CN113604479A - 茶树hak4基因及其在提高植物钾吸收转运效率上的应用 - Google Patents
茶树hak4基因及其在提高植物钾吸收转运效率上的应用 Download PDFInfo
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Abstract
本发明公开了茶树HAK4基因及其在提高植物钾吸收转运效率上的应用。茶树HAK4基因的核苷酸序列如序列表中SEQ ID NO.1所示。茶树HAK4的表达模式与茶树根系响应低钾胁迫相关,CsHAK4在茶树根中特异高表达,将该基因构建的pDR196‑CsHAK4质粒转化钾吸收缺陷型酵母菌株R5421,能够恢复酵母突变体的生长能力。将不含CsHAK4终止密码子的整个ORF融合到带有绿色荧光蛋白的pCAMBIA1305.1载体上。构建CsHAK4::GFP和AtPIP2A::mCherry表达菌株,转染烟草叶表皮细胞。该基因的克隆有助于解析茶树对低钾环境茶树根系吸收和转运K+分子机制,为培育钾养分高效利用茶树新品种提供基因资源和理论基础具有重要意义。
Description
技术领域
本发明涉及基因工程技术领域,具体涉及茶树HAK4基因及其在提高植物钾吸收转运效率上的应用。
背景技术
茶树(Camellia sinensis(L.)O.Kuntze)是重要的叶用经济作物,钾是茶树的“品质元素”。茶树在发育过程需要大量的钾素,茶树重要的次生代谢物的合成、代谢与钾素营养关系密切。茶树是喜钾植物,钾在提高茶叶品质以及抗逆等方面具有不可替代的作用。茶树对K+的吸收、转运和再分配过程都需要钾通道蛋白和钾转运蛋白来完成,其中在茶园钾素营养缺乏时,钾转运蛋白是茶树根系吸收和转运K+主要途径。
发明内容
本发明的目的在于提供茶树HAK4基因及其在提高植物钾吸收转运效率上的应用,丰富了茶树中转运蛋白的研究,为茶树提高钾吸收转运效率上提供了新思路,为实现茶树抗性性状育种提供了理论和实际参考基础。
为实现上述目的,本发明提供如下技术方案:
茶树HAK4基因,所述茶树HAK4基因为HAK/KUP/KT家族类转运蛋白基因,所述茶树HAK4基因的核苷酸序列,如序列表SEQ ID NO.1所示。
所述茶树HAK4基因及其编码的蛋白质,应用于植物响应低钾胁迫过程中的调控作用,提高植物根系对钾的吸收转运效率。
与现有技术相比,本发明的有益效果是:
本发明中,首次克隆并验证了调控茶树响应低钾胁迫的HAK/KUP/KT家族类转运蛋白基因茶树HAK4基因(即CsHAK4),该转录蛋白在茶树响应低钾胁迫过程中实现调控作用,影响茶叶的钾吸收转运效率和品质的形成。本发明还提供了含有CsHAK4基因的重组质粒、转基因工程菌。本发明丰富了茶树中转运蛋白的研究,为茶树在低钾环境中吸收转运K+提供了新思路,为培育钾养分高效利用茶树新品种提供基因资源和理论基础。
附图说明
图1为茶树不同组织中HAK4基因表达差异以及不同时间低钾处理下的表达变化图;
图2为CsHAK4在烟草叶片中的亚细胞定位图;
图3为CsHAK4在酵母异源体系低钾生长表型以及不同胁迫处理下的酵母生长表型分析图;
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明的一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
1、CsHAK4基因的克隆与序列结构分析
茶树HAK4基因(即CsHAK4基因),为HAK/KUP/KT家族类转运蛋白基因,其克隆与序列结构分析,具体如下:
茶树国家级良种舒茶早种植于安徽省庐阳区合肥安徽农业大学农业产业园,取幼嫩根用于RNA的提取。总RNA的抽提采用RNAprep Pure Plant Kit(Tiangen,Beijing,China)试剂盒按照说明操作,用分光度计检测其RNA含量和质量。
反转录生成第一链:取1μg RNA作模板,根据PrimeScript II 1st Strand cDNASynthesis Kit(Takara Biotech,China)试剂盒说明配置,加入Oligo dT Primer(50μM)0.6μl,Random 6mers(50μM)0.4μl,dNTP Mixture(10mM each)1μl,RNase Free dH2O补足至10μl,65℃变性5min,立即在冰上放置。然后在上述反应液中加入5×PrimerScriptbuffer 4μl,RNase Inhibitor(40U)0.5μl,PrimerScript RTase(200U)1μl dH2O补足20μl,42℃温育45min,95℃5min灭活反转录酶。经过优化后,取适量的反转录产物用于随后的PCR。以cDNA第一链作为RT-PCR模板,常规方法做PCR,扩增CsHAK4基因。其中上游引物:(5’-ACTGGAGGAATGTGGGACTG-3’),下游引物:(5’-AATGACGGGATGAAGACGGT-3’)。20μl PCR反应体系为:10×Ex taq buffer 2.5μl,dNTP 2.0μl,Mg2+1.5μl,上、下游引物各1μl,Ex taq 0.2μl,模板1μl,ddH20 15.8μl。
反应程序如下为:98℃10sec,98℃10sec,57℃30sec,72℃2min,72℃10min,35个循环。PCR产物CsHAK4基因经纯化回收后,连接到pGEM-T Easy Vector载体(Promega,Shanghai,China)上得到pGEM-T Easy::CsHAK4质粒,转化大肠杆菌感受态细胞DH5α,送通用公司测序,得到的CsHAK4基因的核苷酸序列如序列表SEQ ID NO.1所示,具体如下:
ATGGCAGAGAGAAGAAGGGAAGCAGAAGCAGAAGCAGAAGCAGCTCAAGAGATGATAGAAGAAGAAGAAGAAGAAGCACTTAATATGAAAGAAAATAAGGTGTCATCGACAAATCTACGTCGTGTGGACTCCCTCGACTTGGAAGCTGGAAAAGTTACTTTTTCTAGAATCCGCAGCCACAATAATAATGTAGATTGGTGGACAACATTGAGTTTGGCATTTCAAAGTGTCGGGGTAATATATGGAGACATAGGGACTTCTCCACTGTATGTGTTTTCAAGCACTTTCCCAAAGGGTATCCAACACACAGAAGATCTTCTGGGAGTGTTGTCTCTCATCATCTACACCATTCTCCTCATTCCCATGCTCAAGTATGTTTTCATTGTCTTGTGGGCCAACGATAATGGTGATGGAGGTACATTTGCACTCTATTCCTTGATATGCCGGTATGCAAAGGTGAGCTTGATACCAAATGAACAGCCAGAGGACAGACAACTATCCAACTACAAGCTTGATCTATCTTCCAACCAATTGAAAAGAGCTCTAAAGATTAAAGAGAAGTTGGAGAATAGCAGAACTGCTCAAATGGTTCTCTTCCTTGTCACCATCTTGGGAACTTCCATGGTCATTGGGGATGGGGTTCTAACTCCATGCATCTCTGTCCTTTCTGCAGTGAGTGGGATCAAGGCCTTGCATCAAGATGCTGTAGTGGGTATTTCAATTGCAATCTTGATAGTTCTTTTTTGTGTTCAAAGATTTGGGACTGATAAAGTGGGATTCTCGTTCGCACCAATCATCATATTGTGGTTCTTGTTCATAAGTGGAATAGGACTCTATAACTTGCTCAAGCATGACGTTGGAGTCTTAAAGGCTTTTAATCCAAAATACATAGTTGATTACTTCAAAAGAAATGGCAAGAAAGGATGGGTATCTCTTGGTGGAGTTGTTCTCTGCATTACAGGGACAGAAGCCATGTTTGCCGATCTAGGGCACTTCAATGTTCGAGCAGTCCAAATTAGTTGCTCTGGAATTGTGTTTCCTGCATTACTAGCTGCCTACAGTGGACAAGCTGCATACCTTGTCAAATTCCCCGGCGATGTGGGTAATACTTTCTACGCCTCGATTCCAGGCAAGTCTAAAATCGAATAATTCATTGTGATTACCCAAACTATCATGCTTACATAATTATCATCATAATATTTTTATTATCGCTATTTTTGCTTTTCATTCATGCTCACCGGTATGCAGGTCCATTATATTGGCCAACATTTGTTGTCGCTGTTGCTGCTGCCATTATCGCTAGCCAAGCTATGATCTCGGGAACATTTTCAATTATCTCCCAGTCCTTAAGTCTAGGTTGTTTTCCGAGGGTTAAAGTTGTCCACACGTCAGCTAAGTACGAGGGTCAAGTTTACATACCGGAGGTCAACTTCTTACTAATGATTGCTTGCGTTATTGTAACCGCTGCCTTCAAGAGCACAACAAATATTGGCAATGCATATGGAATCGCCGTGGTTGCTGTAATGATCATCACAACCTGTATGCTTACTCTTATCATGCTTGTTATATGGAAGACAAGCATATGGTGGATAGTTCTATTCCTCGTGATTTTTGGTTCGATTGAGGGCTTATACCTGTCATCTGTCTTGTTCAAATTCGTTGAGGGCGGTTACCTTCCACTAGCCTTTTCAGTTGTTCTAATGCTGATAATGGGGTTGTGGCATTACGTACACAAAAAGAGATACATGTTTGAGCTCAACAACAAGGTTTCTAGTAATTTTATAAAAGACTTGGCACTGAATCCAAATATTAACAGGGTGCCCGGAATTGCACTTTTATACTCTGAGCTTGTGCAGGGCATTCCTCCAATATTTCGCCATTTCATTACCTATATACCATCCATCCATTCAGTCTTGGTGTTTGTCTCTATCAAGTCTATTCCCACTAGCAAAGTGGTCATGGAGGAGAGATTTTTATTTCGGCTGGTCGAACCAAGAGACTACCACATGTTTCGCTGTGTGGTGAGATATGGATATAATGATGTGATTGAAGGATCAGAGGTGTTTGAAAAGCAACTGATTGAGCACTTGAAGCAGTACCTTCAGCATGAACACTTCATTCACGAAGGACAACAGCCTAATGAAGAAATAGTCGAGCCAGTGAACACTGGAGCCATCCCAGGTGCCGAAGAGGAGATGCAATTTGTCCATAAAGCAATGGAAAATGGTGTTGTTTATTTCTTAGGGGAAGCAGAAGTTCGAGCTGAACAGAACTCATCCCTATTCAAGAAGATTGTTGTCAACTATGCCTATAATTTCCTCAGGAAAAACTTTAGCCGAGGGGAACAAGTATTGGGAATCCCTCGGAGTCGACTCCTTAGGGTTGGAATGACATATGAGATAG
2、CsHAK4基因的表达差异分析
(1)茶树不同组织CsHAK4基因表达
茶树国家级良种舒茶早品种种植于安徽省庐阳区合肥安徽农业大学农业产业园,14个组织器官用来分析基因表达。这14个组织器官包括芽(Bud)、1叶(1st Leaf)、1叶脉(1st Main Vein)、2叶(2nd Leaf)、2叶脉(2nd Main Vein)、3叶(3rd Leaf)、3叶脉(3rdMain Vein)、4叶(4th Leaf)、4叶脉(4th Main Vein)、5叶(5th Leaf)、5叶脉(5th MainVein)、维管束(Vascular Bundle)、2叶和3叶之间的嫩茎(Stem)和根(Root)。同时这些样品用于总RNA的提取以及cDNA第一链合成。反转录产物(cDNA第一条链)稀释5倍作为模板,使用2×AceQ Universal qPCRMaster Mix(Vazyme,Nanjing,China),配制10μl反应体系:1.0μl稀释5倍的逆转录产物,上下游引物各0.4μl(10pmol/μl),5μl 2×AceQUniversal qPCRMaster Mix,3.2μl ddH20,每个反应配3个重复。然后在Bio-radCFX-384仪器上以程序:①95℃5min②95℃10sec,60℃30sec,72℃30sec运行39个循环③从65℃到95℃,以0.1℃/sec绘制熔解曲线。上游引物:(5’-GGAATCGCCGTGGTTGCTGTAA-3’),下游引物:(5’-AGTGGAAGGTAACCGCCCTCAA-3’),以茶树GADPH基因为内参,上游引物:(5’-TTGGCATCGTTGAGGGTCT-3’),下游引物:(5’-CAGTGGGAACACGGAAAGC-3’)通过仪器自带分析软件,计算出CsHAK4在不同组织中的相对表达水平。
(2)茶树在低钾胁迫下的CsHAK4基因表达
2年生茶树扦插苗。茶苗(舒茶早)取自中国安徽省舒城县德昌育木基地。采用大小均匀的茶树幼苗进行水培。在安徽农业大学茶树生物学与利用国家重点实验室的生长温室中,温室设置为温度25℃,光照时间14h,黑暗时间10h,相对湿度设置为70%。将茶树幼苗在全基础营养液中生长1个月,以生长发育良好的根。将茶树扦插苗在含0.513mM钾离子的溶液中生长1个月,然后转移到完全缺钾(Na2SO4和NaH2PO4替代K2SO4和KH2PO4)处理溶液中,处理后,在不同时间点(1h、6h、12h、24h和48h)收集根组织样本,并立即在液氮中速冻,存储于-80℃的超低温冰箱用于分析CsHAK4基因的表达量。RNA的提取和定量PCR方法同上。
图1是茶树不同组织中CsHAK4编码蛋白的表达差异以及不同时间低钾处理下的表达变化。如图1所示,以茶树品种舒茶早为研究材料,不同组织中CsHAK4表达模式具有组织特异性,在低钾处理24h后呈现上调表达模式。由图1可知,通过qRT-PCR检测结果显示CsHAK4表达量与茶树根响应低钾转运相关。CsHAK4在根中表达较高,其它组织表达较低。结合蛋白序列基因注释,暗示CsHAK4可能与茶树根响应低钾转运有关系。
3、CsHAK4基因在烟草叶片中的定位
(1)CsHAK4-pCAMBIA1305.1载体构建
以pGEM-T Easy::CsWRKY29质粒为模板,上游引物:(5’-GGACTAGTATGGCAGAGAGAAGAAGGGA-3’),下游引物:(5’-GGACTAGTATGGCAGAGAGAAGAAGGGA-3’),进行PCR扩增。PCR产物用1.2%的琼脂糖胶电泳条带进行回收。首先将基因PCR回收产物与载体质粒进行双酶切,酶切产物用1.2%的琼脂糖胶电泳条带再进行回收,利用T4酶连技术,加入2μl载体和6μl基因酶切后回收产物,1μl T4 DNA Ligase Mix及1μl T4 DNALigase Buffer,4℃过夜后转化DH5α,送生工公司测序。
(2)农杆菌注射渗透法注射烟草
把CsHAK4-pCAMBIA1305.1载体电击法转化到EHA105农杆菌当中,并通过常规PCR方法鉴定阳性克隆。挑取菌落PCR验证正确的单克隆,接种于5ml液体LB培养基(含50μg/mlrif+和100μg/ml Spec+),培养至OD600=0.8-1.2。取1ml过夜培养的农杆菌,接入100ml液体LB培养基含(50μg/ml rif+和100μg/ml Spec+),28℃200r/min过夜培养。离心收集菌体,用10mM MgCl2和10mM 2-(N-吗啉代)乙磺酸调节PH为5.6的重悬液重悬菌体至OD600=0.4。在菌液中加入100μM的乙酰丁香酮(As),28℃孵育2h,注射前按1:1的比例与P19-GV3101混合菌液。用一次性1ml注射器去掉针头吸取菌液,从烟草叶片下表皮注射,使菌液渗透到整个叶片组织中。注射后的烟草暗处理8-12h,正常温室培养2-3天后,使用激光共聚焦显微镜观察GFP荧光记录并拍照。
图2是CsHAK4在烟草叶片中的亚细胞定位图。如图2所示,其中,GFP:绿色荧光蛋白;AtPIP2A::mCherry:质膜maker基因;Bright Field:pCAMBIA1305.1-CsHAK4的明场图片;Merged:pCAMBIA1305.1-CsHAK4融合图片。由图2可知:通过与EGFP的融合表达,在烟草叶片的细胞质膜中能够检测到CsHAK4的绿色荧光,表明CsHAK4蛋白定位在细胞质膜上。
4、CsHAK4基因在酵母体内功能验证
(1)CsHAK4-pDR196载体构建
以pGEM-T Easy::CsWRKY29质粒为模板,上游引物:(5’-CCGGAATTCATGGCAGAGAGAAGAAGGGAAGCAG-3’),下游引物:(5’-CCGCTCGAGCTATCTCATATGTCATTCCAACCCT-3’),进行PCR扩增。PCR产物用1.2%的琼脂糖胶电泳条带进行回收。首先将基因PCR回收产物与载体质粒进行双酶切,酶切产物用1.2%的琼脂糖胶电泳条带再进行回收,利用T4酶连技术,加入2μl载体和6μl基因酶切后回收产物,1μl T4 DNA Ligase Mix及1μl T4 DNA Ligase Buffer,4℃过夜后转化DH5α,送生工公司测序。
(2)载体转化酵母突变体
酵母突变体G19是盐敏感型菌株,在高盐环境下生长受阻(南京农业大学沈立柯老师馈赠),用来验证所选基因的抗胁迫能力。酵母突变体R5421是钾吸收缺陷型酵母菌,在低钾条件下生长受阻,该菌来自杨天元老师之前博士论文所用(Dr.Richard F.Gaber,Northwestern University赠送),用来验证所选基因的亲和性吸收钾离子的能力。将R5421菌株在YPDA固体培养基(含有100mM K+)上划线,28℃恒温培养箱倒置培养2-3天。挑取长势良好的单菌落于2mL离心管中,加入1mL YPDA液体培养基(含有100mM K+),在28℃摇床,180r/min培养1天。吸取200μl生长良好的菌液转移至30mL的YPDA液体培养基(含有100mM K+),在28℃摇床,180r/min培养12-24h。待菌液摇至OD为0.8-1.2,吸取1mL至1.5mL离心管,12000rpm离心1min收集菌体,去上清,用灭菌水清洗一遍。在管底加入5μl转化的质粒DNA,用枪头吸打混匀。加入500μl PEG mixture,5μl DTT(1M,现配现用),涡旋混匀。室温静置20min,45℃水浴20min,冰浴20min。吸取底部沉淀菌体50μl,涂于SD-U(含有100mM K+)的固体培养基上,28℃恒温培养箱倒置培养2-3天。酵母菌落PCR验证阳性菌落。菌落PCR反应程序:98℃变性10min,94℃预变性5min,{98℃变性10s,60℃退火10s,72℃延伸2min 30s},30个循环,72℃延伸10min,16℃保温1s。
(3)酵母打点功能验证
挑取单菌落于20mL的SD-U(含有100mM K+)固体培养基中,28℃、180r/min的情况下,摇菌至OD值为1.0左右(约48h);吸取4mL菌液,5000rpm离心2min,去上清(无菌操作)后用无菌水重新悬浮菌体,检测OD值;用无菌水稀释菌液至OD值为0.6(各个样品之间OD值相差不要超过0.02);(无菌操作)将菌液依次稀释为100、10-1、10-2、10-3。例如10-1为从原菌液(100)中吸取100μl到新的1.5mL离心管中,并加入900μl无菌水,混匀;10-2为从上一稀释液(10-1)中吸取100μl到新的1.5mL离心管中,并加入900μl无菌水,混匀;10-3为从上一稀释液(10-2)中吸取100μl到新的1.5mL离心管中,并加入900μl无菌水,混匀。(无菌操作)
将配好的固体培养基放在已经画好的网格线上,利用10μl移液器吸取2μl的菌液,并将菌液滴在网格的交叉线上,同一横排点同一菌液的不同浓度,同一纵列点同一浓度的不同菌液。等菌液完全风干后再进行封板,在30℃恒温培养箱中倒置培养。注意打点时,移液枪应垂直于平皿。一般情况下,培养三天即可观察酵母生长情况。
图3是CsHAK4在酵母异源体系低钾生长表型以及不同胁迫处理下的酵母生长表型分析图。如图3所示,将酵母空载体pDR196和pDR196-CsHAK4的质粒转化钾吸收缺陷型酵母菌株R5421和盐敏感酵母菌株G19,在较低钾浓度(0.1mM)下,pDR196酵母转化体的生长完全抑制,CsHAK4转化的酵母突变体却回补了部分生长状态。在较高钠浓度(150mM)下,CsHAK4转化的酵母突变体较pDR196酵母转化体生长受到部分抑制。表明CsHAK4是一种功能性钾离子转运蛋白且部分表现出对盐胁迫敏感。
以上内容仅仅是对本发明结构所作的举例和说明,所属本技术领域的技术人员对所描述的具体实施例做各种各样的修改或补充或采用类似的方式替代,只要不偏离本发明的结构或者超越本权利要求书所定义的范围,均应属于本发明的保护范围。
序列表
<110> 安徽农业大学
<120> 茶树HAK4基因及其在提高植物钾吸收转运效率上的应用
<130> NO
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2403
<212> DNA
<213> 茶树(Camellia sinensis L. O. Kuntze)
<400> 1
atggcagaga gaagaaggga agcagaagca gaagcagaag cagctcaaga gatgatagaa 60
gaagaagaag aagaagcact taatatgaaa gaaaataagg tgtcatcgac aaatctacgt 120
cgtgtggact ccctcgactt ggaagctgga aaagttactt tttctagaat ccgcagccac 180
aataataatg tagattggtg gacaacattg agtttggcat ttcaaagtgt cggggtaata 240
tatggagaca tagggacttc tccactgtat gtgttttcaa gcactttccc aaagggtatc 300
caacacacag aagatcttct gggagtgttg tctctcatca tctacaccat tctcctcatt 360
cccatgctca agtatgtttt cattgtcttg tgggccaacg ataatggtga tggaggtaca 420
tttgcactct attccttgat atgccggtat gcaaaggtga gcttgatacc aaatgaacag 480
ccagaggaca gacaactatc caactacaag cttgatctat cttccaacca attgaaaaga 540
gctctaaaga ttaaagagaa gttggagaat agcagaactg ctcaaatggt tctcttcctt 600
gtcaccatct tgggaacttc catggtcatt ggggatgggg ttctaactcc atgcatctct 660
gtcctttctg cagtgagtgg gatcaaggcc ttgcatcaag atgctgtagt gggtatttca 720
attgcaatct tgatagttct tttttgtgtt caaagatttg ggactgataa agtgggattc 780
tcgttcgcac caatcatcat attgtggttc ttgttcataa gtggaatagg actctataac 840
ttgctcaagc atgacgttgg agtcttaaag gcttttaatc caaaatacat agttgattac 900
ttcaaaagaa atggcaagaa aggatgggta tctcttggtg gagttgttct ctgcattaca 960
gggacagaag ccatgtttgc cgatctaggg cacttcaatg ttcgagcagt ccaaattagt 1020
tgctctggaa ttgtgtttcc tgcattacta gctgcctaca gtggacaagc tgcatacctt 1080
gtcaaattcc ccggcgatgt gggtaatact ttctacgcct cgattccagg caagtctaaa 1140
atcgaataat tcattgtgat tacccaaact atcatgctta cataattatc atcataatat 1200
ttttattatc gctatttttg cttttcattc atgctcaccg gtatgcaggt ccattatatt 1260
ggccaacatt tgttgtcgct gttgctgctg ccattatcgc tagccaagct atgatctcgg 1320
gaacattttc aattatctcc cagtccttaa gtctaggttg ttttccgagg gttaaagttg 1380
tccacacgtc agctaagtac gagggtcaag tttacatacc ggaggtcaac ttcttactaa 1440
tgattgcttg cgttattgta accgctgcct tcaagagcac aacaaatatt ggcaatgcat 1500
atggaatcgc cgtggttgct gtaatgatca tcacaacctg tatgcttact cttatcatgc 1560
ttgttatatg gaagacaagc atatggtgga tagttctatt cctcgtgatt tttggttcga 1620
ttgagggctt atacctgtca tctgtcttgt tcaaattcgt tgagggcggt taccttccac 1680
tagccttttc agttgttcta atgctgataa tggggttgtg gcattacgta cacaaaaaga 1740
gatacatgtt tgagctcaac aacaaggttt ctagtaattt tataaaagac ttggcactga 1800
atccaaatat taacagggtg cccggaattg cacttttata ctctgagctt gtgcagggca 1860
ttcctccaat atttcgccat ttcattacct atataccatc catccattca gtcttggtgt 1920
ttgtctctat caagtctatt cccactagca aagtggtcat ggaggagaga tttttatttc 1980
ggctggtcga accaagagac taccacatgt ttcgctgtgt ggtgagatat ggatataatg 2040
atgtgattga aggatcagag gtgtttgaaa agcaactgat tgagcacttg aagcagtacc 2100
ttcagcatga acacttcatt cacgaaggac aacagcctaa tgaagaaata gtcgagccag 2160
tgaacactgg agccatccca ggtgccgaag aggagatgca atttgtccat aaagcaatgg 2220
aaaatggtgt tgtttatttc ttaggggaag cagaagttcg agctgaacag aactcatccc 2280
tattcaagaa gattgttgtc aactatgcct ataatttcct caggaaaaac tttagccgag 2340
gggaacaagt attgggaatc cctcggagtc gactccttag ggttggaatg acatatgaga 2400
tag 2403
Claims (2)
1.茶树HAK4基因,其特征在于:所述茶树HAK4基因为HAK/KUP/KT家族类转运蛋白基因,所述茶树HAK4基因的核苷酸序列,如序列表SEQ ID NO.1所示。
2.根据权利要求1所述的茶树HAK4基因在提高植物钾吸收转运效率上的应用。
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