RESUMO
The Hieracium and Pilosella (Lactuceae, Asteraceae) genera of closely related hawkweeds contain species with two different modes of gametophytic apomixis (asexual seed formation). Both genera contain polyploid species, and in wild populations, sexual and apomictic species co-exist. Apomixis is known to co-exist with sexuality in apomictic Pilosella individuals, however, apomictic Hieracium have been regarded as obligate apomicts. Here, a developmental analysis of apomixis within 16 Hieracium species revealed meiosis and megaspore tetrad formation in 1 to 7% of ovules, for the first time indicating residual sexuality in this genus. Molecular markers linked to the two independent, dominant loci LOSS OF APOMEIOSIS (LOA) and LOSS OF PARTHENOGENESIS (LOP) controlling apomixis in Pilosella piloselloides subsp. praealta were screened across 20 phenotyped Hieracium individuals from natural populations, and 65 phenotyped Pilosella individuals from natural and experimental cross populations, to examine their conservation, inheritance and association with reproductive modes. All of the tested LOA and LOP-linked markers were absent in the 20 Hieracium samples irrespective of their reproductive mode. Within Pilosella, LOA and LOP-linked markers were essentially absent within the sexual plants, although they were not conserved in all apomictic individuals. Both loci appeared to be inherited independently, and evidence for additional genetic factors influencing quantitative expression of LOA and LOP was obtained. Collectively, these data suggest independent evolution of apomixis in Hieracium and Pilosella and are discussed with respect to current knowledge of the evolution of apomixis.
Assuntos
Apomixia/genética , Asteraceae/genética , Evolução Biológica , Genes de Plantas , Loci Gênicos , Sequência Conservada , DNA de Cloroplastos/genética , DNA de Plantas/genética , Regulação da Expressão Gênica de Plantas , Marcadores Genéticos , Genética Populacional , Haplótipos , Padrões de Herança , Dados de Sequência Molecular , Sementes/genéticaRESUMO
Callose accumulates in the walls of cells undergoing megasporogenesis during embryo sac formation in angiosperm ovules. Deficiencies in callose deposition have been observed in apomictic plants and causal linkages between altered callose deposition and apomictic initiation proposed. In apomictic Hieracium, embryo sacs initiate by sexual and apomictic processes within an ovule, but sexual development terminates in successful apomicts. Callose deposition and the events that lead to sexual termination were examined in different Hieracium apomicts that form initials pre- and post-meiosis. In apomictic plants, callose was not detected in initial cell walls and deficiencies in callose deposition were not observed in cells undergoing megasporogenesis. Multiple initial formation pre-meiosis resulted in physical distortion of cells undergoing megasporogenesis, persistence of callose and termination of the sexual pathway. In apomictic plants, callose persistence did not correlate with altered spatial or temporal expression of a beta-1,3-glucanase gene (HpGluc) encoding a putative callose-degrading enzyme. Expression analysis indicated HpGluc might function during ovule growth and embryo sac expansion in addition to callose dissolution in sexual and apomictic plants. Initial formation pre-meiosis might therefore limit the access of HpGluc protein to callose substrate while the expansion of aposporous embryo sacs is promoted. Callose deposition and dissolution during megasporogenesis were unaffected when initials formed post-meiosis, indicating other events cause sexual termination. Apomixis in Hieracium is not caused by changes in callose distribution but by events that lead to initial cell formation. The timing of initial formation can in turn influence callose dissolution.
Assuntos
Glucanos/metabolismo , Magnoliopsida/metabolismo , beta-Glucosidase/biossíntese , Parede Celular/metabolismo , DNA Complementar , Expressão Gênica , Glucana 1,3-beta-Glucosidase , Glucanos/análise , Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Imuno-Histoquímica , Magnoliopsida/fisiologia , Meiose , Proteínas de Membrana , Morfogênese , Óvulo/crescimento & desenvolvimento , Filogenia , Ploidias , Reprodução , Sementes/crescimento & desenvolvimento , beta-Glucosidase/genéticaRESUMO
The effects of the Agrobacterium rhizogenes rolB oncogene on apomixis were examined in the facultative apomictic plant Hieracium piloselloides because the oncogene has been shown to alter plant growth, morphogenesis and cellular sensitivity to auxin. Introduction of rolB under the control of either its own promoter or the CaMV35S promoter induced ectopic meristem formation from the inflorescence, confirming in planta a meristem-inducing role for this oncogene previously observed only in tissue culture. These ectopic meristems formed vegetative rosettes and floral plant organs. Upon immersion in water these meristems generated roots, suggesting that meristem commitment towards the generation of a specific organ type is a separate and later event that is dependent upon the developmental context. Ovule identity and form was altered in ectopically induced florets in plants expressing the CaMV35S::rolB construct. In contrast to the ovules of untransformed apomictic plants, the sexual process ceased earlier, prior to meiosis, yet surprisingly, apomixis initiated from a greater number of cells, and embryos and endosperm continued to develop in the structurally altered ovules. The alternative possibilities that the effects on reproduction might result from rolB influencing cellular response to auxin, or from alterations in cell signaling caused by changes in ovule morphology that are induced because of the expression of the oncogene are discussed.
Assuntos
Asteraceae/crescimento & desenvolvimento , Proteínas de Bactérias/genética , Meristema/crescimento & desenvolvimento , Estruturas Vegetais/crescimento & desenvolvimento , beta-Glucosidase/genética , Asteraceae/genética , Proteínas de Bactérias/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Ácidos Indolacéticos/administração & dosagem , Meiose/genética , Meristema/genética , Estruturas Vegetais/genética , Regiões Promotoras Genéticas , Reprodução/efeitos dos fármacos , Reprodução/genética , Transdução de Sinais , beta-Glucosidase/metabolismoRESUMO
Apomixis, the asexual formation of seed, has been known in angiosperms for more than a century yet the genetic mechanisms that control this trait remain poorly understood. Most members of the genus Hieracium are apomicts, forming predominantly asexual seed. Some purely sexual forms, however, also exist. In this paper we present a study of the inheritance of apomixis using two apomictic species of Hieracium which utilize very different forms of megagametogenesis. In both cases the progeny inherited apomixis as a monogenic, dominant trait that could be transferred by both haploid and diploid male gametes. In common with observations from other systems, no diploid apomictic progeny were recovered from these crosses. This appears to have been caused by selection against the survival of diploid zygotes, rather than against the mediation of haploid gametes as has been noted in other systems. Crosses between the two apomicts showed that the dominant determinants in the two forms examined were closely linked, possibly allelic. The significance of these data is discussed with respect to current theories on the associative link between gametophytic apomixis and polyploidy.
Assuntos
Asteraceae/genética , Reprodução Assexuada/genética , Reprodução/genética , Sementes/genética , Alelos , Cruzamentos Genéticos , Genes de Plantas , Haploidia , Modelos GenéticosRESUMO
In Arabidopsis, seedless silique development or parthenocarpy can be induced by the application of various plant growth regulators (PGRs) to unfertilized pistils. Ecotype-specific responses were observed in the Arabidopsis ecotypes Columbia and Landsberg relative to the type of PGR and level applied. The parthenocarpic response was greatest in ecotype Landsberg, and comparisons of fruit growth and morphology were studied primarily in this ecotype. Gibberellic acid application (10 micromol pistil(-1)) caused development similar to that in pollinated pistils, while benzyladenine (1 micromol pistil(-1)) and naphthylacetic acid (10 micromol pistil(-1)) treatment produced shorter siliques. Naphthylacetic acid primarily modified mesocarp cell expansion. Arabidopsis mutants were employed to examine potential dependencies on gibberellin biosynthesis (ga1-3, ga4-1, and ga5-1) and perception (spy-4 and gai) during parthenocarpic silique development. Emasculated spy-4 pistils were neither obviously parthenocarpic nor deficient in PGR perception. By contrast, emasculated gai mutants did not produce parthenocarpic siliques following gibberellic acid application, but silique development occurred following pollination or application of auxin and cytokinin. Pollinated gai siliques had decreased cell numbers and morphologically resembled auxin-induced parthenocarpic siliques. This shows that a number of independent and possibly redundant pathways can direct hormone-induced parthenocarpy, and that endogenous gibberellins play a role in regulating cell expansion and promoting cell division in carpels.
Assuntos
Arabidopsis/genética , Giberelinas/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Modelos Biológicos , Mutação , Reguladores de Crescimento de Plantas/fisiologiaRESUMO
Citrus exhibits polyembryonic seed development, an apomictic process in which many maternally derived embryos arise from the nucellus surrounding the developing zygotic embryo. Citrus seed storage proteins were used as markers to compare embryogenesis in developing seeds and somatic embryogenesis in vitro. The salt-soluble, globulin protein fraction (designated citrin) was purified from Citrus sinensis cv Valencia seeds. Citrins separated into two subunits averaging 22 and 33 kD under denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A cDNA clone was isolated representing a citrin gene expressed in seeds when the majority of embryos were at the early globular stage of embryo development. The predicted protein sequence was most related to the globulin seed storage proteins of pumpkin and cotton. Accumulation of 33-kD polypeptides was first detected in polyembryonic Valencia seeds when the majority of embryos were at the globular stage of development. Somatic Citrus embryos cultured in vivo were observed to initiate 33-kD polypeptide accumulation later in embryo development but accumulated these peptides at only 10 to 20% of the level observed in polyembryonic seeds. Therefore, factors within the seed environment must influence the higher quantitative levels of citrin accumulation in nucellar embryos developing in vivo, even though nucellar embryos, like somatic embryos, are not derived from fertilization events.
Assuntos
Citrus/fisiologia , Expressão Gênica , Proteínas de Plantas/biossíntese , Sequência de Aminoácidos , Sequência de Bases , Células Cultivadas , Clonagem Molecular , Primers do DNA , DNA Complementar , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Genes de Plantas , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/isolamento & purificação , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , RNA Mensageiro/isolamento & purificação , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas de Armazenamento de Sementes , Sementes/fisiologia , Especificidade da EspécieRESUMO
We identified two Arabidopsis embryo mutants, designated as raspberry1 and raspberry2, by screening T-DNA-mutagenized Arabidopsis lines. Embryogenesis in these mutants is indistinguishable from that of wild-type plants until the late-globular stage, after which raspberry1 and raspberry2 embryos fail to undergo the transition to heart stage, remain globular shaped, and proliferate an enlarged suspensor region. raspberry1 and raspberry2 embryo-proper regions enlarge during embryogenesis, become highly vacuolate, and display prominent convex, or "raspberry-like" protuberances on their outer cell layers. In situ hybridization studies with several embryo cell-specific mRNA probes indicated that the raspberry1 and raspberry2 embryo-proper regions differentiate tissue layers in their correct spatial contexts and that the regulation of cell-specific genes within these layers is normal. Surprisingly, a similar spatial and temporal pattern of mRNA accumulation occurs within the enlarged suspensor region of raspberry1 and raspberry2 embryos, suggesting that a defect in embryo-proper morphogenesis can cause the suspensor to take on an embryo-proper-like state and differentiate a radial tissue-type axis. We conclude that cell differentiation can occur in the absence of both organ formation and morphogenesis during plant embryogenesis and that interactions occur between the embryo-proper and suspensor regions.
RESUMO
We studied the temporal and spatial regulation of three mRNA sequence sets that are present exclusively, or at elevated levels, in the tobacco anther. One mRNA set accumulates in the tapetum and decays as the tapetum degenerates later in anther development. The second mRNA set accumulates after the tapetal-specific mRNAs, is localized within the stomium and connective, and also decays as these cell types degenerate during anther maturation. The third mRNA sequence set persists throughout anther development and is localized within most anther tissues. A tapetal-specific gene, designated as TA29, was isolated from a tobacco genome library. Runoff transcription studies and experiments with chimeric [beta]-glucuronidase and diphtheria toxin A-chain genes showed that the TA29 gene is regulated primarily at the transcriptional level and that a 122-base pair 5[prime] region can program the tapetal-specific expression pattern. Destruction of the tapetum by the cytotoxic gene had no effect on the differentiation and/or function of surrounding sporophytic tissues but led to the production of male-sterile plants. Together, our studies show that several independent gene expression programs occur during anther development and that these programs correlate with the differentiated state of specific anther cell types.
RESUMO
GV1B is one of five viroids that have recently been purified from grapevines. GV1B has now been sequenced and its 363 nucleotide residues can potentially form the typical rod-like structure of viroids with 67% of nucleotides base-paired. GV1B has highest sequence similarity with grapevine yellow speckle viroid (GYSV; 73%) and has a central sequence which is conserved in GYSV and apple scar skin viroid (ASSV) which have been reported to constitute the ASSV group. Therefore, we have placed GV1B into the ASSV group. GV1B contains a direct repeat sequence at the terminal portions of its T1 and T2 regions. GV1B also contains a sequence of 69 nucleotides in the terminal portion of its T1 region which is almost identical to the corresponding region in tomato planta macho viroid (TPMV). This provides further evidence of the importance of RNA recombination in viroid evolution.
Assuntos
DNA Viral/genética , Vírus de Plantas/genética , Viroides/genética , Sequência de Bases , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Sequências Repetitivas de Ácido NucleicoRESUMO
A scheme for viroid classification is proposed based on the nature of the strictly conserved core sequence present in the central portion of the secondary structure of viroids. In this scheme, all of the known viroids can be classified as potato spindle tuber-type viroids consisting of two viroid groups and avocado sunblotch-type viroids containing one viroid group.
Assuntos
RNA Viral , Viroides/classificação , Sequência de Bases , Dados de Sequência Molecular , Conformação de Ácido Nucleico , RNA Viral/genética , Sequências Reguladoras de Ácido Nucleico , Viroides/genéticaRESUMO
A single stranded circular RNA was isolated from grapevines infected with yellow speckle disease. The RNA which we have called grapevine yellow speckle viroid (GYSV), contains 367 nucleotide residues and has the potential to form the rod-like secondary structure characteristic of viroids. GYSV has 37% sequence homology with the recently described apple scar skin viroid (ASSV; 330 residues) and has some sequence homology with the viroids in the potato spindle tuber viroid (PSTV) group. The sequence of GYSV has characteristics which fit the structural domains described for the PSTV group. However, GYSV lacks the PSTV central conserved sequence. Instead, there is a conserved sequence in the central region of GYSV and ASSV which has the potential to form a stem loop configuration and a stable palindromic structure as does the central conserved region of the PSTV group. These structural features suggest there is a different central conserved region for GYSV and ASSV. The results support the viroid nature of GYSV and its inclusion into a separate viroid group which we suggest should be represented by ASSV.
Assuntos
Vírus de Plantas/ultraestrutura , Viroides/ultraestrutura , Sequência de Bases , DNA Viral/análise , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Vírus de Plantas/genética , Homologia de Sequência do Ácido Nucleico , Viroides/genéticaRESUMO
Analysis of nucleic acids from grapevine tissues by two-dimensional gel electrophoresis demonstrated the presence of two bands of circular RNA. The smaller RNA contained about 300 nucleotide residues and was identified as hop stunt viroid by nucleotide sequencing. The larger RNA band was a mixture of species and contained similar amounts of two components, referred to as RNA 1a and RNA 1b, and in addition a trace amount of citrus exocortis viroid (CEV) which became detectable only after inoculation of the mixture to tomato. The identity of CEV was determined by probe hybridization and nucleotide sequencing. Both RNAs 1a and 1b are distinct from CEV and have estimated sizes larger than those of CEV and other viroids reported so far. RNA 1a preparations were infectious in cucumber and in tomato and the recovered viroid had unique properties. We have provisionally named this viroid Australian grapevine viroid. Evidence for the autonomous replication of RNA 1b was not obtained.
Assuntos
Frutas/microbiologia , RNA Viral/isolamento & purificação , RNA/isolamento & purificação , Viroides/isolamento & purificação , Sequência de Bases , Frutas/análise , Dados de Sequência Molecular , RNA CircularRESUMO
Experiments concerning gene transcription in Xenopus oocytes have revealed that the efficiency of the HSV-TK promoter is dependent upon the nature of the attached gene sequences. Transcriptional efficiency of the TK promoter when attached to its own gene was 30-fold higher than that observed when the promoter was attached either to an avian keratin gene or to the chicken histone H2B gene. Furthermore, attachment of the keratin gene promoter to the TK gene resulted in a 20-fold increase in keratin promoter efficiency. It was found by subsequent experiments that the TK gene is able to exert a stimulatory influence upon attached promoter sequences. This cis effect was shown to be independent of orientation but dependent upon the distance between the DNA sequences involved.
Assuntos
Genes Sintéticos , Genes Virais , Regiões Promotoras Genéticas , Animais , Galinhas/genética , Feminino , Histonas/genética , Queratinas/genética , Oócitos/metabolismo , RNA Mensageiro/biossíntese , Proteínas Recombinantes de Fusão/biossíntese , Simplexvirus/genética , Especificidade da Espécie , Timidina Quinase/genética , Proteínas Virais/genética , Xenopus laevisRESUMO
We have shown that chicken feather keratin genes are transcribed at extremely low levels in Xenopus oocytes. Primer extension reactions using carefully selected primers have shown that none of the detectable transcripts are spliced. When the single intron, located in the 5' untranslated region, was removed from the gene, we observed a 5-fold increase in transcript initiation from the in vivo "cap" site. Significantly, this 5-fold increase was also observed when the feather keratin intron was replaced with a similar sized fragment from pBR322. We conclude that the efficiency of accurate transcriptional initiation of chicken feather keratin genes in the oocyte is affected by DNA sequences within the intron.