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2.
Plants (Basel) ; 11(7)2022 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-35406958

RESUMO

Powdery mildew caused by the fungus Erysiphe necator is a major grape disease worldwide. It attacks foliage and berries and reduces yield and wine quality. Fungicides are mainly used for combating the disease. Fungicide resistance and the global requisite to reduce pesticide deployment encourage the use of environment-friendly alternatives for disease management. Our field experiments showed that the foliar application of the potassium phosphate fertilizer Top-KP+ (1-50-33 NPK) reduced disease incidence on leaves and clusters by 15-65% and severity by 75-90%, compared to untreated vines. Top-KP+ mixed with Nanovatz (containing the micronutrients boron (B) and zinc (Zn)) or with TruPhos Platinum (a mixture containing N, P2O5, K2O, Zn, B, Mg, Fe, Mn, Cu, Mo, and CO) further reduced disease incidence by 30-90% and disease severity by 85-95%. These fertilizers were as effective as the fungicide tebuconazole. Tank mixtures of fertilizers and tebuconazole further enhanced control efficacy in the vineyards. The modes of action of fertilizers in disease control were elucidated via tests with grape seedlings, microscopy, and berry metabolomics. Fertilizers applied preventively to the foliage of grape seedlings inhibited powdery mildew development. Application onto existing mildew colonies plasmolyzed mycelia and conidia and arrested the development of the disease. Berries treated with fertilizers or with a fungicide showed a significant increase in anti-fungal and antioxidant metabolites. Twenty-two metabolites, including non-protein amino acids and carbohydrates, known for their anti-fungal and bioactive effects, were significantly upregulated in grapes treated with fertilizers as compared to grapes treated with a fungicide, suggesting possible indirect activity against the pathogen. Esters and organic acids that contribute to wine quality were also upregulated. We conclude that integrating macro and micronutrients in spray programs in commercial vineyards shall control powdery mildew, reduce fungicide deployment, delay the buildup of fungicide resistance, and may improve wine quality.

3.
Biology (Basel) ; 10(9)2021 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-34571814

RESUMO

Regeneration is usually regarded as a unique plant or some animal species process. In reality, regeneration is a ubiquitous process in all multicellular organisms. It ranges from response to wounding by healing the wounded tissue to whole body neoforming (remaking of the new body). In a larger context, regeneration is one facet of two reproduction schemes that dominate the evolution of life. Multicellular organisms can propagate their genes asexually or sexually. Here I present the view that the ability to regenerate tissue or whole-body regeneration is also determined by the sexual state of the multicellular organisms (from simple animals such as hydra and planaria to plants and complex animals). The above idea is manifested here by showing evidence that many organisms, organs, or tissues show inhibited or diminished regeneration capacity when in reproductive status compared to organs or tissues in nonreproductive conditions or by exposure to sex hormones.

4.
J Fungi (Basel) ; 7(7)2021 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-34356943

RESUMO

Fruit body rot and calyx rot caused by Alternaria alternata f. sp. mali is an important disease of apple worldwide. The disease has recently become severe in cv. Pink Lady apple in Israel to an extent that has never been reported elsewhere in the world. No alternative control measures of the disease except fungicides are known. Here, we show for the first time that dl-ß-aminobutyric acid (BABA) induces resistance against Alternaria fruit rot (AFR) in apple fruits in the laboratory and in the orchard. AFR was inhibited in fruits treated with BABA of 1000 µg/mL. BABA did not inhibit spore germination or mycelial growth of the pathogen in vitro (up to 2000 µg/mL). It was most inhibitory when applied 4 days prior to inoculation of detached fruits. BABA inhibited AFR also curatively when applied at 24 h post inoculation. Five other isomers of aminobutyric acid failed to protect the fruits from rot formation. Three field trials in commercial apple orchards proved that BABA was as protective against AFR as the commercial standard fungicidal mixture of azoxystrobin and difenoconazole. This research suggests that BABA may serve as a resistance inducer in apple against AFR. It can be used as an adequate alternative to the currently used fungicides or integrated in disease management programs to reduce fungicide load and buildup of fungicide resistance.

5.
Plants (Basel) ; 10(7)2021 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-34371642

RESUMO

Macromolecule and cytosolic signal distribution throughout the plant employs a unique cellular and intracellular mechanism called plasmodesmata (PD). Plant viruses spread throughout plants via PD using their movement proteins (MPs). Viral MPs induce changes in plasmodesmata's structure and alter their ability to move macromolecule and cytosolic signals. The developmental distribution of a family member of proteins termed plasmodesmata located proteins number 5 (PDLP5) conjugated to GFP (PDLP5-GFP) is described here. The GFP enables the visual localization of PDLP5 in the cell via confocal microscopy. We observed that PDLP5-GFP protein is present in seed protein bodies and immediately after seed imbibition in the plasma membrane. The effect of three different plant viruses, the tobacco mosaic virus (TMV), tomato brown rugose fruit virus (ToBRFV, tobamoviruses), and tomato yellow leaf curl virus (TYLCV, begomoviruses), on PDLP5-GFP accumulation at the plasmodesmata was tested. In tobacco leaf, TMV and ToBRFV increased PDLP5-GFP amount at the plasmodesmata of cell types compared to control. However, there was no statistically significant difference in tomato leaf. On the other hand, TYLCV decreased PDLP5-GFP quantity in plasmodesmata in all tomato leaf cells compared to control, without any significant effect on plasmodesmata in tobacco leaf cells.

6.
Sci Rep ; 11(1): 13710, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34211083

RESUMO

It is widely known that during the reproductive stage (flowering), plants do not root well. Most protocols of shoot regeneration in plants utilize juvenile tissue. Adding these two realities together encouraged us to study the role of florigen in shoot regeneration. Mature tobacco tissue that expresses the endogenous tobacco florigen mRNA regenerates poorly, while juvenile tissue that does not express the florigen regenerates shoots well. Inhibition of Nitric Oxide (NO) synthesis reduced shoot regeneration as well as promoted flowering and increased tobacco florigen level. In contrast, the addition of NO (by way of NO donor) to the tissue increased regeneration, delayed flowering, reduced tobacco florigen mRNA. Ectopic expression of florigen genes in tobacco or tomato decreased regeneration capacity significantly. Overexpression pear PcFT2 gene increased regeneration capacity. During regeneration, florigen mRNA was not changed. We conclude that florigen presence in mature tobacco leaves reduces roots and shoots regeneration and is the possible reason for the age-related decrease in regeneration capacity.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Florígeno/metabolismo , Nicotiana/crescimento & desenvolvimento , Brotos de Planta/crescimento & desenvolvimento , Pyrus/crescimento & desenvolvimento , Solanum lycopersicum/crescimento & desenvolvimento , Arabidopsis/genética , Arabidopsis/metabolismo , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Óxido Nítrico/metabolismo , Persea/genética , Persea/crescimento & desenvolvimento , Persea/metabolismo , Desenvolvimento Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Brotos de Planta/genética , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo , Pyrus/genética , Pyrus/metabolismo , RNA Mensageiro/genética , Nicotiana/genética , Nicotiana/metabolismo
7.
Physiol Plant ; 172(3): 1630-1640, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33547660

RESUMO

Flavonoids are natural pigments occurring in plants and are present in fruits, leaves, stems, roots, and flowers. Tobacco plants transformed with an MYB regulatory gene from either Solanum chilense (Sc) or S. lycopersicum (Sl) demonstrate that ScANT1 induces a higher level of anthocyanin accumulation in comparison to SlANT1 and that this gene is sufficient to promote increased anthocyanin levels. We compared the aptitude of ScANT1 protein to induce anthocyanin accumulation to that of SlANT1 protein in tobacco plants. We also tested the effect of amino acid substitutions in ScANT1 and SlANT1. We examined these synthetic alleles' effect following the over-expression of additional anthocyanin synthesis regulators, such as the tomato bHLH (SlJAF13) protein. Our results show that the amino acid changes that differentiate ScANT1 from SlANT1 are the main contributors to the advantage that ScANT1 has over SlANT1 in anthocyanin accumulation per transcript unit. We further demonstrated that altering the amino acid composition of SlANT1 could increase anthocyanin accumulation, while reciprocally modifying ScANT1 lowers the anthocyanin level. These results confirm the increased anthocyanin level in tobacco is attributed to the amino acid differences between ScANT1 and SlANT1. We also show that the co-expression of SlJAF13 with SlANT1 in tobacco plants represses the anthocyanin production.


Assuntos
Solanum lycopersicum , Solanum , Alelos , Antocianinas , Regulação da Expressão Gênica de Plantas/genética , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Solanum/genética , Solanum/metabolismo , Nicotiana/genética , Nicotiana/metabolismo
8.
Environ Microbiol ; 23(10): 6019-6037, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-33459475

RESUMO

Plant pathogens usually originate and diversify in geographical regions where hosts and pathogens co-evolve. Erysiphe necator, the causal agent of grape powdery mildew, is a destructive pathogen of grapevines worldwide. Although Eastern US is considered the centre of origin and diversity of E. necator, previous reports on resistant native wild and domesticated Asian grapevines suggest Asia as another possible origin of the pathogen. By using multi-locus sequencing, microsatellites and a novel application of amplicon sequencing (AmpSeq), we show that the population of E. necator in Israel is composed of three genetic groups: Groups A and B that are common worldwide, and a new group IL, which is genetically differentiated from any known group in Europe and Eastern US. Group IL showed distinguished ecological characteristics: it was dominant on wild and traditional vines (95%); its abundance increased along the season; and was more aggressive than A and B isolates on both wild and domesticated vines. The low genetic diversity within group IL suggests that it has invaded Israel from another origin. Therefore, we suggest that the Israeli E. necator population was founded by at least two invasions, of which one could be from a non-East American source, possibly from Asian origin.


Assuntos
Ascomicetos , Vitis , Ascomicetos/genética , Erysiphe , Doenças das Plantas
9.
Plants (Basel) ; 9(9)2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32887438

RESUMO

The essential tea tree oil (TTO) derived from Melaleuca alternifolia plant is widely used as a biopesticide to protect crops from several plant-pathogens. Its activity raised queries regarding its ability to, not only act as a bio-fungicide or bio-bactericide, but also systemically inducing resistance in plants. This was examined by TTO application to banana plants challenged by Fusarium oxysporum f. sp. cubense (Foc, Race 1) causing Fusarium wilt and to tomato plants challenged by Xanthomonas campestris. Parameters to assess resistance induction included: disease development, enzymatic activity, defense genes expression correlated to systemic acquired resistance (SAR) and induced systemic resistance (ISR) and priming effect. Spraying TTO on field-grown banana plants infected with Foc and greenhouse tomato plants infected with Xanthomonas campestris led to resistance induction in both hosts. Several marker genes of salicylic acid, jasmonic acid and ethylene pathways were significantly up-regulated in parallel with symptoms reduction. For tomato plants, we have also recorded a priming effect following TTO treatment. In addition to fungicidal and bactericidal effect, TTO can be applied in more sustainable strategies to control diseases by enhancing the plants ability to defend themselves against pathogens and ultimately diminish chemical pesticides applications.

10.
Mol Plant Pathol ; 21(7): 895-906, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32352631

RESUMO

T2 ribonucleases (RNases) are RNA-degrading enzymes that function in various cellular processes, mostly via RNA metabolism. T2 RNase-encoding genes have been identified in various organisms, from bacteria to mammals, and are most diverse in plants. The existence of T2 RNase genes in almost every organism suggests an important biological function that has been conserved through evolution. In plants, T2 RNases are suggested to be involved in phosphate scavenging and recycling, and are implicated in defence responses to pathogens. We investigated the function of the tomato T2 RNase LE, known to be induced by phosphate deficiency and wounding. The possible involvement of LE in pathogen responses was examined. Expression analysis showed LE induction during fungal infection and by stimuli known to be associated with pathogen inoculation, including oxalic acid and hydrogen peroxide. Analysis of LE-suppressed transgenic tomato lines revealed higher susceptibility to oxalic acid, a cell death-inducing factor, compared to the wild type. This elevated sensitivity of LE-suppressed lines was evidenced by visual signs of necrosis, and increased ion leakage and reactive oxygen species levels, indicating acceleration of cell death. Challenge of the LE-suppressed lines with the necrotrophic pathogen Botrytis cinerea resulted in accelerated development of disease symptoms compared to the wild type, associated with suppressed expression of pathogenesis-related marker genes. The results suggest a role for plant endogenous T2 RNases in antifungal activity.


Assuntos
Botrytis , Endorribonucleases/metabolismo , Doenças das Plantas , Solanum lycopersicum/enzimologia , Solanum lycopersicum/microbiologia , Endorribonucleases/genética , Indução Enzimática , Solanum lycopersicum/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Plantas Geneticamente Modificadas , Espécies Reativas de Oxigênio/metabolismo
11.
Plants (Basel) ; 10(1)2020 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-33383798

RESUMO

Shoot regeneration is a key tool of modern plant biotechnology. While many researchers use this process empirically, very little is known about the early molecular genetic factors and signaling events that lead to shoot regeneration. Using tobacco as a model system, we found that the inductive events required for shoot regeneration occur in the first 4-5 days following incubation on regeneration medium. Leaf segments placed on regeneration medium did not produce shoots if removed from the medium before four days indicating this time frame is crucial for the induction of shoot regeneration. Leaf segments placed on regeneration medium for longer than five days maintain the capacity to produce shoots when removed from the regeneration medium. Analysis of gene expression during the early days of incubation on regeneration medium revealed many changes occurring with no single expression pattern evident among major gene families previously implicated in developmental processes. For example, expression of Knotted gene family members increased during the induction period, whereas transcription factors from the Wuschel gene family were unaltered during shoot induction. Expression levels of genes involved in cell cycle regulation increased steadily on regeneration medium while expression of NAC genes varied. No obvious possible candidate genes or developmental processes could be identified as a target for the early events (first few days) in the induction of shoot regeneration. On the other hand, observations during the early stages of regeneration pointed out that regeneration does not occur from a single cell but a group of cells. We observed that while cell division starts just as leaf segments are placed on regeneration medium, only a group of cells could become shoot primordia. Still, these primordia are not identifiable during the first days.

12.
Plant Methods ; 15: 90, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31404403

RESUMO

BACKGROUND: Characterization and quantification of visual plant traits is often limited to the use of tools and software that were developed to address a specific context, making them unsuitable for other applications. CoverageTool is flexible multi-purpose software capable of area calculation in cm2, as well as coverage area in percentages, suitable for a wide range of applications. RESULTS: Here we present a novel, semi-automated and robust tool for detailed characterization of visual plant traits. We demonstrate and discuss the application of this tool to quantify a broad spectrum of plant phenotypes/traits such as: tissue culture parameters, ground surface covered by annual plant canopy, root and leaf projected surface area, and leaf senescence area ratio. The CoverageTool software provides easy to use functions to analyze images. While use of CoverageTool involves subjective operator color selections, applying them uniformly to full sets of samples makes it possible to provide quantitative comparison between test subjects. CONCLUSION: The tool is simple and straightforward, yet suitable for the quantification of biological and environmental effects on a wide variety of visual plant traits. This tool has been very useful in quantifying different plant phenotypes in several recently published studies, and may be useful for many applications.

13.
Plant Dis ; 102(6): 1072-1080, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30673439

RESUMO

Fruit body rot and calyx rot caused by Alternaria alternata f. sp. mali has recently become a severe disease of Pink Lady apple in Israel. Severe fruit rot caused by A. alternata f. sp. mali has not been reported elsewhere in the world, and control measures are not currently known. Our objective was to determine the peak periods of susceptibility and develop methods to manage the disease by timing fungicide applications according to fruit phenological stage. We determined the relationship between fruit phenological stage and rot susceptibility by (i) monitoring the appearance of first fruit symptoms in the orchard; (ii) inoculating detached and attached fruit in the laboratory and orchard, respectively, at various time intervals after petal fall; and (iii) starting fungicide applications at various time intervals after petal fall. Fruit of Pink Lady acquired susceptibility to the disease at about 115 days after petal fall (DAPF) when reaching a diameter of ≥55 mm. Based on these findings, a new spray strategy was adopted involving a limited number of 4 to 6 foliar sprays of azoxystrobin + difenoconazole or tebuconazole + captan, or their alternation, starting at 115 DAPF. This strategy provided excellent control of both body rot and calyx rot in Pink Lady apple fruit in Israel.


Assuntos
Alternaria/fisiologia , Frutas/microbiologia , Fungicidas Industriais/farmacologia , Malus/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Alternaria/efeitos dos fármacos
14.
Plant Sci ; 241: 164-76, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26706068

RESUMO

Annual and perennial plants represent two different evolutionary strategies based on differential synchronization of their reproductive development. The mobile signal protein FLOWERING LOCUS T (FT) plays a central role in mediating the onset of reproduction in both plant types. Two novel FT-like genes from pear (Pyrus communis)-PcFT1 and PcFT2-were isolated, and their expression profiles were determined for one annual cycle. The effects of PcFT2 on flowering were investigated in annual (tobacco) and perennial (apple) plants by means of grafting and generating transgenic plants. Long-distance graft transmission of PcFT2 in both annual and perennial plants was confirmed using a 35S::PcFT2-YFP construct. Ectopic overexpression of PcFT2 caused early flowering in tobacco but not in apple. Transgenic apples were less sensitive to short-day-induced dormancy, and this phenotype was also observed in wild-type apples grafted onto the transgenic plants. Comparison of PcFT2 protein structure to the paralogous FT proteins from apple and pear showed alterations that could influence protein structure and thus the florigen-activation complex. PcFT2 protein seems to function by promoting flowering as all other FT proteins in the annual plant tobacco while in the perennial plant apple PcFT2 does not promote flowering but delays senescence. This observation may hint to a modified function of FT2 in perennial plants.


Assuntos
Regulação da Expressão Gênica de Plantas , Malus/fisiologia , Nicotiana/crescimento & desenvolvimento , Proteínas de Plantas/genética , Pyrus/genética , Sequência de Aminoácidos , Flores/crescimento & desenvolvimento , Malus/genética , Dados de Sequência Molecular , Filogenia , Dormência de Plantas , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/fisiologia , Pyrus/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Transgenes
15.
PLoS Genet ; 11(10): e1005538, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26448569

RESUMO

Tomato yellow leaf curl virus (TYLCV) is a devastating disease of tomato (Solanum lycopersicum) that can be effectively controlled by the deployment of resistant cultivars. The TYLCV-resistant line TY172 carries a major recessive locus for TYLCV resistance, designated ty-5, on chromosome 4. In this study, the association between 27 polymorphic DNA markers, spanning the ty-5 locus, and the resistance characteristics of individual plants inoculated with TYLCV in 51 segregating recombinant populations were analyzed. These analyses localized ty-5 into a 425 bp region containing two transversions: one in the first exon of a gene encoding the tomato homolog of the messenger RNA surveillance factor Pelota (Pelo), and a second in its proximal promoter. Analyses of susceptible and resistant lines revealed that the relative transcript level of the gene remained unchanged, regardless of whether the plants were infected with TYLCV or not. This suggests that the polymorphism discovered in the coding region of the gene controls the resistance. Silencing of Pelo in a susceptible line rendered the transgenic plants highly resistant, while in the resistant line TY172 had no effect on symptom development. In addition, over-expression of the susceptible allele of the gene in the resistant TY172 line rendered it susceptible, while over-expression of the resistant allele in susceptible plants had no effect. These results confirm that Pelo is the gene controlling resistance at the ty-5 locus. Pelo, implicated in the ribosome recycling-phase of protein synthesis, offers an alternative route to promote resistance to TYLCV and other viruses.


Assuntos
Resistência à Doença/genética , Estudos de Associação Genética , Doenças das Plantas/genética , Proteínas de Plantas/genética , Solanum lycopersicum/genética , Begomovirus/genética , Begomovirus/patogenicidade , Mapeamento Cromossômico , Marcadores Genéticos/genética , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/virologia , Dados de Sequência Molecular , Doenças das Plantas/virologia , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/virologia , Plantas Geneticamente Modificadas , RNA Mensageiro/genética
16.
BMC Res Notes ; 8: 207, 2015 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-26032618

RESUMO

BACKGROUND: In cancer cells in vitro, the glycolytic pathway and the mitochondrial tricarboxylic acid (TCA) cycle are programmed to produce more precursor molecules, and relatively less ATP, than in differentiated cells. We address the questions of whether and where these changes occur in vivo in glioblastomas grown from C6 cells in rat brain. These gliomas show some spatial organization, notably in the upregulation of membrane proton transporters near the rim. RESULTS: We immunolabeled pairs of proteins (as well as DNA) on sections of rat brains containing gliomas, measured the profiles of fluorescence intensity on strips 200 µm wide and at least 3 mm long running perpendicular to the tumor rim, and expressed the intensity in the glioma relative to that outside. On averaged profiles, labeling of a marker of the glycolytic pathway, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), was, as expected, greater in the glioma. Over distances up to 2.5 mm into the glioma, expression of a marker of the TCA cycle, Tom20, a pre-protein receptor on the translocation complex of the mitochondrial outer membrane, was also upregulated. The ratio of upregulation of Tom20 to upregulation of GAPDH was, on average, slightly greater than one. Near the rim (0.4-0.8 mm), GAPDH was expressed less and there was a peak in the mean ratio of 1.16, SEM = 0.001, N = 16 pairs of profiles. An antibody to V-ATPase, which, by pumping protons into vacuoles contributes to cell growth, also indicated upregulation by about 40%. When compared directly with GAPDH, upregulation of V-ATPase was only 0.764, SD = 0.016 of GAPDH upregulation. CONCLUSIONS: Although there was considerable variation between individual measured profiles, on average, markers of the glycolytic pathway, of mitochondria, and of cell proliferation showed coherent upregulation in C6 gliomas. There is a zone, close to the rim, where mitochondrial presence is upregulated more than the glycolytic pathway, in agreement with earlier suggestions that lactate is taken up by cells near the rim.


Assuntos
Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Glioma/metabolismo , Glioma/patologia , Glicólise , Mitocôndrias/metabolismo , Bombas de Próton/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Masculino , Proteínas de Membrana Transportadoras , Proteínas do Complexo de Importação de Proteína Precursora Mitocondrial , Proteínas Mitocondriais/metabolismo , Ratos Wistar , Receptores de Superfície Celular , Receptores Citoplasmáticos e Nucleares/metabolismo , Regulação para Cima , ATPases Vacuolares Próton-Translocadoras/metabolismo
17.
J Integr Plant Biol ; 54(2): 115-30, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22221327

RESUMO

Plant mitochondrial genomes (mtDNAs) are large and undergo frequent recombination events. A common phenotype that emerges as a consequence of altered mtDNA structure is cytoplasmic-male sterility (CMS). The molecular basis for CMS remains unclear, but it seems logical that altered respiration activities would result in reduced pollen production. Analysis of tobacco (Nicotiana tabacum) mtDNAs indicated that CMS-associated loci often contain fragments of known organellar genes. These may assemble with organellar complexes and thereby interfere with normal respiratory functions. Here, we analyzed whether the expression of truncated fragments of mitochondrial genes (i.e. atp4, cox1 and rps3) may induce male sterility by limiting the biogenesis of the respiratory machinery. cDNA fragments corresponding to atp4f, cox1f and rps3f were cloned in-frame to a mitochondrial localization signal and a C-termini HA-tag under a tapetum-specific promoter and introduced to tobacco plants by Agrobacterium-mediated transformation. The constructs were then analyzed for their effect on mitochondrial activity and pollen fertility. Atp4f, Cox1f and Rps3f plants demonstrated male sterility phenotypes, which were tightly correlated with the expression of the recombinant fragments in the floral meristem. Fractionation of native organellar extracts showed that the recombinant ATP4f-HA, COX1f-HA and RPS3f-HA proteins are found in large membrane-associated particles. Analysis of the respiratory activities and protein profiles indicated that organellar complex I was altered in Atp4f, Cox1f and Rps3f plants.


Assuntos
Regulação da Expressão Gênica de Plantas , Genes Mitocondriais/genética , Nicotiana/anatomia & histologia , Nicotiana/genética , Infertilidade das Plantas/genética , Respiração Celular/genética , Espaço Intracelular/metabolismo , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Mutagênese Insercional/genética , Fases de Leitura Aberta/genética , Fenótipo , Folhas de Planta/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Pólen/genética , Transporte Proteico , Proteínas Recombinantes/metabolismo , Recombinação Genética/genética , Nicotiana/citologia
18.
Theor Appl Genet ; 124(2): 295-307, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21947299

RESUMO

Anthocyanins are flavonoid metabolites contributing attractive colors and antioxidant qualities to the human diet. Accordingly, there is a growing interest in developing crops enriched with these compounds. Fruits of the cultivated tomato, Solanum (S.) lycopersicum, do not normally produce high levels of anthocyanins. However, several wild tomato species yield anthocyanin-pigmented fruits, and this trait has been introgressed into the cultivated tomato. Two genes encoding homologous R2R3 MYB transcription factors, termed ANT1 and AN2, were previously genetically implicated in anthocyanin accumulation in tomato fruit peels of the ANTHOCYANIN FRUIT (AFT) genotype originating from S. chilense. Here we compared transgenic tomato plants constitutively over-expressing the S. lycopersicum (35S::ANT1 ( L ) ) or the S. chilense (35S::ANT1 ( C )) allele, and show that each displayed variable levels of purple pigmentation in vegetative as well as reproductive tissues. However, 35S::ANT1 ( C ) was significantly more efficient in producing anthocyanin pigments, attributed to its gene coding-sequence rather than to its transcript levels. These results expand the potential of enhancing anthocyanin levels through engineering coding-sequence polymorphisms in addition to the transcriptional alterations commonly used. In addition, a segregating population obtained from a recombinant genotype revealed that the native ANT1, and not AN2, is fully associated with the AFT phenotype and that ANT1 alone can generate the characteristic phenotype of anthocyanin accumulation in AFT fruits. Our results therefore provide further support to the hypothesis that ANT1 is the gene responsible for anthocyanin accumulation in fruits of the AFT genotype.


Assuntos
Antocianinas/metabolismo , Frutas/metabolismo , Genes de Plantas/genética , Fenótipo , Solanum/genética , Análise de Variância , Primers do DNA/genética , Genótipo , Plantas Geneticamente Modificadas , Plasmídeos/genética , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo Real , Solanum/metabolismo , Especificidade da Espécie
19.
Physiol Plant ; 142(3): 211-23, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21338368

RESUMO

Tomato (Solanum lycopersicum) fruits expressing a yeast S-adenosyl methionine decarboxylase (ySAMdc) gene under control of a ripening-induced promoter show altered phytonutrient content and broad changes in gene expression. Genome-wide transcriptional alterations in pericarp tissues of the ySAMdc-expressing fruits are shown. Consistent with the ySAMdc expression pattern from the ripening-induced promoter, very minor transcriptional alterations were detected at the mature green developmental stage. At the breaker and red stages, altered levels of numerous transcripts were observed with a general tendency toward upregulation in the transgenic fruits. Ontological analysis of up- and downregulated transcript groups revealed various affected metabolic processes, mainly carbohydrate and amino acid metabolism, and protein synthesis, which appeared to be intensified in the ripening transgenic fruits. Other functional ontological categories of altered transcripts represented signal transduction, transcription regulation, RNA processing, molecular transport and stress response, as well as metabolism of lipids, glycans, xenobiotics, energy, cofactors and vitamins. In addition, transcript levels of genes encoding structural enzymes for several biosynthetic pathways showed strong correlations to levels of specific metabolites that displayed altered levels in transgenic fruits. Increased transcript levels of fatty acid biosynthesis enzymes were accompanied by a change in the fatty acid profile of transgenic fruits, most notably increasing ω-3 fatty acids at the expense of other lipids. Thus, SAMdc is a prime target in manipulating the nutritional value of tomato fruits. Combined with analyses of selected metabolites in the overripe fruits, a model of enhanced homeostasis of the pericarp tissue in the polyamine-accumulating tomatoes is proposed.


Assuntos
Adenosilmetionina Descarboxilase/genética , Ácidos Graxos/metabolismo , Frutas/genética , Regulação da Expressão Gênica de Plantas , Genes Fúngicos/genética , Saccharomyces cerevisiae/genética , Solanum lycopersicum/genética , Carotenoides/metabolismo , Respiração Celular/genética , Respiração Celular/efeitos da radiação , Etilenos/metabolismo , Frutas/citologia , Frutas/crescimento & desenvolvimento , Frutas/efeitos da radiação , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Genes de Plantas/genética , Luz , Solanum lycopersicum/citologia , Solanum lycopersicum/efeitos da radiação , Redes e Vias Metabólicas/genética , Redes e Vias Metabólicas/efeitos da radiação , Análise de Sequência com Séries de Oligonucleotídeos , Plantas Geneticamente Modificadas , Poliaminas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/efeitos da radiação , Transcrição Gênica/efeitos da radiação
20.
J Exp Bot ; 61(13): 3627-37, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20566564

RESUMO

Fruits of tomato plants carrying the high pigment-1 mutations hp-1 and hp-1(w) are characterized by an increased number of plastids coupled with enhanced levels of functional metabolites. Unfortunately, hp-1 mutant plants are also typified by light-dependent retardation in seedling and whole-plant growth and development, which limits their cultivation. These mutations were mapped to the gene encoding UV-DAMAGED DNA BINDING PROTEIN 1 (DDB1) and, recently, fruit-specific RNA interference studies have demonstrated an increased number of plastids and enhanced carotenoid accumulation in the transgenic tomato fruits. However, whole-plant overexpression of DDB1, required to substantiate its effects on seedling and plant development and to couple them with fruit phenotypes, has heretofore been unsuccessful. In this study, five transgenic lines constitutively overexpressing normal DDB1 in hp-1 mutant plants were analysed. Eleven-day-old seedlings, representing these lines, displayed up to approximately 73- and approximately 221-fold overexpression of the gene in hypocotyls and cotyledons, respectively. This overexpression resulted in statistically significant reversion to the non-mutant developmental phenotypes, including more than a full quantitative reversion. This reversion of phenotypes was generally accompanied by correlated responses in chlorophyll accumulation and altered expression of selected light signalling genes: PHYTOCHROME A, CRYPTOCHROME 1, ELONGATED HYPOCOTYL 5, and the gene encoding CHLOROPHYLL A/B-BINDING PROTEIN 4. Cumulatively, these results provide the missing link between DDB1 and its effects on tomato plant development.


Assuntos
Expressão Gênica , Genes de Plantas/genética , Solanum lycopersicum , Clorofila/metabolismo , Clorofila A , Regulação da Expressão Gênica de Plantas , Hipocótilo/metabolismo , Luz , Solanum lycopersicum/genética , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/metabolismo , Mutação , Fenótipo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plântula/genética , Plântula/metabolismo , Transdução de Sinais/genética
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