植物印迹基因研究进展

印迹基因的表达受表观机制调控,依据其亲本来源在植物胚乳中表现为单等位基因表达的特殊模式。这些基因在调控胚及其附属结构的发育、控制种子的大小、生殖隔离以及防止无性生殖上发挥着关键作用。随着植物表观遗传学研究的不断深入,目前对于印迹基因的探索已逐渐成为表观遗传学研究的热点。文章介绍了关于印迹基因起源的亲本冲突学说,并以拟南芥的MEA、FIS2、FWA、MPC、PHE1,玉米的FIE1、FIE2等重要印迹基因为例,阐述了有关植物印迹基因的表达调控机制及最新研究进展。     

拟南芥印迹基因MEA

MEDEA(MEA)是第1个在拟南芥胚乳中发现的印迹基因.MEA基因的印迹受甲基转移酶MET1和DNA糖基酶DME之间的拮抗作用调控.MEA基因编码polycomb group(PcG)蛋白,并与FIE、MSI1等其他PcG蛋白形成复合物,维持与细胞增殖相关基因的表达抑制状态.mea突变会启动未受精状态下中央细胞核的复制,而使种子发生败育.目前已发现Ⅰ型MADSbox基因PHERES1是受MEA直接作用的.研究MEA基因对种子发育的调控,不仅有助于阐明原胚及胚乳启动的机制,在无融合生殖研究中也有意义.PcG蛋白在动植物中的强保守性说明,PcG基因印迹在个体发育过程中有调控作用.     

被子植物胚乳自主发生的分子机理研究进展

双受精是被子植物的一个有效的进化策略,增加了胚和种苗成活的机会,其中胚乳在胚以及种子发育中至关重要。然而极少数进行无融合生殖的被子植物,它们的胚乳可以在假受精或不受精的情况下克服基因组印记效应的影响且正常发育。拟南芥胚乳自主发生突变体及相关基因的克隆,使人们可以研究和比较自然和突变植物胚乳自主发生的分子机制。本文着重介绍基因组印记对有性生殖和无融合生殖植物胚乳发育的影响,分析和讨论近年来发现的有关胚乳自主发生的基因（如MEA,FIS2,FIE,MSI1和PHE1）及其可能的作用机理。     

植物胚乳发育的表观遗传学调控

被子植物的种子发育从双受精开始, 产生二倍体的胚和三倍体的胚乳。在种子发育和萌发过程中, 胚乳向胚组织提供营养物质, 因此胚乳对胚和种子的正常生长发育至关重要。开花植物发生基因组印迹的主要器官是胚乳。印迹基因的表达受表观遗传学机制的调控, 包括DNA甲基化和组蛋白H3K27甲基化修饰以及依赖于PolIV的siRNAs (p4-siRNAs)调控。基因组印迹的表观遗传学调控对胚乳的正常发育和种子育性具有不可或缺的重要作用。最新研究显示, 胚乳的整个基因组DNA甲基化水平降低, 而且去甲基化作用可能源于雌配子体的中央细胞。该文综述了种子发育的表观遗传学调控机制, 包括基因组印迹机制以及胚乳基因组DNA甲基化变化研究的最新进展。     

拟南芥的印记基因和印记表达调控

基因组印记是指后代仅表达亲本之一基因拷贝的现象。印记基因的发生是防止孤雌生殖发生的有效手段之一。拟南芥FIS(Fertilisation-independent seed)印记基因mea、fis2和fie在中央细胞分裂抑制和早期胚乳发育调节中发挥重要作用。fis突变体具有两种表型:当受精缺失时二倍体胚乳自主发育,而当受精发生时形成非细胞化的胚乳。FIS多梳蛋白复合体(Polycomb protein complex)包括上述3种FIS蛋白,在目标位点催化组蛋白H3第27位赖氨酸的tri-甲基化(H3K27 tri-methylation)。DME(DEMETER)和AtMET1(Methyltransferase1)参与了mea和fis2的印记表达控制。最近研究结果表明,开花植物中转座子的插入影响邻近基因的表达,是基因组印记进化的主要驱动力量。本文综述了10年来拟南芥中FIS印记基因和相关基因的发现及其调控机理,期望能为水稻、玉米等重要作物中印记基因的研究提供借鉴和参考。     

胁迫诱导植物体细胞胚发生中相关基因及蛋白的研究进展

植物体细胞胚发生是一个复杂的发育过程,体细胞胚发生已成为研究植物胚胎发育过程中生理、生化、分子生物学等方面分子机理的模式系统。胁迫被认为是对体细胞胚的诱导有重要作用的因素。植物生长调节物质如2,4-D、ABA等目前认为是与胚性能力获得有关的胁迫物质。在蛋白和转录水平上对基因表达的分析中已鉴定出一些与体细胞胚发生相关的基因和蛋白。该文主要对近年来国内外有关胁迫诱导体细胞胚发生的相关基因及蛋白的研究进展进行综述。     

小麦×大麦胚胎发育的研究——Ⅲ.小麦×大麦胎胚发育过程中淀粉的积累和动态

我们观察了小麦与大麦杂交胚胎发育过程中,雌蕊各个组成部分发生的淀粉积累和转移的动态。结果如下: 1.胚乳和杂种胚早期发育过程中,子房壁和胚囊中淀粉的积累和动态的趋势与其他学者所作小麦自交的情况基本相同。2.当胚乳细胞充满胚囊而胚没有分化时,子房壁中的淀粉已极少,当胚乳解体时,子房壁中淀粉已几乎消失。可见,子房壁中淀粉的迅速消失与胚乳的迅速败育是平行的。3.胚囊发育的停滞与败育跟子房壁组织中淀粉的积累及对胚囊营养的正常供应有密切关系。4.花柱、珠被、珠心组织及胚囊中的助细胞和反足细胞,在整个杂种胚和胚乳发育过程中,始终不存在淀粉粒。助细胞胚亦和助细胞一样,无淀粉粒的存在。     

小麦×大麦胚胎发育的研究——Ⅲ.小麦×大麦胎胚发育过程中淀粉的积累和动态

我们观察了小麦与大麦杂交胚胎发育过程中,雌蕊各个组成部分发生的淀粉积累和转移的动态。结果如下: 1.胚乳和杂种胚早期发育过程中,子房壁和胚囊中淀粉的积累和动态的趋势与其他学者所作小麦自交的情况基本相同。2.当胚乳细胞充满胚囊而胚没有分化时,子房壁中的淀粉已极少,当胚乳解体时,子房壁中淀粉已几乎消失。可见,子房壁中淀粉的迅速消失与胚乳的迅速败育是平行的。3.胚囊发育的停滞与败育跟子房壁组织中淀粉的积累及对胚囊营养的正常供应有密切关系。4.花柱、珠被、珠心组织及胚囊中的助细胞和反足细胞,在整个杂种胚和胚乳发育过程中,始终不存在淀粉粒。助细胞胚亦和助细胞一样,无淀粉粒的存在。     

miRNA参与植物胚和胚乳发育调控的研究进展

籽粒性状是构成产量的重要基础,是粮食产量的最终体现,也是育种最复杂的性状之一。籽粒主要由胚和胚乳组成,胚乳是积累和贮藏营养物质的场所,主要为胚的萌发和生长提供营养。胚乳细胞的发育、增殖和充实情况决定了籽粒的重量和品质。籽粒发育是一个非常复杂的生物过程,涉及许多基因的时空表达以及转录水平和转录后水平的调控。微小RNA（microRNAs,miRNAs）是一类内源性的非编码小RNA（21～24 nt）,可通过靶向降解和翻译抑制在转录后水平调控植物基因表达。miRNA及其靶基因组成精密的调控网络参与籽粒的发育。基于此,概述了植物miRNAs的生成及作用机制,综述了miRNAs在植物胚和胚乳发育中的调控功能研究进展,以期为进一步鉴定与玉米籽粒发育相关的miRNAs并解析其调控功能提供更好的研究方向。     

植物体细胞胚发生过程中基因表达的研究进展

植物体细胞胚胎发生是一个复杂的发育过程,研究者们通过分析植物体细胞胚发生过程中的基因表达或胚性组织和非胚性组织中基因的差异表达,获得了在体细胞胚发生过程不同时期表达的基因,并分析了这些基因在胚胎发生途径中可能的作用。综述了在植物体细胞胚发生过程中细胞周期相关基因、胁迫和激素应答相关基因、信号转导相关基因、晚期胚胎丰富蛋白基因及与体细胞胚发生相关的胞外蛋白基因表达的研究进展。     

Maintenance of DNA methylation during the Arabidopsis life cycle is essential for parental imprinting

Imprinted genes are expressed predominantly from either their paternal or their maternal allele. To date, all imprinted genes identified in plants are expressed in the endosperm. In Arabidopsis thaliana, maternal imprinting has been clearly demonstrated for the Polycomb group gene MEDEA (MEA) and for FWA. Direct repeats upstream of FWA are subject to DNA methylation. However, it is still not clear to what extent similar cis-acting elements may be part of a conserved molecular mechanism controlling maternally imprinted genes. In this work, we show that the Polycomb group gene FERTILIZATION-INDEPENDENT SEED2 (FIS2) is imprinted. Maintenance of FIS2 imprinting depends on DNA methylation, whereas loss of DNA methylation does not affect MEA imprinting. DNA methylation targets a small region upstream of FIS2 distinct from the target of DNA methylation associated with FWA. We show that FWA and FIS2 imprinting requires the maintenance of DNA methylation throughout the plant life cycle, including male gametogenesis and endosperm development. Our data thus demonstrate that parental genomic imprinting in plants depends on diverse cis-elements and mechanisms dependent or independent of DNA methylation. We propose that imprinting has evolved under constraints linked to the evolution of plant reproduction and not by the selection of a specific molecular mechanism.     

Histone H1 affects gene imprinting and DNA methylation in Arabidopsis

Imprinting, i.e. parent-of-origin expression of alleles, plays an important role in regulating development in mammals and plants. DNA methylation catalyzed by DNA methyltransferases plays a pivotal role in regulating imprinting by silencing parental alleles. DEMETER (DME), a DNA glycosylase functioning in the base-excision DNA repair pathway, can excise 5-methylcytosine from DNA and regulate genomic imprinting in Arabidopsis. DME demethylates the maternal MEDEA (MEA) promoter in endosperm, resulting in expression of the maternal MEA allele. However, it is not known whether DME interacts with other proteins in regulating gene imprinting. Here we report the identification of histone H1.2 as a DME-interacting protein in a yeast two-hybrid screen, and confirmation of their interaction by the in vitro pull-down assay. Genetic analysis of the loss-of-function histone h1 mutant showed that the maternal histone H1 allele is required for DME regulation of MEA, FWA and FIS2 imprinting in Arabidopsis endosperm but the paternal allele is dispensable. Furthermore, we show that mutations in histone H1 result in an increase of DNA methylation in the maternal MEA and FWA promoter in endosperm. Our results suggest that histone H1 is involved in DME-mediated DNA methylation and gene regulation at imprinted loci.     

DEMETER,Goddess of the harvest,activates maternal MEDEA to produce the perfect seed

Parental imprinting regulates processes as diverse as mammalian development and seed size in crop plants. In Arabidopsis, the DNA glycosylase DEMETER regulates early seed development, through activation of the maternal copy of the imprinted MEDEA Polycomb gene. Paternal silencing of MEA appears to be a default condition.     

Imprinting of the MEDEA polycomb gene in the Arabidopsis endosperm.

In flowering plants, two cells are fertilized in the haploid female gametophyte. Egg and sperm nuclei fuse to form the embryo. A second sperm nucleus fuses with the central cell nucleus that replicates to generate the endosperm, which is a tissue that supports embryo development. MEDEA (MEA) encodes an Arabidopsis SET domain Polycomb protein. Inheritance of a maternal loss-of-function mea allele results in embryo abortion and prolonged endosperm production, irrespective of the genotype of the paternal allele. Thus, only the maternal wild-type MEA allele is required for proper embryo and endosperm development. To understand the molecular mechanism responsible for the parent-of-origin effects of mea mutations on seed development, we compared the expression of maternal and paternal MEA alleles in the progeny of crosses between two Arabidopsis ecotypes. Only the maternal MEA mRNA was detected in the endosperm from seeds at the torpedo stage and later. By contrast, expression of both maternal and paternal MEA alleles was observed in the embryo from seeds at the torpedo stage and later, in seedling, leaf, stem, and root. Thus, MEA is an imprinted gene that displays parent-of-origin-dependent monoallelic expression specifically in the endosperm. These results suggest that the embryo abortion observed in mutant mea seeds is due, at least in part, to a defect in endosperm function. Silencing of the paternal MEA allele in the endosperm and the phenotype of mutant mea seeds supports the parental conflict theory for the evolution of imprinting in plants and mammals.     

DEMETER DNA glycosylase establishes MEDEA polycomb gene self-imprinting by allele-specific demethylation

MEDEA (MEA) is an Arabidopsis Polycomb group gene that is imprinted in the endosperm. The maternal allele is expressed and the paternal allele is silent. MEA is controlled by DEMETER (DME), a DNA glycosylase required to activate MEA expression, and METHYLTRANSFERASE I (MET1), which maintains CG methylation at the MEA locus. Here we show that DME is responsible for endosperm maternal-allele-specific hypomethylation at the MEA gene. DME can excise 5-methylcytosine in vitro and when expressed in E. coli. Abasic sites opposite 5-methylcytosine inhibit DME activity and might prevent DME from generating double-stranded DNA breaks. Unexpectedly, paternal-allele silencing is not controlled by DNA methylation. Rather, Polycomb group proteins that are expressed from the maternal genome, including MEA, control paternal MEA silencing. Thus, DME establishes MEA imprinting by removing 5-methylcytosine to activate the maternal allele. MEA imprinting is subsequently maintained in the endosperm by maternal MEA silencing the paternal allele.     

DEMETER,a DNA glycosylase domain protein,is required for endosperm gene imprinting and seed viability in arabidopsis

We isolated mutations in Arabidopsis to understand how the female gametophyte controls embryo and endosperm development. For the DEMETER (DME) gene, seed viability depends only on the maternal allele. DME encodes a large protein with DNA glycosylase and nuclear localization domains. DME is expressed primarily in the central cell of the female gametophyte, the progenitor of the endosperm. DME is required for maternal allele expression of the imprinted MEDEA (MEA) Polycomb gene in the central cell and endosperm. Ectopic DME expression in endosperm activates expression of the normally silenced paternal MEA allele. In leaf, ectopic DME expression induces MEA and nicks the MEA promoter. Thus, a DNA glycosylase activates maternal expression of an imprinted gene in the central cell.     

Identification of new members of Fertilisation Independent Seed Polycomb Group pathway involved in the control of seed development in Arabidopsis thaliana

In higher plants, double fertilisation initiates seed development. One sperm cell fuses with the egg cell and gives rise to the embryo, the second sperm cell fuses with the central cell and gives rise to the endosperm. The endosperm develops as a syncytium with the gradual organisation of domains along an anteroposterior axis defined by the position of the embryo at the anterior pole and by the attachment to the placenta at the posterior pole. We report that ontogenesis of the posterior pole in Arabidopsis thaliana involves oriented migration of nuclei in the syncytium. We show that this migration is impaired in mutants of the three founding members of the FERTILIZATION INDEPENDENT SEED (FIS) class, MEDEA (MEA), FIS2 and FERTILIZATION INDEPENDENT ENDOSPERM (FIE). A screen based on a green fluorescent protein (GFP) reporter line allowed us to identify two new loci in the FIS pathway, medicis and borgia. We have cloned the MEDICIS gene and show that it encodes the Arabidopsis homologue of the yeast WD40 domain protein MULTICOPY SUPRESSOR OF IRA (MSI1). The mutations at the new fis loci cause the same cellular defects in endosperm development as other fis mutations, including parthenogenetic development, absence of cellularisation, ectopic development of posterior structures and overexpression of the GFP marker.