RESUMEN
Two APETALA2 domain transcription factors were characterized first in angiosperms, and, recently, in several gymnosperms. These proteins are involved in several processes, from flowering to embryogenesis in Arabidopsis thaliana. We extrapolated this result to hybrid larch (Larixxmarschlinsii Coaz) resulting from a cross between European (Larix decidua) and Japanese (Larix kaempferi) larches. Somatic embryogenesis is well described and controlled for this Pinaceae. We characterized two-AP2 domain genes: LmAP2L1 and LmAP2L2. Phylogenetic analysis confirmed that LmAP2L1 and LmAP2L2 were orthologous to Norway spruce PaAP2L1 and PaAP2L2 and that L1 forms appeared to be specific to Pinaceae. RT-PCR analysis showed that larch APETALA2 was differentially expressed during late somatic embryogenesis and during the first steps of germination. Whereas LmAP2L2 was constitutively expressed during this process, LmAP2L1 expression appeared only during late somatic embryogenesis, when embryos were able to germinate. Further, LmAP2L1 appeared to be the preferentially expressed form during embryo germination. Thus, LmAP2L1 seems to be a valuable molecular marker for hybrid larch late somatic embryogenesis and could play a role during post-embryonic development.
Asunto(s)
Proteínas de Arabidopsis/química , Desarrollo Embrionario/genética , Regulación de la Expresión Génica de las Plantas , Germinación/genética , Proteínas de Homeodominio/química , Larix/embriología , Larix/genética , Proteínas Nucleares/química , Proteínas de Plantas/genética , Secuencia de Aminoácidos , Secuencia Conservada , Cruzamientos Genéticos , Hibridación Genética , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/metabolismo , Estructura Terciaria de Proteína , Alineación de SecuenciaRESUMEN
In plants, Lewisa type N-glycans may be involved in cell-to-cell communication and recognition. N-glycoproteins harboring Lewisa glycotopes are mainly found in plasma membranes and cell walls. Some can be also involved in cell wall synthesis or the loosening process, and subsequently in cell elongation. In order to determine the potential role(s) of the alpha4-fucosylation during vegetative development, transgenic tobacco plants overexpressing a human Lewis fucosyltransferase (hFUT3), which transfers a fucose residue in a alpha(1,4)-linkage on complex glycans, have been developed. The heterologous enzyme hFUT3 was strongly expressed and fully functional in transgenic tobacco. Transgenic plants showed a delay in growth linked to a reduction of internode length. Furthermore, transgenic seedling roots were significantly shorter than wild-type roots and the length of their epidermis cells was reduced. Strikingly, root growth was completely and specifically restored following gibberellin treatment. Etiolated hypocotyls of hFUT3 overexpressors were also more sensitive to exogenous gibberellin. Furthermore, paclobutrazol, an inhibitor of gibberellin synthesis, induced a similar effect on control and transgenic dark-grown hypocotyls suggesting that gibberellin biosynthesis was probably not altered in seedlings overexpressing hFUT3. Thus, alpha4-fucosylation could act as a possible modulator of conformation and/or functioning of N-glycoproteins involved in the gibberellin-dependent elongation process.
Asunto(s)
Fucosiltransferasas/metabolismo , Giberelinas/farmacología , Nicotiana/enzimología , Envejecimiento , Membrana Celular/enzimología , Pared Celular/enzimología , Fucosiltransferasas/genética , Humanos , Cinética , Plantas Modificadas Genéticamente , Nicotiana/efectos de los fármacos , Nicotiana/genética , Nicotiana/crecimiento & desarrolloRESUMEN
CYCD3;1 expression in Arabidopsis is associated with proliferating tissues such as meristems and developing leaves but not with differentiated tissues. Constitutive overexpression of CYCD3;1 increases CYCD3;1-associated kinase activity and reduces the proportion of cells in the G1-phase of the cell cycle. Moreover, CYCD3;1 overexpression leads to striking alterations in development. Leaf architecture in overexpressing plants is altered radically, with a failure to develop distinct spongy and palisade mesophyll layers. Associated with this, we observe hyperproliferation of leaf cells; in particular, the epidermis consists of large numbers of small, incompletely differentiated polygonal cells. Endoreduplication, a marker for differentiated cells that have exited from the mitotic cell cycle, is inhibited strongly in CYCD3;1-overexpressing plants. Transcript analysis reveals an activation of putative compensatory mechanisms upon CYCD3;1 overexpression or subsequent cell cycle activation. These results demonstrate that cell cycle exit in the G1-phase is required for normal cellular differentiation processes during plant development and suggest a critical role for CYCD3 in the switch from cell proliferation to the final stages of differentiation.
Asunto(s)
Arabidopsis/genética , Ciclo Celular/fisiología , Quinasas Ciclina-Dependientes/metabolismo , Ciclinas/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Diferenciación Celular/fisiología , División Celular/fisiología , Ciclina D3 , Quinasas Ciclina-Dependientes/genética , Ciclinas/genética , Copas de Floración/genética , Copas de Floración/fisiología , Regulación del Desarrollo de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Meristema/genética , Meristema/fisiología , Brotes de la Planta/genética , Brotes de la Planta/fisiología , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Tuber explants of Jerusalem artichoke (Helianthus tuberosus L.) are a model system for cell-cycle re-entry from a quiescent state, involving the activation of division of tuber parenchyma cells in response to exogenous auxin. To enable molecular studies of this system, two cyclin (Heltu;CYCD1;1 and Heltu; CYCD3;1) and two cyclin-dependent kinase (Heltu; CDKA;1 and Heltu;CDKB1;1) genes have been isolated from a Jerusalem artichoke cDNA library and their expression demonstrated during the activation of cell division. It was found that CDKA;1 transcripts are present in quiescent tubers, whereas CYCD1;1, CYCD3;1 and CDKB1;1 transcripts are induced during cell-cycle re-entry as well as during bud growth of whole tubers. Both CYCD1;1 and CYCD3;1 transcripts appear shortly before, or coincident with, the onset of S phase.
Asunto(s)
Quinasas Ciclina-Dependientes/genética , Ciclinas/genética , Helianthus/genética , Ciclina D , Expresión Génica , Helianthus/enzimología , Fase SRESUMEN
The basic pattern of controls that operate during the G1 phase of the plant cell cycle shows much closer similarity to animals than to the yeasts and other fungi. The activity of D-type cyclin (CycD) kinases is induced in response to stimulatory signals, and these phosphorylate the plant homologue of the retinoblastoma tumour susceptibility (Rb) protein. It is likely that Rb phosphorylation results in the activation of genes under the control of E2F transcription factors, including those required for S phase entry. As the initial triggers of the cascade, attention has focused on the CycDs, and a family of 10 genes is present in Arabidopsis, divided into three major and three minor groups. Analysis to date suggests that these groups are functionally distinct.
Asunto(s)
Ciclo Celular/fisiología , Ciclinas/fisiología , Células Vegetales , División Celular , Ciclinas/genética , Fase G1 , Regulación de la Expresión Génica de las Plantas , Levaduras/citologíaRESUMEN
alpha4-Fucosylation represents a final step of protein N- glycosylation. alpha4-fucosylated N-glycans are thought to be involved in cell-to-cell communication and recognition in primates and plants. Nevertheless, in the plant life cycle, the function of alpha4-fucosylation remains largely unknown. To gain an insight into the role of alpha4-fucosylation during development, the study focused on tobacco flowers. It is shown that an increase in alpha(1,4)fucosyltransferase (Fuc-T) activity is only observed during anther development, whereas it remains at a constant but low level (around 20 pmol Fuc h(-1) mg(-1) protein) in the gynoecium and perianth. At least a 4-fold higher activity is detected in mature pollen grains. These data suggest that alpha(1,4)Fuc-T activity is regulated during anther development. Furthermore, alpha(1,4)Fuc-T activity could be required during pollen tube elongation where the activity level peaks at 350 pmol h(-1) mg(-1) protein. Based on enzyme profile and cycloheximide effects on pollen germination and activity, it is hypothesized that the gene encoding alpha4-Fuc-T could be regulated late during pollen development. A potential role of alpha4- fucosylation during pollen tube elongation is also discussed.
Asunto(s)
Fucosiltransferasas/metabolismo , Estructuras de las Plantas/crecimiento & desarrollo , Polen/crecimiento & desarrollo , Comunicación Celular/fisiología , Cicloheximida/farmacología , Activación Enzimática , Glicosilación , Estructuras de las Plantas/metabolismo , Polen/efectos de los fármacos , Reproducción/fisiología , Nicotiana/crecimiento & desarrollo , Nicotiana/metabolismoRESUMEN
Plant development requires stringent controls between cell proliferation and cell differentiation. Proliferation is positively regulated by cyclin dependent kinases (CDKs). Acting in opposition to CDKs are CDK inhibitors (CKIs). The first tobacco CKI (NtKIS1a) identified was shown to inhibit in vitro the kinase activity of CDK/cyclin complexes and to interact with CDK and D-cyclins. However, these features, which are common to other plant and animal CKIs already characterised, did not provide information about the function of NtKIS1a in plants. Thus, to gain insight into the role of NtKIS1a and especially its involvement in cell proliferation during plant development, we generated transgenic Arabidopsis thaliana plants that overexpress NtKIS1a. These plants showed reduced growth with smaller organs that contained larger cells. Moreover, these plants displayed modifications in plant morphology. These results demonstrated that plant organ size and shape, as well as organ cell number and cell size, might be controlled by modulation of the single NtKIS1a gene activity. Since in mammals, D-cyclins control cell cycle progression in a CDK-dependent manner but also play a CDK independent role by sequestering the CKIs p27(Kip1) and p21(Cip1), we tested the significance of cyclin D-CKI interaction within a living plant. With this aim, NtKIS1a and AtCycD3;1 were overexpressed simultaneously in plants by two different methods. Our results demonstrated that overexpression of the CKI NtKIS1a restores essentially normal development in plants overexpressing AtCycD3;1, providing the first evidence of cyclin D-CKI co-operation within the context of a living plant.