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.     

基因调控高等植物开花机制的研究

对高等植物开花过程中基因参与调控的多条遗传途径进行了综述分析,并以拟南芥为例,提炼出在维持营养生长—进人生殖生长—花器官形成的多个重要阶段,各调节因子及各开花途径的互作效应,并归纳出以FLC、SOCl、FT、LFY等关键整合因子为纽带的植物开花基因调控模式.     

转录组分析揭示BAM1/2与SPL调控拟南芥花药发育的功能关系及调控网络

花药的正常发育对于植物正常繁衍和农作物产量都有着至关重要的作用.转录因子SPL及受体蛋白激酶BAM1和BAM2均在调控孢原细胞分化的过程中发挥重要作用.但迄今对其功能关系及下游信号途径仍缺乏深入研究.本文利用高通量转录组测序手段,通过对拟南芥spl、bam1、bam2、bam1bam2突变体花药的转录组对比分析,明确了:1)BAM1和BAM2功能冗余地调控花药绒毡层发育、脂质转运、花粉壁形成等方面,但两者在调控部分基因表达上存在亚功能化;2)SPL与BAM1/2共同调控791个花粉发育、脂类转运和细胞壁形成等过程相关基因的表达,同时BAM1/2特异调控326个脱落酸、水杨酸、茉莉酸信号途径以及水分、防御等胁迫刺激相关基因的表达,而SPL还单独调控3 789个基因的表达,主要参与到生长素合成和信号响应、水分、温度、光等相关的胁迫刺激.以上结果为生殖发育领域相关研究提供了重要参考.     

珙桐AP2/ERF家族相关基因的克隆及功能鉴定

为探究AP2/ERF类转录因子在珙桐开花调控及花器官发育中的作用,本研究通过从实验室珙桐不同发育时期苞片和叶片的转录组数据中筛选出调控珙桐苞片发育相关AP2/ERF家族基因DiAP2-1、DiDREB-1和DiRAV-1,经过生物信息学分析、基因克隆及功能鉴定初步研究了目的基因在珙桐苞片发育过程中的调控机制.亚细胞定位实验显示DiAP2-1主要定位于细胞质,DiDREB-1和DiRAV-1主要定位于细胞核.对目的基因在珙桐中的表达模式分析发现DiAP2-1和DiRAV-1在苞片发育过程中表达量逐渐降低,DiDREB-1只在苞片第三时期高表达,推测DiDREB-1、DiRAV-1属于A类基因,DiAP2-1属于B类基因.表型实验显示DiDREB-1和DiRAV-1转基因拟南芥株系较野生型表现为明显的早花,DiAP2-1同源基因对应的拟南芥突变体与同一时期野生型拟南芥对比表现为花瓣减少且萼片大,角果呈短粗状且败育.对关键开花调控基因表达模式分析发现DiDREB-1和DiRAV-1异源表达后使其他促花因子表达量上调进而使拟南芥花期提前.以上结果表明,AP2/ERF类转录因子DiAP2-1、D...     

榛子SBP基因家族鉴定及其系统进化和表达分析

SBP蛋白是植物所特有的一类转录因子,并广泛存在于植物中.研究发现,该基因家族成员参与响应包括植物生长、发育和抗逆等生物学过程,其功能研究具有重要意义.榛子是一种主要以果实为产品的重要经济林木,且该物种SBP基因家族的研究也未见报道.本研究以生物信息学手段为基础,结合转录组数据,从中筛选得到10个榛子SBP-like基因(SPL基因),并对其编码蛋白理化性质、保守区域和模体进行预测和分析,进而对基因家族进化关系和花发育不同时期表达水平进行了分析.结果表明:榛子的10个SPL基因可以分为5个大组群;均含有该基因家族所特有的C3H和C2CH结构域以及一个核定位信号;同时,基于花发育不同时期的转录组数据分析,部分成员在不同时期表达水平存在显著差异,预示该基因可能与花果发育有关.该研究将有助于榛子SPL基因参与其花果发育分子调控机制的深入研究.     

植物成花转变过程中的分子机理研究

植物由营养状态转入生殖状态是发育过程中的重要一步，其转变过程受遗传因子及环境的双重控制，光周期与温度是影响开花诱导的主要环境因子，而植物的同源异型基因参与植物器官内细胞排列方式形成的调控过程，是控制花分生组织特异性、花序分生组织特异性和花器官特异性的基础。     

甘蓝型油菜MADS-box基因家族APETALA3基因编码区的克隆与序列分析

随着对植物花发育分子机制研究取得的重大进展，通过对双子叶植物拟南芥和金鱼草的花同源异型突变体的研究确定了花器官发育的“ABC模式”，初步揭示了植物花发育的奥秘．在拟南芥中，属于A类基因的有APETALA1(AP1)和APETALA2(AP2)，属于B类基因的有APETALA3(AP3)PISTILATA     

植物激素调控籽粒大小的研究进展

植物种子的大小和品质是影响农作物产量的主要因素之一,研究控制种子籽粒大小发育的相关因素,对提高农作物产量具有重要意义.近年通过分析种子发育缺陷突变体或QTL等分子遗传学的方法,发现许多控制种子发育的重要基因影响着种子的大小和产量.对模式植物拟南芥和水稻的研究发现,许多调控种子发育的功能基因通过整合到植物激素的代谢或信号转导途径起作用,说明植物激素在调控籽粒发育中发挥重要作用,但有关作用的分子机理及其遗传调控网络待阐明.该文以模式植物拟南芥和水稻种子发育研究为例,着重介绍植物激素调控种子籽粒大小调控的研究进展.     

茉莉酸处理对木本油料植物小桐子早期花序芽基因表达的影响

小桐子(Jatropha curcas)是大戟科多年生木本油料植物,其种子含油率较高(30％～40％),是一种潜在的可再生能源植物.然而小桐子雌花较少,种子产量较低,严重限制其推广应用.茉莉酸是一种重要的植物激素,在花发育过程中起着重要作用.为了解茉莉酸在小桐子花发育过程中的作用,应用第二代高通量测序方法,对1.0 mmol/L茉莉酸处理24 h后的小桐子早期花序芽进行转录组测序分析,观测茉莉酸对花发育及相关基因转录的影响.结果显示,外源茉莉酸处理导致早期花序芽中1259个基因上调表达,695个基因下调表达;其中,10个与成花转变相关,8个与花器官发育相关以及18个与茉莉酸合成和信号转导途径相关的拟南芥同源基因表达发生了显著变化,但是花器官表型并未有显著的改变.差异表达基因的GO注释显示,"响应茉莉酸"功能分类基因在上调表达基因群中富集,表明在小桐子中存在可以响应外源茉莉酸处理的信号途径.茉莉酸处理后,与拟南芥花器官发育相关基因同源JcFUL、JcSRS、JcSEP1、JcAGL61、JcWOX1、JcTPR4和JcSEU下调表达,然而小桐子花器官表型没有明显改变,说明在小桐子中这些基因的变化表达不足以改变小桐子花器官表型.实验结果对解析茉莉酸在小桐子花发育调控过程中的作用有一定的参考价值.     

2013-27 科技要闻

植物生长素空间分布和器官形态新发现作为植物发育调控最重要的激素,生长素的含量及其在器官中的分布(空间分布)决定了植物器官的形态建成、株型以及向重性反应等生物学进程。然而,目前对植物生长素在器官中空间分布的调控机制仍缺乏了解。中国科学院植物研究所胡玉欣研究组以拟南芥为材料,通过研究功能获得及缺陷突变体,发现植物特有转录因子IDD14、IDD15和IDD16协     

The war of the whorls: genetic interactions controlling flower development.

The analysis of mutations affecting flower structure has led to the identification of some of the genes that direct flower development. Cloning of these genes has allowed the formulation of molecular models of how floral meristem and organ identity may be specified, and has shown that the distantly related flowering plants Arabidopsis thaliana and Antirrhinum majus use homologous mechanisms in floral pattern formation.     

Functional characterization and mapping of two MADS box genes from peach (Prunus persica)

Previously an AGAMOUS gene homologue PpMADS4 and a FRUITFULL gene homologue PpMADS6 were isolated from peach (Prunus persica), and both genes were shown to express in the developing floral and fruits. To gain insight into their function, the two genes were constitutively expressed in Arabidopsis thaliana and their effects on plant growth and floral organ development were studied in this work. The transgenic plants all displayed early flowering and conversion of inflorescence to floral meristem. However, the two genes had different effects on the floral organ structures in A. thaliana. The transgenic plants overexpressing PpMADS4 displayed homeotic conversion of floral organs, and par- ticularly the perianth abscission was inhibited. The plants overexpressing PpMADS6 showed early flowering, produced higher number of carpels, petals, and stamens than nontransgenic plants, and pod shatter was prevented; significantly, the transgenic plants yielded more than one siliques from a single flower. A SSR molecular marker was developed for PpMADS4, and it was then assigned into the G5 linkage group of Prunus sp. Both PpMADS4 and PpMADS6 genes were located at the same region in the G5 linkage group. Our results showed the potential application of these two MADS box genes for crop and fruit tree improvement.     

簸箕柳SPL基因家族分析

【目的】确定SPL基因家族在不同物种之间的选择性保留和丢失情况,为后续研究被子植物花发育提供参考。【方法】通过在拟南芥(Arabidopsis thaliana)、毛果杨(Populus trichocarpa)、簸箕柳(Salix suchowensis)、葡萄(Vitis vinifera)、番木瓜(Carica papaya)、水稻(Oryza sativa)6种被子植物基因组中查找SPL结构域,寻找6个物种的SPL同源序列。对所找到的SPL序列进行BLASTN比对鉴定同源基因类型。使用自编Perl脚本结合KaKs_Calculator计算SPL同源基因的非同义突变(K_a)以及同义突变(K_s)值,采用共线性分析确定该基因家族的复制和扩张方式。【结果】在6种被子植物基因组中,共发现120个SPL基因。根据种内和种间旁系同源、直系同源基因以及这些同源基因选择压的计算显示:杨柳科SPL同源基因最多,共有旁系同源基因24对,直系同源基因50对; 番木瓜没有旁系同源基因。6个物种中,木本植物比草本植物直系同源基因更多,双子叶植物比单子叶植物直系同源基因更多; 所有旁系同源和直系同源基因的K_a/K_s值均小于1。系统发育树的分析结果与基因同源性分析基本吻合,证明了这两种分析方法具有较高的可靠性。此次研究选取了簸箕柳同一植株上开花和不开花的枝条进行了转录组测序分析,差异表达分析发现了1个SPL基因(willow_GLEAN_10025160)在两种枝条的转录组中表达差异显著(P≤0.01),其在开花枝条中的表达量显著高于未开花枝条,该基因被选为SPL基因家族中参与簸箕柳开花调控的候选基因。【结论】通过对6个物种SPL基因家族的分析,发现6个物种中所有直系同源和旁系同源基因都经历了纯化选择(K_a/K_s<1),基因功能保守。这6个物种除了经历过全基因组复制事件,还发生过大规模的基因丢失或者通过其他方式产生的基因扩张,阐明了它们在进化历史上的复制事件及SPL基因在不同物种中的选择性保留与丢失情况,为进一步研究其在调控簸箕柳开花中的作用提供了有力证据。     

F-box proteins in flowering plants

In eukaryotes, the ubiquitin-mediated protein degradation pathway has been shown to control several key biological processes such as cell division, development, metabolism and immune response. F-box proteins, as a part of SCF (Skp1-Cullin (or Cdc53)-F-box) complex, functioned by interacting with substrate proteins, leading to their subsequent degradation by the 26S proteasome. To date, several F-box proteins identified in Arabidopsis and Antirrhinum have been shown to play important roles in auxin signal transduction, floral organ formation, flowering and leaf senescence. Arabidopsis genome sequence analysis revealed that it encodes over 1000 predicted F-box proteins accounting for about 5% of total predicted proteins. These results indicate that the ubiquitin-mediated protein degradation involving the F-box proteins is an important mechanism controlling plant gene expression. Here, we review the known F-box proteins and their functionsin flowering plants.     

Functional characterization and mapping of two MADS box genes from peach （Prunus persica）

Previously an AGAMOUS gene homologue PpMADS4 and a FRUITFULL gene homologue PpMADS6 were isolated from peach （Prunus persica）, and both genes were shown to express in the developing floral and fruits. To gain insight into their function, the two genes were constitutively expressed in Arabidopsis thaliana and their effects on plant growth and floral organ development were studied in this work. The transgenic plants all displayed early flowering and conversion of inflorescence to floral meristem. However, the two genes had different effects on the floral organ structures in A. thaliana. The transgenic plants overexpressing PpMADS4 displayed homeotic conversion of floral organs, and particularly the perianth abscission was inhibited. The plants overexpressing PpMADS6 showed early flowering, produced higher number of carpels, petals, and stamens than nontransgenic plants, and pod shatter was prevented; significantly, the transgenic plants yielded more than one siliques from a single flower. A SSR molecular marker was developed for PpMADS4, and it was then assigned into the G5 linkage group of Prunus sp. Both PpMADS4 and PpMADS6 genes were located at the same region in the G5 linkage group. Our results showed the potential application of these two MADS box genes for crop and fruit tree improvement.     

Evolution of genetic mechanisms controlling petal development.

Molecular genetic studies in Arabidopsis thaliana and other higher-eudicot flowering plants have led to the development of the 'ABC' model of the determination of organ identity in flowers, in which three classes of gene, A, B and C, are thought to work together to determine organ identity. According to this model, the B-class genes APETALA3 (AP3) and PISTILLATA (PI) act to specify petal and stamen identity. Here we test whether the roles of these genes are conserved throughout the angiosperms by analysing the expression of AP3 and PI orthologues in the lower eudicot subclass Ranunculidae. We show that, although expression of these orthologues in the stamens is conserved, the expression patterns in the petals differ from those found in the higher eudicots. The differences between these expression patterns suggest that the function of AP3 and PI homologues as B-class organ-identity genes is not rigidly conserved among all angiosperms. These observations have important implications for understanding the evolution of both angiosperm petals and the genetic mechanisms that control the identities of floral organs.     

Integration of floral inductive signals in Arabidopsis

Flowering of Arabidopsis is regulated by a daylength-dependent pathway that accelerates flowering in long days and a daylength-independent pathway that ensures flowering in the absence of inductive conditions. These pathways are genetically separable, as there are mutations that delay flowering in long but not short days. Conversely, mutations that block synthesis of the hormone gibberellin abolish flowering in short days, but have on their own only a minor effect in long days. A third pathway, the autonomous pathway, probably acts by modulating the other two pathways. Understanding where and how these pathways are integrated is a prerequisite for understanding why similar environmental or endogenous cues can elicit opposite flowering responses in different plants. In Arabidopsis, floral induction leads ultimately to the upregulation of floral meristem-identity genes such as LEAFY, indicating that floral inductive signals are integrated upstream of LEAFY Here we show that gibberellins activate the LEAFY promoter through cis elements that are different from those that are sufficient for the daylength response, demonstrating that the LEAFY promoter integrates environmental and endogenous signals controlling flowering time.     

一种带有表型标记基因的诱导型抗旱植物表达载体的构建

为了克服组成型表达转录因子基因影响转基因植物性状的缺点,并构建一种具有级联放大作用并带有表型标记的诱导型植物双价表达载体。研究采用PCR方法从拟南芥克隆获得冷诱导转录因子CBF3基因,蜡质合成相关WIN1基因,干旱诱导RD29A基因启动子和冷诱导的LEA14基因启动子,并用CBF3转录因子所调控的下游RD29A基因启动子和LEA14基因启动子分别驱动CBF3基因和W1N1基因表达,构建了双价植物表达载体RD29AP-CBF3／LEA14P—WIN1／pcAMBIA2201。我们预测在转基因植物中,该表达系统可在干旱等逆境信号存在条件下,通过级联放大的方式诱导表达,在增加植物抗逆性的同时,增加叶片表层蜡质的积累,从而易于表型识别。本研究为利用花粉管通道法转化棉花,提高抗逆转基因棉花田间筛选的效率奠定了基础。