RESUMO
Grafting is possible in both animals and plants. Although in animals the process requires surgery and is often associated with rejection of non-self, in plants grafting is widespread, and has been used since antiquity for crop improvement1. However, in the monocotyledons, which represent the second largest group of terrestrial plants and include many staple crops, the absence of vascular cambium is thought to preclude grafting2. Here we show that the embryonic hypocotyl allows intra- and inter-specific grafting in all three monocotyledon groups: the commelinids, lilioids and alismatids. We show functional graft unions through histology, application of exogenous fluorescent dyes, complementation assays for movement of endogenous hormones, and growth of plants to maturity. Expression profiling identifies genes that unify the molecular response associated with grafting in monocotyledons and dicotyledons, but also gene families that have not previously been associated with tissue union. Fusion of susceptible wheat scions to oat rootstocks confers resistance to the soil-borne pathogen Gaeumannomyces graminis. Collectively, these data overturn the consensus that monocotyledons cannot form graft unions, and identify the hypocotyl (mesocotyl in grasses) as a meristematic tissue that allows this process. We conclude that graft compatibility is a shared ability among seed-bearing plants.
Assuntos
Avena , Raízes de Plantas , Brotos de Planta , Transplantes , Triticum , Ascomicetos/patogenicidade , Avena/embriologia , Avena/microbiologia , Hipocótilo , Meristema , Raízes de Plantas/embriologia , Raízes de Plantas/microbiologia , Brotos de Planta/embriologia , Brotos de Planta/microbiologia , Triticum/embriologia , Triticum/microbiologiaRESUMO
Multicellular animals and plants represent independent evolutionary experiments with complex multicellular bodyplans. Differences in their life history, a mobile versus sessile lifestyle, and predominant embryonic versus postembryonic development, have led to the evolution of highly different body plans. However, also many intriguing parallels exist. Extension of the vertebrate body axis and its segmentation into somites bears striking resemblance to plant root growth and the concomittant prepatterning of lateral root competent sites. Likewise, plant shoot phyllotaxis displays similarities with vertebrate limb and digit patterning. Additionally, both plants and animals use complex signalling systems combining systemic and local signals to fine tune and coordinate organ growth across their body. Identification of these striking examples of convergent evolution provides support for the existence of general design principles: the idea that for particular patterning demands, evolution is likely to arrive at highly similar developmental patterning mechanisms. Furthermore, focussing on these parallels may aid in identifying core mechanistic principles, often obscured by the highly complex nature of multiscale patterning processes.
Assuntos
Padronização Corporal/fisiologia , Desenvolvimento Embrionário/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/genética , Brotos de Planta/embriologia , Transdução de Sinais/fisiologia , Animais , Biologia do Desenvolvimento , Camundongos , Modelos Biológicos , Plantas , Biologia de SistemasRESUMO
The degree of shoot branching in Arabidopsis is determined by the activation of axillary buds. Bud activity is regulated by diverse environmental and developmental signals, often mediated via plant hormones, including auxin, strigolactone and cytokinin. The transcription factor BRANCHED1 (BRC1) has been proposed to integrate these regulatory signals. This idea is based on increased branching in brc1 mutants, the effects of bud-regulating hormones on BRC1 expression, and a general correlation between BRC1 expression and bud growth inhibition. These data demonstrate the important role of BRC1 in shoot branching, but here we show that in Arabidopsis this correlation can be broken. Buds lacking BRC1 expression can remain inhibited and sensitive to inhibition by strigolactone. Furthermore, buds with high BRC1 transcript levels can be active. Based on these data, we propose that BRC1 regulates bud activation potential in concert with an auxin transport-based mechanism underpinning bud activity. In the context of strigolactone-mediated bud regulation, our data suggest a coherent feed-forward loop in which strigolactone treatment reduces the probability of bud activation by parallel effects on BRC1 transcription and the shoot auxin transport network.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/embriologia , Arabidopsis/genética , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Brotos de Planta/embriologia , Brotos de Planta/genética , Fatores de Transcrição/genética , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Epistasia Genética/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Lactonas/farmacologia , Mutação/genética , Brotos de Planta/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND: Tissue culture and rapid propagation technology is an important way to solve the difficulties of plant propagation. This experiment aims to explore the appropriate conditions at each stage of the red maple's tissue culture process and to obtain plantlets, thus providing a theoretical basis for the establishment of the red maple's tissue culture system. RESULTS: The results showed that the stem segment is the most suitable explant for inducing embryogenic callus. The MS (Murashige&Skoog) + 0.8 mg/L TDZ (Thidiazuron) + 1.0 mg/L 6-BA (6-Benzylaminopurine) + 0.5 mg/L IAA(Indole-3-acetic acid) + 35 g/L sucrose+ 7.5 g/L semi-fixed medium was the best for callus formation. When selecting type VI callus as embryonic callus induction material, MS + 0.6 mg/L TDZ + 0.5 mg/L 6-BA + 2.0 mg/L IAA + 35 g/L sucrose+ 7.5 g/L semi-fixed medium can get embryonic callus. The optimal medium for adventitious bud induction is MS + 1.0 mg/L TDZ + 3.0 mg/L 6-BA+ 0.2 mg/L NAA (1-Naphthaleneacetic acid) + 1.2 mg/L IAA + 35 g/L sucrose+ 7.5 g/L semi-fixed medium. The induction rate of adventitious roots in MS + 0.6 mg/L TDZ + 1.0 mg/L 6-BA+ 3 mg/L NAA + 35 g/L sucrose+ 7.5 g/L semi-fixed medium was the highest, reaching 76%. CONCLUSIONS: In the course of our research, we found that PGRs play an important role in the callus induction stage, and the effect of TDZ is particularly obvious; The callus cells grow and proliferate according to the "S" growth curve, and can be sub-cultured when the highest growth point is reached to maintain the rapid proliferation of the callus cells and to avoid inactivation of callus caused by tight niche.
Assuntos
Acer/crescimento & desenvolvimento , Câmbio/embriologia , Brotos de Planta/crescimento & desenvolvimento , Acer/embriologia , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/embriologia , RegeneraçãoRESUMO
Purple coneflower (Echinacea purpurea (L.) Moench) is a widely used medicinal and ornamental plant. In the present study, the callus embryogenesis was examined using benzyl adenine (BA) at three levels (3, 4, 5 mg L-1), 1-Naphthalene acetic acid (NAA) at three levels (0.1, 0.2 and 0.5 mg L-1) with or without activated charcoal (1 g L-1), coconut milk (50 ml L-1) and casein hydrolysate (50 mg L-1) in the MS (Murashige and Skoog 1962) medium. The embryogenesis indirectly occurred with the production of callus. The calli were observed in three forms: undifferentiated, embryogenic and organogenic. The embryogenic calli were dark green and coherent with a faster growth rate. The highest embryogenesis (100%) and embryonic regeneration (plantlet production) were obtained in the combined BA + NAA treatments with the activated charcoal, coconut milk and casein hydrolysate. However, the combined treatments of growth regulators failed to produce somatic embryos without the use of coconut milk and casein hydrolysate. The maximum amount of protein, peroxidase and catalase activity of embryogenic calli (2.02, 1.79 and 6.62ΔOD/Min/mg.protein, respectively), and highest percentage of acclimatization success (29.3% of plants) were obtained in the combined treatment of 5 mg L-1 BA + 0.5 mg L-1 NAA + activated charcoal + coconut milk + casein hydrolysate. The highest amount of chlorophyll content (33.3 SPAD value) and growth characteristics of acclimatized plantlets were observed in the media containing 3 mg L-1 BA + 0.1 and 0.2 mg L-1 NAA + 1 g. L-1 combined activated charcoal, coconut milk, casein hydrolysate. The histological studies confirmed the somatic embryogenesis in purple coneflower. Generally, it was found that the somatic embryogenesis of E. purpurea occurs at high levels of BA and low levels of NAA with the addition of coconut milk and casein hydrolysate.
Assuntos
Antioxidantes/farmacologia , Echinacea/química , Echinacea/embriologia , Reguladores de Crescimento de Plantas/farmacologia , Brotos de Planta/efeitos dos fármacos , Técnicas de Embriogênese Somática de Plantas/métodos , Adenina/análogos & derivados , Adenina/farmacologia , Caseínas/farmacologia , Carvão Vegetal/farmacologia , Cocos/química , Meios de Cultura , Echinacea/enzimologia , Ácidos Naftalenoacéticos/farmacologia , Organoides/citologia , Organoides/efeitos dos fármacos , Organoides/embriologia , Organoides/crescimento & desenvolvimento , Brotos de Planta/embriologia , Brotos de Planta/crescimento & desenvolvimento , Plantas Medicinais/químicaRESUMO
Agrobacterium-mediated genetic transformation is well established in the model grass Brachypodium distachyon. However, most protocols employ immature embryos because of their better regenerative capacity. A major problem associated with the immature embryo system is that they are available only during a limited time window of growing plants. In this study, we have developed an optimized Agrobacterium-mediated genetic transformation protocol that utilizes mature embryos. We have adopted seed shearing and photoautotrophic rooting (PR) in callus induction and root regeneration, respectively, with evident significant improvement in these aspects. We have also revealed that the newly developed chemical inducer Fipexide (FPX) had the ability to induce callus, shoots, and roots. By comparison, we have demonstrated that FPX shows higher efficiency in shoot generation than other frequently used chemicals in our mature embryo-based system. In addition, we demonstrated that the age of embryogenetic callus severely affects the transformation efficiency (TE), with the seven-week-old embryogenetic callus having the highest TE reaching 52.6%, which is comparable with that in immature embryo transformation. The new methodologies reported here will advance the development and utilization of Brachypodium as a new model system for grass genomics.
Assuntos
Brachypodium/genética , Sementes/genética , Técnicas de Cultura de Tecidos/métodos , Agrobacterium/fisiologia , Brachypodium/efeitos dos fármacos , Brachypodium/embriologia , Piperazinas/farmacologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/embriologia , Raízes de Plantas/genética , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/embriologia , Brotos de Planta/genética , Plantas Geneticamente Modificadas , Regeneração/efeitos dos fármacos , Regeneração/genética , Sementes/efeitos dos fármacos , Sementes/embriologia , Transformação GenéticaRESUMO
Background and Aims: Setaria viridis is being promoted as a model C4 photosynthetic plant because it has a small genome (~515 Mb), a short life cycle (~60 d) and it can be transformed. Unlike other C4 grasses such as maize, however, there is very little information about how C4 leaf anatomy (Kranz anatomy) develops in S. viridis. As a foundation for future developmental genetic studies, we provide an anatomical and ultrastructural framework of early shoot development in S. viridis, focusing on the initiation of Kranz anatomy in seed leaves. Methods: Setaria viridis seeds were germinated and divided into five stages covering development from the dry seed (stage S0) to 36 h after germination (stage S4). Material at each of these stages was examined using conventional light, scanning and transmission electron microscopy. Key Results: Dry seeds contained three embryonic leaf primordia at different developmental stages (plastochron 1-3 primordia). The oldest (P3) leaf primordium possessed several procambial centres whereas P2 displayed only ground meristem. At the tip of P3 primordia at stage S4, C4 leaf anatomy typical of the malate dehydrogenase-dependent nicotinamide dinucleotide phosphate (NADP-ME) subtype was evident in that vascular bundles lacked a mestome layer and were surrounded by a single layer of bundle sheath cells that contained large, centrifugally located chloroplasts. Two to three mesophyll cells separated adjacent vascular bundles and one mesophyll cell layer on each of the abaxial and adaxial sides delimited vascular bundles from the epidermis. Conclusions: The morphological trajectory reported here provides a foundation for studies of gene regulation during early leaf development in S. viridis and a framework for comparative analyses with other C4 grasses.
Assuntos
Folhas de Planta/embriologia , Setaria (Planta)/embriologia , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Floema/ultraestrutura , Folhas de Planta/anatomia & histologia , Folhas de Planta/ultraestrutura , Brotos de Planta/anatomia & histologia , Brotos de Planta/embriologia , Brotos de Planta/ultraestrutura , Sementes/crescimento & desenvolvimento , Setaria (Planta)/anatomia & histologia , Setaria (Planta)/ultraestrutura , Xilema/ultraestruturaRESUMO
BACKGROUND: Vegetative propagation of Fragaria sp. is traditionally carried out using stolons. This system of propagation, in addition to being slow, can spread plant diseases, particularly serious being viral. In vitro culture of meristems and the establishment of micropropagation protocols are important tools for solving these problems. In recent years, considerable effort has been made to develop in vitro propagation of the commercial strawberry in order to produce virus-free plants of high quality. These previous results can serve as the basis for developing in vitro-based propagation technologies in the less studied species Fragaria chiloensis. RESULTS: In this context, we studied the cultivation of meristems and establishment of a micropropagation protocol for F. chiloensis. The addition of polyvinylpyrrolidone (PVP) improved the meristem regeneration efficiency of F. chiloensis accessions. Similarly, the use of 6-benzylaminopurine (BAP) in the culture media increased the average rate of multiplication to 3-6 shoots per plant. In addition, the use of 6-benzylaminopurine (BAP), had low levels (near zero) of explant losses due to oxidation. However, plant height as well as number of leaves and roots were higher in media without growth regulators, with average values of 0.5 cm, 9 leaves and 4 roots per plant. CONCLUSIONS: For the first time in Chilean strawberry, meristem culture demonstrated to be an efficient tool for eliminating virus from infected plants, giving the possibility to produce disease free propagation material. Also, the addition of PVP into the basal MS medium improved the efficiency of plant recovery from isolated meristems. Farmers can now access to high quality plant material produced by biotech tools which will improve their technological practices.
Assuntos
Compostos de Benzil/farmacologia , Fragaria/embriologia , Meristema/crescimento & desenvolvimento , Brotos de Planta/embriologia , Purinas/farmacologia , Regeneração/efeitos dos fármacos , Chile , Meios de Cultura , Fragaria/efeitos dos fármacos , Meristema/efeitos dos fármacos , Brotos de Planta/efeitos dos fármacosRESUMO
Stem cells are responsible for organogenesis, but it is largely unknown whether and how information from stem cells acts to direct organ patterning after organ primordia are formed. It has long been proposed that the stem cells at the plant shoot apex produce a signal, which promotes leaf adaxial-abaxial (dorsoventral) patterning. Here we show the existence of a transient low auxin zone in the adaxial domain of early leaf primordia. We also demonstrate that this adaxial low auxin domain contributes to leaf adaxial-abaxial patterning. The auxin signal is mediated by the auxin-responsive transcription factor MONOPTEROS (MP), whose constitutive activation in the adaxial domain promotes abaxial cell fate. Furthermore, we show that auxin flow from emerging leaf primordia to the shoot apical meristem establishes the low auxin zone, and that this auxin flow contributes to leaf polarity. Our results provide an explanation for the hypothetical meristem-derived leaf polarity signal. Opposite to the original proposal, instead of a signal derived from the meristem, we show that a signaling molecule is departing from the primordium to the meristem to promote robustness in leaf patterning.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/embriologia , Proteínas de Ligação a DNA/metabolismo , Ácidos Indolacéticos/metabolismo , Organogênese Vegetal/fisiologia , Folhas de Planta/enzimologia , Brotos de Planta/embriologia , Fatores de Transcrição/metabolismo , Arabidopsis/citologia , Arabidopsis/genética , Folhas de Planta/citologia , Brotos de Planta/citologiaRESUMO
Plants, similarly to animals, form polarized axes during embryogenesis on which cell differentiation and organ patterning programs are orchestrated. During Arabidopsis embryogenesis, establishment of the shoot and root stem cell populations occurs at opposite ends of an apical-basal axis. Recent work has identified the PLETHORA (PLT) genes as master regulators of basal/root fate, whereas the master regulators of apical/shoot fate have remained elusive. Here we show that the PLT1 and PLT2 genes are direct targets of the transcriptional co-repressor TOPLESS (TPL) and that PLT1/2 are necessary for the homeotic conversion of shoots to roots in tpl-1 mutants. Using tpl-1 as a genetic tool, we identify the CLASS III HOMEODOMAIN-LEUCINE ZIPPER (HD-ZIP III) transcription factors as master regulators of embryonic apical fate, and show they are sufficient to drive the conversion of the embryonic root pole into a second shoot pole. Furthermore, genetic and misexpression studies show an antagonistic relationship between the PLT and HD-ZIP III genes in specifying the root and shoot poles.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/embriologia , Arabidopsis/metabolismo , Padronização Corporal/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/antagonistas & inibidores , Proteínas de Arabidopsis/genética , Padronização Corporal/genética , Diferenciação Celular/genética , Linhagem da Célula/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Proteínas de Homeodomínio , Zíper de Leucina , Raízes de Plantas/citologia , Raízes de Plantas/embriologia , Brotos de Planta/citologia , Brotos de Planta/embriologia , Células-Tronco/citologia , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/genéticaRESUMO
In Arabidopsis, SEUSS (SEU) and SEUSS-LIKE 2 (SLK2) are components of the LEUNIG (LUG) repressor complex that coordinates various aspects of post-embryonic development. The complex also plays a critical role during embryogenesis, as seu slk2 double mutants have small, narrow cotyledons and lack a shoot apical meristem (SAM). Here we show that seu slk2 double mutant embryos exhibit delayed cotyledon outgrowth and that this is associated with altered PIN-FORMED1 (PIN1) expression and localisation during the early stages of embryogenesis. These observations suggest that SEU and SLK2 promote the transition to bilateral symmetry by modulating auxin distribution in the embryonic shoot. This study also shows that loss of SAM formation in seu slk2 mutants is associated with reduced expression of the class I KNOX (KNOXI) genes SHOOTMERISTEMLESS (STM), BREVIPEDICELLUS and KNAT2. Furthermore, elevating STM expression in seu slk2 mutant embryos was sufficient to restore SAM formation but not post-embryonic activity, while both SAM formation and activity were rescued when SLK2 expression was restored in either the cotyledons or boundary regions. These results demonstrate that SEU and SLK2 function redundantly to promote embryonic shoot development and likely act through a non-cell autonomous pathway to promote KNOXI activity.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/embriologia , Proteínas de Homeodomínio/metabolismo , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Cotilédone/embriologia , Cotilédone/genética , Cotilédone/metabolismo , Flores/embriologia , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Homeodomínio/genética , Meristema/embriologia , Meristema/genética , Meristema/metabolismo , Mutação , Fenótipo , Proteínas de Plantas/genética , Brotos de Planta/embriologia , Brotos de Planta/genética , Brotos de Planta/metabolismo , Fatores de Transcrição/genéticaRESUMO
Development is often coordinated by biologically active mobile compounds that move between cells or organs. Arabidopsis mutants with defects in the BYPASS1 (BPS1) gene overproduce an active mobile compound that moves from the root to the shoot and inhibits growth. Here, we describe two related Arabidopsis genes, BPS2 and BPS3. Analyses of single, double and triple mutants revealed that all three genes regulate production of the same mobile compound, the bps signal, with BPS1 having the largest role. The triple mutant had a severe embryo defect, including the failure to properly establish provascular tissue, the shoot meristem and the root meristem. Aberrant expression of PINFORMED1, DR5, PLETHORA1, PLETHORA2 and WUSCHEL-LIKE HOMEOBOX5 were found in heart-stage bps triple-mutant embryos. However, auxin-induced gene expression, and localization of the PIN1 auxin efflux transporter, were intact in bps1 mutants, suggesting that the primary target of the bps signal is independent of auxin response. Thus, the bps signal identifies a novel signaling pathway that regulates patterning and growth in parallel with auxin signaling, in multiple tissues and at multiple developmental stages.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/embriologia , Arabidopsis/genética , Ácidos Indolacéticos/metabolismo , Transdução de Sinais/fisiologia , Arabidopsis/anatomia & histologia , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Meristema/anatomia & histologia , Meristema/embriologia , Fenótipo , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/embriologia , Brotos de Planta/anatomia & histologia , Brotos de Planta/embriologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Plântula/anatomia & histologia , Plântula/embriologiaRESUMO
Human pharmaceuticals have been detected in the terrestrial environment at µg to mg kg(-1) concentrations. Repeated application of sewage sludge (biosolids) and increasing reclaimed wastewater use for irrigation could lead to accumulation of these novel contaminants in soil systems. Despite this, potential phytotoxicological effects on higher plants have rarely been evaluated. These studies aimed to test effects upon germination, development, growth and physiology of two crop plants, namely radish (Raphanus sativus Spakler 3) and lettuce (Lactuca sativa All Year Around), after exposure to different, but structurally related non-steroidal anti-inflammatory drugs (NSAIDs) at environmentally relevant concentrations. A range of biological endpoints comprising biomass, length, water content, specific root and shoot length, root to shoot ratio, daily progress of stages of cell elongation and organ emergence (primary root, hypocotyl elongation, cotyledon emergence, cotyledon opening, and no change), as well as photosynthetic measurements were evaluated. Compounds from the fenamic acid class were found to affect R. sativus root endpoints (root length and water content), while ibuprofen affected early root development of L. sativa. In general, phytotoxicological effects on root endpoints demonstrated that impacts upon higher plants are not only compound specific, but also differ between plant species. It was found that the usage of a wide range of biological endpoints (all simple, cost-effective and ecologically relevant) were beneficial in detecting differences in plant responses to NSAID exposure. Due to paucity and discrepancy within the few previously available phytotoxicological studies with pharmaceuticals, it is now essential to allocate time and resources to consider development of suitable chronic toxicity tests, and some suggestions regarding this are presented.
Assuntos
Anti-Inflamatórios não Esteroides/toxicidade , Lactuca/efeitos dos fármacos , Raphanus/efeitos dos fármacos , Anti-Inflamatórios não Esteroides/química , Relação Dose-Resposta a Droga , Germinação/efeitos dos fármacos , Lactuca/embriologia , Lactuca/crescimento & desenvolvimento , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/embriologia , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/embriologia , Brotos de Planta/crescimento & desenvolvimento , Raphanus/embriologia , Raphanus/crescimento & desenvolvimento , Sementes/efeitos dos fármacos , Sementes/embriologia , Sementes/crescimento & desenvolvimentoRESUMO
Protein phosphorylation is a key molecular switch used to transmit information in biological signalling networks. The output of these signalling circuits is governed by the counteracting activities of protein kinases and phosphatases that determine the direction of the switch. Whereas many kinases have been functionally characterized, it has been difficult to ascribe precise cellular roles to plant phosphatases, which are encoded by enlarged gene families that may provide a high degree of genetic redundancy. In this work we have analysed the role in planta of catalytic subunits of protein phosphatase 2A (PP2A), a family encoded by five genes in Arabidopsis. Our results indicate that the two members of subfamily II, PP2A-C3 and PP2A-C4, have redundant functions in controlling embryo patterning and root development, processes that depend on auxin fluxes. Moreover, polarity of the auxin efflux carrier PIN1 and auxin distribution, determined with the DR5(pro) :GFP proxy, are affected by mutations in PP2A-C3 and PP2A-C4. Previous characterization of mutants in putative PP2A regulatory subunits had established a link between this class of phosphatases and PIN dephosphorylation and subcellular distribution. Building on those findings, the results presented here suggest that PP2A-C3 and PP2A-C4 catalyse this reaction and contribute critically to the establishment of auxin gradients for proper plant development.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Ácidos Indolacéticos/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Proteína Fosfatase 2/metabolismo , Arabidopsis/embriologia , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Transporte Biológico , Padronização Corporal , Domínio Catalítico , Técnicas de Inativação de Genes , Proteínas de Membrana Transportadoras/genética , Meristema/embriologia , Meristema/enzimologia , Meristema/genética , Meristema/fisiologia , Mutação , Fenótipo , Fosforilação , Raízes de Plantas/embriologia , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Brotos de Planta/embriologia , Brotos de Planta/enzimologia , Brotos de Planta/genética , Brotos de Planta/fisiologia , Plantas Geneticamente Modificadas , Proteína Fosfatase 2/genética , Transporte Proteico , Proteínas Recombinantes de Fusão , Plântula/embriologia , Plântula/enzimologia , Plântula/genética , Plântula/fisiologia , Transdução de SinaisRESUMO
BACKGROUND: Asymmetric cell divisions are formative divisions that generate daughter cells of distinct identity. These divisions are coordinated by either extrinsic ('niche-controlled') or intrinsic regulatory mechanisms and are fundamentally important in plant development. SCOPE: This review describes how asymmetric cell divisions are regulated during development and in different cell types in both the root and the shoot of plants. It further highlights ways in which omics and modelling approaches have been used to elucidate these regulatory mechanisms. For example, the regulation of embryonic asymmetric divisions is described, including the first divisions of the zygote, formative vascular divisions and divisions that give rise to the root stem cell niche. Asymmetric divisions of the root cortex endodermis initial, pericycle cells that give rise to the lateral root primordium, procambium, cambium and stomatal cells are also discussed. Finally, a perspective is provided regarding the role of other hormones or regulatory molecules in asymmetric divisions, the presence of segregated determinants and the usefulness of modelling approaches in understanding network dynamics within these very special cells. CONCLUSIONS: Asymmetric cell divisions define plant development. High-throughput genomic and modelling approaches can elucidate their regulation, which in turn could enable the engineering of plant traits such as stomatal density, lateral root development and wood formation.
Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Divisão Celular Assimétrica , Genômica , Arabidopsis/embriologia , Magnoliopsida/embriologia , Magnoliopsida/genética , Magnoliopsida/crescimento & desenvolvimento , Modelos Genéticos , Raízes de Plantas/embriologia , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/embriologia , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimentoRESUMO
One of the most fundamental events in plant ontogeny is the specification of the shoot and root apical meristem (SAM and RAM) in embryogenesis. In Arabidopsis, the restricted expression of class III homeodomain leucine zipper (HD-ZIP III) transcription factors (TFs) at the central-apical domain of early embryos is required for the correct specification of the SAM and RAM. Because the expression of HD-ZIP III TFs is suppressed by microRNA165/166 (miR165/6), elucidation of the sites of miR165/6 production and their activity range is a key to understanding the molecular basis of SAM and RAM specification in embryogenesis. Here, we present a comprehensive reporter analysis of all nine Arabidopsis MICRORNA165/166 (MIR165/6) genes during embryogenesis. We show that five MIR165/6 genes are transcribed in a largely conserved pattern in embryos, with their expression being preferentially focused at the basal-peripheral region of embryos. Our analysis also indicated that MIR165/6 transcription does not depend on SCARECROW (SCR) function in early embryos, in contrast to its requirement in post-embryonic roots. Furthermore, by observing the expression pattern of the miR-resistant PHBmu-GFP (green fluorescent protein) reporter, in either the presence or absence of the MIR165Amu transgene, which targets PHBmu-GFP, we obtained data that indicate a non-cell-autonomous function for miR165 in early embryos. These results suggest that miR165, and possibly miR166 as well, has the capacity to act as a positional cue from the basal-peripheral region of early embryos, and remotely controls SAM and RAM specification with their non-cell-autonomous function.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Homeodomínio/genética , Meristema/genética , MicroRNAs/genética , Arabidopsis/citologia , Arabidopsis/embriologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Fluorescência Verde , Proteínas de Homeodomínio/metabolismo , Zíper de Leucina , Meristema/citologia , Meristema/embriologia , Meristema/metabolismo , MicroRNAs/metabolismo , Família Multigênica , Mutação , Especificidade de Órgãos , Raízes de Plantas/citologia , Raízes de Plantas/embriologia , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Brotos de Planta/citologia , Brotos de Planta/embriologia , Brotos de Planta/genética , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , RNA de Plantas/genética , RNA de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , TransgenesRESUMO
BACKGROUND: Among the many commercial opportunities afforded by somatic embryogenesis (SE), it is the ability to clonally propagate individual plants with rare or elite traits that has some of the most significant implications. This is particularly true for many long-lived species, such as conifers, but whose long generation times pose substantive challenges, including increased recalcitrance for SE as plants age. Identification of a clonal line of somatic embryo-derived trees whose shoot primordia have remained responsive to SE induction for over a decade, provided a unique opportunity to examine the molecular aspects underpinning SE within shoot tissues of adult white spruce trees. RESULTS: Microarray analysis was used to conduct transcriptome-wide expression profiling of shoot explants taken from this responsive genotype following one week of SE induction, which when compared with that of a nonresponsive genotype, led to the identification of four of the most differentially expressed genes within each genotype. Using absolute qPCR to expand the analysis to three weeks of induction revealed that differential expression of all eight candidate genes was maintained to the end of the induction treatment, albeit to differing degrees. Most striking was that both the magnitude and duration of candidate gene expression within the nonresponsive genotype was indicative of an intense physiological response. Examining their putative identities further revealed that all four encoded for proteins with similarity to angiosperm proteins known to play prominent roles in biotic defense, and that their high-level induction over an extended period is consistent with activation of a biotic defense response. In contrast, the more temperate response within the responsive genotype, including induction of a conifer-specific dehydrin, is more consistent with elicitation of an adaptive stress response. CONCLUSIONS: While additional evidence is required to definitively establish an association between SE responsiveness and a specific physiological response, these results suggest that biotic defense activation may be antagonistic, likely related to the massive transcriptional and metabolic reprogramming that it elicits. A major issue for future work will be to determine how and if suppressing biotic defense activation could be used to promote a physiological state more conducive to SE induction.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Picea/embriologia , Picea/fisiologia , Sequência de Aminoácidos , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Picea/química , Picea/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brotos de Planta/embriologia , Brotos de Planta/genética , Brotos de Planta/metabolismo , Alinhamento de SequênciaRESUMO
Plant development is a continuous process, mainly due to the presence of stem cell niches within the root and shoot. The interplay between a host of transcription factors determines whether the cells within the meristem maintain their stem cell state, differentiate into leaves or form secondary meristems, which develop into shoots and flowers. Several recent studies provide new insight into how transcription factors and phytohormones interact within meristems to control cell proliferation and differentiation.
Assuntos
Diferenciação Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica de Plantas/genética , Sementes/genética , Sementes/metabolismo , Hormônios/genética , Hormônios/metabolismo , Meristema/citologia , Meristema/embriologia , Meristema/metabolismo , Folhas de Planta/citologia , Folhas de Planta/embriologia , Folhas de Planta/metabolismo , Brotos de Planta/citologia , Brotos de Planta/embriologia , Brotos de Planta/metabolismo , Sementes/citologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
In plant shoot meristems, cells with indeterminate fate are separated from determinate organ founder cells by morphological boundaries. Organ founder cells are selected at sites of auxin accumulation. Auxin is channeled between cells via efflux carrier proteins, but influx carriers are needed to concentrate auxin in the outer meristem layer. The genetic programmes executed by organs and meristems are established by mutual repression of transcription factors, involving the sequestration of enhancer elements into DNA loops. Boundary cells play a dual role in separating and maintaining meristem and organ domains, and express unique genes that reduce cell division and auxin efflux carrier activity, but activate meristematic gene expression. Boundary positions depend on signals emitted from indeterminate cells at the meristem center.
Assuntos
Meristema/embriologia , Meristema/fisiologia , Brotos de Planta/embriologia , Estruturas Vegetais/embriologia , Comunicação Celular/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Modelos Biológicos , Morfogênese/genética , Morfogênese/fisiologiaRESUMO
Local efflux-dependent auxin gradients and maxima mediate organ and tissue development in plants. Auxin efflux is regulated by dynamic expression and subcellular localization of the PIN auxin-efflux proteins, which appears to be established not only through a self-organizing auxin-mediated polarization mechanism, but also through other means, such as cell fate determination and auxin-independent mechanisms. Here, we show that the Arabidopsis thaliana NO VEIN (NOV) gene, encoding a novel, plant-specific nuclear factor, is required for leaf vascular development, cellular patterning and stem cell maintenance in the root meristem, as well as for cotyledon outgrowth and separation. nov mutations affect many aspects of auxin-dependent development without directly affecting auxin perception. NOV is required for provascular PIN1 expression and region-specific expression of PIN7 in leaf primordia, cell type-specific expression of PIN3, PIN4, and PIN7 in the root, and PIN2 polarity in the root cortex. NOV is specifically expressed in developing embryos, leaf primordia, and shoot and root apical meristems. Our data suggest that NOV function underlies cell fate decisions associated with auxin gradients and maxima, thus establishing cell type-specific PIN expression and polarity. We propose that NOV mediates the acquisition of competence to undergo auxin-dependent coordinated cell specification and patterning, thereby eliciting context-dependent auxin-mediated developmental responses.