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烟草NCED3基因的克隆及其干旱胁迫表达分析 被引量:18
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作者 牛志强 刘国顺 +5 位作者 师婷婷 张松涛 贾宏昉 张洪映 崔红 杨永霞 《中国烟草学报》 EI CAS CSCD 北大核心 2015年第3期100-106,共7页
为揭示9-顺式环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase,NCED)在烟草生长发育过程中的功能,通过同源克隆方法获得烟草品种K326类胡萝卜素降解关键基因NCED3两个c DNA全长序列。序列分析表明:K326 NCED3-1和NCED3-2基... 为揭示9-顺式环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase,NCED)在烟草生长发育过程中的功能,通过同源克隆方法获得烟草品种K326类胡萝卜素降解关键基因NCED3两个c DNA全长序列。序列分析表明:K326 NCED3-1和NCED3-2基因分别包含一个1842bp和1830bp的开放读码框(ORF),各编码613个和609个氨基酸。预测蛋白质分子量分别为68177.8 Da和67754.2Da,理论等电点(p I)各为7.66和7.28。跨膜区预测、亲水性和信号肽分析表明很可能是定位于线粒体膜上的亲水性跨膜蛋白。二级结构预测模型显示此蛋白结构以β折叠和无规则卷曲为主。蛋白三级结构预测分析表明NCED3-1与NCED3-2空间结构极为相似。PEG6000胁迫分析表明,干旱胁迫可以诱导NCED3基因表达以及内源ABA的积累。且在处理12h之前,NCED3表达与ABA累积速度均呈现升高趋势,之后逐渐降低。研究结果为解析NCED3在烟草抗旱方面的功能提供一定的研究基础。 展开更多
关键词 烟草 nced3基因 克隆 序列分析 表达 ABA
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利用TAIL-PCR方法克隆拟南芥干旱主效基因NCED3 被引量:3
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作者 刘浩 苏凡 王棚涛 《河南大学学报(自然科学版)》 CAS 北大核心 2009年第6期621-625,共5页
利用远红外成像技术筛选得到一株T-DNA插入的干旱敏感突变体,并成功利用TAIL-PCR技术克隆到该突变基因为NCED3,该基因编码植物体内ABA合成的关键限速酶,突变体在干旱胁迫下不能合成ABA而表现出不耐干旱的特性.最后通过PCR及RT-PCR方法... 利用远红外成像技术筛选得到一株T-DNA插入的干旱敏感突变体,并成功利用TAIL-PCR技术克隆到该突变基因为NCED3,该基因编码植物体内ABA合成的关键限速酶,突变体在干旱胁迫下不能合成ABA而表现出不耐干旱的特性.最后通过PCR及RT-PCR方法验证了TAIL-PCR结果的可靠性. 展开更多
关键词 拟南芥 干旱突变体 TAIL-PCR技术 nced3基因
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亚洲棉NCED3基因克隆及其抗旱功能分析 被引量:2
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作者 蔡肖 王海涛 +5 位作者 李兴河 甄军波 刘琳琳 刘迪 迟吉娜 张建宏 《核农学报》 CAS CSCD 北大核心 2022年第9期1713-1722,共10页
9-顺式-环氧类胡萝卜素双加氧酶(NCED)是脱落酸(ABA)合成过程中的关键限速酶,被证实广泛参与植物的生长发育进程及对非生物胁迫的应答。为了挖掘、鉴定棉花抗旱相关的NCEDs基因,本研究克隆了亚洲棉(Gossypium arboreum)GaNCED3基因,利... 9-顺式-环氧类胡萝卜素双加氧酶(NCED)是脱落酸(ABA)合成过程中的关键限速酶,被证实广泛参与植物的生长发育进程及对非生物胁迫的应答。为了挖掘、鉴定棉花抗旱相关的NCEDs基因,本研究克隆了亚洲棉(Gossypium arboreum)GaNCED3基因,利用实时荧光定量PCR(qRT-PCR)技术分析了干旱胁迫下该基因的表达特性;并遗传转化拟南芥(Arabidopsis thaliana),研究了该基因在干旱应答中的功能。结果显示,GaNCED3基因开放阅读框(ORF)全长为1794 bp,其编码的蛋白质包含597个氨基酸。该基因在干旱胁迫处理后上调表达,在处理3 h的根中表达量最高,是对照的27.6倍。在萌发期进行甘露醇模拟的干旱处理,超表达GaNCED3的转基因拟南芥种子萌发率和绿苗率均高于野生型。在幼苗期进行自然干旱处理后,转基因拟南芥植株叶片丙二醛含量显著低于对照(P<0.05),游离脯氨酸积累量显著高于对照(P<0.05)。本研究初步证明了外源超表达GaNCED3基因使植株抗旱能力增强,为进一步研究GaNCED3干旱应答的分子机制提供了理论依据,为抗旱育种提供了候选基因资源。 展开更多
关键词 棉花 干旱 nced3 9-顺式-环氧类胡萝卜素双加氧酶 基因功能
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白菜型冬油菜NCED3基因、启动子的克隆及其表达分析 被引量:3
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作者 武军艳 马骊 +7 位作者 方彦 张亚宏 马学才 刘丽君 牛早霞 李学才 李爱国 孙万仓 《干旱地区农业研究》 CSCD 北大核心 2020年第5期64-70,共7页
从2个不同抗寒性的冬油菜品种中克隆了NCED3基因及其启动子序列,并分析其在叶片和根中的表达,研究NCED3基因在冬油菜中的作用机理。结果表明,陇油6号的NCED3基因开放阅读框(ORF)长度为1794 bp,编码597个氨基酸,分子量65.74 kD,理论等电... 从2个不同抗寒性的冬油菜品种中克隆了NCED3基因及其启动子序列,并分析其在叶片和根中的表达,研究NCED3基因在冬油菜中的作用机理。结果表明,陇油6号的NCED3基因开放阅读框(ORF)长度为1794 bp,编码597个氨基酸,分子量65.74 kD,理论等电点为5.81;天油2号的ORF与其长度相同,分子量为65.78 kD,理论等电点为5.94。2个蛋白都是亲水蛋白,具有疏水峰。根据预测,启动子具有生物过程中常见的顺式作用元件如CAAT-box等、分生组织表达CAT-box相关的顺式作用元件、-30 TATA box附近的核心启动子元件等,还有ABRE、TGA-element、CGTCA-motif、TGACG-element等激素响应元件,此外,还鉴定出低温响应元件(LTR)。2个品种启动子序列的相似度为99.38%,只有1个不同的顺式作用元件,即circadian,推测其与昼夜节律相关。低温、PEG及ABA处理后NCED3在叶片和根中的表达均高于对照,增加范围为0.07~6.48,且均于8 h达到峰值。与根系的表达特性相比,胁迫处理后叶片的基因表达均在2 h显著升高,天油2号的升高幅度(1.54~6.00)均大于陇油6号(0.04~1.95)。 展开更多
关键词 nced3基因 白菜型冬油菜 抗寒性 启动子克隆 序列分析 胁迫 基因相对表达量
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Cloning and molecular evolution of 9-cis-epoxycarotenoid dioxygenase gene (NCED3) in six species of Glycyrrhiza L 被引量:1
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作者 Li Wang Guangxi Ren +2 位作者 Mingyang Chen Jing Chen Chunsheng Liu 《Journal of Traditional Chinese Medical Sciences》 2017年第4期357-365,共9页
Objective:To understand the evolutionary relationship of NCED3 in Glycyrrhiza L.and find genetic evidence of its local adaptation.Meanwhile,lay the foundation for relieving the demand and subsequent starvation of Glyc... Objective:To understand the evolutionary relationship of NCED3 in Glycyrrhiza L.and find genetic evidence of its local adaptation.Meanwhile,lay the foundation for relieving the demand and subsequent starvation of Glycyrrhiza resources.Methods:NCED3 were isolated from six species of Glycyrrhiza L.by rapid amplification of cDNA ends and a homology cloning strategy.The basic properties and evolutionary relationships of NCED3 were analyzed by bioinformatics.Results:Six NCED3 genes formed two clusters;they were under purifying selection and were highly conserved in evolution (Ka/Ks≤).There was very little functional divergence among the NCED3s (Alpha value 0.071).The contents of ABA in the six species were divided into two groups.Conclusion:This study provides the first analysis of the evolution of NCED3s in six species of Glycyrrhiza.Six NCED3 genes were slow evolutionary rates in six NCED3s.Different genetic relationships to Glycyrrhiza adapted to various environments.The changes in ABA content were consistent with the evolution of NCED3.Whether there are direct relationships between NCED3 and environmental adaptation,the answer is uncertain,and we hope to show more positive results in the further study of NCED3 in Glycyrrhiza L. 展开更多
关键词 GLYCYRRHIZA nced3 Evolution Abscisic acid
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NCED3基因的持续诱导及ABA合成与代谢的协同调控在拟南芥ABA信号积累中的作用 被引量:7
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作者 任慧波 范意娟 +6 位作者 魏开发 高志晖 李桂芬 刘静 李冰冰 胡建芳 贾文锁 《科学通报》 EI CAS CSCD 北大核心 2007年第1期59-66,共8页
ABA作为逆境信号在植物抗逆特别是抗旱中起着重要的作用.由于ABA生物合成是ABA信号产生的根本基础,因此ABA合成关键酶基因NCED3的启动一直被认为是操纵ABA信号产生的惟一机制.本研究报道了ABA信号累积中ABA代谢和合成的协同操纵机制.结... ABA作为逆境信号在植物抗逆特别是抗旱中起着重要的作用.由于ABA生物合成是ABA信号产生的根本基础,因此ABA合成关键酶基因NCED3的启动一直被认为是操纵ABA信号产生的惟一机制.本研究报道了ABA信号累积中ABA代谢和合成的协同操纵机制.结果表明,水分胁迫可导致拟南芥叶片中ABA水平急剧增加,且在长期干旱胁迫情况下,ABA累积的最高水平始终处于一个相对稳定的状态.无论是胁迫还是非胁迫状态下,ABA代谢都呈现指数递减规律,且其代谢的半衰期没有太大的变化,这意味着干旱条件下ABA的绝对代谢速率将随ABA水平上升而急剧加快,由此可以推断ABA信号的产生是一个由多酶共同操纵的系统控制,且NCED3的持续诱导是ABA信号稳定积累的前提.进一步研究表明,干旱可诱导一系列ABA合成酶的基因表达,其中包括NCED3,AAO3和ABA3等.伴随ABA的持续积累,NCED3,AAO3和ABA3的基因始终处于诱导表达状态.ABA代谢研究和基因表达分析结果相互印证,共同揭示ABA信号的产生机制是一个由多酶共同参与,且以ABA合成关键酶基因持续诱导为前提的操纵机制,其中ABA代谢在ABA信号的操纵中起着重要的作用. 展开更多
关键词 ABA代谢 ABA积累 nced3 水分胁迫 拟南芥
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Roles of a sustained activation of NCED3 and the synergistic regulation of ABA biosynthesis and catabolism in ABA signal production in Arabidopsis 被引量:12
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作者 REN HuiBo FAN YiJian GAO ZhiHui WEI KaiFa LI GuiFen LIU Jing CHEN Lin LI BingBing HU JianFang JIA WenSuo 《Chinese Science Bulletin》 SCIE EI CAS 2007年第4期484-491,共8页
ABA, acting as a stress signal, plays crucial roles in plant resistance to water stress. Because ABA signal production is based on ABA biosynthesis, the regulation of NCED, a key enzyme in the ABA biosynthesis pathway... ABA, acting as a stress signal, plays crucial roles in plant resistance to water stress. Because ABA signal production is based on ABA biosynthesis, the regulation of NCED, a key enzyme in the ABA biosynthesis pathway, is normally thought of as the sole factor controlling ABA signal production. Here we demonstrate that ABA catabolism in combination with a synergistic regulation of ABA biosynthesis plays a crucial role in governing ABA signal production. Water stress induced a significant accumulation of ABA, which exhibited different patterns in detached and attached leaves. ABA catabolism followed a temporal trend of exponential decay for both basic and stress ABA, and there was little difference in the catabolic half-lives of basic ABA and stress ABA. Thus, the absolute rate of ABA catabolism, i.e. the amount of ABA catabolized per unit time, increases with increased ABA accumulation. From the dynamic processes of ABA biosynthesis and catabolism, it can be inferred that stress ABA accumulation may be governed by a synergistic regulation of all the steps in the ABA biosynthesis pathway. Moreover, to maintain an elevated level of stress ABA sustained activation of NCED3 should be required. This inference was supported by further findings that the genes encoding major enzymes in the ABA biosynthesis pathway, e.g. NCED3, AAO3 and ABA3 were all activated by water stress, and with ABA accumulation progressing, the expressions of NCED3, AAO3 and ABA3 remained activated. Data on ABA catabolism and gene expression jointly indicate that ABA signal production is controlled by a sustained activation of NCED3 and the synergistic regulation of ABA biosynthesis and catabolism. 展开更多
关键词 拟南芥 ABA信号 ABA生物合成 分解代谢 协同调节 nced3 持续致活 水胁迫 干旱
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Nucleotide Variation in the NCED3 Region of Arabidopsis thaliana and its Association Study with Abscisic Acid Content under Drought Stress 被引量:10
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作者 Gang-Ping Hao Xiu-Hai Zhang +2 位作者 Yong-Qin Wang Zhong-Yi Wu Cong-Lin Huang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2009年第2期175-183,共9页
Drought tolerance is a comprehensive quantitative trait that is being understood further at the molecular genetic level. Abscisic acid (ABA) is the main drought-induced hormone that regulates the expression of many ... Drought tolerance is a comprehensive quantitative trait that is being understood further at the molecular genetic level. Abscisic acid (ABA) is the main drought-induced hormone that regulates the expression of many genes related to drought responses. 9-cis-epoxycarotenoid dioxygenase (NCED3) is thought to be a key enzyme in ABA biosynthesis. In this paper, we measured the ABA content increase under drought stress, and sequenced and compared the sequence of AtNCED3 among 22 Arabidopsis thaliana accessions. The results showed that the fold of ABA content increase under drought stress was highly variable among these accessions. High density single nucleotide polymorphism (SNP) and insertion/deletion (indel) were found in the AtNCED3 region, on average one SNP per 87.4 bp and one indel per 502 bp. Nucleotide diversity was significantly lower in the coding region than that in non-coding regions. The results of an association study with ANOVA analysis suggested that the 274th site (P←→S) and the 327th site (P←→R) amino acid variations might be the cause of ABA content increase of 163av accession under drought stress. 展开更多
关键词 abscisic acid Arabidopsis thaliana association study drought stress nced3 region nucleotide variation.
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