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1.
Planta ; 254(4): 80, 2021 Sep 21.
Artículo en Inglés | MEDLINE | ID: mdl-34546416

RESUMEN

MAIN CONCLUSION: A greater rate of phloem unloading and storage in the stem, not a higher rate of sugar production by photosynthesis or sugar export from leaves, is the main factor that results in sugar accumulation in sweet dwarf sorghum compared to grain sorghum. At maturity, the stem internodes of sweet sorghum varieties accumulate high concentrations of fermentable sugars and represent an efficient feedstock for bioethanol production. Although stem sugar accumulation is a heritable trait, additional factors that drive sugar accumulation in sorghum have not been identified. To identify the constraints on stem sugar accumulation in sweet sorghum, we used a combination of carbon-11 (11C) radiotracer, physiological and biochemical approaches, and compared a grain sorghum and sweet dwarf sorghum line that have similar growth characteristics including height. Photosynthesis did not increase during development or differ between the sorghum lines. During the developmental transition to the reproductive stage, export of 11C from leaves approximately doubled in both sorghum lines, but 11C export in the sweet dwarf line did not exceed that of the grain sorghum. Defoliation to manipulate relative sink demand did not result in increased photosynthetic rates, indicating that the combined accumulation of C by all sink tissues was limited by the maximum photosynthetic capacity of source leaves. Nearly 3/4 of the 11C exported from leaves was transported to the lower stem in sweet sorghum within 2 h, whereas in grain sorghum nearly 3/4 of the 11C was in the panicle. Accordingly, the transcripts of several sucrose transporter (SUT) genes were more abundant in the stem internodes of the sweet dwarf line compared to the grain sorghum. Overall, these results indicate that sugar accumulation in sweet sorghum stems is influenced by the interplay of different sink tissues for the same sugars, but is likely driven by elevated sugar phloem unloading and uptake capacity in mature stem internodes.


Asunto(s)
Sorghum , Carbono , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Sorghum/metabolismo , Azúcares
2.
J Exp Bot ; 71(15): 4512-4530, 2020 07 25.
Artículo en Inglés | MEDLINE | ID: mdl-32161967

RESUMEN

Seasonal nitrogen (N) cycling in Populus, involves bark storage proteins (BSPs) that accumulate in bark phloem parenchyma in the autumn and decline when shoot growth resumes in the spring. Little is known about the contribution of BSPs to growth or the signals regulating N remobilization from BSPs. Knockdown of BSP accumulation via RNAi and N sink manipulations were used to understand how BSP storage influences shoot growth. Reduced accumulation of BSPs delayed bud break and reduced shoot growth following dormancy. Further, 13N tracer studies also showed that BSP accumulation is an important factor in N partitioning from senescing leaves to bark. Thus, BSP accumulation has a role in N remobilization during N partitioning both from senescing leaves to bark and from bark to expanding shoots once growth commences following dormancy. The bark transcriptome during BSP catabolism and N remobilization was enriched in genes associated with auxin transport and signaling, and manipulation of the source of auxin or auxin transport revealed a role for auxin in regulating BSP catabolism and N remobilization. Therefore, N remobilization appears to be regulated by auxin produced in expanding buds and shoots that is transported to bark where it regulates protease gene expression and BSP catabolism.


Asunto(s)
Populus , Ácidos Indolacéticos , Nitrógeno , Radioisótopos de Nitrógeno , Proteínas de Plantas/genética , Brotes de la Planta , Populus/genética , Estaciones del Año , Árboles
3.
Proc Natl Acad Sci U S A ; 111(16): 6092-7, 2014 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-24711430

RESUMEN

For almost a century the plant hormone auxin has been central to theories on apical dominance, whereby the growing shoot tip suppresses the growth of the axillary buds below. According to the classic model, the auxin indole-3-acetic acid is produced in the shoot tip and transported down the stem, where it inhibits bud growth. We report here that the initiation of bud growth after shoot tip loss cannot be dependent on apical auxin supply because we observe bud release up to 24 h before changes in auxin content in the adjacent stem. After the loss of the shoot tip, sugars are rapidly redistributed over large distances and accumulate in axillary buds within a timeframe that correlates with bud release. Moreover, artificially increasing sucrose levels in plants represses the expression of BRANCHED1 (BRC1), the key transcriptional regulator responsible for maintaining bud dormancy, and results in rapid bud release. An enhancement in sugar supply is both necessary and sufficient for suppressed buds to be released from apical dominance. Our data support a theory of apical dominance whereby the shoot tip's strong demand for sugars inhibits axillary bud outgrowth by limiting the amount of sugar translocated to those buds.


Asunto(s)
Carbohidratos/farmacología , Ácidos Indolacéticos/farmacología , Pisum sativum/crecimiento & desarrollo , Brotes de la Planta/crecimiento & desarrollo , Carbono/metabolismo , Flores/efectos de los fármacos , Flores/fisiología , Flores/efectos de la radiación , Luz , Modelos Biológicos , Pisum sativum/efectos de los fármacos , Pisum sativum/efectos de la radiación , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/fisiología , Hojas de la Planta/efectos de la radiación , Proteínas de Plantas/metabolismo , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/efectos de la radiación , Transducción de Señal/efectos de los fármacos , Transducción de Señal/efectos de la radiación , Sacarosa/farmacología
4.
Plant Cell ; 25(7): 2714-30, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23903318

RESUMEN

Salicylic acid (SA) has long been implicated in plant responses to oxidative stress. SA overproduction in Arabidopsis thaliana leads to dwarfism, making in planta assessment of SA effects difficult in this model system. We report that transgenic Populus tremula × alba expressing a bacterial SA synthase hyperaccumulated SA and SA conjugates without negative growth consequences. In the absence of stress, endogenously elevated SA elicited widespread metabolic and transcriptional changes that resembled those of wild-type plants exposed to oxidative stress-promoting heat treatments. Potential signaling and oxidative stress markers azelaic and gluconic acids as well as antioxidant chlorogenic acids were strongly coregulated with SA, while soluble sugars and other phenylpropanoids were inversely correlated. Photosynthetic responses to heat were attenuated in SA-overproducing plants. Network analysis identified potential drivers of SA-mediated transcriptome rewiring, including receptor-like kinases and WRKY transcription factors. Orthologs of Arabidopsis SA signaling components NON-EXPRESSOR OF PATHOGENESIS-RELATED GENES1 and thioredoxins were not represented. However, all members of the expanded Populus nucleoredoxin-1 family exhibited increased expression and increased network connectivity in SA-overproducing Populus, suggesting a previously undescribed role in SA-mediated redox regulation. The SA response in Populus involved a reprogramming of carbon uptake and partitioning during stress that is compatible with constitutive chemical defense and sustained growth, contrasting with the SA response in Arabidopsis, which is transient and compromises growth if sustained.


Asunto(s)
Fotosíntesis , Populus/genética , Populus/metabolismo , Ácido Salicílico/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Ontología de Genes , Redes Reguladoras de Genes , Calor , Liasas/genética , Liasas/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Oxidación-Reducción , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Populus/crecimiento & desarrollo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/genética , Transcriptoma
5.
BMC Plant Biol ; 15: 273, 2015 Nov 09.
Artículo en Inglés | MEDLINE | ID: mdl-26552889

RESUMEN

BACKGROUND: Although important aspects of whole-plant carbon allocation in crop plants (e.g., to grain) occur late in development when the plants are large, techniques to study carbon transport and allocation processes have not been adapted for large plants. Positron emission tomography (PET), developed for dynamic imaging in medicine, has been applied in plant studies to measure the transport and allocation patterns of carbohydrates, nutrients, and phytohormones labeled with positron-emitting radioisotopes. However, the cost of PET and its limitation to smaller plants has restricted its use in plant biology. Here we describe the adaptation and optimization of a commercial clinical PET scanner to measure transport dynamics and allocation patterns of (11)C-photoassimilates in large crops. RESULTS: Based on measurements of a phantom, we optimized instrument settings, including use of 3-D mode and attenuation correction to maximize the accuracy of measurements. To demonstrate the utility of PET, we measured (11)C-photoassimilate transport and allocation in Sorghum bicolor, an important staple crop, at vegetative and reproductive stages (40 and 70 days after planting; DAP). The (11)C-photoassimilate transport speed did not change over the two developmental stages. However, within a stem, transport speeds were reduced across nodes, likely due to higher (11)C-photoassimilate unloading in the nodes. Photosynthesis in leaves and the amount of (11)C that was exported to the rest of the plant decreased as plants matured. In young plants, exported (11)C was allocated mostly (88 %) to the roots and stem, but in flowering plants (70 DAP) the majority of the exported (11)C (64 %) was allocated to the apex. CONCLUSIONS: Our results show that commercial PET scanners can be used reliably to measure whole-plant C-allocation in large plants nondestructively including, importantly, allocation to roots in soil. This capability revealed extreme changes in carbon allocation in sorghum plants, as they advanced to maturity. Further, our results suggest that nodes may be important control points for photoassimilate distribution in crops of the family Poaceae. Quantifying real-time carbon allocation and photoassimilate transport dynamics, as demonstrated here, will be important for functional genomic studies to unravel the mechanisms controlling phloem transport in large crop plants, which will provide crucial insights for improving yields.


Asunto(s)
Botánica/métodos , Carbono/metabolismo , Tomografía de Emisión de Positrones/métodos , Sorghum/metabolismo , Transporte Biológico , Fotosíntesis
6.
Plant Cell Physiol ; 55(9): 1669-78, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-25016610

RESUMEN

4-Coumarate:CoA ligase (4CL) catalyzes the formation of hydroxycinnamoyl-CoA esters for phenylpropanoid biosynthesis. Phylogenetically distinct Class I and Class II 4CL isoforms occur in angiosperms, and support lignin and non-lignin phenylpropanoid biosynthesis, respectively. In contrast, the few experimentally characterized gymnosperm 4CLs are associated with lignin biosynthesis and belong to the conifer-specific Class III. Here we report a new Pinus taeda isoform Pinta4CL3 that is phylogenetically more closely related to Class II angiosperm 4CLs than to Class III Pinta4CL1. Like angiosperm Class II 4CLs, Pinta4CL3 transcript levels were detected in foliar and root tissues but were absent in xylem, and recombinant Pinta4CL3 exhibited a substrate preference for 4-coumaric acid. Constitutive expression of Pinta4CL3 in transgenic Populus led to significant increases of hydroxycinnamoyl-quinate esters at the expense of hydroxycinnamoyl-glucose esters in green tissues. In particular, large increases of cinnamoyl-quinate in transgenic leaves suggested in vivo utilization of cinnamic acid by Pinta4CL3. Lignin was unaffected in transgenic Populus, consistent with Pinta4CL3 involvement in biosynthesis of non-structural phenylpropanoids. We discuss the in vivo cinnamic acid utilization activity of Pinta4CL3 and its adaptive significance in conifer defense. Together with phylogenetic inference, our data support an ancient origin of Class II 4CLs that pre-dates the angiosperm-gymnosperm split.


Asunto(s)
Coenzima A Ligasas/metabolismo , Regulación de la Expresión Génica de las Plantas , Pinus/enzimología , Populus/enzimología , Propanoles/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Coenzima A Ligasas/genética , Ácidos Cumáricos/metabolismo , Expresión Génica , Regulación Enzimológica de la Expresión Génica , Isoenzimas , Lignina/metabolismo , Datos de Secuencia Molecular , Filogenia , Pinus/genética , Hojas de la Planta/química , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/química , Raíces de Plantas/enzimología , Raíces de Plantas/genética , Populus/química , Populus/genética , Propionatos , Análisis de Secuencia de ADN , Xilema/química , Xilema/enzimología , Xilema/genética
7.
Plant Cell Environ ; 37(11): 2613-22, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-24762051

RESUMEN

Upon attack by leaf herbivores, many plants reallocate photoassimilates below ground. However, little is known about how plants respond when the roots themselves come under attack. We investigated induced resource allocation in maize plants that are infested by the larvae Western corn rootworm Diabrotica virgifera virgifera. Using radioactive (11) CO(2), we demonstrate that root-attacked maize plants allocate more new (11) C carbon from source leaves to stems, but not to roots. Reduced meristematic activity and reduced invertase activity in attacked maize root systems are identified as possible drivers of this shoot reallocation response. The increased allocation of photoassimilates to stems is shown to be associated with a marked thickening of these tissues and increased growth of stem-borne crown roots. A strong quantitative correlation between stem thickness and root regrowth across different watering levels suggests that retaining photoassimilates in the shoots may help root-attacked plants to compensate for the loss of belowground tissues. Taken together, our results indicate that induced tolerance may be an important strategy of plants to withstand belowground attack. Furthermore, root herbivore-induced carbon reallocation needs to be taken into account when studying plant-mediated interactions between herbivores.


Asunto(s)
Adaptación Fisiológica , Carbono/metabolismo , Escarabajos/fisiología , Herbivoria/fisiología , Raíces de Plantas/fisiología , Zea mays/parasitología , Animales , Biomasa , Metabolismo de los Hidratos de Carbono , Meristema/fisiología , Raíces de Plantas/crecimiento & desarrollo , Brotes de la Planta/fisiología , Tallos de la Planta/fisiología , Agua , Zea mays/fisiología , beta-Fructofuranosidasa/metabolismo
8.
J Exp Bot ; 65(15): 4191-200, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24803501

RESUMEN

The diversity of phenylpropanoids offers a rich inventory of bioactive chemicals that can be exploited for plant improvement and human health. Recent evidence suggests that glycosylation may play a role in the partitioning of phenylpropanoid precursors for a variety of downstream uses. This work reports the functional characterization of a stress-responsive glycosyltransferase, GT1-316 in Populus. GT1-316 belongs to the UGT84A subfamily of plant glycosyltransferase family 1 and is designated UGT84A17. Recombinant protein analysis showed that UGT84A17 is a hydroxycinnamate glycosyltransferase and able to accept a range of unsubstituted and substituted cinnamic and benzoic acids as substrates in vitro. Overexpression of GT1-316 in transgenic Populus led to plant-wide increases of hydroxycinnamoyl-glucose esters, which were further elevated under N-limiting conditions. Levels of the two most abundant flavonoid glycosides, rutin and kaempferol-3-O-rutinoside, decreased, while levels of other less abundant flavonoid and phenylpropanoid conjugates increased in leaves of the GT1-316-overexpressing plants. Transcript levels of representative phenylpropanoid pathway genes were unchanged in transgenic plants, supporting a glycosylation-mediated redirection of phenylpropanoid carbon flow as opposed to enhanced phenylpropanoid pathway flux. The metabolic response of N-replete transgenic plants overlapped with that of N-stressed wild types, as the majority of phenylpropanoid derivatives significantly affected by GT1-316 overexpression were also significantly changed by N stress in the wild types. These results suggest that UGT84A17 plays an important role in phenylpropanoid metabolism by modulating biosynthesis of hydroxycinnamoyl-glucose esters and their derivatives in response to developmental and environmental cues.


Asunto(s)
Ácidos Cumáricos/metabolismo , Glicosiltransferasas/metabolismo , Hidroxibenzoatos/metabolismo , Populus/enzimología , Estrés Fisiológico , Familia de Multigenes , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Populus/genética
9.
Plant Cell Physiol ; 54(6): 1016-25, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23531845

RESUMEN

Metabolism and phloem transport of carbohydrates are interactive processes, yet each is often studied in isolation from the other. Carbon-11 ((11)C) has been successfully used to study transport and allocation processes dynamically over time. There is a need for techniques to determine metabolic partitioning of newly fixed carbon that are compatible with existing non-invasive (11)C-based methodologies for the study of phloem transport. In this report, we present methods using (11)C-labeled CO2 to trace carbon partitioning to the major non-structural carbohydrates in leaves-sucrose, glucose, fructose and starch. High-performance thin-layer chromatography (HPTLC) was adapted to provide multisample throughput, raising the possibility of measuring different tissues of the same individual plant, or for screening multiple plants. An additional advantage of HPTLC was that phosphor plate imaging of radioactivity had a much higher sensitivity and broader range of sensitivity than radio-HPLC detection, allowing measurement of (11)C partitioning to starch, which was previously not possible. Because of the high specific activity of (11)C and high sensitivity of detection, our method may have additional applications in the study of rapid metabolic responses to environmental changes that occur on a time scale of minutes. The use of this method in tandem with other (11)C assays for transport dynamics and whole-plant partitioning makes a powerful combination of tools to study carbohydrate metabolism and whole-plant transport as integrated processes.


Asunto(s)
Metabolismo de los Hidratos de Carbono , Carbohidratos/química , Plantas/metabolismo , Transporte Biológico , Radioisótopos de Carbono , Cromatografía en Capa Delgada , Floema/metabolismo , Hojas de la Planta/metabolismo , Almidón/metabolismo
10.
Ecol Lett ; 15(1): 55-64, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22070646

RESUMEN

The most valuable organs of plants are often particularly rich in essential elements, but also very well defended. This creates a dilemma for herbivores that need to maximise energy intake while minimising intoxication. We investigated how the specialist root herbivore Diabrotica virgifera solves this conundrum when feeding on wild and cultivated maize plants. We found that crown roots of maize seedlings were vital for plant development and, in accordance, were rich in nutritious primary metabolites and contained higher amounts of the insecticidal 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and the phenolic compound chlorogenic acid. The generalist herbivores Diabrotica balteata and Spodoptera littoralis were deterred from feeding on crown roots, whereas the specialist D. virgifera preferred and grew best on these tissues. Using a 1,4-benzoxazin-3-one-deficient maize mutant, we found that D. virgifera is resistant to DIMBOA and other 1,4-benzoxazin-3-ones and that it even hijacks these compounds to optimally forage for nutritious roots.


Asunto(s)
Escarabajos/fisiología , Zea mays/metabolismo , Adaptación Fisiológica , Animales , Conducta Alimentaria , Valor Nutritivo , Fenoles/metabolismo , Raíces de Plantas/metabolismo
11.
Nat Plants ; 8(2): 171-180, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-35194203

RESUMEN

Phloem transport of photoassimilates from leaves to non-photosynthetic organs, such as the root and shoot apices and reproductive organs, is crucial to plant growth and yield. For nearly 90 years, evidence has been generally consistent with the theory of a pressure-flow mechanism of phloem transport. Central to this hypothesis is the loading of osmolytes, principally sugars, into the phloem to generate the osmotic pressure that propels bulk flow. Here we used genetic and light manipulations to test whether sugar import into the phloem is required as the driving force for phloem sap flow. Using carbon-11 radiotracer, we show that a maize sucrose transporter1 (sut1) loss-of-function mutant has severely reduced export of carbon from photosynthetic leaves (only ~4% of the wild type level). Yet, the mutant remarkably maintains phloem pressure at ~100% and sap flow speeds at ~50-75% of those of wild type. Potassium (K+) abundance in the phloem was elevated in sut1 mutant leaves. Fluid dynamic modelling supports the conclusion that increased K+ loading compensated for decreased sucrose loading to maintain phloem pressure, and thereby maintained phloem transport via the pressure-flow mechanism. Furthermore, these results suggest that sap flow and transport of other phloem-mobile nutrients and signalling molecules could be regulated independently of sugar loading into the phloem, potentially influencing carbon-nutrient homoeostasis and the distribution of signalling molecules in plants encountering different environmental conditions.


Asunto(s)
Floema , Zea mays , Hojas de la Planta/genética , Plantas , Azúcares , Zea mays/genética
12.
J Chem Ecol ; 36(3): 286-97, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-20177744

RESUMEN

Salicylate-containing phenolic glycosides (PGs) are abundant and often play a dominant role in plant-herbivore interactions of Populus and Salix species (family Salicaceae), but the biosynthetic pathway to PGs remains unclear. Cinnamic acid (CA) is thought to be a precursor of the salicyl moiety of PGs. However, the origin of the 6-hydroxy-2-cyclohexen-on-oyl (HCH) moiety found in certain PGs, such as salicortin, is not known. HCH is of interest because it confers toxicity and antifeedant properties against herbivores. We incubated Populus nigra leaf tissue with stable isotope-labeled CA, benzoates, and salicylates, and measured isotopic incorporation levels into both salicin, the simplest PG, and salicortin. Labeling of salicortin from [13C6]-CA provided the first evidence that HCH, like the salicyl moiety, is a phenylpropanoid derivative. Benzoic acid and benzaldehyde also labeled both salicyl and HCH, while benzyl alcohol labeled only the salicyl moiety in salicortin. Co-administration of unlabeled benzoates with [13C6]-CA confirmed their contribution to the biosynthesis of the salicyl but not the HCH moiety of salicortin. These data suggest that benzoate interconversions may modulate partitioning of phenylpropanoids to salicyl and HCH moieties, and hence toxicity of PGs. Surprisingly, labeled salicyl alcohol and salicylaldehyde were readily converted to salicin, but did not result in labeled salicortin. Co-administration of unlabeled salicylates with labeled CA suggested that salicyl alcohol and salicylaldehyde may have inhibited salicortin biosynthesis. A revised metabolic grid model of PG biosynthesis in Populus is proposed, providing a guide for functional genomic analysis of the PG biosynthetic pathway.


Asunto(s)
Glicósidos/biosíntesis , Populus/metabolismo , Aldehídos/química , Aldehídos/farmacología , Animales , Benzaldehídos/química , Benzaldehídos/farmacología , Ácido Benzoico/química , Ácido Benzoico/farmacología , Alcoholes Bencílicos/química , Alcoholes Bencílicos/metabolismo , Isótopos de Carbono/metabolismo , Glucósidos/biosíntesis , Glucósidos/química , Salicilatos/química , Salicilatos/farmacología
13.
BMC Plant Biol ; 9: 151, 2009 Dec 29.
Artículo en Inglés | MEDLINE | ID: mdl-20040108

RESUMEN

BACKGROUND: Phenylpropanoid-derived phenolic glycosides (PGs) and condensed tannins (CTs) comprise large, multi-purpose non-structural carbon sinks in Populus. A negative correlation between PG and CT concentrations has been observed in several studies. However, the molecular mechanism underlying the relationship is not known. RESULTS: Populus cell cultures produce CTs but not PGs under normal conditions. Feeding salicyl alcohol resulted in accumulation of salicins, the simplest PG, in the cells, but not higher-order PGs. Salicin accrual reflected the stimulation of a glycosylation response which altered a number of metabolic activities. We utilized this suspension cell feeding system as a model for analyzing the possible role of glycosylation in regulating the metabolic competition between PG formation, CT synthesis and growth. Cells accumulated salicins in a dose-dependent manner following salicyl alcohol feeding. Higher feeding levels led to a decrease in cellular CT concentrations (at 5 or 10 mM), and a negative effect on cell growth (at 10 mM). The competition between salicin and CT formation was reciprocal, and depended on the metabolic status of the cells. We analyzed gene expression changes between controls and cells fed with 5 mM salicyl alcohol for 48 hr, a time point when salicin accumulation was near maximum and CT synthesis was reduced, with no effect on growth. Several stress-responsive genes were up-regulated, suggestive of a general stress response in the fed cells. Salicyl alcohol feeding also induced expression of genes associated with sucrose catabolism, glycolysis and the Krebs cycle. Transcript levels of phenylalanine ammonia lyase and most of the flavonoid pathway genes were reduced, consistent with down-regulated CT synthesis. CONCLUSIONS: Exogenous salicyl alcohol was readily glycosylated in Populus cell cultures, a process that altered sugar utilization and phenolic partitioning in the cells. Using this system, we identified candidate genes for glycosyltransferases that may mediate the glycosylation, and for transporters that mediate the subcellular compartmentalization of sugars and phenolic glycosides. The suspension cells appear to represent a facile system for dissecting the regulation of phenolic carbon partitioning, and in turn, its effects on growth in Populus.


Asunto(s)
Glicósidos/metabolismo , Populus/metabolismo , Taninos/metabolismo , Alcoholes Bencílicos/metabolismo , Células Cultivadas , Medios de Cultivo , Etiquetas de Secuencia Expresada , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Glucósidos , Glicosilación , Análisis de Secuencia por Matrices de Oligonucleótidos , Populus/citología , Populus/genética , Populus/crecimiento & desarrollo , ARN de Planta/genética
14.
Methods Mol Biol ; 2014: 163-176, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31197795

RESUMEN

Noninvasive techniques to measure phloem transport of carbon will be crucial to efforts to engineer improved crop yields, which are highly dependent on carbon partitioning. Phloem, which is buried in the interior of the plant, is highly sensitive to tissue damage. Here we describe nondestructive methods using carbon-11, fed to leaves as 11CO2, as a tracer to track export of recently fixed carbon from leaves, transport speed through the phloem, and distribution or partitioning throughout the plant.


Asunto(s)
Radioisótopos de Carbono/metabolismo , Carbono/metabolismo , Floema/metabolismo , Transporte Biológico , Dióxido de Carbono , Radioisótopos de Carbono/análisis , Proteínas de Transporte de Membrana/metabolismo , Fotosíntesis , Hojas de la Planta/metabolismo
15.
Biosensors (Basel) ; 9(4)2019 Oct 13.
Artículo en Inglés | MEDLINE | ID: mdl-31614897

RESUMEN

The emerald ash borer (EAB) has been the most destructive and costly nonnative insect to threaten the health of ash (Fraxinus) species in North America for at least the past 25 years. The development of methods for detecting visually-hidden EAB galleries at early stages of infestation would provide a useful tool to more effectively facilitate the planning and implementation of targeted EAB pest-suppression and management activities. We tested the efficacy of using a dual-technology electronic-nose (e-nose)/gas chromatograph device as a means for detection of EAB infestations in green ash trees in different EAB-decline classes by analysis of VOC emissions in sapwood. We found significant differences in VOC profiles for trees from the four decline classes. The VOC composition, quantities, and types of volatile metabolites present in headspace volatiles varied considerably across sample types, and resulted in distinct e-nose smellprint patterns that were characteristic of each unique chemical composition. In addition, specific VOC metabolites were identified as potential healthy and EAB-infestation biomarkers, indicative of the health states of individual trees. Few significant differences in major bark phenolic compounds were found between ash decline classes using LC-MS. The e-nose was effective in discriminating between uninfested and EAB-infested trees based on sapwood VOC emissions.


Asunto(s)
Nariz Electrónica , Fraxinus/química , Control de Plagas/métodos , Compuestos Orgánicos Volátiles/análisis , Animales , Cromatografía de Gases , Cromatografía Líquida de Alta Presión , Escarabajos , Fenoles/análisis , Fenoles/química , Madera/química
16.
Plant Physiol Biochem ; 143: 1-10, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31473400

RESUMEN

Internal nitrogen (N) cycling is crucial to N use efficiency. For example, N may be remobilized from older, shaded leaves to young leaves near the apex that receive more direct sunlight, where the N can be used more effectively for photosynthesis. Yet our understanding of the mechanisms and regulation of N transport is limited. To identify relevant transporters in Arabidopsis, fifteen transporter knockout mutants were screened for defects in leaf N export using nitrogen-13 (13N) administered as 13NH3 gas to leaves. We found that three nitrate/peptide transporter family (NPF) genes were necessary for normal leaf N export under low N but not adequate soil N availability, including AtNPF7.1, which has not been previously characterized. High-throughput phenotyping revealed altered leaf area and chlorophyll fluorescence relative to wild-type plants. High AtNPF7.1 expression in flowers and large flower stalks of Atnpf7.1 mutants in low N suggests that AtNPF7.1 influences leaf N export via sink-to-source feedback, perhaps via a role in sensing plant internal N-status. We also identified previously unreported phenotypes for the mutants of the other two NPF transporters that indicate possible roles in N sensing networks.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Nitrógeno/metabolismo , Proteínas de Transporte de Anión/genética , Proteínas de Transporte de Anión/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Transportadores de Nitrato , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo
17.
Plant Sci ; 270: 268-277, 2018 May.
Artículo en Inglés | MEDLINE | ID: mdl-29576080

RESUMEN

Nutrient accumulation, one of the major ecosystem services provided by forests, is largely due to the accumulation and retention of nutrients in trees. This review focuses on seasonal cycling of nitrogen (N), often the most limiting nutrient in terrestrial ecosystems. When leaves are shed during autumn, much of the N may be resorbed and stored in the stem over winter, and then used for new stem and leaf growth in spring. A framework exists for understanding the metabolism and transport of N in leaves and stems during winter dormancy, but many of the underlying genes remain to be identified and/or verified. Transport of N during seasonal N cycling is a particularly weak link, since the physical pathways for loading and unloading of amino N to and from the phloem are poorly understood. Short-day photoperiod followed by decreasing temperatures are the environmental cues that stimulate dormancy induction, and nutrient remobilization and storage. However, beyond the involvement of phytochrome, very little is known about the signal transduction mechanisms that link environmental cues to nutrient remobilization and storage. We propose a model whereby nutrient transport and sensing plays a major role in source-sink transitions of leaves and stems during seasonal N cycling.


Asunto(s)
Ciclo del Nitrógeno , Nitrógeno/metabolismo , Árboles/metabolismo , Transporte Biológico , Ecosistema , Floema/metabolismo , Hojas de la Planta/metabolismo , Tallos de la Planta/metabolismo , Estaciones del Año
18.
Front Plant Sci ; 7: 1207, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27563305

RESUMEN

Movement of nitrogen to the plant tissues where it is needed for growth is an important contribution to nitrogen use efficiency. However, we have very limited knowledge about the mechanisms of nitrogen transport. Loading of nitrogen into the xylem and/or phloem by transporter proteins is likely important, but there are several families of genes that encode transporters of nitrogenous molecules (collectively referred to as N transporters here), each comprised of many gene members. In this study, we leveraged publicly available microarray data of Arabidopsis to investigate the gene networks of N transporters to elucidate their possible biological roles. First, we showed that tissue-specificity of nitrogen (N) transporters was well reflected among the public microarray data. Then, we built coexpression networks of N transporters, which showed relationships between N transporters and particular aspects of plant metabolism, such as phenylpropanoid biosynthesis and carbohydrate metabolism. Furthermore, genes associated with several biological pathways were found to be tightly coexpressed with N transporters in different tissues. Our coexpression networks provide information at the systems-level that will serve as a resource for future investigation of nitrogen transport systems in plants, including candidate gene clusters that may work together in related biological roles.

19.
Elife ; 52016 06 02.
Artículo en Inglés | MEDLINE | ID: mdl-27253062

RESUMEN

Long distance transport in plants occurs in sieve tubes of the phloem. The pressure flow hypothesis introduced by Ernst Münch in 1930 describes a mechanism of osmotically generated pressure differentials that are supposed to drive the movement of sugars and other solutes in the phloem, but this hypothesis has long faced major challenges. The key issue is whether the conductance of sieve tubes, including sieve plate pores, is sufficient to allow pressure flow. We show that with increasing distance between source and sink, sieve tube conductivity and turgor increases dramatically in Ipomoea nil. Our results provide strong support for the Münch hypothesis, while providing new tools for the investigation of one of the least understood plant tissues.


Asunto(s)
Ipomoea nil/metabolismo , Floema/metabolismo , Transporte Biológico , Ipomoea nil/crecimiento & desarrollo , Presión Osmótica
20.
Mol Plant Pathol ; 15(3): 275-83, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24128370

RESUMEN

Several genes in the Agrobacterium tumefaciens transferred (T)-DNA encode proteins that are involved in developmental alterations, leading to the formation of tumours in infected plants. We investigated the role of the protein encoded by the Atu6002 gene, the function of which is completely unknown. Atu6002 expression occurs in Agrobacterium-induced tumours, and is also activated on activation of plant cell division by growth hormones. Within the expressing plant cells, the Atu6002 protein is targeted to the plasma membrane. Interestingly, constitutive ectopic expression of Atu6002 in transgenic tobacco plants leads to a severe developmental phenotype characterized by stunted growth, shorter internodes, lanceolate leaves, increased branching and modified flower morphology. These Atu6002-expressing plants also display impaired response to auxin. However, auxin cellular uptake and polar transport are not significantly inhibited in these plants, suggesting that Atu6002 interferes with auxin perception or signalling pathways.


Asunto(s)
Agrobacterium/genética , Proteínas Bacterianas/metabolismo , ADN Bacteriano/genética , Interacciones Huésped-Patógeno , Ácidos Indolacéticos/metabolismo , Transporte Biológico , Flores/anatomía & histología , Flores/crecimiento & desarrollo , Proteínas Fluorescentes Verdes/metabolismo , Proteínas de la Membrana/metabolismo , Datos de Secuencia Molecular , Fenotipo , Células Vegetales/metabolismo , Hojas de la Planta/anatomía & histología , Hojas de la Planta/crecimiento & desarrollo , Tumores de Planta/microbiología , Transducción de Señal , Fracciones Subcelulares/metabolismo , Factores de Tiempo , Nicotiana/crecimiento & desarrollo
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