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Alleviation by abscisic acid of Al toxicity in rice bean is not associated with citrate efflux but depends on ABI5-mediated signal transduction pathways 被引量:9
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作者 Wei Fan Jia Meng Xu +6 位作者 Pei Wu Zhi Xin Yang He Qiang Lou Wei Wei Chen Jian Fen Jin Shao Jian Zheng Jian Li Yang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2019年第2期140-154,共15页
Under conditions of aluminum(Al) toxicity,which severely inhibits root growth in acidic soils, plants rapidly alter their gene expression to optimize physiological fitness for survival. Abscisic acid(ABA) has been sug... Under conditions of aluminum(Al) toxicity,which severely inhibits root growth in acidic soils, plants rapidly alter their gene expression to optimize physiological fitness for survival. Abscisic acid(ABA) has been suggested as a mediator between Al stress and gene expression, but the underlying mechanisms remain largely unknown. Here,we investigated ABA-mediated Al-stress responses, using integrated physiological and molecular biology approaches.We demonstrate that Al stress caused ABA accumulation in the root apex of rice bean(Vigna umbellata [Thunb.] Ohwi &Ohashi), which positively regulated Al tolerance. However,this was not associated with known Al-tolerance mechanisms. Transcriptomic analysis revealed that nearly one-third of the responsive genes were shared between the Al-stress and ABA treatments. We further identified a transcription factor, ABI5, as being positively involved in Al tolerance. Arabidopsis abi5 mutants displayed increased sensitivity to Al, which was not related to the regulation of AtALMT1 and AtMATE expression. Functional categorization of ABI5-mediated genes revealed the importance of cell wall modification and osmoregulation in Al tolerance, a finding supported by osmotic stress treatment on Al tolerance. Our results suggest that ABA signal transduction pathways provide an additional layer of regulatory control over Al tolerance in plants. 展开更多
关键词 Alleviation by abscisic acid of Al toxicity in rice bean IS not associated with CITRATE EFFLUX BUT depends on abi5-mediated sign ABA Figure abi
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黄瓜逆境胁迫响应基因CsABI5的克隆与表达分析
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作者 陈岳 张微微 +2 位作者 姚丹青 潘明 费国强 《山东农业科学》 北大核心 2024年第8期30-36,共7页
脱落酸不敏感蛋白5(abscisic acid insensitive 5,ABI5)属于bZIP转录因子家族,广泛参与植物生长发育和逆境胁迫响应。本研究采用同源克隆的方法获得了黄瓜CsABI5基因,对其进行了生物信息学分析和组织特异性表达分析,并分析了其在盐胁迫... 脱落酸不敏感蛋白5(abscisic acid insensitive 5,ABI5)属于bZIP转录因子家族,广泛参与植物生长发育和逆境胁迫响应。本研究采用同源克隆的方法获得了黄瓜CsABI5基因,对其进行了生物信息学分析和组织特异性表达分析,并分析了其在盐胁迫和ABA处理下的表达情况。结果表明,CsABI5基因的CDS为1230 bp,编码409个氨基酸,其编码蛋白的分子量为44.88 kDa,理论等电点为9.53,亲水性平均系数为-0.603,属于亲水性碱性蛋白。多序列比对显示该蛋白与其他物种的ABI5蛋白具有较高的相似性,C端均含有典型的保守bZIP结构域。系统发育树显示葫芦科的ABI5聚为一支,CsABI5与甜瓜的CmABI5亲缘关系最近。实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)测定发现,CsABI5在黄瓜营养器官和生殖器官中均有表达,在果刺中相对表达量最高。四叶期幼苗受100 mmol/L NaCl和100μmol/L ABA处理后CsABI5表达量显著上调,均在处理6 h升至最高。本研究结果可为抗逆黄瓜品种的选育提供参考。 展开更多
关键词 黄瓜 abi5蛋白 盐胁迫 生物信息学分析 表达特性
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甘蓝型油菜转录因子BnaABI5表达特征分析及互作蛋白鉴定
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作者 奥倩倩 陆方筱 +5 位作者 杨柳卿 李春 翟增康 贾东晔 江元清 杨博 《遗传》 CAS CSCD 北大核心 2024年第9期737-749,共13页
甘蓝型油菜(Brassica napus L.)是重要的油料作物之一,但是其种植效益在西北地区经常受到干旱等环境逆境的影响。脱落酸(abscisic acid,ABA)信号通路在植物对非生物胁迫的响应与耐受中具有重要作用,而ABFs/AREBs(ABA-responsive element... 甘蓝型油菜(Brassica napus L.)是重要的油料作物之一,但是其种植效益在西北地区经常受到干旱等环境逆境的影响。脱落酸(abscisic acid,ABA)信号通路在植物对非生物胁迫的响应与耐受中具有重要作用,而ABFs/AREBs(ABA-responsive element binding factors/ABA-responsive element binding proteins)是调控ABA响应基因表达的核心转录因子。为了解析油菜响应逆境的关键转录因子,本研究首先对甘蓝型油菜转录因子BnaABI5(abscisic acid insensitive 5)进行了亚细胞定位、逆境响应及组织器官表达的检测、转录活性分析以及与激酶BnaMPKs(mitogen-activated protein kinases)的互作筛选。结果显示:BnaABI5-GFP融合蛋白定位于细胞核,该基因响应干旱逆境且主要在油菜的根中表达,并且在酵母体系中呈现出一定的转录激活活性;随后利用烟草瞬时表达体系发现BnaABI5能够激活靶标基因EM6(early methionine-labeled 6)的启动子活性。通过双分子荧光互补(bimolecular fluorescence complementation,BiFC)及酵母双杂交(yeast two-hybrid,Y2H)实验,发现BnaABI5与BnaMPK6及BnaMPK13互作。本研究初步探索了转录因子BnaABI5的表达特征及与BnaMPKs的互作,对于深入理解BnaABI5的功能具有一定的指导意义。 展开更多
关键词 甘蓝型油菜 abi5 MPK 互作蛋白
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The NAC Transcription Factor ANAC089 Modulates Seed Vigor through the ABI5-VTC2 Module in Arabidopsis thaliana
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作者 Yuan Tian Lulu Zhi +1 位作者 Ping Li Xiangyang Hu 《Phyton-International Journal of Experimental Botany》 SCIE 2024年第6期1101-1116,共16页
Seed viability is an essential feature for genetic resource conservation as well as sustainable crop production.Long-term storage induces seed viability deterioration or seed aging,accompanied by the accumulation of t... Seed viability is an essential feature for genetic resource conservation as well as sustainable crop production.Long-term storage induces seed viability deterioration or seed aging,accompanied by the accumulation of toxic reactive oxygen species(ROS)to suppress seed germination.Controlled deterioration treatment(CDT)is a gen-eral approach for mimicking seed aging.The transcription factor ANAC089 was previously reported to modulate seed primary germination.In this study,we evaluated the ability of ANAC089 to control seed viability during aging.Compared with that in the wild-type line,the mutation of ANAC089 significantly increased H_(2)O_(2),thereby reducing seed viability after CDT,while the overexpression of ANAC089 reduced H_(2)O_(2) and improved seed long-evity,indicating a critical role for ANAC089 in maintaining seed viability through H_(2)O_(2) signaling.A series of stu-dies have shown that ANAC089 targets and negatively regulates the level of ABI5,an important transmitter of abscisic acid(ABA)signals,to affect seed viability after CDT.Furthermore,ABI5 negatively regulated the expres-sion of VTC2,which is involved in the biosynthesis of the antioxidant ascorbic acid and H_(2)O_(2) scavenging.As a result,ANAC089 attenuates the generation of H_(2)O_(2),thereby enhancing seed viability through the ABI5-VTC2 module during the seed aging process.Taken together,our results reveal a novel mechanism by which ANAC089 enhances seed viability by coordinating ABI5 and VTC2 expression,ultimately preventing the overac-cumulation of H_(2)O_(2),which would have led to reduced seed viability. 展开更多
关键词 ARabiDOPSIS seed aging ANAC089 abi5 VTC2 H_(2)O_(2)
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拟南芥ABF1及ABI5在不同处理下的表达模式分析
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作者 陈欣 薛才 朱培源 《中国食品工业》 2024年第14期124-126,共3页
本文以7d龄哥伦比亚野生型拟南芥为研究对象,探究在不同NaCl、山梨醇、ABA、蔗糖处理下ABF1及ABI5的表达模式。qRT-PCR结果表明,与对照组相比,ABF1及ABI5的mRNA含量随NaCl浓度的增高逐渐降低,100mM及150mM NaCl处理下,ABF1的mRNA含量降... 本文以7d龄哥伦比亚野生型拟南芥为研究对象,探究在不同NaCl、山梨醇、ABA、蔗糖处理下ABF1及ABI5的表达模式。qRT-PCR结果表明,与对照组相比,ABF1及ABI5的mRNA含量随NaCl浓度的增高逐渐降低,100mM及150mM NaCl处理下,ABF1的mRNA含量降至对照组的0.11和0.08倍,而ABI5的mRNA含量分别降至对照组的0.01和0.02倍;50mM的山梨醇处理下,ABF1及ABI5的mRNA含量增至对照组的3.7和1.32倍,100mM和150mM的山梨醇处理下,ABF1的mRNA含量分别增至3.32和1.72倍,ABI5的mRNA含量分别增至2.87和1.42倍;在50mM的蔗糖处理下,ABF1的mRNA含量降至对照组的0.91倍,此时ABI5的mRNA含量增至1.32倍,100mM和150mM的蔗糖处理下,ABF1的mRNA含量降至对照组0.57和0.15倍,ABI5的mRNA含量降至0.67和0.38倍。在10、30及50μM ABA处理下,ABF1的mRNA含量分别增至对照组的5.32、4.14及4.6倍,ABI5的mRNA含量分别增至8.53、7.2及11.5倍;在4℃和16℃处理下,ABF1的mRNA含量为对照组的0.86和3.02倍,而ABI5的mRNA含量为2.92和1.75倍。这些结果均表明ABF1及ABI5响应高盐、低温和ABA等过程。 展开更多
关键词 ABF1 abi5 拟南芥 非生物胁迫 基因表达
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拟南芥ABI5亚家族基因表达特性的比较研究 被引量:5
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作者 王新文 王世伟 +2 位作者 苟琦敏 王永刚 马建忠 《西北植物学报》 CAS CSCD 北大核心 2021年第7期1101-1108,共8页
拟南芥ABI5亚家族转录因子AtDPBFs(DC3 promoter binding factors)/ABFs(ABRE binding factors)由9个高度同源的成员组成,它们参与了植物生长发育过程中种子成熟和休眠、开花时间、根的生长、ABA信号转导和逆境响应等过程。该研究采用qR... 拟南芥ABI5亚家族转录因子AtDPBFs(DC3 promoter binding factors)/ABFs(ABRE binding factors)由9个高度同源的成员组成,它们参与了植物生长发育过程中种子成熟和休眠、开花时间、根的生长、ABA信号转导和逆境响应等过程。该研究采用qRT-PCR方法分析了拟南芥ABI5亚家族9个基因在不同组织和生长阶段、以及苗期对ABA、高盐、高渗和低温等响应下mRNA的相对含量变化,以明确9个基因的生物学功能及其异同。结果显示:(1)在拟南芥种子刚萌发时有6个基因较5 d龄幼苗均极显著大量表达;在营养生长阶段,未检测到At5G42910基因的信号,其余8个基因均随幼苗生长mRNA相对含量也相应增加,且AtDPBF2和AtDPBF4的增加幅度最大;13 d龄幼苗中ABF3的相对含量最高,约为ABI5的59倍。(2)在生殖生长阶段,茎和根中9个基因均少量表达,而在花和种子中均大量表达,且ABI5表达量最高,分别约为根中表达量的122倍和730倍;叶片中ABF2表达量最高,果荚中AtDPBF2表达量最高,两者分别约为根中表达量的10倍和12倍;At5G42910基因在花和果荚中高表达,约为其他器官表达量的5倍。(3)100 mmol·L^(-1) NaCl、50μmol·L^(-1)山梨醇、10μmol·L^(-1)ABA分别处理8 d后,各处理幼苗生长发育较对照明显减缓,且随着处理浓度上升抑制效应进一步加剧。(4)20μmol·L^(-1) ABA处理13 d龄拟南芥幼苗后,ABF1和ABI5基因的mRNA相对含量增加最高,二者的增幅均接近30倍;100 mmol·L^(-1)山梨醇处理使ABF1和ABI5基因的mRNA相对含量增加最高,两者的增幅分别接近120倍和30倍;100 mmol·L^(-1) NaCl处理导致大多数ABI5亚家族基因表达下调,但仅ABF3基因有较明显的上调;4℃低温处理使ABF1和ABI5基因的mRNA相对含量分别上升了约110倍和25倍。研究表明,在营养生长阶段ABI5亚家族8个成员的表达逐渐增加,进入生殖生长阶段后各基因的表达大幅上升且存在组织差异性,花和种子中9个基因均大量表达而根和茎中少量表达,叶片中ABFs基因发挥主要作用,在种子形成及成熟过程中ABI5和AtDPBF2基因发挥主要作用;在非生物胁迫响应方面,ABF1和ABI5主要响应ABA和渗透胁迫,ABF3主要响应盐胁迫,应对冷胁迫时除ABF1外,ABI5也发挥重要作用,推测At5G42910基因在种子形成以及成熟过程中具有重要作用。 展开更多
关键词 拟南芥 abi5亚家族 转录因子 基因表达 ABA 逆境
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拟南芥ABI5基因对BR胁迫响应及其对下胚轴生长的调节作用 被引量:3
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作者 林建辉 刘自广 +6 位作者 张楠 严容 张楠 何鑫淼 王文涛 刘娣 吴娟 《遗传》 CAS CSCD 北大核心 2021年第9期901-909,共9页
植物脱落酸不敏感蛋白5(abscisic acid-insensitive 5,ABI5)是种子中大量表达的碱性亮氨酸拉链类型(basic leucine zipper,bZIP)转录因子,在调节种子萌发和幼苗早期生长的脱落酸(abscisic acid,ABA)信号中起着核心作用。油菜素内酯(bras... 植物脱落酸不敏感蛋白5(abscisic acid-insensitive 5,ABI5)是种子中大量表达的碱性亮氨酸拉链类型(basic leucine zipper,bZIP)转录因子,在调节种子萌发和幼苗早期生长的脱落酸(abscisic acid,ABA)信号中起着核心作用。油菜素内酯(brassinosteroid,BR)是一种新型植物内源激素,具有调节植物生长发育和逆境胁迫响应等诸多生理功能。近期研究发现,油菜素内酯胁迫条件下,BR信号通路中BIN2(BRASSINOSTEROID INSENSITIVE2)和BES1(BRI1-EMS-SUPPRESSOR 1)通过抑制ABI5表达,促进拟南芥(Arabidopsis thaliana)种子萌发。为进一步探究BR胁迫下ABI5功能,本研究分析了种子萌发期ABI5表达特性,鉴定出拟南芥ABI5基因缺失突变体abi5-1并对BR胁迫下其功能进行解析。结果表明:ABI5在拟南芥干种子中大量表达并响应萌发期BR胁迫;正常条件下,abi5-1与野生型幼苗下胚轴无明显差异;BR胁迫下,abi5-1幼苗下胚轴明显长于野生型。本研究结果揭示了ABI5调控BR胁迫下拟南芥下胚轴生长,为深入了解ABI5调节植物发育的分子机制提供了依据。 展开更多
关键词 abi5 油菜素内酯 拟南芥 下胚轴
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植物ABI5转录因子的研究进展 被引量:1
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作者 李鲁华 王忠妮 +3 位作者 王文新 王怀玉 任明见 徐如宏 《山东农业大学学报(自然科学版)》 北大核心 2019年第4期537-541,共5页
植物脱落酸不敏感蛋白5(Abscisic acid-insensitive ,ABI5)为碱性亮氨酸拉链类型(Basic leucine zipper,bZIP)的转录因子,在ABA信号途径中起着重要作用。目前已经报道的ABI5功能研究主要是以模式植物拟南芥(Arabidopsis thaliana)为研... 植物脱落酸不敏感蛋白5(Abscisic acid-insensitive ,ABI5)为碱性亮氨酸拉链类型(Basic leucine zipper,bZIP)的转录因子,在ABA信号途径中起着重要作用。目前已经报道的ABI5功能研究主要是以模式植物拟南芥(Arabidopsis thaliana)为研究对象,而在小麦(Triticum aestivum)、水稻(Oryza sativa)等农作物中的研究报道极少。本文概述了植物中ABI5在种子发育、植物生长(特别是蔗糖信号途径和蛋白质泛素化过程)、花青素积累等生物学过程以及植物对干旱、低氮等逆境胁迫响应方面的最新研究进展,阐明在农作物中开展ABI5分子网络调控解析工作的重要性,以期为选育具有种子萌发可控(如抗穗发芽)、抗逆性强(如低氮)以及高产量等优良性状的作物品种提供理论基础,对作物分子育种具有指导意义。 展开更多
关键词 abi5 转录因子 研究进展
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拟南芥AFP4的克隆、原核表达和纯化及其与ABI5的互作
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作者 邓子兵 邱梁堃 马建忠 《浙江农业学报》 CSCD 北大核心 2018年第12期2072-2080,共9页
拟南芥ABI5相互作用蛋白AFP4(ABI five binding protein 4)是植物中的一个小分子蛋白,其表达特性与ABI5相似,受ABA诱导。以拟南芥Arabidopsis thaliana L.(Heyn)cv.Columbia为材料,通过PCR扩增了AFP4基因的完整编码序列。扩增片段插入... 拟南芥ABI5相互作用蛋白AFP4(ABI five binding protein 4)是植物中的一个小分子蛋白,其表达特性与ABI5相似,受ABA诱导。以拟南芥Arabidopsis thaliana L.(Heyn)cv.Columbia为材料,通过PCR扩增了AFP4基因的完整编码序列。扩增片段插入到原核表达载体pET-32a(+)的多克隆位点Eco RⅠ和XhoⅠ酶切位点之间和酵母双杂交载体pGBKT7的多克隆位点Eco RⅠ和PstⅠ酶切位点之间。将ABI5基因的编码序列插入到酵母双杂交载体pGADT7的多克隆位点Eco RⅠ和PstⅠ酶切位点之间。重组质粒测序结果表明,所克隆的AFP4和ABI5编码区序列分别与NCBI数据库收录的AFP4基因(Gen Bank登录号NM_111081.2)和ABI5基因(Gen Bank登录号NM_129185.3)完全一致。重组原核表达载体含有AFP4片段的重组融合蛋白的理论分子量为53.4 ku。将重组质粒转化至大肠埃希菌表达菌株BL21 Star(DE3)中诱导表达,并经Ni-NTA亲和层析柱分离纯化、SDS-PAGE分析和Western blot鉴定。将含有AFP4和ABI5的酵母双杂交重组质粒共同转化酿酒酵母AH109中进行酵母双杂交。结果表明,重组融合蛋白在大肠埃希菌E.coli BL21 Star(DE3)中表达的适宜条件为:IPTG浓度为0.4 mmol·L^(-1)、25℃下诱导表达6 h。在上述条件下,重组融合蛋白占细胞破碎后上清液总蛋白的41.6%。经Ni-NTA亲和层析柱纯化后,AFP4融合蛋白在SDS-PAGE分析时呈现单一条带。该条带经过抗6xHis标签肽的抗体分析,呈阳性。该条带经酵母双杂交分析结果表明,AFP4和ABI5在酵母细胞中可以相互作用。 展开更多
关键词 拟南芥 AFP4 原核表达 WESTERNBLOT 酵母双杂交 abi5
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S-Nitrosoglutathion Reductase Activity Modulates the Thermotolerance of Seeds Germination by Controlling ABI5 Stability under High Temperature 被引量:4
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作者 Wenjie Wei Yulan Hu +4 位作者 Wenjuan Yang Xiaoli Li Jiali Wei Xiangyang Hu Ping Li 《Phyton-International Journal of Experimental Botany》 SCIE 2021年第4期1075-1087,共13页
Seed germination or dormancy status is strictly controlled by endogenous phytohormone and exogenous environment signals.Abscisic acid(ABA)is the important phytohormone to suppress seed germination.Ambient high tempera... Seed germination or dormancy status is strictly controlled by endogenous phytohormone and exogenous environment signals.Abscisic acid(ABA)is the important phytohormone to suppress seed germination.Ambient high temperature(HT)also suppressed seed germination,or called as secondary seed dormancy,through upregulating ABI5,the essential component of ABA signal pathway.Previous result shows that appropriate nitric oxide(NO)breaks seed dormancy through triggering S-nitrosoglutathion reductase(GSNOR1)-dependent S-nitrosylation modification of ABI5 protein,subsequently inducing the degradation of ABI5.Here we found that HT induced the degradation of GSNOR1 protein and reduced its activity,thus accumulated more reactive nitrogen species(RNS)to damage seeds viability.Furthermore,HT increased the S-nitrosylation modification of GSNOR1 protein,and triggered the degradation of GSNOR1,therefore stabilizing ABI5 to suppress seed germination.Consistently,the ABI5 protein abundance was lower in the transgenic line overexpressing GSNOR1,but higher in the gsnor mutant after HT stress.Genetic analysis showed that GSNOR1 affected seeds germination through ABI5 under HT.Taken together,our data reveals a new mechanism by which HT triggers the degradation of GSNOR1,and thus stabilizing ABI5 to suppress seed germination,such mechanism provides the possibility to enhance seed germination tolerance to HT through genetic modification of GNSOR1. 展开更多
关键词 Seed germination ambient high temperature GSNOR1 S-nitrosoglutathion abi5
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The MdABI5 transcription factor interacts with the MdNRT1.5/MdNPF7.3 promoter to fine-tune nitrate transport from roots to shoots in apple 被引量:1
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作者 Ya-Jing Liu Ning Gao +6 位作者 Qi-Jun Ma Jiu-Cheng Zhang Xun Wang Jing Lu Yu-Jin Hao Xiao-Fei Wang Chun-Xiang You 《Horticulture Research》 SCIE 2021年第1期3131-3143,共13页
Nitrate is a major nitrogen resource for plant growth and development and acts as both a crucial nutrient and a signaling molecule for plants;hence,understanding nitrate signaling is important for crop production.Absc... Nitrate is a major nitrogen resource for plant growth and development and acts as both a crucial nutrient and a signaling molecule for plants;hence,understanding nitrate signaling is important for crop production.Abscisic acid(ABA)has been demonstrated to be involved in nitrate signaling,but the underlying mechanism is largely unknown in apple.In this study,we found that exogenous ABA inhibited the transport of nitrate from roots to shoots in apple,and the transcription of the nitrate transporter MdNRT1.5/MdNPF7.3 was noticeably reduced at the transcriptional level by ABA,which inhibited the transport of nitrate from roots to shoots.Then,it was found that the ABA-responsive transcription factor MdABI5 bound directly to the ABRE recognition site of the MdNRT1.5 promoter and suppressed its expression.Overexpression of MdABI5 inhibited ABA-mediated transport of nitrate from roots to shoots.Overall,these results demonstrate that MdABI5 regulates the transport of nitrate from roots to shoots partially by mediating the expression of MdNRT1.5,illuminating the molecular mechanism by which ABA regulates nitrate transport in apple. 展开更多
关键词 abi5 nitrate ROOTS
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利用CRISPR/Cas9技术编辑MODD增强水稻休眠性
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作者 郭乃辉 张文忠 +1 位作者 圣忠华 胡培松 《中国农业科学》 CAS CSCD 北大核心 2024年第2期227-235,共9页
【目的】休眠是水稻重要的农艺性状。适当的休眠可以抑制水稻的穗发芽现象,是确保产量和品质的关键因素。然而,水稻休眠调控的基因及其调控网络仍需进一步研究。已知基因MODD编码未知功能的蛋白,负向调控水稻脱落酸信号和抗旱性,但其调... 【目的】休眠是水稻重要的农艺性状。适当的休眠可以抑制水稻的穗发芽现象,是确保产量和品质的关键因素。然而,水稻休眠调控的基因及其调控网络仍需进一步研究。已知基因MODD编码未知功能的蛋白,负向调控水稻脱落酸信号和抗旱性,但其调控水稻休眠的功能未知。研究MODD在调控水稻休眠中的功能,有助于完善水稻休眠调控网络,同时为抗穗发芽遗传育种提供新的理论基础和种质资源。【方法】根据RGAP数据库公布的基因序列,构建MODD的CRISPR-Cas9敲除载体,通过农杆菌介导的遗传转化方法转化中花11(ZH11)愈伤组织,从而获得水稻转基因植株;利用PCR扩增、测序技术及qRT-PCR技术筛选并鉴定MODD敲除纯合系;根据2个突变系(KO-1和KO-2)的CDS得到突变系的氨基酸序列,然后,用DNAMAN对比ZH11和2个突变系(KO-1和KO-2)的蛋白序列;利用Linux系统筛选出MODD在水稻中的同源基因;取开花后35d的种子,调查ZH11和敲除系的发芽率;利用酵母单杂和LUC试验验证MODD的上游基因。【结果】查找到水稻中有6个MODD的同源基因,分别为LOC_Os07g41160、LOC_Os03g30570、LOC_Os03g53630、LOC_Os04g35430、LOC_Os03g17050和LOC_Os06g01170;成功构建了敲除载体,并转入ZH11中,获得2个纯合突变系(KO-1和KO-2);qRT-PCR结果表明,2个突变系(KO-1和KO-2)的MODD表达量显著降低;蛋白序列分析表明,KO-1和KO-2的移码突变造成了蛋白翻译的提前终止;发芽率结果显示,2个突变系(KO-1和KO-2)的发芽率在吸水第3天比ZH11分别显著降低了15%和15%;之后差异逐渐扩大,在第6天差异达到最大,比ZH11分别显著降低35%和35%;2个突变系(KO-1和KO-2)的穗发芽现象显著低于ZH11;酵母单杂试验结果表明,在酵母中,ABI5可以结合MODD的启动子区域,并且进一步把结合范围缩小至300 bp以内;LUC结果显示,加入ABI5的荧光值是单独加NONE空载荧光值的2.6倍,说明ABI5可以激活MODD的表达。【结论】敲除MODD可以增加种子的休眠,MODD可能通过ABA信号途径调控种子休眠。 展开更多
关键词 水稻 休眠 abi5 MODD
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天山云杉ABI5基因的克隆与原核表达活性测定
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作者 范晓聪 王佳琰 +4 位作者 阮晓 奚雨欣 杨丽 吴庆飞 王强 《分子植物育种》 CAS 北大核心 2024年第9期2898-2904,共7页
ABI5又名植物脱落酸不敏感蛋白5(Abscisic acid-insensitive 5),作为ABA信号通路中重要的转录因子,协调参与了种子发育和萌发、植物生长、逆境胁迫等方面的调控。目前的研究多是以拟南芥为对象,而在天山云杉等木本植物上报道极少。本研... ABI5又名植物脱落酸不敏感蛋白5(Abscisic acid-insensitive 5),作为ABA信号通路中重要的转录因子,协调参与了种子发育和萌发、植物生长、逆境胁迫等方面的调控。目前的研究多是以拟南芥为对象,而在天山云杉等木本植物上报道极少。本研究以天山云杉的cDNA为模板进行了PCR扩增,得到了天山云杉ABI5序列,经测序发现其全长为1071 bp,共编码了352个氨基酸。将ABI5基因连接pET-24a原核表达载体,得到了重组质粒pET-24a-ABI5。对IPTG浓度、诱导时间、培养基类型和孵育温度进行了优化,得到了最适培养条件为采用LB培养基,IPTG终浓度为0.2 mmol/L,20℃下诱导8 h。通过SDS-PAGE检测纯化产物发现大部分蛋白以包涵体形式存在,对部分包涵体蛋白进行了复性并利用孔雀石绿显色反应进行了活性的验证。 展开更多
关键词 天山云杉 abi5 分子克隆 原核表达 包涵体复性
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OsNAC5 orchestrates OsABI5 to fine-tune cold tolerance in rice 被引量:1
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作者 Ruiqing Li Yue Song +8 位作者 Xueqiang Wang Chenfan Zheng Bo Liu Huali Zhang Jian Ke Xuejing Wu Liquan Wu Ruifang Yang Meng Jiang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2024年第4期660-682,共23页
Due to its tropical origins,rice(Oryza sativa)is susceptible to cold stress,which poses severe threats to production.OsNAC5,a NAC-type transcription factor,participates in the cold stress response of rice,but the deta... Due to its tropical origins,rice(Oryza sativa)is susceptible to cold stress,which poses severe threats to production.OsNAC5,a NAC-type transcription factor,participates in the cold stress response of rice,but the detailed mechanisms remain poorly understood.Here,we demonstrate that OsNAC5 positively regulates cold tolerance at germination and in seedlings by directly activating the expression of ABSCISIC ACID INSENSITIVE 5(OsABI5).Haplotype analysis indicated that single nucleotide polymorphisms in a NAC-binding site in the OsABI5 promoter are strongly associated with cold tolerance.OsNAC5 also enhanced OsABI5 stability,thus regulating the expression of cold-responsive(COR)genes,enabling fine-tuned control of OsABI5 action for rapid,precise plant responses to cold stress.DNA affinity purification sequencing coupled with transcriptome deep sequencing identified several OsABI5 target genes involved in COR expression,including DEHYDRATION-RESPONSIVE ELEMENT BINDING FACTOR 1A(OsDREB1A),OsMYB20,and PEROXIDASE 70(OsPRX70).In vivo and in vitro analyses suggested that OsABI5 positively regulates COR gene transcription,with marked COR upregulation in OsNAC5-overexpressing lines and downregulation in osnac5 and/or osabi5 knockout mutants.This study extends our understanding of cold tolerance regulation via OsNAC5 through the OsABI5-CORs transcription module,which may be used to ameliorate cold tolerance in rice via advanced breeding. 展开更多
关键词 ABA signaling abi5 cold tolerance DREB NAC5 RICE
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The BHLH Transcriptional Factor PIF4 Competes with the R2R3-MYB Transcriptional Factor MYB75 to Fine-Tune Seeds Germination under High Glucose Stress 被引量:2
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作者 Xiaoli Li Shiyan Lu +4 位作者 Yaru Yang Wenjie Wei Jiali Wei Xiaojun Yuan Ping Li 《Phyton-International Journal of Experimental Botany》 SCIE 2021年第5期1387-1400,共14页
It is known that the high level of sugar including glucose suppresses seed germination through ABA signal.ABI5 is an essential component to mediate ABA-dependent seed germination inhibition,but underlying mechanism ne... It is known that the high level of sugar including glucose suppresses seed germination through ABA signal.ABI5 is an essential component to mediate ABA-dependent seed germination inhibition,but underlying mechanism needs more investigation.Previous study demonstrated the PIF4 activated the expression of ABI5 to suppress seed germination in darkness.Here we reported that PIF4 also mediated the seed germination inhibition through ABI5 under high concentration of glucose treatment.Furthermore,we found that PIF4 interacted with PAP1,the central factor to control anthocyanin biosynthesis.Such interaction was confirmed in vitro and in planta.Biochemical and physiological analysis revealed that PAP1 bond the promoter of ABI5 to suppress its expression,thus enhanced seed germination under high concentration of glucose treatment.Specially,PAP1 competed with PIF4 to antagonize the activation of PIF4 on ABI5 expression,thus promoted seed germination under high glucose treatment.Given these,we uncover a novel role for PIF4 and PAP1 in controlling seed germination under high glucose treatment,and reveal their antagonistic mechanism by which coordinates ABI5 expression to control seed germination in response to the glucose signal. 展开更多
关键词 Seeds germination GLUCOSE MYB75 PIF4 abi5
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The GATA transcription factor TaGATA1 recruits demethylase TaELF6-A1 and enhances seed dormancy in wheat by directly regulating TaABI5 被引量:3
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作者 Xuening Wei Yuyan Li +4 位作者 Xiuliang Zhu Xin Liu Xingguo Ye Miaoping Zhou Zengyan Zhang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2023年第5期1262-1276,共15页
Seed dormancy is an important agronomic trait in crops, and plants with low dormancy are prone to preharvest sprouting(PHS) under high-temperature and humid conditions. In this study,we report that the GATA transcript... Seed dormancy is an important agronomic trait in crops, and plants with low dormancy are prone to preharvest sprouting(PHS) under high-temperature and humid conditions. In this study,we report that the GATA transcription factor TaGATA1 is a positive regulator of seed dormancy by regulating TaABI5 expression in wheat.Our results demonstrate that TaGATA1 overexpression significantly enhances seed dormancy and increases resistance to PHS in wheat. Gene expression patterns, abscisic acid(ABA) response assay, and transcriptome analysis all indicate that TaGATA1 functions through the ABA signaling pathway. The transcript abundance of TaABI5, an essential regulator in the ABA signaling pathway,is significantly elevated in plants overexpressing TaGATA1. Chromatin immunoprecipitation assay(ChIP) and transient expression analysis showed that TaGATA1 binds to the GATA motifs at the promoter of TaABI5 and induces its expression.We also demonstrate that TaGATA1 physically interacts with the putative demethylase TaELF6-A1, the wheat orthologue of Arabidopsis ELF6.ChIP–qPCR analysis showed that H3K27me3 levels significantly decline at the TaABI5 promoter in the TaGATA1-overexpression wheat line and that transient expression of TaELF6-A1 reduces methylation levels at the TaABI5 promoter, increasing TaABI5 expression. These findings reveal a new transcription module, including TaGATA1–TaELF6-A1–TaABI5, which contributes to seed dormancy through the ABA signaling pathway and epigenetic reprogramming at the target site. TaGATA1 could be a candidate gene for improving PHS resistance. 展开更多
关键词 ABSCISIC ACID INSENSITIVE5(abi5) common wheat DEMETHYLASE GATA transcription factor preharvest sprouting seed dormancy
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The DELLA-ABI4-HY5 module integrates light and gibberellin signals to regulate hypocotyl elongation 被引量:2
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作者 Haibo Xiong Dandan Lu +9 位作者 Zhiyuan Li Jianghao Wu Xin Ning Weijun Lin Zechen Bai Canhui Zheng Yang Sun Wei Chi Lixin Zhang Xiumei Xu 《Plant Communications》 SCIE CSCD 2023年第5期223-233,共11页
Plant growth is coordinately controlled by various environmental and hormonal signals,of which light and gibberellin(GA)signals are two critical factors with opposite effects on hypocotyl elongation.Although interacti... Plant growth is coordinately controlled by various environmental and hormonal signals,of which light and gibberellin(GA)signals are two critical factors with opposite effects on hypocotyl elongation.Although interactions between the light and GA signaling pathways have been studied extensively,the detailed regulatory mechanism of their direct crosstalk in hypocotyl elongation remains to be fully clarified.Previously,we reported that ABA INSENSITIVE 4(ABI4)controls hypocotyl elongation through its regulation of cellelongation-related genes,but whether it is also involved in GA signaling to promote hypocotyl elongation is unknown.In this study,we showthat promotion of hypocotyl elongation by GA is dependent on ABI4 activation.DELLAs interact directly with ABI4 and inhibit its DNA-binding activity.In turn,ABI4 combined with ELONGATED HYPOCOTYL 5(HY5),a key positive factor in light signaling,feedback regulates the expression of the GA2ox GA catabolism genes and thus modulates GA levels.Taken together,our results suggest that the DELLA-ABI4-HY5 module may serve as a molecular link that integrates GA and light signals to control hypocotyl elongation. 展开更多
关键词 light signal GA signal hypocotyl elongation DELLA abi4 HY5
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ABI5–FLZ13 module transcriptionally represses growth-related genes to delay seed germination in response to ABA 被引量:1
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作者 Chao Yang Xibao Li +13 位作者 Shunquan Chen Chuanliang Liu Lianming Yang Kailin Li Jun Liao Xuanang Zheng Hongbo Li Yongqing Li Shaohua Zeng Xiaohong Zhuang Pedro L.Rodriguez Ming Luo Ying Wang Caiji Gao 《Plant Communications》 SCIE CSCD 2023年第6期277-295,共19页
The bZIP transcription factor ABSCISIC ACID INSENSITIVE5(ABI5)is a master regulator of seed germination and post-germinative growth in response to abscisic acid(ABA),but the detailed molecularmechanism by which it rep... The bZIP transcription factor ABSCISIC ACID INSENSITIVE5(ABI5)is a master regulator of seed germination and post-germinative growth in response to abscisic acid(ABA),but the detailed molecularmechanism by which it represses plant growth remains unclear.In this study,we used proximity labeling to map the neighboring proteome of ABI5 and identified FCS-LIKE ZINC FINGER PROTEIN 13(FLZ13)as a novel ABI5 interaction partner.Phenotypic analysis of flz13 mutants and FLZ13-overexpressing lines demonstrated that FLZ13 acts as a positive regulator of ABA signaling.Transcriptomic analysis revealed that both FLZ13 and ABI5 downregulate the expression of ABA-repressed and growth-related genes involved in chlorophyll biosynthesis,photosynthesis,and cell wall organization,thereby repressing seed germination and seedling establishment in response to ABA.Further genetic analysis showed that FLZ13 and ABI5 function together to regulate seed germination.Collectively,our findings reveal a previously uncharacterized transcriptional regulatorymechanismby which ABA mediates inhibition of seed germination and seedling establishment. 展开更多
关键词 ABA abi5 FLZ gene expression seed germination
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Tonoplast targeting of VHA-a3 relies on a Rab5-mediated but Rab7-independent vacuolar trafficking route 被引量:2
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作者 qiang-Nan Feng Yan Zhang Sha Li 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2017年第4期230-233,共4页
Summary Vacuolar trafficking routes and their regu- lators have recently drawn lots of attention in plant cell biology. A recent study reported the discovery of a plant-specific vacuolar trafficking route, i.e., a dir... Summary Vacuolar trafficking routes and their regu- lators have recently drawn lots of attention in plant cell biology. A recent study reported the discovery of a plant-specific vacuolar trafficking route, i.e., a direct ER- to-vacuole route, through analysis of VHA-a3 subcellular targeting, a key component for the tonoplast V- ATPases. Our recent findings showed that VHA-a3 targets to the tonoplast through a Rab5-mediated but Rab7-independent pathway, shedding new lights on the unconventional vacuolar trafficking route in plant cells. 展开更多
关键词 Rab DN Tonoplast targeting of VHA-a3 relies on a Rab5-mediated but Rab7-independent vacuolar trafficking route Figure RFP BFA YFP TGN
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CDK5-mediated phosphorylation of CP190 may regulate locomotor activity in adult female Drosophila
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作者 Chin-Tong Ong Wei-Qi Lin 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2018年第3期177-181,共5页
The three-dimensional organization of the genome plays a crucial role in regulating gene expression patterns in metazoans(Ong and Corces,2014).The nuclear architectural proteins are known to facilitate the formation... The three-dimensional organization of the genome plays a crucial role in regulating gene expression patterns in metazoans(Ong and Corces,2014).The nuclear architectural proteins are known to facilitate the formation of topological domains within the genome through mediating inter-and intra-chromosomal interactions.In vertebrate,CCCTC-binding factor(CTCF)is the main architectural protein that mediates long-range chromosomal interactions among its DNA binding sites through a process that is stabilized by cohesin (Parelho et al., 2008; Wendt et al., 2008). 展开更多
关键词 CP CDK5-mediated phosphorylation of CP190 may regulate locomotor activity in adult female Drosophila
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