm离心10分钟,取上清。取100μL浸提液和葡萄糖标准液加入400μLH2O和4mL蒽酮硫酸溶液,在沸水浴中加热7分钟。冷却至室温后,放置10分钟,在波长620nm处,以空白试剂作为参比液,测定吸光度。
7.实时荧光定量
提取拟南芥RNA、并进行反转录cDNA。参考实施例2。以拟南芥actin为内参,进行荧光定量分析。引物序列见表3:
表3
结果如图2所示,与野生型株系相比,过表达株系根的长度更长,侧根更多,生长状况更好,其中未添加铵(0mM NH4 +)的过表达植株生长状况较4、10mM NH4 +下的植株稍好。
如图3所示,在铵浓度为4mM时,过表达植株叶绿素的含量对比野生型植株显著增加。过表达植株对比野生型植株的可溶性糖含量在铵浓度为4mM时显著下降。而游离氨基酸的含量,在0、4、10mM的铵浓度下CsAMT1.3过表达植株都显著高于野生型,且游离氨基酸的含量随着铵浓度的增加而增加,说明CsAMT1.3可能促进了铵的同化和氨基酸的合成。
如图4所示,在4mM NH4 +的处理下,所有与碳氮代谢相关的基因都有不同水平的上升。其中上调最明显的是磷酸烯醇丙酮酸羧化酶基因PEPC和谷氨酸合成酶基因GLT。其中PEPC是调节植物碳氮代谢过程的关键基因,GLT则是铵同化过程中的限速酶,该基因可以反映铵同化的强弱,说明转基因植株能够加快植物的碳氮代谢,还能促进铵的同化。
实施例4 CsAMT1.3的酵母功能互补实验
1.酵母表达载体CsAMT1.3-pYES2的构建
根据pYES2载体序列和酶切位点Hind III,设计同源重组引物:
F:5’-actatagggaatattaagcttATGGAGGCATCATGGGAAGC-3’;
R:5’-tccgagctcggtaccaagcttTCATTGGTTTTGCAGCCTCAT-3’。
以‘龙井43’cDNA为模板,扩增CsAMT1.3基因。并参考实施例3,进行目的片段和线性载体的连接,并获得载体。
2.pYES2空载体和CsAMT1.3-pYES2载体转入缺陷型酵母
1)将质粒和鲑鱼精混合。
2)加入100μL酵母感受态细胞,振荡混匀。
3)加入600μL PEG/LiAc,用手剧烈振荡,30℃恒温,100rpm培养1小时。
4)加入70μL DMSO,缓缓倒置混匀。42℃水浴15分钟,然后迅速插入冰中冷却2分钟。
5)室温离心2500rpm,1分钟,弃掉上清。用0.5mL 1×TE重悬沉淀细胞。最后取100μL涂Arg平板。30度倒置培养3-4天。取单克隆进行验证。
3.酵母培养
将验证好的空载和CsAMT1.3的酵母单克隆在YNB液体培养基中30℃生长,待OD600达到0.6-0.8左右时收获菌体。用无菌水调整菌体OD600浓度达到2.0。然后按照1、10、100、1000的比例稀释菌体待用。各取5μL混匀的酵母悬浮液点于互补固体培养基上。以NH4Cl作为唯一氮源,30mmol/L NH4Cl为对照,pH 5.8,3天后观察并拍照酵母的生长状况。NH4Cl生长培养基配方见表4。
表4
酵母菌株31019b是一个被敲除了全部内源铵吸收系统的铵吸收缺失突变体,不能在低于5mmol/L铵态氮作为唯一氮源的培养基上生长。如图5所示,在以30mmol/L NH4Cl为唯一氮源的固体培养基上,转化了pYES2空载体和CsAMT1.3-pYES2的酵母菌株均能正常生长。但在以0.3和1mmol/L NH4Cl为唯一氮源的培养基上,转化了pYES2的空载体的不能正常生长,而CsAMT1.3-pYES2的酵母菌株则能正常生长。而且随着铵浓度的不断升高,CsAMT1.3-pYES2/31019b重组酵母生长得越好。说明CsAMT1.3具有吸收铵的功能,能够介导铵的吸收。
序列表
<110> 新昌中国大佛龙井研究院
<120> 茶树CsAMT1.3基因在调控植物氮代谢中的应用
<160> 28
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1389
<212> DNA
<213> 茶(Camellia sinensis)
<400> 1
atggaggcat catgggaagc cagcgtcacc gattcaatca acaccatata cctccttttc 60
tccgcctacc tagccttcgt aatgcagctc ggcttcgcca tgctctgtgc cggctctgtc 120
cgagccaaga acgccatgaa cataatgctc accaacgtcg ttgacgccgt tgtcggcagc 180
atctcctact acctcttcgg cttcgctttc gctttcggcg acggctccag ttccaaccca 240
ttcattggca ctgaattttt tgctctcaaa gacatccccg atagttcata tgactacagt 300
tactttctct accaatgggc tttcgctatc gctgtctctg gcatcactag cggctcgata 360
gccgagcgaa cccagttcag cgcctacctc gttttctcat tttttttaac ggggttcgtt 420
tacccggtgg tggttcactg ggtttggtca tcgagtggct ggcttagtcc gagctcaact 480
agtttgattt ttagttcggg tgcaattgac tttgctggga gtggagtggt gcatttagtg 540
ggtggaattg ctgggctttg gggctcattg atagaaggcc caagggtggg ccggttcgat 600
gcctttggca agcccgtgcc aatgcgaggc cacaatgcaa ctcttgtggt gctagggacc 660
ttcttgttgt ggtttgggtg gttcgggttc aatccaggtt cgttcgacaa aattcttgtt 720
gcgtaccctg acacatccaa tcaaggaaac tggaccggaa ttggccgaac agcggtcacg 780
accgcacttg ccggttcaac agccggactc gtaacactat tcggccgacg gttactagtg 840
ggccactggg atgcattaga tgtgtgcaat gggcttcttg gtgggtttgt cgcgatcaca 900
tcaggctgcg cagtggtgga accatgggct gccattgtat gcgggttttt ttcggcgtgg 960
gtattgattg ggctgaatat cttggccctg aaactgaaat tcgatgaccc cttagaggca 1020
acccaattac atggtgggtg tggggcttgg ggtctgcttt ttacaggcct gtttgcaaaa 1080
gaggaattca ttgttcaggc ctataattcg ggtgagactg gagtggttcg accctatggg 1140
cttttattgg gtgggggatg gggcctacta ggagcccaag tgattgaggt attggttatt 1200
gtgggctggg ttagtataac aatgggccca cttttctatg cactacacag gctcgagatt 1260
ttgaggatat ctgttgatga ggaagttgca gggcttgaca tctctagcca tggaggttat 1320
gcctatactg ctcatccaga ggacacccca cgctattatg ctgattacat gaggctgcaa 1380
aaccaatga 1389
<210> 2
<211> 462
<212> PRT
<213> 茶(Camellia sinensis)
<400> 2
Met Glu Ala Ser Trp Glu Ala Ser Val Thr Asp Ser Ile Asn Thr Ile
1 5 10 15
Tyr Leu Leu Phe Ser Ala Tyr Leu Ala Phe Val Met Gln Leu Gly Phe
20 25 30
Ala Met Leu Cys Ala Gly Ser Val Arg Ala Lys Asn Ala Met Asn Ile
35 40 45
Met Leu Thr Asn Val Val Asp Ala Val Val Gly Ser Ile Ser Tyr Tyr
50 55 60
Leu Phe Gly Phe Ala Phe Ala Phe Gly Asp Gly Ser Ser Ser Asn Pro
65 70 75 80
Phe Ile Gly Thr Glu Phe Phe Ala Leu Lys Asp Ile Pro Asp Ser Ser
85 90 95
Tyr Asp Tyr Ser Tyr Phe Leu Tyr Gln Trp Ala Phe Ala Ile Ala Val
100 105 110
Ser Gly Ile Thr Ser Gly Ser Ile Ala Glu Arg Thr Gln Phe Ser Ala
115 120 125
Tyr Leu Val Phe Ser Phe Phe Leu Thr Gly Phe Val Tyr Pro Val Val
130 135 140
Val His Trp Val Trp Ser Ser Ser Gly Trp Leu Ser Pro Ser Ser Thr
145 150 155 160
Ser Leu Ile Phe Ser Ser Gly Ala Ile Asp Phe Ala Gly Ser Gly Val
165 170 175
Val His Leu Val Gly Gly Ile Ala Gly Leu Trp Gly Ser Leu Ile Glu
180 185 190
Gly Pro Arg Val Gly Arg Phe Asp Ala Phe Gly Lys Pro Val Pro Met
195 200 205
Arg Gly His Asn Ala Thr Leu Val Val Leu Gly Thr Phe Leu Leu Trp
210 215 220
Phe Gly Trp Phe Gly Phe Asn Pro Gly Ser Phe Asp Lys Ile Leu Val
225 230 235 240
Ala Tyr Pro Asp Thr Ser Asn Gln Gly Asn Trp Thr Gly Ile Gly Arg
245 250 255
Thr Ala Val Thr Thr Ala Leu Ala Gly Ser Thr Ala Gly Leu Val Thr
260 265 270
Leu Phe Gly Arg Arg Leu Leu Val Gly His Trp Asp Ala Leu Asp Val
275 280 285
Cys Asn Gly Leu Leu Gly Gly Phe Val Ala Ile Thr Ser Gly Cys Ala
290 295 300
Val Val Glu Pro Trp Ala Ala Ile Val Cys Gly Phe Phe Ser Ala Trp
305 310 315 320
Val Leu Ile Gly Leu Asn Ile Leu Ala Leu Lys Leu Lys Phe Asp Asp
325 330 335
Pro Leu Glu Ala Thr Gln Leu His Gly Gly Cys Gly Ala Trp Gly Leu
340 345 350
Leu Phe Thr Gly Leu Phe Ala Lys Glu Glu Phe Ile Val Gln Ala Tyr
355 360 365
Asn Ser Gly Glu Thr Gly Val Val Arg Pro Tyr Gly Leu Leu Leu Gly
370 375 380
Gly Gly Trp Gly Leu Leu Gly Ala Gln Val Ile Glu Val Leu Val Ile
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Val Gly Trp Val Ser Ile Thr Met Gly Pro Leu Phe Tyr Ala Leu His
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Arg Leu Glu Ile Leu Arg Ile Ser Val Asp Glu Glu Val Ala Gly Leu
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Asp Ile Ser Ser His Gly Gly Tyr Ala Tyr Thr Ala His Pro Glu Asp
435 440 445
Thr Pro Arg Tyr Tyr Ala Asp Tyr Met Arg Leu Gln Asn Gln
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<210> 3
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
atggaggcat catgggaag 19
<210> 4
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
tcattggttt tgcagcctc 19
<210> 5
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
ttggcatcgt tgagggtct 19
<210> 6
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
cagtgggaac acggaaagc 19
<210> 7
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
ggaagccagc gtcaccgatt c 21
<210> 8
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
agcgaagccg aagaggtagt a 21
<210> 9
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
acgggggact cttgaccatg gggatggagg catcatggga agc 43
<210> 10
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
aactagtcag atctaccatg gtcattggtt ttgcagcctc 40
<210> 11
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
agctcctgga atccatgaaa 20
<210> 12
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
cggtgatctc tttgctcata c 21
<210> 13
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
caatgaggga agaaggcggt 20
<210> 14
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
cgcaacaccc caaaggaaag 20
<210> 15
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
tatcggtttg atcaggttat gtg 23
<210> 16
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
cgattccata ccatggcact tc 22
<210> 17
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
tgctgctggt atgagtggtg 20
<210> 18
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
gagcaagttg gctgttggtg 20
<210> 19
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
gggaggaaga gaaggtgaa 19
<210> 20
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
agtgtgaaag ctcccatccg 20
<210> 21
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
gttccctccg tgtcacagtt 20
<210> 22
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
attggaggac gcataccgtg 20
<210> 23
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
ggaggaactt taggccaccc 20
<210> 24
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
ctgcaagatc acgtccctca 20
<210> 25
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
tatttgttcg tccggtgggg 20
<210> 26
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
agatcacgtc ggtgagag 18
<210> 27
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
actataggga atattaagct tatggaggca tcatgggaag c 41
<210> 28
<211> 42
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
tccgagctcg gtaccaagct ttcattggtt ttgcagcctc at 42
Claims (9)
1.茶树CsAMT1.3基因在调控植物氮代谢中的应用,其特征在于,所述CsAMT1.3基因的编码序列如SEQ ID NO.1所示,所述氮代谢包括对铵态氮的吸收和同化。
2.如权利要求1所述的应用,其特征在于,所述CsAMT1.3基因编码蛋白的氨基酸序列如SEQ ID NO.2所示。
3.如权利要求1所述的应用,其特征在于,所述应用包括:利用生物学技术手段过表达所述CsAMT1.3基因,获得促进氮代谢的转基因植株。
4.如权利要求3所述的应用,其特征在于,所述CsAMT1.3基因过表达,促进植物对铵态氮的吸收。
5.如权利要求3所述的应用,其特征在于,所述CsAMT1.3基因过表达,转基因植株根长增加,促进侧根分枝。
6.如权利要求3所述的应用,其特征在于,所述转基因植株通过上调磷酸烯醇丙酮酸羧化酶编码基因和谷氨酸合成酶编码基因的表达,加快碳氮代谢,促进铵态氮同化。
7.如权利要求6所述的应用,其特征在于,所述CsAMT1.3基因过表达,促进转基因植株合成叶绿素、氨基酸。
8.如权利要求1所述的应用,其特征在于,所述植物为茶树、拟南芥。
9.如权利要求8所述的应用,其特征在于,所述CsAMT1.3基因过表达促进茶叶对铵态氮的吸收和同化,改良茶叶品质。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102121019A (zh) * | 2010-01-08 | 2011-07-13 | 安徽师范大学 | 一种茶树谷氨酸脱氢酶基因CsGDH及其编码蛋白 |
CN110078804A (zh) * | 2019-04-09 | 2019-08-02 | 中国农业科学院茶叶研究所 | 一种提高植物和微生物耐低氮胁迫能力的蛋白质及其基因 |
CN112708603A (zh) * | 2021-01-28 | 2021-04-27 | 中国科学院遗传与发育生物学研究所 | 水稻are2基因在植物氮代谢调控中的应用 |
CN113512550A (zh) * | 2021-06-11 | 2021-10-19 | 中国科学院华南植物园 | 茶树CsHAC1基因和蛋白及其应用 |
CN114606245A (zh) * | 2022-04-24 | 2022-06-10 | 安徽农业大学 | 茶树CsVAAT3基因及其应用 |
-
2022
- 2022-06-24 CN CN202210738575.8A patent/CN114934055A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102121019A (zh) * | 2010-01-08 | 2011-07-13 | 安徽师范大学 | 一种茶树谷氨酸脱氢酶基因CsGDH及其编码蛋白 |
CN110078804A (zh) * | 2019-04-09 | 2019-08-02 | 中国农业科学院茶叶研究所 | 一种提高植物和微生物耐低氮胁迫能力的蛋白质及其基因 |
CN112708603A (zh) * | 2021-01-28 | 2021-04-27 | 中国科学院遗传与发育生物学研究所 | 水稻are2基因在植物氮代谢调控中的应用 |
CN113512550A (zh) * | 2021-06-11 | 2021-10-19 | 中国科学院华南植物园 | 茶树CsHAC1基因和蛋白及其应用 |
CN114606245A (zh) * | 2022-04-24 | 2022-06-10 | 安徽农业大学 | 茶树CsVAAT3基因及其应用 |
Non-Patent Citations (3)
Title |
---|
DANDAN TANG 等: "Preferential assimilation of NH4+ over NO3− in tea plant associated with genes involved in nitrogen transportation, utilization and catechins biosynthesis", PLANT SCIENCE, vol. 291, pages 5 - 6 * |
YU WANG 等: "Genome-wide identification, characterization, and expression analysis of the ammonium transporter gene family in tea plants (Camellia sinensis L.)", PHYSIOLOGIA PLANTARUM, vol. 174, no. 1, pages 2 - 3 * |
张文婧 等: "茶树CsAMT1s亚家族基因的克隆与表达", 应用与环境生物学报,, vol. 28, no. 1, pages 57 - 66 * |
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