CN115010796A - 一种赖草生长素输出载体及其编码基因与应用 - Google Patents
一种赖草生长素输出载体及其编码基因与应用 Download PDFInfo
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- CN115010796A CN115010796A CN202210426568.4A CN202210426568A CN115010796A CN 115010796 A CN115010796 A CN 115010796A CN 202210426568 A CN202210426568 A CN 202210426568A CN 115010796 A CN115010796 A CN 115010796A
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Abstract
本发明具体涉及一种赖草生长素输出载体及其编码基因与应用,属于植物基因工程技术领域。本发明首次发现了赖草的生长素输出载体,其氨基酸序列如SEQ ID NO.2所示,编码基因的核苷酸序列如SEQ ID NO.1所示。本发明的赖草生长素输出载体与拟南芥AtPIN1的功能相近,在赖草中是影响根茎生长素运输及根发育的关键转运子。赖草生长素输出载体编码基因的过表达促进了转基因拟南芥中生长素的极性运输和根的发育,并促进了生长素的积累,这些结果表明,赖草生长素输出载体对赖草根系发育和生长素运输具有重要影响,通过利用赖草生长素输出载体调控根系发育的功能,有助于实现调控赖草形态结构、根系发育并最终提高赖草品质的目标。
Description
技术领域
本发明涉及植物基因工程技术领域,具体涉及一种赖草生长素输出载体及其编码基因与应用。
背景技术
赖草是典型的复合型分蘖克隆植物,通过根茎逐级分蘖进行无性繁殖的根茎类禾草,是草原牧草种群及群落结构的重要组成部分,很多优良牧草都具有地下根茎,通过不断分蘖形成纵横交错的地下根茎网,以及与之对应的地上子代分株,该特点赋予了赖草很强的水平拓展迁移能力和抗扰动能力,以及极强的防止风沙、稳固沙地的能力,所以种植赖草对防止土地荒漠化、提高环境恶劣地区物种的多样性、生态环境的保护和恢复等方面都具有极为重要的作用。大赖草(Leymus racemosus)、灰色赖草(Leymus cinereus)常被作为农、牧业良种繁育及麦类作物育种时的重要远缘杂交材料,所以发掘和应用赖草属基因库资源愈来愈受到人们的重视。因此,选择赖草根茎作为研究对象,研究其调控根系分蘖、生长方向性基因及生长机理,具有特殊的代表性。
侧枝(禾本科植物中称作分蘖)的起始和生长是决定株型发育的重要因素,植物激素在调控分蘖产生方面具有不可替代的作用。生长素是一种重要的植物激素,调节多种植物的生理和发育过程。生长素梯度分布受生长素外排载体调控,对植物生长至关重要。大量研究表明,PIN基因家族在调控生长素运输中起着至关重要的作用。目前,PIN基因家族已在多种植物中被鉴定,如拟南芥、水稻和杨树。人们从拟南芥中先后分离了AtPIN1、AtPIN2、AtPIN3、AtPIN4和AtPIN7等基因,其中,AtPIN1定位于根或地上部分的维管组织及胚胎中,参与根系和分蘖等多种器官的发育调控;AtPIN2仅在根中表达,其转录定位于根部因重力刺激发生弯曲的区域,在根毛的发生区域也观测到AtPIN2的大量表达;AtPIN3的分布较广,在根分生组织的中柱鞘细胞(pericycle)、根冠柱(columella)以及幼芽的内皮层(endodemis)细胞中都可以观测到其表达;AtPIN4在根静止中心(quiescent center)区域表达,对维持分生组织中生长素的浓度梯度至关重要,此外,它还在胚胎的极性建成时大量表达;同样AtPIN7在根冠和胚胎中表达,它在维持生长素的向基式浓度梯度时起作用。(刘士平等,高等植物的PIN基因家族,植物生理学通讯,2009,45(8):833-841)
然而,目前赖草中的生长素输出载体的具体结构还不清楚,其具体功能也尚不明确。在赖草种植与培育中,草原土地沙漠化抑制根系发育,导致水分和肥料吸收减少,而生长素输出载体对生长素的运输与根系的发育起着重要作用。因此,挖掘赖草生长素输出的关键基因,开发其在赖草中的功能与应用,可以更好的改善赖草根系品质,使之可以适应复杂严峻的生态环境,改善土地荒漠化,是行业内研发人员积极探索的课题之一。
发明内容
针对现有技术的不足,本发明提供了一种赖草生长素输出载体及其编码基因与应用,赖草生长素输出载体参与根茎分蘖及侧生分生组织分化,为解决赖草分子设计育种技术提供了理论基础。
本发明采用的技术方案如下:
一种赖草生长素输出载体,其氨基酸序列如SEQ ID NO.2所示。
上述赖草生长素输出载体的编码基因的核苷酸序列如SEQ ID NO.1所示。
本发明中,通过对赖草进行转录组测序,构建赖草转录组序列信息数据库,根据与拟南芥AtPIN1基因的同源性,得到一个1761bp的开放阅读框,将其确定为调控赖草生长素输出的关键候选基因。由于转基因赖草技术不够成熟,所以将该基因转化至拟南芥中,构建转基因拟南芥,并对其进行功能验证,确定其为赖草生长素输出载体的编码基因,具有调控生长素运输的功能。
一种含有上述赖草生长素输出载体的编码基因的重组质粒。
根据本发明优选的,所述重组质粒的载体为pEASY-blunt或pCAMBIA1300-GFP。
一种含有上述赖草生长素输出载体的编码基因或上述重组质粒的重组菌。
根据本发明优选的,所述重组菌的宿主为大肠杆菌Trans1-T1或大肠杆菌DH5α或农杆菌GV3101。
上述赖草生长素输出载体在促进植株分蘖或调控植株顶芽转向中的应用。
根据本发明优选的,所述植株为拟南芥或赖草。
根据本发明优选的,所述应用是将赖草生长素输出载体的编码基因转化入植株中,实现生长素输出载体的编码基因在植株中的过表达。
上述赖草生长素输出载体在与外源激素协同促进植株分蘖中的应用。
根据本发明优选的,所述植株为拟南芥或赖草。
根据本发明优选的,所述外源激素为GA3或IAA。
本发明中,构建生长素输出载体编码基因的过表达载体,转化进宿主中,构建转基因拟南芥,发现生长素输出载体参与调控主根的生长和发育,也影响侧根的发生和形态建成;另外,通过重力响应实验和向光性实验,发现生长素输出载体可以响应重力信号调控根系的方向性生长以及响应光信号调控根系的向光性生长,表明赖草生长素输出载体可以调控植株的顶芽转向;本发明还研究了外源激素GA3和IAA对转基因拟南芥根系的影响,发现生长素输出载体会受到外源激素GA3和IAA的诱导,进一步调控拟南芥侧根的发育,表明赖草生长素输出载体可以与外源激素协同作用,共同促进植株分蘖及在侧生分生组织分化中发挥作用。
有益效益:
1、本发明首次发现了赖草中的生长素输出载体,其氨基酸序列如SEQ ID NO.2所示,编码基因的核苷酸序列如SEQ ID NO.1所示。
2、本发明通过对赖草生长素输出载体编码基因的克隆与鉴定、基因的表达分析及基因的遗传转化等手段分析验证其功能。赖草生长素输出载体与拟南芥AtPIN1的功能相近,在赖草中是影响根茎生长素运输及根发育的关键转运子。赖草生长素输出载体的编码基因在多个赖草部位中检测到,且在垂直根茎顶芽中含量较高,这可能是由于赖草生长素输出载体介导的生长素从芽到根的运输。赖草生长素输出载体编码基因的过表达促进了转基因拟南芥中生长素的极性运输和根的发育,并促进了生长素的积累,因此,赖草生长素输出载体参与了生长素的运输,并与其他生长素输出载体蛋白协同调节生长素的稳态,这种转运途径使生长素处于相对平衡和稳定的状态。这些结果表明,赖草生长素输出载体对赖草根系发育和生长素运输具有重要影响,研究赖草生长素输出载体蛋白可以更好的改善赖草根系分蘖习性,使之可以适应更加复杂严峻的生态环境,改善土地荒漠化,为未来赖草种植与培育提供了潜在的应用前景。
3、本发明公开了赖草生长素输出载体在促进植株分蘖或调控植株顶芽转向中的应用,以及赖草生长素输出载体在与外源激素协同促进植株分蘖中的应用,通过利用赖草生长素输出载体调控根系发育的功能,以及生长素输出载体与其他外源激素的协同作用,有助于实现调控赖草形态结构、根系发育并最终提高赖草品质的目标。
附图说明
图1为LsPIN1蛋白的跨膜区域分析图;
图2为赖草总RNA凝胶电泳图;图中,泳道M为标准品,条带大小从上到下依次为5000bp、3000bp、2000bp、1500bp、1000bp、750bp、500bp、250bp,泳道1为赖草水平根茎顶芽样品,泳道2为赖草垂直根茎顶芽样品,泳道3为赖草叶片样品;
图3为LsPIN1基因在赖草水平根茎顶芽和垂直根茎顶芽中的相对表达量;
图4为扩增LsPIN1基因琼脂糖凝胶电泳图;图中,泳道M为标准品,泳道1为赖草水平根茎顶芽扩增样品,泳道2为赖草垂直根茎顶芽扩增样品,泳道3为赖草叶片扩增样品;
图5为SpeI和XbaI双酶切后的琼脂糖凝胶电泳图;图中,左图为质粒pEASY-LsPIN1的双酶切电泳图,1-6泳道均为质粒pEASY-LsPIN1的酶切样品;右图为表达载体pCAMBIA1300-GFP的双酶切电泳图,两个样品泳道均为表达载体pCAMBIA1300-GFP的酶切样品;
图6为重组载体pCAMBIA1300-GFP-LsPIN1转化大肠杆菌阳性菌落PCR验证的琼脂糖凝胶电泳图;图中,泳道1-8及泳道12、14、15、17扩增得到目的基因,验证为阳性转化子;
图7为重组载体pCAMBIA1300-GFP-LsPIN1转化农杆菌GV3101的阳性菌落PCR验证的琼脂糖凝胶电泳图;图中,泳道1-6均扩增得到目的基因,验证为阳性转化子;
图8为LsPIN1蛋白的亚细胞定位分析图;
图9为LsPIN1转基因拟南芥T1代种子阳性苗筛选照片;
图10为T1代LsPIN1转基因拟南芥阳性苗PCR扩增验证的凝胶电泳检测图;
图11为LsPIN1转基因拟南芥和野生型拟南芥的表型和数据统计;其中,图a为拟南芥垂直培养7天后的形态照片,图b为拟南芥主根长度柱状图,图c为拟南芥侧根数量柱状图;
图12为GA3激素处理LsPIN1转基因拟南芥表型和数据统计;其中,图a为拟南芥垂直培养7天后的形态照片,图b为拟南芥主根长度柱状图,图c为拟南芥侧根数量柱状图;
图13为IAA激素处理LsPIN1转基因拟南芥表型和数据统计;其中,图a为拟南芥垂直培养7天后的形态照片,图b为拟南芥主根长度柱状图,图c为拟南芥侧根数量柱状图;
图14为LsPIN1转基因拟南芥逆时针旋转135°重力刺激后向地性生长结果;其中,图a为拟南芥的整体形态照片,图b为拟南芥根系的局部形态照片,图c为拟南芥根系旋转角度折线图;
图15为LsPIN1转基因拟南芥微重力刺激响应图;其中,左图为处理四天和处理七天的拟南芥的形态照片,右图为拟南芥根系弯转角度柱状体;
图16为LsPIN1转基因拟南芥向光性响应照片;
图17为DR5::GUS-LsPIN1转基因拟南芥在主根和侧根根尖的生长素积累图。
具体实施方式
以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为常规生化试剂商店购买得到的。以下实施例中的定量试验,均设置三次重复实验,结果取平均值。
实施例1:赖草LsPIN1基因序列的鉴定
利用转录组测序技术测定赖草的转录组序列信息,构建赖草转录组序列信息数据库,在所有的赖草转录组序列信息中,为了鉴定赖草基因组中调控赖草生长素输出的候选基因,以拟南芥AtPIN1基因(UniProtKB:Q9C6B8)为模板,在赖草的转录组序列中比对寻找相似序列,发现在赖草的转录组序列中有一个1761bp的ORF(开放阅读框),其与AtPIN1基因的同源性为67.79%,命名为LsPIN1,其核苷酸序列如SEQ ID NO.1所示。
LsPIN1基因编码的蛋白质序列包含587个氨基酸,其氨基酸序列如SEQ ID NO.2所示,其分子质量为64223.44Da,等电点PI值9.04。LsPIN1蛋白为疏水性蛋白质,其跨膜区域分析图如图1所示,LsPIN1蛋白存在两个跨膜区域(氨基酸残基1-151和437-587氨基酸残基)可能暴露在质膜的外表面,表明了其结构和功能与膜蛋白具有相关性。
实施例2:赖草LsPIN1基因重组载体的构建
以赖草为材料,提取总RNA,并反转成cDNA,进行组织部位分析,设计相应引物进行PCR,琼脂糖凝胶电泳检测后,回收目的条带,与pEASY-bLunt载体连接,转入大肠杆菌Trans1-T1感受态细胞,测序并分析,挑取阳性克隆进行质粒抽提,加入酶切位点后与载体pCAMBIA1300-GFP同时双酶切,在T4连接酶的作用下连接后,转入大肠杆菌感受态细胞,挑取阳性克隆进行质粒提取,转入农杆菌GV3101中,待拟南芥适龄后,通过农杆菌介导法进行转化,观察T3代表型,进行统计分析,分析其功能。
1.总RNA提取
以赖草水平根茎顶芽、垂直根茎顶芽和叶片为材料,按照植物总RNA提取试剂盒(天根生化)的操作步骤进行提取。所用试剂和耗材均无RNA酶。利用超微量分光光度计Nano-300检测水平根茎顶芽、垂直根茎顶芽和叶的RNA浓度分别为891.2ng/μL、985.5ng/μL、906.3ng/μL,A260/A280在2.0左右,并用1%琼脂糖凝胶电泳检测如图2所示,总RNA的28S、18S、5S条带均清晰无降解。说明RNA未出现明显降解,质量较好,可满足后续反转录实验的要求。
2.cDNA的获得
按照反转录试剂盒(诺唯赞)的说明书,将上述提取的赖草水平根茎顶芽、垂直根茎顶芽和叶片的RNA分别反转录合成cDNA,cDNA产物用于qPCR和PCR。
3.LsPIN1基因组织表达分析
为了分析LsPIN1基因在赖草水平根茎顶芽和垂直根茎顶芽中的表达模式,对其进行荧光定量PCR(qPCR)分析。以上述获得的用于qPCR的cDNA为模板,测定LsPIN1基因的表达量,以18S为内参基因,以水平顶芽LsPIN1基因的表达量为对照,即将水平顶芽LsPIN1基因的表达量设定为1。
其中,用于qPCR的引物序列为:
LsPIN1-RT-F:5’-TCCTGCACGTCGCCATCGTCCAG-3’(SEQ ID NO.3),
LsPIN1-RT-R:5’-GCAGGATGTAGTAGACGAGCGTG-3’(SEQ ID NO.4);
根据荧光定量PCR试剂盒的要求配制反应体系后,在预变性95℃,10min;变性95℃,15s,60℃退火1min,循环数40的程序下扩增,并根据ABI 7500荧光定量PCR仪推荐程序进行溶解曲线分析。每个样品进行三次生物学重复。结果如图3所示:LsPIN1基因在赖草水平根茎顶芽和垂直根茎顶芽中均有表达,且LsPIN1基因在垂直根茎顶芽中表达量比水平根茎顶芽表达量高。
4.LsPIN1基因的克隆
(1)根据LsPIN1基因的全长序列和所用表达载体的限制性酶切位点,设计包含整个ORF(去除终止密码子)的特异性引物,在引物的5’端和3’端分别加上酶切位点,引物序列为:
LsPIN1-F-XbaI:5’-GCTCTAGAATGATCACGGGCACGGACTTC-3’(SEQ ID NO.5),
LsPIN1-R-SpeI:5’-GGACTAGTCAGGCCGAGCAGGATGTAGTAG-3’(SEQ ID NO.6);
(2)利用上述获得的用于PCR的cDNA第一链与LsPIN1的特异性引物,用2×PhantaMax高保真酶(诺唯赞)进行扩增,扩增反应条件为:预变性95℃,3min;95℃变性15sec,56℃退火15sec,72℃延伸1kb/min,35个循环后72℃继续延伸5min;
(3)用1%琼脂糖凝胶电泳检测,如图4所示,扩增片段均与目的基因大小一致,参照胶回收试剂盒(天根生化)说明书切胶回收。
5.连接pEASY-bLunt载体与Trans1-T1感受态细胞的转化
将4μL胶回收产物与1μL克隆载体pEASY-bLunt载体轻轻混匀后,25℃反应30min,在超净工作台中将全部连接产物转移至50μL大肠杆菌Trans1-T1感受态细胞中(感受态细胞-80℃冰箱拿出,冰上融化),将其缓慢混匀,冰浴30min,42℃水浴热激30秒后,立即置于冰上2分钟;在超净台中加入250μL的LB液体培养基,37℃、220rpm震荡培养1小时;4500rpm离心1min,保留150μL上清液,轻弹悬浮菌体后将其全部涂布在含有100μg/mL卡那霉素(Kan)的LB固体培养基中。用封口膜将培养皿封好后,在37℃培养箱中倒置、过夜培养。
挑取培养好的单菌落接种至3mL含100μg/mL卡那霉素的LB液体培养基中,37℃,220rpm摇菌,以LsPIN1-F-XbaI和LsPIN1-R-SpeI为引物进行PCR扩增验证,PCR反应程序为:预变性95℃,3min;95℃变性15sec,56℃退火15sec,72℃延伸1kb/min,35个循环后72℃继续延伸5min;4℃保存;用1%琼脂糖凝胶电泳检测为阳性克隆后,送测序公司进行检测,将测序结果与LsPIN1基因序列进行比对,测序正确的菌落提取质粒pEASY-LsPIN1。
6.赖草LsPIN1基因过表达载体的构建
采用双酶切的方法,将携带目的基因LsPIN1的质粒pEASY-LsPIN1与表达载体pCAMBIA1300-GFP,分别用SpeI和XbaI在PCR仪中37℃进行双酶切1h,用1%的琼脂糖凝胶进行电泳检测,如图5所示,根据条带大小判断LsPIN1基因与pCAMBIA1300-GFP载体已被正确切开。切胶回收目的条带后,利用T4连接酶将酶切后的目的基因与表达载体以3:1的浓度,25℃连接2h,转入DH5α大肠杆菌感受态细胞,获得重组载体pCAMBIA1300-GFP-LsPIN1,挑取单菌落PCR验证阳性菌,如图6所示,将与目的基因一致的阳性单菌落摇菌培养进行测序,测序结果与LsPIN1基因序列比对结果完全一致,说明pCAMBIA1300-GFP-LsPIN1过表达载体构建成功。从阳性转化子中提取质粒,利用冻融法转入农杆菌GV3101感受态细胞,挑取单菌落进行菌落PCR检测,目的条带一致,如图7所示,证明成功转入农杆菌,得到农杆菌阳性转化子,摇菌保存。
实施例3:赖草LsPIN1基因功能的验证
1.LsPIN1亚细胞定位
首先利用在线预测软件https://wolfpsort.hgc.jp/对LsPIN1蛋白进行亚细胞定位预测,结果显示,LsPIN1蛋白可能定位于细胞膜。利用注射烟草下表皮的方法进行亚细胞定位验证,将实施例2构建得到的农杆菌阳性转化子摇菌活化,用烟草悬浮液将农杆菌菌体重悬至OD600为0.9~1.0,室温静置2h后转入烟草下表皮细胞,在烟草表皮细胞进行瞬时转化,激光共聚焦显微镜观察LsPIN1蛋白在烟草表皮细胞中的定位情况。结果如图8所示,绿色荧光蛋白GFP在整个细胞中都有表达,LsPIN1蛋白定位于细胞膜中。
2.利用农杆菌介导的遗传转化方法获得转基因拟南芥
在超净工作台中,将拟南芥种子放入到1mL的7.5%次氯酸钠和0.01%的Triton-X100溶液消毒液中,200rpm振荡10min;灭菌ddH2O反复冲洗5~6次,将种子点播于MS培养基上,晾干后密封,放入4℃冰箱避光培养2~3天进行春化,随后22℃、日照16h/黑暗8h、光强度40μmoL·m-2·s-1的条件下,培养2周后移栽到基质中,每盆移栽九株野生型拟南芥,覆膜一周后揭开,先培养3-4周后浇灌花多多营养液促进其抽薹开花,侵染拟南芥前一天剪掉果荚,浇足水。
将实施例2得到农杆菌阳性转化子接种于YEP培养基(含100ng/mL卡那霉素和100ng/mL利福平抗生素)中,28℃,220rpm振荡培养至菌液OD600至1.2~1.5,将其转移至50mL灭菌离心管中,5000rpm离心10min,收集沉淀菌体,弃液体培养基,再将菌体沉淀用100μmol/L乙酰丁香酮溶液重悬,将菌体重悬至OD600为1.0左右,在无菌的平板中倒入上述重悬的农杆菌菌悬液作为浸染液,采用花浸法将拟南芥全部花序浸泡在侵染液中40~60sec,全部侵染完成后黑暗培养一天,再继续正常生长培养,一周后再次进行侵染,约侵染3~4次,待果荚成熟后收取T1代种子。
3.转化植株的筛选和初步检查
将收获的T1代种子在24℃晾晒一周后,消毒点播于MS固体培养基(含100mg/L潮霉素)中进行T1代转基因拟南芥的筛选,筛选结果如图9所示,发现有些拟南芥根系变长,叶片增大,株高强壮,说明可能获得阳性苗。将阳性苗移栽到营养土中,待其生长一周后用CTAB法提取拟南芥DNA验证是否为阳性苗。用提取的DNA为模板,LsPIN1基因的特异性引物LsPIN1-F-XbaI和LsPIN1-R-SpeI进行PCR验证。验证结果如图10显示,样本均扩增出目的片段条带,说明成功获得T1代转基因拟南芥。将T1代LsPIN1转基因拟南芥继续培养,通过遗传分离比筛选出T3代纯合阳性植株。
4.LsPIN1基因在拟南芥中异位表达对根发育的影响
以野生型拟南芥为对照组,以LsPIN1转基因拟南芥为处理组,将野生型拟南芥和LsPIN1转基因拟南芥的种子分别播种在含有MS培养基的培养皿上,4℃春化3天后,转入温度22℃、日照16h黑暗8h的环境下培养7天,然后将幼苗转移到方形(13×13cm)MS培养基中,垂直培养7天。从转移第1天开始,每天拍照并统计可见的主根长度和侧根数量。对照组和处理组均为三个生物学重复。使用Image J软件测量根系长度,Excel 2019做初步的数据统计并进行柱形图的绘制,使用stst来进行单因素方差分析以及显著性差异分析(p<0.05)。结果如图11所示,通过对各株系主根、侧根进行表型观察可知,LsPIN1转基因拟南芥(35S::LsPIN1)与野生型拟南芥(Col-0)相比,主根明显变长,侧根数目增多,LsPIN1转基因拟南芥比野生型拟南芥主根变长了约20%,侧根数目增多了约40%。表明LsPIN1基因参与调控主根的生长和发育,也影响侧根的发生和形态建成。
5.外源激素GA3(赤霉素)、IAA(吲哚乙酸)对LsPIN1转基因拟南芥根系的影响
以野生型拟南芥为对照组,以LsPIN1转基因拟南芥为处理组,将在MS培养基培养了7天的野生型拟南芥和LsPIN1转基因拟南芥幼苗转移到分别含有0.02mg/LGA3和0.005mg/LIAA的方形(13×13cm)MS培养基中,垂直培养7天。从转移第1天开始,每天拍照并统计可见的主根长度和侧根数量。对照组和处理组均为三个生物学重复。使用Image J软件测量根系长度,Excel 2019做初步的数据统计并进行柱形图的绘制,使用stst来进行单因素方差分析以及显著性差异分析(p<0.05)。结果如图12、13所示,LsPIN1转基因拟南芥(35S::LsPIN1)的主根长度与野生型拟南芥(Col-0)相比无明显变化,但侧根数目差异显著,在GA3的刺激下LsPIN1转基因拟南芥的侧根数量增加10%,在IAA的刺激下LsPIN1转基因拟南芥的侧根数量增加13%。说明LsPIN1基因会受到外源激素GA3和IAA的诱导,并进一步调控拟南芥侧根的发育。
6.LsPIN1蛋白对生长素运输及株系生长的影响
为了验证LsPIN1蛋白是否通过调控生长素的运输影响根系的发育,进行了重力响应和向光性试验。重力响应实验分为向重力实验和微重力实验。
在向重力实验中,将野生型拟南芥和LsPIN1转基因拟南芥的种子在MS培养皿上正常培养5天后,将幼苗转移到方形(13×13cm)MS培养基中,并对拟南芥进行135°逆时针旋转,经过0,2,3,5天的连续刺激,结果如图14所示,与野生型拟南芥(Col-0)相比,LsPIN1转基因拟南芥(35S::LsPIN1)的主根、侧根弯曲严重;用Image J软件测量根系旋转角度发现,在相同温度条件下,野生型拟南芥对重力刺激的响应比LsPIN1转基因拟南芥慢,根系转向差异显著,说明LsPIN1的过表达影响了根的向地性生长。
在微重力实验中,将野生型拟南芥和LsPIN1转基因拟南芥的种子在MS培养皿上正常培养5天后,将幼苗转移到方形(13×13cm)MS培养基中,在微重力低速旋转离心机上(2档速4rpm/min)放置转移到方形(13×13cm)MS培养基的拟南芥,进行7天的微重力刺激,结果如图15所示,LsPIN1转基因拟南芥(35S::LsPIN1)与野生型拟南芥(Col-0)相比,LsPIN1转基因拟南芥对各个方向进行响应,而野生型拟南芥表型不明显,说明LsPIN1基因响应重力信号调控根系的方向性生长。
在向光性实验中,将4日龄的拟南芥转移到24h黑暗环境下生长3天,再用单侧光培养12h,观察野生型拟南芥和LsPIN1转基因拟南芥的下胚轴是否会向光弯曲。用LUX meterTES-1332A(TES测试仪器)测定光照强度,在光照强度为4500LUX(约为60.8μmol/㎡/sec)的条件下完成向光性弯曲试验。单侧白光照射12h用数码相机拍照。然后利用Image J软件测量下胚轴的弯曲角度。对照组和处理组均为三个生物学重复。结果如图16所示,相较于野生型拟南芥(Col-0),LsPIN1转基因拟南芥(35S::LsPIN1,处理组)的下胚轴向光弯曲更严重,说明LsPIN1基因影响拟南芥根系的向光性生长。
7.LsPIN1蛋白对生长素积累的影响
过表达LsPIN1基因对生长素转运的影响表明根系生长素的积累可能也会受到影响,因此,将LsPIN1转化到生长素响应基因的DR5::GUS转基因植株中,研究生长素的积累。将DR5::GUS拟南芥和DR5::GUS-LsPIN1转基因拟南芥在MS培养基中培育两周,待其长出主根和侧根后将拟南芥浸泡在丙酮中固定植株内部细胞,再放进装有GUS染色液的离心管中抽取真空,37℃浸染3-12h后用70%乙醇进行脱色,脱色完毕后(约7-10天)在显微镜下对不同遗传背景的GUS植株进行观察,获得DR5::GUS-LsPIN1转基因拟南芥生长素积累的定位情况。结果如图17所示,与DR5::GUS野生型相比,DR5::GUS-LsPIN1转基因拟南芥的主根、侧根的根尖GUS活性有显著差异。说明LsPIN1的异位表达有助于拟南芥生长素的积累。
综上,本发明发现了赖草生长素输出载体,其编码基因LsPIN1基因的过表达促进了LsPIN1转基因拟南芥中生长素的极性运输和根的发育,并促进了生长素的积累。基于上述研究可知,LsPIN1蛋白作为赖草生长素输出载体,定位于细胞膜中,可以通过膜结构来完成生长素的跨膜运输,对于赖草根系发育和生长素运输具有重要影响。
SEQUENCE LISTING
<110> 济南大学
<120> 一种赖草生长素输出载体及其编码基因与应用
<160> 6
<170> PatentIn version 3.5
<210> 1
<211> 1764
<212> DNA
<213> Leymus secalinus (Georgi) Tzvel.
<400> 1
atgatcacgg gcacggactt ctaccacgtg atgacggcgg tggtgccgct gtacgtggcg 60
atgatcctcg cctacggctc cgtcaagtgg tggggcatct tcacgccgga ccagtgctcc 120
gggatcaacc gcttcgtcgc gctcttcgcc gtgccgctgc tctccttcca cttcatctcc 180
accaacaacc cctacaccat gaacctgcgc ttcatcgccg ccgacacgct gcagaagctc 240
atgatgctcg ccatgctcac cgcctggagc cacctctccc gcagcggcag cctcgagtgg 300
accatcacgc tcttctccct ctccacgctg cccaacacgc tcgtcatggg catcccgctg 360
ctcaagggca tgtacggcga cttctccggc agcctcatgg tgcagatcgt cgtgctgcag 420
tgcatcatct ggtacacgct catgctcttc atgttcgagt accgcggcgc caggatcctc 480
atcacggagc agttccccga caccgccggc gccatcgcct ccatcgccgt cgacccggac 540
gtcatgtcgc tcgacggcag gagggacatg atcgagacgg aggcagaggt gaaggaggac 600
ggcaagatac acgtcacggt gcgccgctcc aacgcgtccc gctccgacat ctactccagg 660
cgctccatgg gcttctccag caccacgccg cgccccagca acctcaccaa cgccgagatc 720
tactcgctgc agtcgtcgcg gaaccccacg cccaggggct ccagcttcaa ccacaccgac 780
ttctactcca tggtcggccg gagctccaac tttggcgccg ccgacgcgta cggcatccgc 840
accggcgcca cgccgcgccc gtctaactac gaggaggacg cgcccaagcc caagcacccc 900
gcgcccgggg cgggacacta cccggcgcct aacccggcgg tggccgcagc gcccaaggga 960
cccaagaagg cggccgcgaa cgggcaggcc aagggcgagg acctccacat gttcgtctgg 1020
agctcgagcg cgtcgccggt gtcggacgtg ttcggcggcg gcgcaccaga ctacaacgac 1080
gccgcggccg ccaagtcccc gcgcaaaatg gatggagcaa aggacaggga ggactacgtg 1140
gagcgagacg acttcagctt cgggaacagg ggcgcgctgg acagggacgc ggaggccggc 1200
gacgagaagg ccatgacggc ggacccgaac aatgcgatga gcgcggggcc gacggcgatg 1260
ccgccgacga gcgtgatgac gcggctgatc ctgatcatgg tgtggcgcaa gctcatccgc 1320
aaccccaaca cctactccag cctcatcggc ctcatctggt cgctcgtctg cttcaggtgg 1380
aacttcacga tgccggcaat cgtcctggga tccatctcca tcctgtcgga tgcagggcta 1440
ggaatggcca tgttcagcct cggtctgttc atggcgctgc agccgcggat catcgcgtgc 1500
gggaacaagg tggcgacgta cgccatggcc gtgcggttcc tcgccggccc ggccgtcatg 1560
accgccgcct ccttcgccgt gggcctccgc ggcacgctcc tgcacgtcgc catcgtccag 1620
gcagcgctgc cgcagggcat tgtccccttc gtcttcgcaa aggagtacag cgtgcaccct 1680
gacattctca gcacagcggt catatttggc atgctcatcg ccctgccgat cacgctcgtc 1740
tactacatcc tgctcggcct gtga 1764
<210> 2
<211> 587
<212> PRT
<213> Leymus secalinus (Georgi) Tzvel.
<400> 2
Met Ile Thr Gly Thr Asp Phe Tyr His Val Met Thr Ala Val Val Pro
1 5 10 15
Leu Tyr Val Ala Met Ile Leu Ala Tyr Gly Ser Val Lys Trp Trp Gly
20 25 30
Ile Phe Thr Pro Asp Gln Cys Ser Gly Ile Asn Arg Phe Val Ala Leu
35 40 45
Phe Ala Val Pro Leu Leu Ser Phe His Phe Ile Ser Thr Asn Asn Pro
50 55 60
Tyr Thr Met Asn Leu Arg Phe Ile Ala Ala Asp Thr Leu Gln Lys Leu
65 70 75 80
Met Met Leu Ala Met Leu Thr Ala Trp Ser His Leu Ser Arg Ser Gly
85 90 95
Ser Leu Glu Trp Thr Ile Thr Leu Phe Ser Leu Ser Thr Leu Pro Asn
100 105 110
Thr Leu Val Met Gly Ile Pro Leu Leu Lys Gly Met Tyr Gly Asp Phe
115 120 125
Ser Gly Ser Leu Met Val Gln Ile Val Val Leu Gln Cys Ile Ile Trp
130 135 140
Tyr Thr Leu Met Leu Phe Met Phe Glu Tyr Arg Gly Ala Arg Ile Leu
145 150 155 160
Ile Thr Glu Gln Phe Pro Asp Thr Ala Gly Ala Ile Ala Ser Ile Ala
165 170 175
Val Asp Pro Asp Val Met Ser Leu Asp Gly Arg Arg Asp Met Ile Glu
180 185 190
Thr Glu Ala Glu Val Lys Glu Asp Gly Lys Ile His Val Thr Val Arg
195 200 205
Arg Ser Asn Ala Ser Arg Ser Asp Ile Tyr Ser Arg Arg Ser Met Gly
210 215 220
Phe Ser Ser Thr Thr Pro Arg Pro Ser Asn Leu Thr Asn Ala Glu Ile
225 230 235 240
Tyr Ser Leu Gln Ser Ser Arg Asn Pro Thr Pro Arg Gly Ser Ser Phe
245 250 255
Asn His Thr Asp Phe Tyr Ser Met Val Gly Arg Ser Ser Asn Phe Gly
260 265 270
Ala Ala Asp Ala Tyr Gly Ile Arg Thr Gly Ala Thr Pro Arg Pro Ser
275 280 285
Asn Tyr Glu Glu Asp Ala Pro Lys Pro Lys His Pro Ala Pro Gly Ala
290 295 300
Gly His Tyr Pro Ala Pro Asn Pro Ala Val Ala Ala Ala Pro Lys Gly
305 310 315 320
Pro Lys Lys Ala Ala Ala Asn Gly Gln Ala Lys Gly Glu Asp Leu His
325 330 335
Met Phe Val Trp Ser Ser Ser Ala Ser Pro Val Ser Asp Val Phe Gly
340 345 350
Gly Gly Ala Pro Asp Tyr Asn Asp Ala Ala Ala Ala Lys Ser Pro Arg
355 360 365
Lys Met Asp Gly Ala Lys Asp Arg Glu Asp Tyr Val Glu Arg Asp Asp
370 375 380
Phe Ser Phe Gly Asn Arg Gly Ala Leu Asp Arg Asp Ala Glu Ala Gly
385 390 395 400
Asp Glu Lys Ala Met Thr Ala Asp Pro Asn Asn Ala Met Ser Ala Gly
405 410 415
Pro Thr Ala Met Pro Pro Thr Ser Val Met Thr Arg Leu Ile Leu Ile
420 425 430
Met Val Trp Arg Lys Leu Ile Arg Asn Pro Asn Thr Tyr Ser Ser Leu
435 440 445
Ile Gly Leu Ile Trp Ser Leu Val Cys Phe Arg Trp Asn Phe Thr Met
450 455 460
Pro Ala Ile Val Leu Gly Ser Ile Ser Ile Leu Ser Asp Ala Gly Leu
465 470 475 480
Gly Met Ala Met Phe Ser Leu Gly Leu Phe Met Ala Leu Gln Pro Arg
485 490 495
Ile Ile Ala Cys Gly Asn Lys Val Ala Thr Tyr Ala Met Ala Val Arg
500 505 510
Phe Leu Ala Gly Pro Ala Val Met Thr Ala Ala Ser Phe Ala Val Gly
515 520 525
Leu Arg Gly Thr Leu Leu His Val Ala Ile Val Gln Ala Ala Leu Pro
530 535 540
Gln Gly Ile Val Pro Phe Val Phe Ala Lys Glu Tyr Ser Val His Pro
545 550 555 560
Asp Ile Leu Ser Thr Ala Val Ile Phe Gly Met Leu Ile Ala Leu Pro
565 570 575
Ile Thr Leu Val Tyr Tyr Ile Leu Leu Gly Leu
580 585
<210> 3
<211> 23
<212> DNA
<213> 人工序列
<400> 3
tcctgcacgt cgccatcgtc cag 23
<210> 4
<211> 23
<212> DNA
<213> 人工序列
<400> 4
gcaggatgta gtagacgagc gtg 23
<210> 5
<211> 29
<212> DNA
<213> 人工序列
<400> 5
gctctagaat gatcacgggc acggacttc 29
<210> 6
<211> 30
<212> DNA
<213> 人工序列
<400> 6
ggactagtca ggccgagcag gatgtagtag 30
Claims (10)
1.一种赖草生长素输出载体,其氨基酸序列如SEQ ID NO.2所示。
2.权利要求1所述的赖草生长素输出载体的编码基因的核苷酸序列如SEQ ID NO.1所示。
3.一种含有权利要求2所述的赖草生长素输出载体的编码基因的重组质粒。
4.如权利要求3所述的重组质粒,其特征在于,所述重组质粒的载体为pEASY-blunt或pCAMBIA1300-GFP。
5.一种含有权利要求2所述的赖草生长素输出载体的编码基因或权利要求3所述的重组质粒的重组菌。
6.如权利要求5所述的重组菌,其特征在于,所述重组菌的宿主为大肠杆菌Trans1-T1或大肠杆菌DH5α或农杆菌GV3101。
7.权利要求1所述的赖草生长素输出载体在促进植株分蘖或调控植株顶芽转向中的应用。
8.如权利要求7所述的应用,其特征在于,所述植株为拟南芥或赖草;
优选的,所述应用是将赖草生长素输出载体的编码基因转化入植株中,实现生长素输出载体的编码基因在植株中的过表达。
9.权利要求1所述的赖草生长素输出载体在与外源激素协同促进植株分蘖中的应用。
10.如权利要求9所述的应用,其特征在于,所述植株为拟南芥或赖草;
优选的,所述外源激素为GA3或IAA。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009105492A2 (en) * | 2008-02-19 | 2009-08-27 | Ceres, Inc. | Transgenic plants having altered nitrogen use efficiency characteristics |
CN101787074A (zh) * | 2009-01-22 | 2010-07-28 | 哈尔滨师范大学 | 羊草水孔蛋白及其编码基因与应用 |
CN108949786A (zh) * | 2018-06-29 | 2018-12-07 | 山东农业大学 | 拟南芥e3泛素连接酶编码基因atl27在调控植物耐盐性能中的应用 |
-
2022
- 2022-04-21 CN CN202210426568.4A patent/CN115010796B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009105492A2 (en) * | 2008-02-19 | 2009-08-27 | Ceres, Inc. | Transgenic plants having altered nitrogen use efficiency characteristics |
CN101787074A (zh) * | 2009-01-22 | 2010-07-28 | 哈尔滨师范大学 | 羊草水孔蛋白及其编码基因与应用 |
CN108949786A (zh) * | 2018-06-29 | 2018-12-07 | 山东农业大学 | 拟南芥e3泛素连接酶编码基因atl27在调控植物耐盐性能中的应用 |
Non-Patent Citations (3)
Title |
---|
SKUPA P, M ET AL: "Characterization of tobacco cell lines transformed with the AtPIN5 gene from the auxin efflux carrier family of arabidopsis", COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY PART A MOLECULAR AND INTEGRATIVE PHYSIOLOGY, vol. 150 * |
张荣华;桂意云;韦金菊;区惠平;刘昔辉;: "作物分蘖特性研究进展", 园艺与种苗, no. 07 * |
杜利霞;赵凯?;王凯鑫;刘伟;钟华;夏方山;朱慧森;: "6-BA引发对赖草种子萌发、贮藏物质和内源激素的影响", 草地学报, no. 03 * |
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