RESUMEN
The ZINC FINGER-HOMEODOMAIN (ZHD) protein family is a plant-specific family of transcription factors containing two conserved motifs: a non-canonical C5H3 zinc finger domain (ZF) and a DNA-binding homeodomain (HD). The MINI ZINC FINGER (MIF) proteins belong to this family, but were possibly derived from the ZHDs by losing the HD. Information regarding the function of ZHD and MIF proteins is scarce. However, different studies have shown that ZHD/MIF proteins play important roles not only in plant growth and development, but also in response to environmental stresses, including drought and pathogen attack. Here we review recent advances relative to ZHD/MIF functions in multiple species, to provide new insights into the diverse roles of these transcription factors in plants. Their mechanism of action in relation to their ability to interact with other proteins and DNA is also discussed. We then propose directions for future studies to understand better their important roles and pinpoint strategies for potential applications in crop improvement.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Dedos de Zinc , Proteínas de Homeodominio/genética , Proteínas de Plantas/metabolismo , Plantas/genética , Plantas/metabolismo , Estrés Fisiológico/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Dedos de Zinc/genéticaRESUMEN
Plant architecture is central in determining crop yield. In the short-day species strawberry, a crop vegetatively propagated by daughter-plants produced by stolons, fruit yield is further dependent on the trade-off between sexual reproduction (fruits) and asexual reproduction (daughter-plants). Both are largely dependent on meristem identity, which establishes the development of branches, stolons and inflorescences. Floral initiation and plant architecture are modulated by the balance between two related proteins, FLOWERING LOCUS T (FT) and TERMINAL FLOWER 1 (TFL1). We explored in woodland strawberry the role of the uncharacterised FveFT2 and FveFT3 genes and of the floral repressor FveTFL1 through gene expression analyses, grafting and genetic transformation (overexpression and gene editing). We demonstrate the unusual properties of these genes. FveFT2 is a nonphotoperiodic florigen permitting short-day (SD) flowering and FveTFL1 is the long-hypothesised long-day systemic antiflorigen that contributes, together with FveFT2, to the photoperiodic regulation of flowering. We additionally show that FveFT3 is not a florigen but promotes plant branching when overexpressed, that is likely to be through changing axillary meristem fate, therefore resulting in a 3.5-fold increase in fruit yield at the expense of stolons. We show that our findings can be translated into improvement of cultivated strawberry in which FveFT2 overexpression significantly accelerates flowering.
Asunto(s)
Florigena , Fragaria , Florigena/metabolismo , Flores/genética , Flores/metabolismo , Fragaria/genética , Regulación de la Expresión Génica de las Plantas , Meristema/genética , Meristema/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Reproducción , Estaciones del AñoRESUMEN
Ascorbate is a major antioxidant buffer in plants. Several approaches have been used to increase the ascorbate content of fruits and vegetables. Here, we combined forward genetics with mapping-by-sequencing approaches using an ethyl methanesulfonate (EMS)-mutagenized Micro-Tom population to identify putative regulators underlying a high-ascorbate phenotype in tomato fruits. Among the ascorbate-enriched mutants, the family with the highest fruit ascorbate level (P17C5, up to 5-fold wild-type level) had strongly impaired flower development and produced seedless fruit. Genetic characterization was performed by outcrossing P17C5 with cv. M82. We identified the mutation responsible for the ascorbate-enriched trait in a cis-acting upstream open reading frame (uORF) involved in the downstream regulation of GDP-l-galactose phosphorylase (GGP). Using a specific CRISPR strategy, we generated uORF-GGP1 mutants and confirmed the ascorbate-enriched phenotype. We further investigated the impact of the ascorbate-enriched trait in tomato plants by phenotyping the original P17C5 EMS mutant, the population of outcrossed P17C5 × M82 plants, and the CRISPR-mutated line. These studies revealed that high ascorbate content is linked to impaired floral organ architecture, particularly anther and pollen development, leading to male sterility. RNA-seq analysis suggested that uORF-GGP1 acts as a regulator of ascorbate synthesis that maintains redox homeostasis to allow appropriate plant development.
Asunto(s)
Solanum lycopersicum , Ácido Ascórbico , Fertilidad , Frutas/genética , Solanum lycopersicum/genética , Polen/genéticaRESUMEN
In angiosperms, the gynoecium is the last structure to develop within the flower due to the determinate fate of floral meristem (FM) stem cells. The maintenance of stem cell activity before its arrest at the stage called FM termination affects the number of carpels that develop. The necessary inhibition at this stage of WUSCHEL (WUS), which is responsible for stem cell maintenance, involves a two-step mechanism. Direct repression mediated by the MADS domain transcription factor AGAMOUS (AG), followed by indirect repression requiring the C2H2 zinc-finger protein KNUCKLES (KNU), allow for the complete termination of floral stem cell activity. Here, we show that Arabidopsis thaliana MINI ZINC FINGER2 (AtMIF2) and its homolog in tomato (Solanum lycopersicum), INHIBITOR OF MERISTEM ACTIVITY (SlIMA), participate in the FM termination process by functioning as adaptor proteins. AtMIF2 and SlIMA recruit AtKNU and SlKNU, respectively, to form a transcriptional repressor complex together with TOPLESS and HISTONE DEACETYLASE19. AtMIF2 and SlIMA bind to the WUS and SlWUS loci in the respective plants, leading to their repression. These results provide important insights into the molecular mechanisms governing (FM) termination and highlight the essential role of AtMIF2/SlIMA during this developmental step, which determines carpel number and therefore fruit size.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas Portadoras/metabolismo , Flores/metabolismo , Meristema/metabolismo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/metabolismo , Acetilación , Arabidopsis/genética , Secuencia de Bases , Proteínas de Unión al ADN , Flores/genética , Frutas , Regulación de la Expresión Génica de las Plantas , Sitios Genéticos , Meristema/genética , Especificidad de Órganos/genética , Fenotipo , Unión Proteica , Homología de Secuencia de AminoácidoRESUMEN
Asexual and sexual reproduction occur jointly in many angiosperms. Stolons (elongated stems) are used for asexual reproduction in the crop species potato (Solanum tuberosum) and strawberry (Fragaria spp), where they produce tubers and clonal plants, respectively. In strawberry, stolon production is essential for vegetative propagation at the expense of fruit yield, but the underlying molecular mechanisms are unknown. Here, we show that the stolon deficiency trait of the runnerless (r) natural mutant in woodland diploid strawberry (Fragaria vesca) is due to a deletion in the active site of a gibberellin20-oxidase (GA20ox) gene, which is expressed primarily in the axillary meristem dome and primordia and in developing stolons. This mutation, which is found in all r mutants, goes back more than three centuries. When FveGA20ox4 is mutated, axillary meristems remain dormant or produce secondary shoots terminated by inflorescences, thus increasing the number of inflorescences in the plant. The application of bioactive gibberellin (GA) restored the runnering phenotype in the r mutant, indicating that GA biosynthesis in the axillary meristem is essential for inducing stolon differentiation. The possibility of regulating the runnering-flowering decision in strawberry via FveGA20ox4 provides a path for improving productivity in strawberry by controlling the trade-off between sexual reproduction and vegetative propagation.
Asunto(s)
Diploidia , Flores/fisiología , Fragaria/enzimología , Fragaria/genética , Oxigenasas de Función Mixta/metabolismo , Arabidopsis/genética , Biocatálisis , Segregación Cromosómica/genética , Cruzamientos Genéticos , Eliminación de Gen , Regulación de la Expresión Génica de las Plantas , Sitios Genéticos , Giberelinas/metabolismo , Homocigoto , Meristema/fisiología , Fenotipo , Plantas Modificadas Genéticamente , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
In grafted plants, rootstocks assure the mineral nutrition of the scion and modify its development. In this study, we show that two grapevine rootstock genotypes have different shoot branching architectures when cultivated as cuttings and that this trait is transmitted to the scion when grafted. Shoot branching plasticity in response to nitrogen supply was also studied. As strigolactones are known to have a role in the regulation of shoot development in response to nutrient availability, their involvement in the control of scion architecture by the rootstock was investigated. Functional characterization of putative grapevine strigolactone biosynthetic genes in Arabidopsis mutants or grapevine cell suspensions showed similar functions to those of Arabidopsis. Both rootstocks produced strigolactone-like compounds; the quantity produced in response to nitrogen treatments differed between the two rootstock genotypes and correlated with the expression of putative strigolactone biosynthetic genes. Exudation of strigolactone-like compounds by both rootstocks was closely related to the developmental pattern of the scion in grafted plants. These results suggest that differential regulation of strigolactone biosynthesis in response to nitrogen availability may contribute to the control of scion development conferred by each rootstock genotype.
Asunto(s)
Lactonas/metabolismo , Nitrógeno/metabolismo , Vitis/metabolismo , Disponibilidad Biológica , Raíces de Plantas/metabolismoRESUMEN
GDP-D-mannose epimerase (GME, EC 5.1.3.18) converts GDP-D-mannose to GDP-L-galactose, and is considered to be a central enzyme connecting the major ascorbate biosynthesis pathway to primary cell wall metabolism in higher plants. Our previous work demonstrated that GME is crucial for both ascorbate and cell wall biosynthesis in tomato. The aim of the present study was to investigate the respective role in ascorbate and cell wall biosynthesis of the two SlGME genes present in tomato by targeting each of them through an RNAi-silencing approach. Taken individually SlGME1 and SlGME2 allowed normal ascorbate accumulation in the leaf and fruits, thus suggesting the same function regarding ascorbate. However, SlGME1 and SlGME2 were shown to play distinct roles in cell wall biosynthesis, depending on the tissue considered. The RNAi-SlGME1 plants harbored small and poorly seeded fruits resulting from alterations of pollen development and of pollination process. In contrast, the RNAi-SlGME2 plants exhibited vegetative growth delay while fruits remained unaffected. Analysis of SlGME1- and SlGME2-silenced seeds and seedlings further showed that the dimerization state of pectin rhamnogalacturonan-II (RG-II) was altered only in the RNAi-SlGME2 lines. Taken together with the preferential expression of each SlGME gene in different tomato tissues, these results suggest sub-functionalization of SlGME1 and SlGME2 and their specialization for cell wall biosynthesis in specific tomato tissues.
Asunto(s)
Ácido Ascórbico/biosíntesis , Carbohidrato Epimerasas/metabolismo , Pared Celular/metabolismo , Solanum lycopersicum/enzimología , Carbohidrato Epimerasas/fisiología , Pared Celular/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Germinación/fisiología , Isoenzimas/metabolismo , Isoenzimas/fisiología , Solanum lycopersicum/crecimiento & desarrollo , Solanum lycopersicum/metabolismo , Polen/metabolismoRESUMEN
Tomato (Solanum lycopersicum Mill.) represents a model species for all fleshy fruits due to its biological cycle and the availability of numerous genetic and molecular resources. Its importance in human nutrition has made it one of the most valuable worldwide commodities. Tomato fruit size results from the combination of cell number and cell size, which are determined by both cell division and expansion. As fruit growth is mainly driven by cell expansion, cells from the (fleshy) pericarp tissue become highly polyploid according to the endoreduplication process, reaching a DNA content rarely encountered in other plant species (between 2C and 512C). Both cell division and cell expansion are under the control of complex interactions between hormone signalling and carbon partitioning, which establish crucial determinants of the quality of ripe fruit, such as the final size, weight, and shape, and organoleptic and nutritional traits. This review describes the genes known to contribute to fruit growth in tomato.
Asunto(s)
Frutas/genética , Regulación de la Expresión Génica de las Plantas/genética , Solanum lycopersicum/genética , Ciclo Celular , División Celular , Proliferación Celular , Tamaño de la Célula , Endorreduplicación , Frutas/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica/genética , Solanum lycopersicum/crecimiento & desarrollo , PoliploidíaRESUMEN
Strawberry (Fragaria sp.) stands as an interesting model for studying flowering behaviour and its relationship with asexual plant reproduction in polycarpic perennial plants. Strawberry produces both inflorescences and stolons (also called runners), which are lateral stems growing at the soil surface and producing new clone plants. In this study, the flowering and runnering behaviour of two cultivated octoploid strawberry (Fragaria × ananassa Duch., 2n = 8× = 56) genotypes, a seasonal flowering genotype CF1116 and a perpetual flowering genotype Capitola, were studied along the growing season. The genetic bases of the perpetual flowering and runnering traits were investigated further using a pseudo full-sibling F1 population issued from a cross between these two genotypes. The results showed that a single major quantitative trait locus (QTL) named FaPFRU controlled both traits in the cultivated octoploid strawberry. This locus was not orthologous to the loci affecting perpetual flowering (SFL) and runnering (R) in Fragaria vesca, therefore suggesting different genetic control of perpetual flowering and runnering in the diploid and octoploid Fragaria spp. Furthermore, the FaPFRU QTL displayed opposite effects on flowering (positive effect) and on runnering (negative effect), indicating that both traits share common physiological control. These results suggest that this locus plays a major role in strawberry plant fitness by controlling the balance between sexual and asexual plant reproduction.
Asunto(s)
Fragaria/metabolismo , Fragaria/fisiología , Proteínas de Plantas/metabolismo , Reproducción/fisiología , Proteínas de Plantas/genética , Sitios de Carácter Cuantitativo/genética , Reproducción/genéticaRESUMEN
The size of tomato fruit results from the combination of cell number and cell size, which are respectively determined by the cell division and cell expansion processes. As fruit growth is mainly sustained by cell expansion, the development of fleshy pericarp tissue is characterized by numerous rounds of endoreduplication inducing a spectacular increase in DNA ploidy and mean cell size. Although a clear relationship exists between endoreduplication and cell growth in plants, the exact role of endoreduplication has not been clearly elucidated. To decipher the molecular basis of endoreduplication-associated cell growth in fruit, we investigated the putative involvement of the tomato cyclin-dependent kinase inhibitor SlKRP1. We studied the kinetics of pericarp development in tomato fruit at the morphological and cytological levels, and demonstrated that endoreduplication is directly proportional to cell and fruit diameter. We established a mathematical model for tissue growth according to the number of divisions and endocycles. This model was tested in fruits where we managed to decrease the extent of endoreduplication by over-expressing SlKRP1 under the control of a fruit-specific promoter expressed during early development. Despite the fact that endoreduplication was affected, we could not observe any morphological, cytological or metabolic phenotypes, indicating that determination of cell and fruit size can be, at least conditionally, uncoupled from endoreduplication.
Asunto(s)
Ciclo Celular , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/metabolismo , Frutas/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/genética , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/genética , Frutas/citología , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/crecimiento & desarrollo , Modelos Biológicos , Proteínas de Plantas/genética , Regiones Promotoras Genéticas , ARN de Planta/genéticaRESUMEN
Understanding the mechanisms underlying differentiation of inflorescence and flower meristems is essential towards enlarging our knowledge of reproductive organ formation and to open new prospects for improving yield traits. Here, we show that SlDOF9 is a new modulator of floral differentiation in tomato. CRISPR/Cas9 knockout strategy uncovered the role of SlDOF9 in controlling inflorescence meristem and floral meristem differentiation via the regulation of cell division genes and inflorescence architecture regulator LIN. Tomato dof9-KO lines have more flowers in both determinate and indeterminate cultivars and produce more fruit upon vibration-assisted fertilization. SlDOF9 regulates inflorescence development through an auxin-dependent ARF5-DOF9 module that seems to operate, at least in part, differently in Arabidopsis and tomato. Our findings add a new actor to the complex mechanisms underlying reproductive organ differentiation in flowering plants and provide leads towards addressing the diversity of factors controlling the transition to reproductive organs.
Asunto(s)
Arabidopsis , Solanum lycopersicum , Arabidopsis/genética , Flores , Regulación de la Expresión Génica de las Plantas , Ácidos Indolacéticos/metabolismo , Inflorescencia , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Mutación , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Tomato fruit growth is characterized by the occurrence of numerous rounds of DNA endo-reduplication in connection with cell expansion and final fruit size determination. Endo-reduplication is an impairment of mitosis that originates from the selective degradation of M phase-specific cyclins via the ubiquitin-mediated proteolytic pathway, requiring the E3 ubiquitin ligase anaphase promoting complex/cyclosome (APC/C). Two types of APC/C activators, namely CCS52 and CDC20 proteins, exist in plants. We report here the molecular characterization of such APC/C activators during fruit development, and provide an in planta functional analysis of SlCCS52A, a gene that is specifically associated with endo-reduplication in tomato. Altering SlCCS52A expression in either a negative or positive manner had an impact on the extent of endo-reduplication in fruit, and fruit size was reduced in both cases. In SlCCS52A over-expressing fruits, endo-reduplication was initially delayed, accounting for the altered final fruit size, but resumed and was even enhanced at 15 days post anthesis (dpa), leading to fruit growth recovery. This induction of growth mediated by endo-reduplication had a considerable impact on nitrogen metabolism in developing fruits. Our data contribute to unravelling of the physiological role of endo-reduplication in growth induction during tomato fruit development.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Frutas/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/genética , Complejos de Ubiquitina-Proteína Ligasa/metabolismo , Ciclosoma-Complejo Promotor de la Anafase , Ciclo Celular , Proteínas de Ciclo Celular/genética , ADN de Plantas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/metabolismo , Proteínas de Plantas/genética , Mapeo de Interacción de Proteínas , Transformación Genética , Complejos de Ubiquitina-Proteína Ligasa/genéticaRESUMEN
BACKGROUND: Endoreduplication is the major source of endopolyploidy in higher plants. The process of endoreduplication results from the ability of cells to modify their classical cell cycle into a partial cell cycle where DNA synthesis occurs independently from mitosis. Despite the ubiquitous occurrence of the phenomenon in eukaryotic cells, the physiological meaning of endoreduplication remains vague, although several roles during plant development have been proposed, mostly related to cell differentiation and cell size determination. SCOPE: Here recent advances in the knowledge of endoreduplication and fruit organogenesis are reviewed, focusing on tomato (Solanum lycopersicum) as a model, and the functional analyses of endoreduplication-associated regulatory genes in tomato fruit are described. CONCLUSIONS: The cyclin-dependent kinase inhibitory kinase WEE1 and the anaphase promoting complex activator CCS52A both participate in the control of cell size and the endoreduplication process driving cell expansion during early fruit development in tomato. Moreover the fruit-specific functional analysis of the tomato CDK inhibitor KRP1 reveals that cell size and fruit size determination can be uncoupled from DNA ploidy levels, indicating that endoreduplication acts rather as a limiting factor for cell growth. The overall functional data contribute to unravelling the physiological role of endoreduplication in growth induction of fleshy fruits.
Asunto(s)
Frutas/crecimiento & desarrollo , Frutas/genética , Duplicación de Gen/genética , Solanum lycopersicum/crecimiento & desarrollo , Solanum lycopersicum/genética , Quinasas Ciclina-Dependientes/genética , Quinasas Ciclina-Dependientes/metabolismo , Frutas/enzimología , Solanum lycopersicum/enzimología , Modelos BiológicosRESUMEN
[This corrects the article DOI: 10.3389/fpls.2019.00403.].
RESUMEN
⢠Cyclin-dependent kinase (CDK) inhibitors (kip-related proteins, KRPs) play a major role in the regulation of plant cell cycle in antagonizing its progression, and are thus regulators of development. The primary sequence of KRPs is characterized by the existence of conserved motifs, for which we have limited information on their functional significance. ⢠We performed a functional analysis of various domains present in KRPs from tomato. A series of deletion mutants of SlKRP1 was generated and used in transient expression assays to define the relevance of conserved protein domains in subcellular and subnuclear localizations. Specific interactions of SlKRP1 and its deletion variants with cell cycle proteins were investigated using two-hybrid assays and bimolecular fluorescent complementation. ⢠Plant KRPs are distributed into two phylogenetic subgroups according to the presence of conserved motifs. Members of subgroup 1 represented by SlKRP1 share 6 conserved motifs whose function in protein localization and protein-protein interactions could be identified. A new interaction motif was localized in the central part of SlKRP1 that targets SlCDKA1 and SlCYCD3;1 to the nucleus. ⢠Our results bring new insights to the functional role of particular domains in KRPs relative to subcellular localization or proteolytic degradation.
Asunto(s)
Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/química , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Mapeo de Interacción de Proteínas/métodos , Solanum lycopersicum/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Núcleo Celular/metabolismo , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/clasificación , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/genética , Modelos Biológicos , Filogenia , Proteínas de Plantas/clasificación , Proteínas de Plantas/genética , Unión Proteica , Transporte de Proteínas , Fracciones Subcelulares/metabolismoRESUMEN
Plants have the ability to form new organs as a result of indeterminate growth ensured by specific regions of pluripotent cells, called meristems. Flowers are produced by the activity of floral meristems which differ from vegetative meristems in their determinate fate. Transcriptional complexes associating C-, D- and E-type MADS box proteins are responsible for flower determinacy by controlling the stem cell population within the floral meristem. We report here that the INHIBITOR OF MERISTEM ACTIVITY (IMA) gene encoding a mini zinc finger (MIF) protein from tomato (Solanum lycopersicum) regulates the processes of flower and ovule development. IMA inhibits cell proliferation during floral termination, controls the number of carpels during floral development and acts as a repressor of the meristem organizing centre gene WUSCHEL. Underexpression of IMA prevents the formation of gametophytic tissue, which is replaced by an undetermined sporophytic tissue. Molecular analyses of MADS box gene expression revealed that IMA participates in the termination of floral meristems and initiation of ovule primordia by activating D-type gene expression. Taken together, our data indicate that both termination of floral meristems and differentiation of nucelli during ovule development require a similar mechanism involving the repression of WUSCHEL and the activation of D-class genes.
Asunto(s)
Meristema/crecimiento & desarrollo , Proteínas de Plantas/fisiología , Proteínas Represoras/fisiología , Solanum lycopersicum/crecimiento & desarrollo , División Celular/genética , División Celular/fisiología , Proliferación Celular , Flores/genética , Flores/crecimiento & desarrollo , Flores/metabolismo , Frutas/genética , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Marcadores Genéticos , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Meristema/genética , Meristema/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Proteínas Represoras/química , Proteínas Represoras/genética , Dedos de ZincRESUMEN
Parthenocarpy arises when an ovary develops into fruit without pollination/fertilization. The mechanisms involved in genetic parthenocarpy have attracted attention because of their potential application in plant breeding and also for their elucidation of the mechanisms involved in early fruit development. We have isolated and characterized a novel small parthenocarpic fruit and flower (spff) mutant in the tomato (Solanum lycopersicum) cultivar Micro-Tom. This plant showed both vegetative and reproductive phenotypes including dwarfism of floral organs, male sterility, delayed flowering, altered axillary shoot development, and parthenocarpic production of small fruits. Genome-wide single nucleotide polymorphism array analysis coupled with mapping-by-sequencing using next generation sequencing-based high-throughput approaches resulted in the identification of a candidate locus responsible for the spff mutant phenotype. Subsequent linkage analysis and RNA interference-based silencing indicated that these phenotypes were caused by a loss-of-function mutation of a single gene (Solyc04g077010), which encodes a receptor-like protein kinase that was expressed in vascular bundles in young buds. Cytological and transcriptomic analyses suggested that parthenocarpy in the spff mutant was associated with enlarged ovarian cells and with elevated expression of the gibberellin metabolism gene, GA20ox1. Taken together, our results suggest a role for Solyc04g077010 in male organ development and indicate that loss of this receptor-like protein kinase activity could result in parthenocarpy.
RESUMEN
Despite a very complex structure, the sugar composition of the rhamnogalacturonan II (RG-II) pectic fraction is extremely conserved. Among its constituting monosaccharides is the seldom-observed eight-carbon sugar 3-deoxy-D-manno-octulosonic acid (Kdo), whose phosphorylated precursor is synthesized by Kdo-8-P synthase. As an attempt to alter specifically the RG-II structure in its sugar composition and assess the consequences on the function of RG-II in cell wall and its relationship with growth, Arabidopsis null mutants were sought in the genes encoding Kdo-8-P synthase. Here, the isolation and characterization of one null mutant for the isoform 1 (AtkdsA1-S) and two distinct null mutants for the isoform 2 of Arabidopsis Kdo-8-P synthase (AtkdsA2-V and AtkdsA2-S) are described. Evidence is provided that AtkdsA2 gene expression is preferentially associated with plantlet organs displaying a meristematic activity, and that it accounts for 75% of the mRNAs to be translated into Kdo-8-P synthase. Furthermore, this predominant expression of AtKDSA2 over AtKDSA1 was confirmed by quantification of the cytosolic Kdo content in the mutants, in a variety of ecotypes. The inability to identify a double knockout mutant originated from pollen abortions, due to the inability of haploid pollen of the AtkdsA1- AtkdsA2- genotype to form an elongated pollen tube properly and perform fertilization.
Asunto(s)
Aldehído-Liasas/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Pectinas/metabolismo , Polen/crecimiento & desarrollo , Polen/metabolismo , Azúcares Ácidos/metabolismo , Fosfatos de Azúcar/metabolismo , Aldehído-Liasas/química , Aldehído-Liasas/genética , Arabidopsis/enzimología , Arabidopsis/genética , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Secuencia de Bases , Regulación de la Expresión Génica de las Plantas , Pectinas/química , Polen/enzimología , Polen/genética , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismoRESUMEN
Tomato (Lycopersicon esculentum cv. Micro-Tom) plants infected by the stolbur phytoplasma (isolate PO) display floral abnormalities, including sepal hypertrophy, virescence, phyllody, and aborted reproductive organs, which are reminiscent of those observed in Arabidopsis thaliana mutants affected in flower development genes. Semiquantitative reverse transcription-polymerase chain reaction and in situ RNA hybridization were used to compare expressions of meristem and flower development genes in healthy and stolbur phytoplasma-infected tomatoes. In infected plants, FALSIFLORA (FA), controlling the identity of the inflorescence meristem, was up-regulated, whereas LeWUSCHEL (LeWUS) and LeCLAVATA1 (LeCLV1), regulating the meristem development, and LeDEFICIENS (LeDEF), responsible for the organ (petals and stamens) identity within the flower, were down-regulated regardless of the development stage of the flower bud. In contrast, expression of TAG1, which regulates stamen and carpel identities and negatively controls LeWUS, was up-regulated at the early stages and down-regulated at the late stages. In situ RNA hybridization analyses revealed that TAG1 transcripts were restricted to the same floral meristem territories in healthy and infected tomatoes, indicating that tissue-specific expression of TAG1 was not affected by the stolbur phytoplasma infection. Taken together, these data indicate that flower malformations of stolbur phytoplasma-infected tomatoes are associated with early changes in the expression of key flower development genes. The possible mechanisms by which the multiplication of stolbur phytoplasma in tomato sieve tubes deregulates floral development are discussed.
Asunto(s)
Flores/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Phytoplasma/patogenicidad , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Solanum lycopersicum/genética , Solanum lycopersicum/microbiología , Infecciones Bacterianas , Flores/genética , Flores/microbiología , Solanum lycopersicum/crecimiento & desarrollo , Modelos Biológicos , Phytoplasma/fisiología , Proteínas de Plantas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
The interactions between Plum pox virus (PPV), a member of the Potyvirus genus, and Prunus host plants are, up to now, poorly understood. In the current paper, fluorescence stereomicroscopy, in situ hybridisation and immunogold detection were performed in order to evaluate the virus transport and cellular distribution. The behavior of PPV in several susceptible (cv. "Moniqui" and "Screara") and resistant apricot genotypes (cv. "Harlayne", "Henderson", "Harcot", "Goldrich", "Stella" and "Stark Early Orange") were compared. Viral RNA was detected by in situ hybridisation in stem tissues close to the inoculation point, irrespective of the resistance status of the variety. Systemic infection was evidenced by virus immunodetection and by fluorescence detection of a GFP-tagged PPV in distant leaf sections. The signal obtained by in situ hybridisation colocalised with the fluorescence produced by GFP-tagged PPV in the same plant material but did not colocalise with the signal obtained by immunostaining. Intensity of the PPV infection in susceptible apricot cultivars varied depending on genotypes. The behavior of PPV in systemic leaves was clearly distinct between susceptible and resistant cultivars. While PPV was spreading widely around the major and minor veins in susceptible leaves, in the resistant apricot genotypes it was restricted to isolated spots consisting of few cells embedded in the mesophyll tissue. In summary, differences in the ability of PPV to systemically infect susceptible and resistant apricot cultivars were evident but nevertheless, long-distance transport of PPV occured in resistant apricot scions.