一株拟南芥晚花突变体的分子分析

植物的开花时间直接控制植物营养生长期和生育期的长短,在相当程度上决定着繁育的成功与否,也与植物的产量和质量性状密切相关.拟南芥花期改变突变体是植物开花调控机制研究重要的植物材料.研究利用前期研究中获得的一株稳定遗传的、晚花表型的T-DNA插入拟南芥突变体pex2为材料,对其进行开花相关表型分析、T-DNA插入位点鉴定及插入位点基因转录分析等研究,结果表明:pex2突变体中存在单一的T-DNA插入,该插入位点位于PEX2基因下游约1 800 bp处,且该位点的插入引起了PEX2全长转录产物的缺失.该研究为进一步探索pex2突变体晚花机制及拟南芥PEX2基因与晚花表型之间的相关性研究,提供了研究材料.     

拟南芥开花时间的分子遗传调控

拟南芥开花主要受四种遗传途径调控,即:光周期途径,春化途径,自主途径和赤霉素途径。各种途径之间通过CO,FLC,FT,SOC1等主效基因的相互作用,最终调节花特异性基因AP1和LFY的表达,从而调控拟南芥的开花时间,其中各种突变体的研究揭示了相关基因的功能。这些主效基因中,CO基因对光周期途径是特异的,FLC基因对开花起抑制作用。其它因素如环境、基因染色质结构的改变、酶等对拟南芥开花时间的影响有待进一步的研究。     

CONSTANS-LIKE 7基因对拟南芥开花的调控分析

为研究CONSTANS-LIKE 7(COL7)在调控植物开花方面的功能,以定量PCR,GUS染色等方法,研究光及生物钟对COL7表达的影响,以及COL7对拟南芥开花的影响.实验结果显示:光及生物钟参与调控COL7的表达;过表达COL7在长日照条件下抑制CONSTANS(CO)以及FLOWERING LOCUS T(FT)的表达,进而抑制拟南芥开花;col7突变体不论是在长日照下还是短日照下都没有明显的开花表型,说明COL7在调控拟南芥开化方面可能与其家族基因中的其它成员存在功能冗余.     

Fibrillins突变体对拟南芥开花时间的影响

本研究所使用突变体为fbn8和fbn1a为研究对象,以杂交的方法获得fbn1a/fbn8双重突变体.对Fibrillins突变体在不同的环境下FBN1a、FBN8对开花时间调节的可能机制进行初步探究.研究发现,Fibrillins突变体在长日照、短日照条件下均为早花,并且fbn1a/fbn8双重突变体早花现象更为明显.说明FBN1a、FBN8有可能共同在开花途径中起到作用.通过连续对突变体开花基因的检测发现,和单突变体相比,fbn1a/fbn8双重突变体中的FT、SOC1、CO的基因表达量都大幅度增加,其中FT增加的更为明显,FLC减少也更为明显.而与野生型相比较,这种现象单突变体fbn1a又要比fbn8更为明显.最后,通过检测了植株体内H2O2含量的变化发现,与野生型比较fbn8、fbn1a和fbn1a/fbn8体内H2O2含量升高程度为20%、30%、50%.由实验结果推测Fibrillins可能通过影响体内H2O2含量变化来参与开花时间的改变.     

Fibrillins突变体对拟南芥开花时间影响

植物的开花受到多条途径的控制,包括春化途径、赤霉素途径、自主途径和光周期途径。以拟南芥Columbia（Col.）生态型和拟南芥Fibrillins突变体(本研究所使用突变体为fbn8和fbn1a)为研究对象,以杂交的方法获得fbn1a /fbn8双重突变体。文章对Fibrillins突变体在不同的环境下FBN1a、 FBN8对开花时间调节的可能机制进行初步探究。本研究发现,Fibrillins突变体在长日照、短日照条件下均为早花,并且fbn1a /fbn8双重突变体早花现象更为明显。说明FBN1a、FBN8有可能共同在开花途径中起到作用。通过连续对突变体开花基因的检测发现,和单突变体相比,fbn1a /fbn8双重突变体中的FT、SOC1、CO的基因表达量都大幅度增加,其中FT增加的更为明显,FLC减少也更为明显。而与野生型相比较,这种现象单突变体fbn1a又要比fbn8更为明显。最后,通过检测了植株体内H2O2含量的变化发现,与野生型比较fbn8、fbn1a和 fbn1a /fbn8体内H2O2含量升高程度为20%、30%、50%。由实验结果推测Fibrillins可能通过影响体内H2O2含量变化来参与开花时间的改变。     

一株晚花和镉敏感拟南芥突变体的分离与鉴定

为了进一步挖掘拟南芥响应Cd胁迫的分子机制,从化学诱导型拟南芥突变体库(约6 000株系)中筛选获得Cd敏感突变体,通过测量经Cd处理后拟南芥植株的根长变化获得Cd响应突株系,然后单株收种并鉴定,最终获得一株对Cd胁迫敏感的突变体。Thermal asymmetric interlaced-PCR(TAIL-PCR)发现这株突变体T-DNA的插入定位于基因At1g35820和At1g35830之间。进一步分析表明这个突变体也是FLC(Flowering locus C)依赖的晚花突变体,将这株突变体命名为fcr(flowering and cadmium related gene)。FLC在拟南芥开花调控网络中起枢纽作用,并且是开花正调控基因Suppressor of overexpression of co 1 (SOCI)的抑制因子。在该突变体中FLC的表达明显高于野生型植株,而FLC下游基因SOCI的表达明显低于野生型植株,但FLC上游基因的表达变化不明显。进一步实验发现,突变体fcr中调节环境因素和春化作用的基因High expression of osmotically responsive gene 1(HOS1)表达量显著增加。通过以上结果我们推测,突变体fcr中的晚开花和Cd敏感表型可能是HOS1基因过量表达所致。     

SPL转录因子调控植物花发育及其分子机制研究进展

SPL(squamosa promoter-binding protein-like)转录因子是植物所特有的一类基因家族,广泛存在于绿色植物中,在植物生长发育中具有重要作用。花发育是植物生殖发育中最为重要的一个过程,涉及不同发育方式的转变,即开花决定、花的发端和花器官发生与发育。简要综述了SPL基因的结构与功能并着重阐述了SPL基因在植物花发育过程中的分子机制及生物学功能。最后总结出: SPL转录因子可直接或间接通过参与光周期途径,赤霉素途径及年龄途径来调控植物的开花时间; SPL基因可通过直接激活下游花分生组织特异基因,如LEAFY(LFY),从而调控植物的成花转变; SPL基因可通过与下游花器官特征基因相互作用来调控花器官及其育性的发育,如调控花序、花柄的长度与外形及花器官的大小; SPL基因可调控植物大小孢子发生及雌雄配子体发育。据拟南芥的相关研究结果,初步构绘出拟南芥开花调控中的分子机制。     

水稻成花素基因Hd3a在ABA调控种子萌发中的功能研究

Hd3a(Heading date 3a)是拟南芥成花素基因FT(Flowering Locus T)的同源基因，作为水稻开花的强诱导因子，对短日照条件下水稻的开花具有重要的促进作用。Hd3a在水稻发育中是否具有其他功能，目前的认识仍非常有限。在我们的研究中，通过对Hd3a基因组序列进行生物信息学分析，发现Hd3a的启动子序列包含响应脱落酸(Abscisic Acid, ABA)信号的ABRE(ABA-Responsive Element)元件。荧光素酶发光反应和RT-qPCR分析都证实了Hd3a的转录表达随外源ABA增加而减少。通过不同浓度的ABA处理水稻野生型和hd3a缺失突变体种子，结果表明hd3a缺失突变体种子萌发明显被抑制，暗示了Hd3a缺失导致种子萌发中对ABA敏感性增强，同时hd3a突变体中ABA信号途径响应基因ABI5(ABA Insensitive 5)、ABL1(ABI5-Like1)的表达量升高，表明Hd3a可负调控ABA信号。综上所述，Hd3a可以响应ABA信号调控水稻种子萌发，这些研究结果对于深入理解水稻Hd3a基因的生物学功能具有重要意义。     

拟南芥18srRNA参与隐花色素介导的信号传导途径的研究

通过筛选以拟南芥蓝光受体隐花色素双突变体cry1cry2为遗传背景的激活标签突变体库,得到一株SCC98-D株系,该突变体具有早开花,下胚轴变短,并具有花瓣数目增加等表型,彻底恢复了cry1cry2的晚开花和长下胚轴表型.通过Tail-PCR的方法克隆得到SCC98-D突变体中激活标签插入位点的侧翼序列,通过对该侧翼序列测序发现插入位点在18srRNA的1751bp处,运用RT-PCR方法分析插入位点周围基因的表达,发现只有18srRNA的表达被激活,由此证实了18srRNA参与隐花色素介导的信号传导途径.     

Adaptation of photoperiodic control pathways produces short-day flowering in rice

The photoperiodic control of flowering is one of the important developmental processes of plants because it is directly related to successful reproduction. Although the molecular genetic analysis of Arabidopsis thaliana, a long-day (LD) plant, has provided models to explain the control of flowering time in this species, very little is known about its molecular mechanisms for short-day (SD) plants. Here we show how the photoperiodic control of flowering is regulated in rice, a SD plant. Overexpression of OsGI, an orthologue of the Arabidopsis GIGANTEA (GI) gene in transgenic rice, caused late flowering under both SD and LD conditions. Expression of the rice orthologue of the Arabidopsis CONSTANS (CO) gene was increased in the transgenic rice, whereas expression of the rice orthologue of FLOWERING LOCUS T (FT) was suppressed. Our results indicate that three key regulatory genes for the photoperiodic control of flowering are conserved between Arabidopsis, a LD plant, and rice, a SD plant, but regulation of the FT gene by CO was reversed, resulting in the suppression of flowering in rice under LD conditions.     

FKF1 is essential for photoperiodic-specific light signalling in Arabidopsis

Adaptation to seasonal change is a crucial component of an organism's survival strategy. To monitor seasonal variation, organisms have developed the capacity to measure day length (photoperiodism). Day-length assessment involves the photoperiodic control of flowering in Arabidopsis thaliana, whereby the coincidence of light and high expression of CONSTANS (CO) induces the expression of FLOWERING LOCUS T (FT), leading to flowering in long-day conditions. Although controlling CO expression is clearly a key step in day-length discrimination, the mechanism that generates day-length-dependent CO expression remains unknown. Here we show that the clock-controlled FLAVIN-BINDING, KELCH REPEAT, F-BOX (FKF1) protein has an essential role in generating the diurnal CO peak and that this function is dependent on light. We show that a recombinant FKF1 LIGHT, OXYGEN OR VOLTAGE (LOV) domain binds the chromophore flavin mononucleotide and undergoes light-induced photochemistry, indicating that FKF1 may function as a photoperiodic blue-light receptor. It is likely that the circadian control of FKF1 expression and the light regulation of FKF1 function coincide to control the daytime CO waveform precisely, which in turn is crucial for day-length discrimination by Arabidopsis.     

CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis

Flowering is often triggered by exposing plants to appropriate day lengths. This response requires an endogenous timer called the circadian clock to measure the duration of the day or night. This timer also controls daily rhythms in gene expression and behavioural patterns such as leaf movements. Several Arabidopsis mutations affect both circadian processes and flowering time; but how the effect of these mutations on the circadian clock is related to their influence on flowering remains unknown. Here we show that expression of CONSTANS (CO), a gene that accelerates flowering in response to long days, is modulated by the circadian clock and day length. Expression of a CO target gene, called FLOWERING LOCUS T (FT), is restricted to a similar time of day as expression of CO. Three mutations that affect circadian rhythms and flowering time alter CO and FT expression in ways that are consistent with their effects on flowering. In addition, the late flowering phenotype of such mutants is corrected by overexpressing CO. Thus, CO acts between the circadian clock and the control of flowering, suggesting mechanisms by which day length regulates flowering time.     

拟南芥雄性不育基因ds254的定位及其表达模式

通过Ac／Ds转座子系统的诱变，得到拟南芥突变体DS254．它的发育进程比野生型慢，植株比野生型矮小，连座叶柄比野生型的短，花苞数目比野生型的多，分枝的数量比野生型的多，且雄性不育．遗传分析表明突变体属于隐性单基因突变，稳定遗传，并且与Ds是紧密连锁的。可能是Ds插入直接引起的突变．TAIL—PCR的方法将该基因定位在第4条染色体上．Ds上携带的GUS基因表迭的初步分析表明，GUS基因在突变体的花药和连座叶中表达，说明该基因也可能在这些部位表达．突变体的表型分析结果说明该基因可能在植物的许多发育过程中起作用．     

拟南芥BLI基因调控植物衰老的研究

BLISTER(BLI)蛋白是多种应激反应的调节因子,可以促进植物对环境的适应性.BLI蛋白通过抑制IRE 1蛋白以维持植物的正常生长,缺失BLI蛋白的拟南芥植株出现多种发育缺陷表型.文章以拟南芥bli突变体为实验材料,通过细胞生物学及遗传学方法检测其衰老相关表征,发现bli突变体植株早期出现叶片黄化、叶绿素含量下降、...     

Control of flowering and storage organ formation in potato by FLOWERING LOCUS T

Seasonal fluctuations in day length regulate important aspects of plant development such as the flowering transition or, in potato (Solanum tuberosum), the formation of tubers. Day length is sensed by the leaves, which produce a mobile signal transported to the shoot apex or underground stems to induce a flowering transition or, respectively, a tuberization transition. Work in Arabidopsis, tomato and rice (Oryza sativa) identified the mobile FLOWERING LOCUS T (FT) protein as a main component of the long-range 'florigen', or flowering hormone, signal. Here we show that expression of the Hd3a gene, the FT orthologue in rice, induces strict short-day potato types to tuberize in long days. Tuber induction is graft transmissible and the Hd3a-GFP protein is detected in the stolons of grafted plants, transport of the fusion protein thus correlating with tuber formation. We provide evidence showing that the potato floral and tuberization transitions are controlled by two different FT-like paralogues (StSP3D and StSP6A) that respond to independent environmental cues, and show that an autorelay mechanism involving CONSTANS modulates expression of the tuberization-control StSP6A gene.