大豆GmAP1基因的功能初探

以大豆天隆1号为材料,根据Glycine max Wm82.a2.v1中大豆基因组的预测序列,克隆出一个AP1同源基因,命名为GmAP1。该基因编码区CDS长度为711 bp,编码一个含236个氨基酸的蛋白质。对蛋白质序列的结构分析结果显示GmAP1蛋白符合MADS-box基因家族特征。为了初步分析GmAP1的功能,利用实时定量RT-PCR分析GmAP1在不同花器官中表达,并且在拟南芥中过表达该基因。在不同花器官中,GmAP1表达量不同,在花萼中表达量较高;花瓣次之。GmAP1过表达植株表现出早花、植株矮小以及花器官(花瓣、雄蕊和雌蕊)数量增多等表型。结果表明,大豆的GmAP1是一个功能保守的AP1基因,在花的发生和花器官发育中起着重要作用。     

植物AP1基因研究进展(综述)

AP1(APETALA1)基因属于植物花分生组织特征基因和花器官形态特征基因,在控制植物花分生组织特性与花器官的形成过程中起着重要的作用。本文综述了近年来植物AP1基因结构、功能、表达调节及其与物种进化关系研究的新进展,并对其在果树上的应用研究进行分析和展望。     

香蕉MuMADS1基因的生物信息学分析

通过筛选用采后0h和48h的香蕉果实构建的果实成熟的SSH(抑制差减杂交)文库,得到一条命名为MuMADS1长度为888bp的片段。通过互联网数据库及生物信息学分析工具对香蕉MuMADS1基因及其编码蛋白进行理化性质预测、序列与结构分析和功能预测。结果表明:MuMADS1基因编码蛋白分子式为C1171H1879N351O367S7,属于亲水的不稳定蛋白;保守结构域分析含有保守的MADS盒和半保守的K-box盒;二级结构主要是以α螺旋为主;具有多种磷酸化位点和核定位信号;同源性比较发现与许多植物的花器官决定基因具有较高的相似性,推测它们可能为同源基因,具有相似的生物学功能。     

APETALA1启动子驱动AtlPT4在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥（Arabidopsis thaliana）花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1（AP1）特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶（isopentyl transferase,IPT）基因IPT4,研究细胞分裂素对花和花器官发育的影响。在pAP1∷IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现,在pAP1∷IPT4转基因植株中,花分生组织特征决定基因LEAFY（LFY）与花器官特征决定基因AP1、PISTILLATA（PI）和AGAMOUS（AG）的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1∷IPT4影响其花和花器官的正常发育。     

APETALA1启动子驱动AtIPT4在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥（Arabidopsis thaliana）花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1（AP1）特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶（isopentyl transferase,IPT）基因IPT4,研究细胞分裂素对花和花器官发育的影响。在pAP1∷IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现,在pAP1∷IPT4转基因植株中,花分生组织特征决定基因LEAFY（LFY）与花器官特征决定基因AP1、PISTILLATA（PI）和AGAMOUS（AG）的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1∷IPT4影响其花和花器官的正常发育。     

棉花MADS框基因GhMADS3的克隆及其组成型表达对烟草花器官决定的影响

MADS框基因在植物花器官发育中发挥着关键性作用。为研究棉花花器官发育的机理,以徐州142花蕾为材料,利用EST数据库资料,通过EST序列整合,克隆出了一个MADS域蛋白的编码区段,GenBank登录号为AY083173。该片段(GhMADS3)包含一个732 bp的开放阅读框,推导的氨基酸序列(244氨基酸)与可可,黄瓜,烟草,矮牵牛,金鱼草等的AG亚家族基因的序列相似性高。进化树重建分析将GhMADS3基因归入MADS框基因AG亚家族C功能分支的euAG分支。RT-PCR分析显示,该基因在雄蕊和心皮中表达,在根、茎、叶等营养器官,萼片,花瓣,花器官变异体chv1(所有花器官均变为苞叶状器官)的花蕾中不表达。将GhMADS3与35S启动子融合构建成嵌合基因转化烟草,转基因烟草植株花朵出现萼片(轮1)向心皮,花瓣(轮2)向雄蕊的转变,花器官表现明显的白化倾向。同时,在轮1观察到丝状结构的出现,该结构在此前类似的研究中尚无报道。这些结果说明,实验中克隆了一个有生物学功能的棉花的AG亚家族MADS框基因,该基因可能在棉花花器官发育中有重要的功能。     

毛白杨PtSEP3-1基因启动子的克隆分析及其表达载体构建

SEP(SEPALLATA)类基因属于花器官发育ABCDE模型中的E类基因,拟南芥中的研究表明该类基因可能具有控制花器官形态发育以及激活其它类型基因的功能,是一类花发育过程中的关键基因。因此,研究杨树SEP类基因启动子表达特性对于杨树的开花调控研究具有重要意义。本文根据毛白杨SEP3基因和毛果杨基因组序列设计引物,通过PCR获得了PtSEP3-1基因上游2000bp的序列。序列分析结果表明该序列具有启动子的基本元件TATA-box和CAAT-box,还包含大量光响应元件ACE、Box I和Box4等,此外还有脱落酸响应元件ABRE,赤霉素响应元件GARE-motif以及胁迫响应元件HSE、TC-richrepeats等。进一步构建了一个以PtSEP3-1启动子驱动GUS基因的植物表达载体pPtSEP3-1protest,为该启动子的功能鉴定奠定了基础。     

APETALA1 启动子驱动AtIPT4 在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥(Arab idopsis thal iana)花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1(AP1)特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶(isopentyl trans ferase, IPT)基因IPT4, 研究细胞分裂素对花和花器官发育的影响。在pAP1::IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现, 在pAP1::IPT4转基因植株中, 花分生组织特征决定基因LEAFY (LFY)与花器官特征决定基因AP1、PISTILLATA (PI )和AGAMOUS (AG)的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1::IPT4影响其花和花器官的正常发育。     

被子植物花器官发育的模型演变和分子调控

基于模式植物拟南芥(Arabidopsis thaliana)和金鱼草(Antirrhinum majus)花器官突变体研究提出的四聚体模型揭示了花同源异型蛋白的相互作用方式;进一步提出的核小体拟态模型,解释了花同源蛋白四聚体调控目标靶基因的分子机理。被子植物花器官形态多样化与MADS-box基因的表达模式和功能分化密切相关。多年生被子植物花发育的高通量转录组分析表明,多种基因参与调控花器官发育过程。本文重点综述了被子植物花器官发育的模型演变、MADS-box基因结构和基因重复、miRNA调控以及相关转录组分析的最新研究成果,并对花器官发育的研究前景进行了展望。     

麝香百合LLGLO1基因的克隆和表达

用RACE方法克隆的麝香百合花发育的GLOBOSA（GLO）类B功能基因LLGLO1,与其他多种单子叶植物的GLO类基因高度同源,且C区具有典型的PI结构基序。通过RT—PCR检测,百合不同组织中的LLGLO1基因表达模式与郁金香的GLO类基因相似。即主要集中在百合第一、二、三轮花器官中表达,心皮和茎中有微量表达,而且随着心皮的成熟,其在心皮中的表达量逐渐增加,但在百合叶片中则未检测到LLGLO1的表达,因此认为LLGLO1在百合花器官中呈特异性表达。LLGLO1在百合第一轮花器官中的表达支持了van Tunen对ABC模型的修正。     

Poppy APETALA1/FRUITFULL orthologs control flowering time, branching, perianth identity, and fruit development

Several MADS box gene lineages involved in flower development have undergone duplications that correlate with the diversification of large groups of flowering plants. In the APETALA1 gene lineage, a major duplication coincides with the origin of the core eudicots, resulting in the euFUL and the euAP1 clades. Arabidopsis FRUITFULL (FUL) and APETALA1 (AP1) function redundantly in specifying floral meristem identity but function independently in sepal and petal identity (AP1) and in proper fruit development and determinacy (FUL). Many of these functions are largely conserved in other core eudicot euAP1 and euFUL genes, but notably, the role of APETALA1 as an "A-function" (sepal and petal identity) gene is thought to be Brassicaceae specific. Understanding how functional divergence of the core eudicot duplicates occurred requires a careful examination of the function of preduplication (FUL-like) genes. Using virus-induced gene silencing, we show that FUL-like genes in opium poppy (Papaver somniferum) and California poppy (Eschscholzia californica) function in axillary meristem growth and in floral meristem and sepal identity and that they also play a key role in fruit development. Interestingly, in opium poppy, these genes also control flowering time and petal identity, suggesting that AP1/FUL homologs might have been independently recruited in petal identity. Because the FUL-like gene functional repertoire encompasses all roles previously described for the core eudicot euAP1 and euFUL genes, we postulate subfunctionalization as the functional outcome after the major AP1/FUL gene lineage duplication event.     

FON1, an Arabidopsis gene that terminates floral meristem activity and controls flower organ number.

A novel gene that regulates floral meristem activity and controls floral organ number was identified in Arabidopsis and is designated FON1 (for FLORAL ORGAN NUMBER1). The fon1 mutants exhibit normal vegetative development and produce normal inflorescence meristems and immature flowers before stage 6. fon1 flowers become visibly different from wild-type flowers at stage 6, when the third-whorl stamen primordia have formed. The fon1 floral meristem functions longer than does that of the wild type: after the outer three-whorl organ primordia have initiated, the remaining central floral meristem continues to produce additional stamen primordia interior to the third whorl. Prolonged fon1 floral meristem activity also results in an increased number of carpels. The clavata (clv) mutations are known to affect floral meristem activity. We have analyzed the clv1 fon1, clv2 fon1, and clv3 fon1 double mutants. These double mutants all have similar phenotypes, with more stamens and carpels than either fon1 or clv single mutants. This indicates that FON1 and CLV genes function in different pathways to control the number of third- and fourth-whorl floral organs. In addition, to test for possible interactions between FON1 and other floral regulatory genes, we have constructed and analyzed the relevant double mutants. Our results suggest that FON1 does not interact with TERMINAL FLOWER1, APETALA1, APETALA2, or UNUSUAL FLORAL ORGAN. In contrast, normal LEAFY function is required for the expression of fon1 phenotypes. In addition, FON1 and AGAMOUS both seem to affect the domain of APETALA3 function, which also affects the formation of stamen-carpel chimera due to fon1 mutations. Finally, genetic analysis suggests that FON1 interacts with SUPERMAN, which also regulates floral meristem activity.     

Floral patterning in Lotus japonicus

Floral patterning in Papilionoideae plants, such as pea (Pisum sativum) and Medicago truncatula, is unique in terms of floral organ number, arrangement, and initiation timing as compared to other well-studied eudicots. To investigate the molecular mechanisms involved in the floral patterning in legumes, we have analyzed two mutants, proliferating floral meristem and proliferating floral organ-2 (pfo-2), obtained by ethyl methanesulfonate mutagenesis of Lotus japonicus. These two mutants showed similar phenotypes, with indeterminate floral structures and altered floral organ identities. We have demonstrated that loss of function of LjLFY and LjUFO/Pfo is likely to be responsible for these mutant phenotypes, respectively. To dissect the regulatory network controlling the floral patterning, we cloned homologs of the ABC function genes, which control floral organ identity in Arabidopsis (Arabidopsis thaliana). We found that some of the B and C function genes were duplicated. RNA in situ hybridization showed that the C function genes were expressed transiently in the carpel, continuously in stamens, and showed complementarity with the A function genes in the heterogeneous whorl. In proliferating floral meristem and pfo-2 mutants, all B function genes were down-regulated and the expression patterns of the A and C function genes were drastically altered. We conclude that LjLFY and LjUFO/Pfo are required for the activation of B function genes and function together in the recruitment and determination of petals and stamens. Our findings suggest that gene duplication, change in expression pattern, gain or loss of functional domains, and alteration of key gene functions all contribute to the divergence of floral patterning in L. japonicus.     

A novel role of BELL1-like homeobox genes, PENNYWISE and POUND-FOOLISH, in floral patterning

Flowers are determinate shoots comprised of perianth and reproductive organs displayed in a whorled phyllotactic pattern. Floral organ identity genes display region-specific expression patterns in the developing flower. In Arabidopsis, floral organ identity genes are activated by LEAFY (LFY), which functions with region-specific co-regulators, UNUSUAL FLORAL ORGANS (UFO) and WUSCHEL (WUS), to up-regulate homeotic genes in specific whorls of the flower. PENNYWISE (PNY) and POUND-FOOLISH (PNF) are redundant functioning BELL1-like homeodomain proteins that are expressed in shoot and floral meristems. During flower development, PNY functions with a co-repressor complex to down-regulate the homeotic gene, AGAMOUS (AG), in the outer whorls of the flower. However, the function of PNY as well as PNF in regulating floral organ identity in the central whorls of the flower is not known. In this report, we show that combining mutations in PNY and PNF enhance the floral patterning phenotypes of weak and strong alleles of lfy, indicating that these BELL1-like homeodomain proteins play a role in the specification of petals, stamens and carpels during flower development. Expression studies show that PNY and PNF positively regulate the homeotic genes, APETALA3 and AG, in the inner whorls of the flower. Moreover, PNY and PNF function in parallel with LFY, UFO and WUS to regulate homeotic gene expression. Since PNY and PNF interact with the KNOTTED1-like homeodomain proteins, SHOOTMERISTEMLESS (STM) and KNOTTED-LIKE from ARABIDOPSIS THALIANA2 (KNAT2) that regulate floral development, we propose that PNY/PNF-STM and PNY/PNF-KNAT2 complexes function in the inner whorls to regulate flower patterning events.