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Molecular Plant 2014年第11期7卷 1653-1669页

作者： Lin-Hui Yu Zi-Qing Miao Guo-Feng Qi Jie Wu Xiao-Teng Cai Jie-Li Mao Cheng-Bin Xiang School of Life Sciences University of Science and Technology of China Hefei Anhui Province 230027 China

Plant root system morphology is dramatically influenced by various environmental cues. The adaptation of root system architecture to environmental constraints, which mostly depends on the formation and growth of lateral roots, is an important agronomic trait. Lateral root development is regulated by the external signals coordinating closely with intrinsic signaling pathways. mads-box transcription factors are known key regulators of the transition to flowering and flower development. However, their functions in root development are still poorly understood. Here we report that AGL21, an AGL17-clade mads-box gene, plays a crucial role in lateral root development. AGL21 was highly expressed in root, particularly in the root central cylinder and lateral root primordia. AGL21 overexpression plants produced more and longer lateral roots while ag121 mutants showed impaired lateral root development, especially under nitrogen-deficient conditions. AGL21 was induced by many plant hormones and environmental stresses, suggesting a function of this gene in root system plasticity in response to various signals. Furthermore, AGL21 was found positively regulating auxin accumulation in lateral root primordia and lateral roots by enhancing local auxin biosynthesis, thus stimulating lateral root initiation and growth. We propose that AGL21 may be involved in various environmental and physiological signals-mediated lateral root development and growth.

关键词： mads root system architecture lateral root AGL21 auxin nitrate sulfate.

灰毡毛忍冬mads-box家族基因SVP克隆及表达分析

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中草药 2023年第16期54卷 5350-5357页

作者：龙丽君刘思思曾慧杰李昌珠张岗马英姿李依民中南林业科技大学生命科学与技术学院湖南长沙410004 湖南省林业科学院省部共建木本油料资源利用国家重点实验室湖南长沙410004 陕西中医药大学陕西省中医药管理局“秦药”研发重点实验室陕西西安712046 陕西中医药大学省部共建特色秦药资源研究开发国家重点实验室(培育) 陕西咸阳712083

目的克隆获得灰毡毛忍冬Lonicera macranthoides mads-box家族基因SHORT VEGETATIVE PHASE(SVP),并对其进行生物信息学、时空表达特异性和蛋白原核表达等特性分析。方法根据灰毡毛忍冬转录组数据设计特异性引物,通过PCR技术克隆Lm SVP... 详细信息

目的克隆获得灰毡毛忍冬Lonicera macranthoides mads-box家族基因SHORT VEGETATIVE PHASE(SVP),并对其进行生物信息学、时空表达特异性和蛋白原核表达等特性分析。方法根据灰毡毛忍冬转录组数据设计特异性引物,通过PCR技术克隆Lm SVP基因全长;运用生物信息学方法分析其编码蛋白的序列特征;利用实时荧光定量技术(reverse-transcription polymerase chain reaction,q RT-PCR)分别检测该基因在灰毡毛忍冬2个品种“龙花”和“金翠蕾”叶片、花早期和晚期中的表达水平;最后构建p TOPO-D1-Lm SVP原核表达载体,转化至大肠杆菌BL21(DE3)中进行蛋白表达。结果Lm SVP基因全长1472bp,开放阅读框(open reading frame,ORF)长720 bp,编码239个氨基酸,蛋白质相对分子质量为26712.63。进化树分析表明,Lm SVP蛋白与中华猕猴桃、小粒咖啡的SVP蛋白亲缘关系最近。q RT-PCR结果显示,Lm SVP基因在“龙花”和“金翠蕾”品种中的表达均存在组织特异性表达,其在叶中的表达量显著高于花中;而在花发育不同时期,Lm SVP基因表达量没有明显差异。Lm SVP在大肠杆菌中成功表达出蛋白,蛋白表达结果显示其相对分子质量约为26710,与预测相符合。结论通过对Lm SVP基因的全长克隆、序列分析和表达特性鉴定,为进一步研究该基因在调控花蕾型灰毡毛忍冬蕾期长及花冠不开放的功能提供实验基础。

关键词：灰毡毛忍冬 mads SVP 基因克隆表达分析原核表达

Arabidopsis mads-Box Transcription Factor AGL21 Acts as Environmental Surveillance of Seed Germination by Regulating ABI5 Expression

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Molecular Plant 2017年第6期10卷 834-845页

作者： Lin-Hui Yu Jie Wu Zi-Sheng Zhang Zi-Qing Miao Ping-Xia Zhao Zhen Wang Cheng-Bin Xiang School of Life Sciences University of Science and Technology of China Hefei Anhui Province 230027 China Shanghai Center for Plant Stress Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China

Seed germination is a crucial checkpoint for plant survival under unfavorable environmental conditions. Ab- scisic acid （ABA） signaling plays a vital role in integrating environmental information to regulate seed germination. It has been well known that MCMI/AGAMOUS/DEFICIENS/SRF （mads）-box transcription factors are key regulators of seed and flower development in Arabidopsis. However, little is known about their functions in seed germination. Here we report that mads-box transcription factor AGL21 is a negative regulator of seed germination and post-germination growth by controlling the expression of ABA-INSENSITIVE 5 （ABIb＇） in Arabidopsis. The AGL21-overexpressing plants were hypersensitive to ABA, salt, and osmotic stresses during seed germination and early post-germination growth, whereas ag121 mutants were less sensitive. We found that AGL21 positively regulated ABI5 expression in seeds. Consistently, genetic analyses showed that AGL21 is epistatic to ABI5 in controlling seed germination. Chromatin immunoprecipitation assays further demonstrated that AGL21 could directly bind to the ABI5 promoter in plant cells. Moreover, we found that AGL21 responded to multiple environmental stresses and plant hormones during seed germination. Taken together, our results suggest that AGL21 acts as a surveillance integrator that incorporates environmental cues and endogenous hormonal signals into ABA signaling to regulate seed germination and early post-germination growth.

关键词： mads AGL21 seed germination ABA ABI5

燕子花花期调控相关mads-box转录因子筛选及功能验证

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燕子花花期调控相关MADS-box转录因子筛选及功能验证

作者：李凤仪东北林业大学

学位级别：硕士

燕子花(Iris laevigata)为鸢尾科(Iridaceae)鸢尾属(Iris)多年生草本,花色、花型独特,观赏价值较高,然而其花的观赏期较短,严重影响了其推广应用。通过基因工程调控植物提前或延迟开花是延长群体花期的有效方法,花期调控相关基因资源的... 详细信息

燕子花(Iris laevigata)为鸢尾科(Iridaceae)鸢尾属(Iris)多年生草本,花色、花型独特,观赏价值较高,然而其花的观赏期较短,严重影响了其推广应用。通过基因工程调控植物提前或延迟开花是延长群体花期的有效方法,花期调控相关基因资源的挖掘非常必要。mads-box转录因子家族中的许多成员参与植株开花时间调控,但目前燕子花中相关研究少见报道。本研究从燕子花花被片的转录组中进行了开花时间相关的mads-box转录因子筛选,克隆了具有互作关系的Il SEP3(Gen Bank登录号:ON398430)、Il SVP(Gen Bank登录号:ON398431)基因并在拟南芥中异源表达进行了花期调控功能的验证;同时筛选了燕子花特异性的内参基因,为燕子花功能基因的时空定量表达研究奠定了基础。主要研究成果如下:1.首次从燕子花花被片的转录组数据鉴定出41个mads-box转录因子,通过系统进化分析确定其中8个为I型mads-box基因(5个属于Mα组,3个属于Mβ组);33个基因为II型mads-box基因,其中2个为MIKC*型,31个基因为MIKC型(包括SEP-like、AGL6-like、SQUA-like、AG-like、TM3-like、DEF+GLO-like及STmads11-like 7个进化支)。通过基因功能预测,选择了两个可能与开花时间相关的目的基因,分别命名为Il SEP3及Il SVP。2.通过PCR扩增出Il SEP3、Il SVP基因,两基因序列分别长720bp及705bp,编码239个、234个氨基酸,蛋白分子量为27.64k Da、26.26k Da,理论等电点为8.99、6.24,为不稳定的亲水蛋白,不存在跨膜结构及信号肽;蛋白二级结构分析发现Il SEP3蛋白中α-螺旋占比51.88%,延伸链占比9.21%,β-转角占比5.02%,随机卷曲占比33.89%;Il SVP蛋白中α-螺旋占比56.84%,延伸链占比11.97%,β-转角占比3.42%,随机卷曲占比27.78%;成功构建蛋白三级结构模型;Il SEP3及Il SVP蛋白均以丝氨酸磷酸化位点为主,亚细胞定位预测蛋白定位于细胞核;系统进化分析发现,Il SEP3与鸢尾科香雪兰属杂交种及番红花亲缘关系最近,Il SVP与番红花亲缘关系最近。通过同源序列功能分析推测Il SEP3及Il SVP可能调节开花时间、花器官发育及花粉发育等过程。3.构建p GBKT7-Il SEP3载体及p GADT7-Il SVP载体,进行酵母双杂交,发现Il SEP3蛋白与ISVP蛋白能发生相互作用。4.确定Il SEP3是调控开花提前的基因,Il SVP对开花时间调控不明显:构建p CAMBIA1300-Il SEP3-GFP及p CAMBIA1300-Il SVP-GFP植物过表达载体,异源转化拟南芥获得转基因T3代株系,经表型观察及与开花相关基因q RT-PCR检测发现Il SEP3可能通过多成花途径直接或间接促进FT及SOC1等基因表达使植株提前开花,与对照相比花期提前约3 d。转Il SVP基因株系的开花时间与对照相比无显著变化。此外农杆菌瞬时转化本氏烟草,确定Il SEP3及Il SVP蛋白均定位于细胞核,与预测结果一致。5.筛选了植物中常用的8个内参基因,在燕子花11个不同的组织(4类样品组)中进行了表达稳定性比较,使用内参基因稳定性分析软件ge Norm、Norm Finder及Best Keeper分析获得8个基因在4类样品组中的综合排名,结合ge Norm评估的各样品组适用内参基因数量,确定所有样品组(11个组织同时比较)中最适内参基因为PP2A、AP-4及CYP;不同开花时期样品组(花苞期、初花期、盛花期及末花期的花)中最适内参基因为PP2A、GAPDH;不同花器官样品组(盛花期的外花被、内花被、雄蕊、雌蕊及子房)中最适内参基因为GAPDH及PP2A;不同组织部位样品组(叶、花葶、盛花期的花)中最适内参基因为AP-4及PP2A。

关键词：燕子花花期调控 mads 内参基因

Genome-wide identification of the CONSTANS-LIKE(COL)family and mechanism of fruit senescence regulation by PpCOL8 in sand pear(Pyrus pyrifolia)

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Journal of Integrative Agriculture 2024年第4期23卷 1222-1237页

作者： Yue Xu Shurui Song Huiying Wang Xilong Cao Xinran Zhao Wenli Wang Liyue Huo Yawei Li Misganaw Wassie Bin Lu Liang Chen Haiyan Shi College of Horticulture Hebei Agricultural UniversityBaoding 071001China Wuhan Botanical Garden Chinese Academy of SciencesWuhan 430074China

Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and ***,the industrial development of pear fruit is significantly limited by its short shelf *** acid(SA),a well-known phytohormone,can delay fruit senescence and improve shelf ***,the mechanism by which SA regulates CONSTANS-LIKE genes(COLs)during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly *** this study,22 COL genes were identified in sand pear,including four COLs(Pp COL8,Pp COL9a,Pp COL9b,and Pp COL14)identified via transcriptome analysis and 18 COLs through genome-wide *** COL genes were divided into three subgroups according to the structural domains of the COL *** COL8,with two B-box motifs and one CCT domain,belonged to the first *** contrast,the other three Pp COLs,Pp COL9a,Pp COL9b,and Pp COL14,with similar conserved protein domains and gene structures,were assigned to the third *** four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit ***,the expression of Pp COL8 was inhibited by exogenous SA treatment,while SA up-regulated the expression of Pp COL9a and Pp ***,Pp COL8 interacts with Pp mads,a mads-box protein preferentially expressed in fruit,and SA up-regulated its *** the production of ethylene and the content of malondialdehyde(MDA)were increased in Pp COL8-overexpression sand pear fruit,the antioxidant enzyme(POD and SOD)activity and the expression of Pp POD1 and Pp SOD1 in the sand pear fruits were down-regulated,which showed that Pp COL8 promoted sand pear fruit *** contrast,the corresponding changes were the opposite in Pp mads-overexpression sand pear fruits,suggesting that Pp mads delayed sand pear fruit *** co-transformation of Pp COL8 and Pp mads also delayed sand pear fruit *** re

关键词： Pyrus pyrifolia CONSTANS-LIKE gene salicylic acid fruit senescence mads

桃中两个mads box基因的克隆与表达分析

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Acta Genetica Sinica 2004年第9期31卷 908-918页

作者：吴凡徐勇常凤启李学东马荣才北京农业生物技术研究中心北京100089 首都师范大学生命科学学院北京100037

为研究李属 (Prunussp .)果树生殖调控的相关基因 ,对国际公共数据库中的李属植物的EST(expressedse quencetags)序列进行了电子拼接 ,获得了 8个madsbox基因的cDNA序列 ,并利用PCR技术从桃中克隆出其中的两个cDNA ,分别命名为Ppmads4和... 详细信息

为研究李属 (Prunussp .)果树生殖调控的相关基因 ,对国际公共数据库中的李属植物的EST(expressedse quencetags)序列进行了电子拼接 ,获得了 8个madsbox基因的cDNA序列 ,并利用PCR技术从桃中克隆出其中的两个cDNA ,分别命名为Ppmads4和Ppmads6 ,在GenBank中的登录号为AY70 5 972和AY70 5 973。Ppmads4基因长85 0bp ,包含一个 732bp的开放阅读框 ,编码 2 4 3个氨基酸。Ppmads6基因长 1190bp ,包含 1个 76 8bp的开放阅读框 ,编码 2 5 6个氨基酸。Ppmads4和Ppmads6在序列上分别与拟南芥中的AGAMOUS基因和矮牵牛中的PFG基因高度同源。RT PCR分析表明 ,Ppmads4基因在桃的花瓣、心皮、果实及果仁中表达 ,应属于控制花器官发育的C类madsbox基因。Ppmads6基因在桃的叶、萼片、花瓣、心皮及果实中表达 ,应属于调控植物由营养生长向生殖生长过渡的A类madsbox基因。

关键词：桃花发育 mads box 幼年期成熟期

Structural and functional analysis of a mads box containing genomic DNA sequence cloned from rice

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Science China(Life Sciences) 1998年第6期41卷 561-568页

作者：孟征刘国振许智宏唐广王宁陈润生朱立煌 Institute of Genetics Chinese Academy of Sciences Beijing China

A putative mads box gene (Rgmads1) was cloned by screening a rice genomic library with a heterogeneous mads box probe derived from \%Antirrhinum majus squamosa\% gene. Sequencing and Southern analysis showed that Rgmads1 contains a highly conserved mads box sequence, whereas the rice genome contains numbers of mads box genes. A chimera gene of Rgmads1 under the control of CaMV 35S promoter was constructed and used to transform \%Arabidopsis.\% Some transformants exhibited abnormal phenotypes in their flowers, which were reflected in the floral structure, number and setting position. Therefore, Rgmads1 is likely a member of rice mads box gene family and may be involved in the floral establishment and function control in the rice developmental process.

关键词： rice mads box genes gene cloning homeotic gene.

桃(Prunus persica)中2个mads box基因功能的初步鉴定和遗传作图

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科学通报 2008年第5期53卷 537-543页

作者：徐勇张林马荣才首都师范大学生命科学学院

我们在以前的研究中克隆了2个桃madsbox基因,Ppmads4和Ppmads6,它们分别为AGAMOUS(AG)和FRUITFULL(FUL)的同源基因,本研究把它们进一步转入拟南芥中分析它们的功能.转基因结果表明,这2个基因都能导致拟南芥早花,二级花序减少,出现顶端花... 详细信息

我们在以前的研究中克隆了2个桃madsbox基因,Ppmads4和Ppmads6,它们分别为AGAMOUS(AG)和FRUITFULL(FUL)的同源基因,本研究把它们进一步转入拟南芥中分析它们的功能.转基因结果表明,这2个基因都能导致拟南芥早花,二级花序减少,出现顶端花,但Ppmads4与Ppmads6对拟南芥花器官有截然不同的影响.Ppmads4引起转基因植株花器官同源异型转变:萼片心皮化,花瓣雄蕊化;Ppmads6转基因植株表现为花瓣、雄蕊和心皮花器官数目的增加.它们对果实发育也产生不同的影响:Ppmads4转基因植株的角果在伸长期花的外2轮萼片和花瓣不脱落;Ppmads6转基因植株的角果成熟后不开裂,并可诱导从一朵花中长出多个果角.这些结果表明,Ppmads4在拟南芥中可模拟AG的功能,而Ppmads6与拟南芥FUL功能相似.Ppmads6对花器官数目的影响暗示FUL同源基因具有调节花器官数目的新功能.通过RT-PCR分析花器官发育和控制顶端分生组织的基因表达,结果表明,Ppmads6诱导产生超数花器官可能是通过控制CLV-WUS通路的基因来实现的.此外,将Ppmads4转化为一个SSR标记,并将其定位在桃属植物的G5连锁群上,与Ppmads6基因位于同一染色体区段上.最后,对Ppmads4和Ppmads6在农作物及果树育种上的潜在应用价值进行了讨论.

关键词： mads box基因花器官果实桃(Prunus perscia) 遗传定位

甜樱桃mads box基因的克隆与表达分析

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园艺学报 2011年第8期38卷 1462-1468页

作者：林苗苗赵长竹姜建福顾红陈锦永方金豹中国农业科学院郑州果树研究所中国农业科学院果树生长发育与品质控制重点开放实验室郑州450009

以甜樱桃'拉宾斯'(Prunus avium L.'Lapins')为试材,采用RT-PCR结合RACE技术,克隆获得1个与花发育相关的mads box基因,命名为Pamads3,其在GenBank中的登录号为HQ229605。Pamads3基因全长1095bp,包含1个723bp的开放阅读框,推断其编码240个氨基酸。序列分析表明:Pamads3蛋白与拟南芥中的SEP蛋白高度同源。组织特异性表达显示Pamads3基因在花瓣、雄蕊、心皮中表达。实时定量RT-PCR分析表明,花芽露绿期的离体枝条经过15和25℃处理后,其雌蕊中Pamads3基因在25℃的表达量高于在15℃的表达量。

关键词：甜樱桃花发育 mads box 克隆表达

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百合花发育相关mads box基因的克隆

园艺学报 2004年第3期31卷 332-336页

作者：张云刘青林欧阳青蔡文启中国农业大学观赏园艺与园林系北京100094 中国科学院微生物研究所北京100080

利用RT PCR的方法 ,从百合中克隆得到了 3个接近全长的花发育相关madsbox基因片段Lf mads1～ 3。其中Lfmads1和Lfmads3以往未见报道 ,分别与多种作物的AG、SEP同源基因具有较高的同源性 ;Lfmads2与已发表的Lmads2对应区段的氨基酸同源... 详细信息

利用RT PCR的方法 ,从百合中克隆得到了 3个接近全长的花发育相关madsbox基因片段Lf mads1～ 3。其中Lfmads1和Lfmads3以往未见报道 ,分别与多种作物的AG、SEP同源基因具有较高的同源性 ;Lfmads2与已发表的Lmads2对应区段的氨基酸同源性高达 98 99% 。

关键词：百合花发育 mads box基因克隆