类基因RT-PCR表达模式研究

CYC 类基因的分子系统学研究已经在苦苣苔科Gesneriaceae中展开, 但是还缺乏对这些基因表达和功能的研究。因此, 我们选择苦苣苔科大叶石上莲Oreocharis benthamii作为实验材料, 分离出了CYC类基因的两个拷贝, 经过分子系统学分析这两个基因分别属于苦苣苔科GCYC1和GCYC2两个分支, 故命名为ObCYC1和ObCYC2。分区的RT-PCR实验结果显示这两个基因拥有不同的时间空间表达模式。ObCYC1与模式植物金鱼草Antirrhinum majus中的CYC基因类似, 集中在花冠背部区域表达, 这与它们拥有保守的功能区TCP和R相一致。但是, ObCYC1与CYC表达模式仍有区别, 即, 和CYC相比ObCYC1在花冠背部区域的表达提前减弱。这可能和大叶石上莲花冠微弱的两侧对称性相关。另外, 由于大叶石上莲的背部花瓣较两侧和腹部花瓣小, 因此, 在功能上ObCYC1可能起抑制背部花瓣生长作用而CYC基因则促进背部花瓣生长。与ObCYC1不同, ObCYC2的保守功能区有更多的氨基酸位点变化, 而且在RT-PCR实验中也没有检测到它的表达。因此, 需要开展更深入的实验研究分析ObCYC2的基本功能, 这将有助于了解GCYC2类基因在苦苣苔科及其近缘科中的功能和进化途径。     

苦苣苔科(Gesneriaceae)植物研究进展

苦苣苔科植物具有极高的分类学、进化系统学、植物地理学等方面的研究价值。对我国的苦苣苔科植物的分布等进行了简要阐述,并对其绝灭和濒危机制、植物形态系统学、花粉形态学、核型分析、DNA序列分析、基因克隆和种群遗传结构以及观赏价值等方面进行了综述分析。     

广西苦苣苔科一新属——方鼎苣苔属

报道了在中国广西发现的苦苣苔科一新属即方鼎苣苔属Paralagarosolen Y. G. Wei 和一新种方鼎苣苔P. fangianum Y. G. Wei。方鼎苣苔属与细筒苣苔属Lagarosolen W. T. Wang近缘,它们的共同特征是花筒细筒状,不肿胀,柱头2;不同点是方鼎苣苔属叶基部有时盾状,聚伞花序具1朵花,花冠裂片顶端圆钝,蒴果宽卵状椭圆球形。     

台闽苣苔(苦苣苔科)花部器官的形态发生

在扫描电镜下对台闽苣苔 (T. oldhamii (Hemsl.) Solereder)进行了花部器官形态发生的观察,为探索该类群的个体发育、类群间的系统发育关系和进化趋势提供依据.研究发现该属植物萼片、花冠和雄蕊发生式样均为五数花类型,它们各自来源于花原基上分化出来的萼片原基、花冠原基和雄蕊原基;花冠与雄蕊的两侧对称性与花冠上唇生长稍快和退化雄蕊原基发育迟滞相关;萼片原基的发生和发育的顺序是不一致的:萼片原基发生的式样为近轴中原基-远轴2原基-2侧原基,发育式样则为近轴中萼片-2侧萼片-远轴2萼片,花蕾时为镊合状排列.花冠裂片原基的发生和发育式样是一致的,即远轴中裂原基(下唇中裂片)-远轴2侧裂原基(下唇2侧裂片)-近轴2裂原基(上唇2裂片).花蕾期卷迭式为覆瓦状排列,从外向内:下唇中裂片-下唇2侧裂片-上唇2裂片或下唇2侧裂片-上唇2裂片-下唇中裂片.雄蕊原基与花冠裂片原基互生,前方雄蕊原基在发生上稍迟于后方雄蕊原基,后者与退化雄蕊原基几乎同时发生,但较小,并与近轴心皮(或柱头上唇)对生.将该属与玄参科(Scrophulariaceae)的地黄属( Rehmannia )、苦苣苔科(Gesneriaceae)的异叶苣苔属( Whytockia)和尖舌苣苔属(Rhynchoglossum )的花部器官比较发现,这四个属在这方面呈现出多样性和交叉.过去一直按子房室数和胎座类型划分玄参科(子房2室、中轴胎座)和苦苣苔科(子房1室、侧膜胎座)这一做法受到了质疑.     

非洲紫罗兰两侧与辐射对称花中两个CYC类完整基因的分离和序列分析

在已知GCYC基因部分序列基础上, 通过改进的mTAIL-PCR方法克隆非洲紫罗兰Saintpaulia ionantha两侧对称栽培种中CYC类基因的5′未知序列, 并进而从两侧与辐射对称栽培种中分离得到苦苣苔科Gesneriaceae中第一组完整基因: SiCYC1A与SiCYC1B。对以上基因的核酸和氨基酸序列比较发现, SiCYC1A与SiCYC1B序列同源性很高, 均含有完整的功能调控区域(即TCP domain和R domain)并与模式植物金鱼草Antirrhinum majus中CYC基因同源。因此, 这两个基因应具有正常功能, 是功能上互补的冗余基因。令人意外的是在辐射对称花栽培品种中的这两个基因和两侧对称花栽培品种中对应基因的序列完全相同。经过对金鱼草以及相关类群辐射对称花突变体中CYC类基因序列的比较分析, 推论在非洲紫罗兰中, SiCYC1A与SiCYC1B基因可能受上游未知的共同调控因子调控, 该调控因子的改变是导致栽培品种中花对称性发生变化的主要原因。另外, 对改进后的TAIL-PCR(mTAIL-PCR)的方法和过程进行了详细叙述, 并对其技术特征和优势开展了简单的论述。     

得与失:苦苣苔科新的属级界定与分类系统——中国该科植物之变迁(英文)

过去20年对苦苣苔科植物的分子系统学研究已经极大地拓展了对这个科的种间关系与属一级之界定的理解。该文提供了一个苦苣苔科植物新分类系统,与传统的经典形态学分类系统比较,这一新系统被认为能更好地反映科下分类单元彼此之间的自然关系。众多传统意义上的属被分割、合并或者重新定义,这些巨大的分类变动正在影响着中国的苦苣苔科植物系统分类。此外,基于最近采集的材料,一些新属得以建立,一方面说明在中国还有相当多的野外工作需要做;而另一方面则再次证明了中国的苦苣苔科植物多样性是如此之丰富。在此,针对目前已经完成的工作和分类结论,对中国苦苣苔科植物的新分类系统进行了总结与概述,并深入地讨论了部分传统分类学上存在困惑的疑难属。     

腺花蛛毛苣苔——中国苦苣苔科蛛毛苣苔属一新记录种

报道了中国苦苣苔科蛛毛苣苔属一新记录种——腺花蛛毛苣苔(Paraboea glanduli flora Barnett)并提供了形态描述和野外生态照片,讨论了该种与近缘种类间的区别.该种为亚灌木状草本,无基生叶,聚伞花序2～4枝成对着生于茎顶叶腋,花冠密被腺毛,在该属中较为特殊.     

与先生之缘

正我任职于广西植物研究所园林园艺研究中心,对于专科专属植物的研究主要集中在苦苣苔科和荨麻科(楼梯草属)上,王文采院士是研究这两个科的泰斗,对我的研究工作给予了极大的指导和帮助,这也让地处一南一北的我们缔结下奇妙的缘分。华南苦苣苔科植物研究苦苣苔科分为苦苣苔亚科和大岩桐亚科,具有重要的观赏、药用和科研价值。中国苦苣苔科植物全部隶属于苦苣苔亚科,华南地区则是世界该亚科植物分布和特有中心之一,属、种分别约占     

广西苦苣苔科稀有珍贵植物——弥勒苣苔

首次报道广西苦苣苔科植物一新记录属:弥勒苣苔属.该属接近金盏苣苔属,但不同在于弥勒苣苔属花冠上唇4浅裂,下唇不分裂,2对雄蕊分别着生于花冠中部及其上方,雌蕊具一个柱头.弥勒苣苔属为中国特有的单型属,仅弥勒苣苔一种,分布于云南东南部和广西西部.该种在广西首次记录,凭证标本存放于广西植物标本馆(IBK).     

中国广西蛛毛苣苔属(苦苣苔科)一新种——桂林蛛毛苣苔

描述了采自中国广西的苦苣苔科新种桂林蛛毛苣苔Paraboea guilinensis L. Xu & Y. G. Wei。新种体态及叶形接近厚叶蛛毛苣苔P. crassifolia (Hemsl.) Burtt,与后者不同在于叶革质,花序梗、花梗及花萼均无毛,花冠明显二唇形,蒴果不旋扭。     

Significance of consensus CYC-binding sites found in the promoters of both ChCYC and ChRAD genes in Chirita heterotricha (Gesneriaceae)

CYC-like genes are widely conserved in controlling floral dorsoventral asymmetry (zygomorphy) through persistent expression in corresponding domains in core eudicots. To understand how CYC-like gene expression is maintained during flower development, we selected Chirita heterotricha as a material and isolated the promoter sequences of the ChCYCIC and ChCYCID genes, homologs of CYC, by inverse polymerase chain reaction. Further promoter analyses led to the identification of a putative cis-regulatory element in each promoter matching the consensus DNA binding site for Antirrhinum CYC protein: GGCCCCTC at-165 for ChCYC1C, and GGCCCCCC at-163 for ChCYCID. This indicates that both the ChCYCIC and ChCYC1D genes have probably evolved autoregulatory loops to sustain their expression in developing flowers. We also isolated the coding and promoter sequences of the ChRAD gene, a homolog of Antirrhinum RAD. Promoter analysis showed that the ChRAD gene promoter also contained a putative CYC-binding site (GGCCCAC at -134). Therefore, ChRAD is likely a direct target of the ChCYC1 genes, which is similar to Antirrhinum RAD. These results imply that the establishment of floral zygomorphy in Chirita may have been achieved by the evolution of an autoregulatory loop for CYC-like genes,which was probably accompanied by simultaneous co-option of the RAD-like gene into their regulatory network.     

Floral zygomorphy, the recurring evolution of a successful trait

The flowers of the primitive angiosperm plants were radially symmetrical (actinomorphic). Flowers with bilateral symmetry (zygomorphic) evolved in several clades independently as an adaptation to specialized methods of pollination and played an important role in the diversification of flowering plants. In the model species Antirrhinum majus (snapdragon), the related genes CYCLOIDEA (CYC) and DICHOTOMA (DICH) are key in the development of this trait. This raises the question of whether they played a role in the evolution of floral bilateral symmetry. To address this, the evolution of CYC in relation to the evolution of zygomorphy is being investigated. Phylogenetic and functional analyses of CYC-like genes are being carried out in groups either closely related to Antirhinum or in families where zygomorphy evolved as an independent event. In addition, the origin of zygomorphy is being studied by comparing the function of CYC-like genes in species with zygomorphic flowers with their function in species with radially symmetrical flowers.     

Evolution of GCYC, a Gesneriaceae homolog of CYCLOIDEA, within Gesnerioideae (Gesneriaceae)

Through recent advances in molecular developmental biology it has become clear that similar morphological traits may sometimes arise from different genetic bases. The molecular developmental biology of floral symmetry has been examined recently in detail and several genes important in controlling floral symmetry in diverse Asteridae have been identified. One of the most important among these is the floral symmetry gene CYCLOIDEA (CYC). We compared GCYC (the Gesneriaceae homolog of CYC) sequences in Gesneriaceae genera with the typical bilaterally symmetric flowers and genera with radial or near radial symmetry. Parsimony, Bayesian and maximum likelihood analyses of GCYC sequences among members of Gesnerioideae are mostly congruent with previous phylogenetic hypotheses, but suggest two unexpected generic positions: Diastema as sister to Gesneria, and Bellonia within Gloxinieae. In order to evaluate whether these results might be artifactual we obtained new gene sequences from chloroplast and nuclear ribosomal regions. These data disagree with GCYC regarding the placement of Diastema, but agree with GCYC regarding Bellonia. We did not find any mutations in GCYC that could explain the shift in symmetry and there were no consistent differences in molecular evolution between taxa with bilaterally or radially symmetric flowers. Likewise taxa with radial floral symmetry are not sister to each other showing that the loss of bilateral symmetry has occurred multiple times in parallel. Further investigations of GCYC expression will be necessary to determine if any of these independent events involved changes in the regulation of GCYC.     

Diversification and co-option of RAD-like genes in the evolution of floral asymmetry

To understand how changes in gene regulatory networks lead to novel morphologies, we have analysed the evolution of a key target gene, RAD, controlling floral asymmetry. In Antirrhinum, flower asymmetry depends on activation of RAD in dorsal regions of the floral meristem by the upstream regulators CYC and DICH. We show that Arabidopsis, a species with radially symmetric flowers, contains six RAD-like genes, reflecting at least three duplications since the divergence of Antirrhinum and Arabidopsis. Unlike the situation in Antirrhinum, none of the Arabidopsis RAD-like genes are activated in dorsal regions of the flower meristem. Rather, the RAD-like genes are expressed in distinctive domains along radial or ab-adaxial axes, consistent with a range of developmental roles. Introduction of a RAD genomic clone from Antirrhinum into Arabidopsis leads to a novel expression pattern that is distinct from the expression pattern of RAD in Antirrhinum and from the endogenous RAD-like genes of Arabidopsis. Nevertheless, RAD is able to influence developmental targets in Arabidopsis, as ectopic expression of RAD has developmental effects in this species. Taken together, our results suggest that duplication and divergence of RAD-like genes has involved a range of cis- and trans-regulatory changes. It is possible that such changes led to the coupling of RAD to CYC regulation in the Antirrhinum lineage and hence the co-option of RAD had a role in the generation of flower dorsoventral asymmetry.