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Cold-night responses in grapevine inflorescences
... However, the impact of chilling stress on starch and trehalose accumulation in inflorescences depends on cultivars. For example, starch level was increased in cold-treated flowers in "Pinot noir", while in "Gewurztraminer" the starch content was not affected (Sawicki et al. 2015). ...
... Almost all the VvWRKY genes (55 out of 59) could respond to at least one specific abiotic stress (Guo et al. 2014), and cold stress Sucrose, glucose, fructose, γ-aminobutyrate, alanine, and lysine "Gewurztraminer" (V. vinifera) Sawicki et al. (2015) Starch, γ-aminobutyrate, alanine, and lysine "Pinot noir" (V. vinifera) led to rapid up-regulation of VvWRKY genes . ...
Grapevine (Vitis ssp.) is a deciduous perennial fruit crop, and the canes and buds of grapevine should withstand low temperatures annually during winter. However, the widely cultivated Vitis vinifera is cold-sensitive and cannot survive the severe winter in regions with extremely low temperatures, such as viticulture regions in northern China. By contrast, a few wild Vitis species like V. amurensis and V. riparia exhibit excellent freezing tolerance. However, the mechanisms underlying grapevine cold tolerance remain largely unknown. In recent years, much progress has been made in elucidating the mechanisms, owing to the advances in sequencing and molecular biotechnology. Assembly of grapevine genomes together with resequencing and transcriptome data enable researchers to conduct genomic and transcriptomic analyses in various grapevine genotypes and populations to explore genetic variations involved in cold tolerance. In addition, a number of pivotal genes have been identified and functionally characterized. In this review, we summarize recent major advances in physiological and molecular analyses of cold tolerance in grapevine and put forward questions in this field. We also discuss the strategies for improving the tolerance of grapevine to cold stress. Understanding grapevine cold tolerance will facilitate the development of grapevines for adaption to global climate change.
... Grapevine (Vitis vinifera L.) is naturally affected by the flower abscission depending on both physiological factors such as the carbon nutrition [1] and cultivars [2], and environmental factors [3] such as chilling or heat [4][5][6][7]. Under optimal growth conditions, the intensity of flower drop represents a specific trait of each cultivar (cv.). ...
... Consequently, sugar contents fluctuate differently according to the metabolism of the cultivar and inflorescences become a sink for carbohydrates from BBCH57 stage. It was already reported that the leaf photosynthetic rates are lower in GW than in PN [41] and that inflorescences have shown fluctuations in the photosynthetic activity during the flower development [6,26,27]. ...
p>In order to further understand the relationships between flower development and sugar metabolism in grapevine, the fluctuations of both starch and sucrose contents were compared with the activity of their related enzymes, in the inflorescences, from the appearance of flower buds until the fruit set. The measurements were carried out on GW and PN cvs., differing in their sensitivity to the flower abscission. The meiosis stage, which is a crucial step for the achievement of sexual reproduction, was particularly screened. Results indicate that the main differences in carbohydrate metabolism occur during meiosis. In the inflorescences of both cvs., variations of enzyme activities can be correlated with their differences in sugar contents. Starch fluctuations were mediated by the activity of amylases (alpha- and beta-) rather than by starch synthase. Changes of sucrose were correlated with the activity of Starch Synthase degradation, both cytoplasmic and wall-bounded invertases but not with the Sucrose Phosphate Synthase activity. Finally, the significant increase of sucrose degrading enzyme activities, such as Starch Synthase degradation, cytoplasmic invertase, and wall-bound invertase, observed after the flower separating stage was interpreted as the first sign of the strong physiological modifications occurred in the ovaries between fertilization and the fruit formation.</p
... This also suggests that cold stress causes relatively more damage to cell membranes and worsens the overall mechanical damage incurred by Z. bungeanum plants. Chl has an important function in the cold response in terms of cell integrity, as it preserves membrane permeability, boosts carbohydrate and protein synthesis, and raises the cell sap content (Sawicki et al., 2015). However, under low-temperature stress, the photosynthetic rate of plants will arguably decrease, which directly affects the structure and activity of the photosynthetic tissues (Panda et al., 2020), resulting in greater cell electrolyte extravasation and reduced photosynthesis (da Silva et al., 2021). ...
... In our study, sheath blades in the middle of bamboo shoots had a relatively strong photosynthetic capacity. Diurnal changes in gas exchange parameters are related to changes in environmental conditions, such as temperature, light, and CO 2 concentration [24][25][26][27]. Our results indicate that sheath blades have a photosynthetic capacity during daytime and have midday depression. ...
Culm sheaths play an important role in supporting and protecting bamboo shoots during the growth and development period. The physiological and molecular functions of bamboo sheaths during the growth of bamboo shoots remain unclear. In this study, we investigated the morphological anatomy of culm sheaths, photosynthesis in sheath blades, storage and distribution of sugars, and the transcriptome of the sheath. Respiration in the base of the culm sheath was higher than that in the sheath blades; chloroplasts matured with the development of the sheath blades, the fluorescence efficiency Fv/Fm value increased from 0.3 to 0.82; and sucrose and hexose accumulated in the sheath blade and the culm sheath. The sucrose, glucose, and fructose contents of the middle sheath blades were 10.66, 5.73, and 8.84 mg/g FW, respectively. Starches accumulated in parenchymal cells close to vascular bundles. Genes related to the plant hormone signaling pathway and sugar catabolism were highly expressed in the culm sheath base. These findings provide a research basis for further understanding the possible role of bamboo sheaths in the growth and development of bamboo shoots.
... GC-MS based metabolite analyses of the inflorescences of two Vitis vinifera cultivars, i.e. Pinot noir and Gewürztraminer, demonstrated that levels of four metabolites including alanine and lysine were enhanced as a result of overnight exposure to chilling (Sawicki et al., 2015). The presence of increased levels of methionine in the flower buds of P. pyrifolia cv. ...
Flowering is one of the most important physiological processes of plants that ensures continuity of genetic flow from one generation to the next and also maintains food security. Therefore, impact of various climate-related abiotic stresses on flowering have been assessed to evaluate the long-term impact of global climate change. In contrast to the enormous volume of research that has been conducted at the genetic, transcriptional, post-transcriptional, and protein level, much less attention has been paid to understand the role of various metabolites in flower induction and floral organ development during normal growth or in stressed environmental condition. This review article aims at summarizing information on various primary (e.g., carbohydrates, lipids, fatty acid derivatives, protein and amino acids) and secondary metabolites (e.g., polyamines, phenolics, neuro-indoles, phenylpropanoid, flavonoids and terpenes) that have so far been identified either during flower induction or in individual floral organs implying their possible role in organ development. Specialized metabolites responsible for flower colour, scent and shape to support plant-pollinator interaction have been extensively reviewed by many research groups and hence are not considered in this article. Many of the metabolites discussed here may be used as metabolomarkers to identify tolerant crop genotypes. Several agrochemicals have been successfully used to release endodormancy in temperate trees. Along the same line, a strategy that combines metabolite profiling, screening of small-molecule libraries, and structural alteration of selected compounds has been proposed in order to identify novel lead compounds that can regulate flowering time when applied exogenously.
... Vineyard B experienced warmer temperature conditions than Vineyard C (Figure 3) and may have reached a threshold for Tmin during the summer. The impact of a high Tmin on yield during this period could be explained by a poorer carbohydrate export from the grapevine leaf, which could affect the photosynthetic activity during the day (Sawicki et al., 2015;Tombesi et al., 2018). A period of negative correlation of Tmin to yield was also found for Vineyard A after veraison. ...
Climate influence on grapevine physiology is prevalent and this influence is expected to increase with climate change. Climate influence on grapevine physiology can vary depending on the terroir. A better understanding of these local terroir variations is likely to be achieved with analyses that use local data; i.e., farm/vineyard data. Thus, the challenge lies in exploiting farm data to enable grape growers to understand their own terroir and consequently adapt their practices to the local conditions. In such a context, this article proposes an analytical process to site-specifically study climate influence on grapevine physiology by focusing on time series of the weather data often contained in farm data sets. This article focuses on temperature and precipitation influence on yield in the form of a case study. The analytical process includes the Extended Growing Degree Days (eGDD) and the Bayesian functional Linear regression with Sparse Steps functions (BLiSS) methods in order to detect site-specific periods of strong climate influence on grapevine yield. It uses data from three commercial vineyards situated in the Bordeaux region (France), California (USA) and Israel. In general, the periods of climate influence on grapevine yield detected for the three vineyards identified the same stages of yield development, which have already been studied in the scientific literature. However, some vineyard differences were observed, including: i) different periods of influence associated with a given stage of yield development between the vineyards, ii) different influential weather variables between the three vineyards for a given period, and iii) differing duration of the period of influence associated with a given stage of yield development between the vineyards. These results show the potential of the proposed analytical process for analysing the time series of farm weather data in order to extract site-specific climate indicators of grapevine yield.
... In grapes, the data about starch accumulation in fruit is scarce. There is one study about AGPase expression in inflorescences (Sawicki et al., 2015), but not in berries. What about in other fleshy fruit species? ...
... These results can be explained because changes in external CO 2 may not adequately reflect CO 2 uptake in photosynthesis and in particular will neglect the amount of internally generated and photosynthetically refixed CO 2 by phosphoenolpyruvate carboxylase (PEPC; Blanke and Lenz 1989). Concerning grapevine inflorescence, previous experiments highlighted an induction of the PEPc gene expression between the female meiosis and the beginning of the flowering process (Sawicki et al. 2015c). Finally, a decrease in the expression of both subunits of Rubisco encoding genes (namely RbcS) as well as genes encoding some proteins of the photosynthetic chain at DS5 was observed, which could be correlated with the drop in P N measured at DS6 (Figs. 1, 5). ...
Using measures of gas exchanges and photosynthetic chain activity, we found some differences between grapevine inflorescence and leaf in terms of photosynthetic activity and photosynthesis regulations. Generally, the leaf showed the higher net photosynthesis (P
N) and lower dark respiration than that of the inflorescence until the beginning of the flowering process. The lower (and negative) PN indicated prevailing respiration over photosynthesis and could result from a higher metabolic activity rather than from a lower activity of the photosynthetic apparatus. Considerable differences were observed between both organs in the functioning and regulation of PSI and PSII. Indeed, in our conditions, the quantum yield efficiency and electron transport rate of PSI and PSII were higher in the inflorescence compared to that of the leaf; nevertheless, protective regulatory mechanisms of the photosynthetic chain were clearly more efficient in the leaf. This was in accordance with the major function of this organ in grapevine but it highlighted also that inflorescence seems to be included in the whole carbon balance of a plant. During inflorescence development, the global PSII activity decreased and PSI regulation tended to be similar to the leaf, where photosynthetic activity and regulations remained more stable. Finally, during flowering, cyclic electron flow (CEF) around PSI was activated in parallel to the decline in the thylakoid linear electron flow. Inflorescence CEF was double compared to the leaf; it might contribute to photoprotection, could promote ATP synthesis and the recovery of PSII.
Low temperature is a critical environmental factor limiting plant productivity, especially in northern vineyards. To clarify the impact of this stress on grapevine flower, we used the Vitis array based on Roche-NimbleGen technology to investigate the gene expression of flowers submitted to a cold night. Our objectives were to identify modifications in the transcript levels after stress and during recovery. Consequently, our results confirmed some mechanisms known in grapes or other plants in response to cold stress, notably, (1) the pivotal role of calcium/calmodulin-mediated signaling; (2) the over-expression of sugar transporters and some genes involved in plant defense (especially in carbon metabolism), and (3) the down-regulation of genes encoding galactinol synthase (GOLS), pectate lyases, or polygalacturonases. We also identified some mechanisms not yet known to be involved in the response to cold stress, i.e., (1) the up-regulation of genes encoding G-type lectin S-receptor-like serine threonine-protein kinase, pathogen recognition receptor (PRR5), or heat-shock factors among others; (2) the down-regulation of Myeloblastosis (MYB)-related transcription factors and the Constans-like zinc finger family; and (3) the down-regulation of some genes encoding Pathogen-Related (PR)-proteins. Taken together, our results revealed interesting features and potentially valuable traits associated with stress responses in the grapevine flower. From a long-term perspective, our study provides useful starting points for future investigation.
The metabolism of carbohydrates in developing rice endosperm was characterized by a comparison of levels of activities of 33 major enzymes between the endosperm and green leaves of rice. Activities of ADPglucose pyrophosphorylase, starch synthase and branching enzyme (Q-en-zyme), compared on the basis of soluble protein content, were markedly higher in endosperm than in green leaves. The high levels of Q-enzyme may be responsible for the efficient production of starch in the rice endosperm. The measurement of levels of metabolic intermediates and the localization of key enzymes in isolated amyloplasts from rice endosperm support the view that sucrose is metabolized in the cytoplasm via the pathway: sucrose→UDPglucose→hexose-P→FBP→triose-P. Triose-P then enters the amyloplast, where it is converted to G1P via FBP and, finally, G1P is converted to starch by the concerted reactions of ADPglucose pyrophosphorylase, starch synthase and Q-enzyme.
In plants, flowering is a crucial process for reproductive success and continuity of the species through time. Fruit production requires the perfect development of reproductive structures. Abscission, a natural process, can occur to facilitate shedding of no longer needed, infected, or damaged organs. If stress occurs during flower development, abscission can intervene at flower level, leading to reduced yield. Flower abscission is a highly regulated developmental process simultaneously influenced and activated in response to exogenous (changing environmental conditions, interactions with microorganisms) and endogenous (physiological modifications) stimuli. During climate change, plant communities will be more susceptible to environmental stresses, leading to increased flower and fruit abscission, and consequently a decrease in fruit yield. Understanding the impacts of stress on the reproductive phase is therefore critical for managing future agricultural productivity. Here, current knowledge on flower/fruit abscission is summarized by focusing specifically on effects of environmental stresses leading to this process in woody plants. Many of these stresses impair hormonal balance and/or carbohydrate metabolism, but the exact mechanisms are far from completely known. Hormones are the abscission effectors and the auxin/ethylene balance is of particular importance. The carbohydrate pathway is the result of complex regulatory processes involving the balance between photosynthesis and mobilization of reserves. Hormones and carbohydrates together participate in complex signal transduction systems, especially in response to stress. The available data are discussed in relation to reproductive organ development and the process of abscission.
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The 'omic' data such as genomic data, transcriptomic data, proteomic data and single nucleotide polymorphism data have been rapidly growing. The omic data are large-scale and high-throughput data. Such data challenge traditional statistical methodologies and require multiple tests. Several multiple-testing procedures such as Bonferroni procedure, Benjamini-Hochberg (BH) procedure and Westfall-Young procedure have been developed, among which some control family-wise error rate and the others control false discovery rate (FDR). These procedures are valid in some cases and cannot be applied to all types of large-scale data. To address this statistically challenging problem in the analysis of the omic data, we propose a general method for generating a set of multiple-testing procedures. This method is based on the BH theorems. By choosing a C-value, one can realize a specific multiple-testing procedure. For example, by setting C = 1.22, our method produces the BH procedure. With C < 1.22, our method generates procedures of weakly controlling FDR, and with C > 1.22, the procedures strongly control FDR. Those with C = G (number of genes or tests) and C = 0 are, respectively, the Bonferroni procedure and the single-testing procedure. These are the two extreme procedures in this family. To let one choose an appropriate multiple-testing procedure in practice, we develop an algorithm by which FDR can be correctly and reliably estimated. Simulated results show that our method works well for an accurate estimation of FDR in various scenarios, and we illustrate the applications of our method with three real datasets.
Availability and implementation:
Our program is implemented in Matlab and is available upon request.
A newly developed fluorescence measuring system is employed for the recording of chlorophyll fluorescence induction kinetics (Kautsky-effect) and for the continuous determination of the photochemical and non-photochemical components of fluorescence quenching. The measuring system, which is based on a pulse modulation principle, selectively monitors the fluorescence yield of a weak measuring beam and is not affected even by extremely high intensities of actinic light. By repetitive application of short light pulses of saturating intensity, the fluorescence yield at complete suppression of photochemical quenching is repetitively recorded, allowing the determination of continuous plots of photochemical quenching and non-photochemical quenching. Such plots are compared with the time courses of variable fluorescence at different intensities of actinic illumination. The differences between the observed kinetics are discussed. It is shown that the modulation fluorometer, in combination with the application of saturating light pulses, provides essential information beyond that obtained with conventional chlorophyll fluorometers.
Grapevine flower development and fruit set are influenced by cold nights in the vineyard. To investigate the impact of cold stress on carbon metabolism in the inflorescence, we exposed the inflorescences of fruiting cuttings to chilling and freezing temperatures overnight and measured fluctuations in photosynthesis and sugar content. Whatever the temperature, after the stress treatment photosynthesis was modified in the inflorescence, but the nature of the alteration depended on the intensity of the cold stress. At 4°C, photosynthesis in the inflorescence was impaired through non-stomatal limitations, whereas at 0°C it was affected through stomatal limitations. A freezing night (-3°C) severely deregulated photosynthesis in the inflorescence, acting primarily on photosystem II. Cold nights also induced accumulation of sugars. Soluble carbohydrates increased in inflorescences exposed to -3°C, 0°C and 4°C, but starch accumulated only in inflorescences of plants treated at 0 and -3°C. These results suggest that inflorescences are able to cope with cold temperatures by adapting their carbohydrate metabolism using mechanisms that are differentially induced according to stress intensity.
An improved method is introduced for the determination of the quantum yield of photosystem I. The new method employs saturating light pulses with steep rise characteristics to distinguish, in a given physiological state, centers with an open acceptor side from centers with a reduced acceptor side. The latter do not contribute to PSI quantum yield (I). Oxidation of P700 is measured by a rapid modulation technique using the absorbance change around 830 nm. The quantum yield I is calculated from the amplitude of the rapid phase of absorbance change (A; 830 nm) upon application of a saturation pulse in a given state, divided by the maximal A (830 nm) which is induced by a saturation pulse with far-red background illumination. Using this technique, I can be determined even under conditions of acceptor-side limitation, as for example in the course of a dark-light induction period or after elimination of CO2 from the gas stream. Thus determined I values display a close-to-linear relationship with those for the quantum yield of PSII (II) calculated from chlorophyll fluorescence parameters. It is concluded that the proposed method may provide new information on the activity of the PSI acceptor side and thus help to separate the effects of acceptorside limitation from those of cyclic PSI, whenever a non-linear relationship between II and the P700-reduction level is observed.
We investigated the physiological effect of night chilling (CN) on potted seedlings of two tropical tree species, Calophyllum polyanthum and Linociera insignis, in Xishuangbanna, southwest China. Seedlings grown under 8, 25, and 50 % daylight for five months were moved to a 4–6 °C
cold storage house for three consecutive nights, and returned to the original shaded sites during the day. CN resulted in
strong suppression of photosynthesis and stomatal conductance for L. insignis, and reduced photorespiration rates, carboxylation efficiency, and maximum photochemical efficiency of photosystem 2 (PS2)
at dawn and midday for both species. CN increased dawn and midday rates of non-photochemical quenching, and the contents of
malondialdehyde and H2O2 for both species. CN also induced inactivation or destruction of PS2 reaction centres. The impacts of CN on tropical seedlings
increased with the number of CN. Shading could significantly mitigate the adverse effects of CN for both species. After 3-d-recovery,
gas exchange and fluorescence parameters for both species returned to pre-treatment levels in most cases. Thus CN induced
mainly stomatal limitation of photosynthesis for L. insignis, and non-stomatal limitation for C. polyanthum. C. polyanthum was more susceptible to CN than L. insignis. Fog, which often occurs in Xishuangbanna, could be beneficial to chilling sensitive tropical seedlings in this area through
alleviating photoinhibition or photodamage by reducing sunlight.
The functional activities of the photosynthetic apparatus of two grapevine genotypes (Vitis vinifera L. cvs. Müller-Thurgau and Lagrein) were investigated after low night temperature (LNT) treatment for 7 d. LNT caused important
reductions of the net photosynthetic rate (PN) of Lagrein plants due to non-stomatal components. These non-stomatal effects were not evident in Müller-Thurgau. At LNT
treatment, the contents of photosynthetic pigments decreased significantly in Lagrein, but in Müller-Thurgau the contents
of chlorophyll (Chl) remained unchanged whereas the contents of carotenoids (Car) increased. An increase and decrease of Chl
a/b was shown in Mü ller-Thurgau and Lagrein stressed plants, respectively. RuBPC activity and content of soluble proteins were
also significantly reduced in Lagrein. Under LNT treatment, photosystem (PS) 2 was markedly more inhibited in Lagrein than
in Müller-Thurgau. The decrease in PS 2 activity in Lagrein was mostly due to the marked loss of D1, 47, 43, 33, 28-25, 23
and 17 kDa proteins determined by immunological and SDS-PAGE studies.
Apart from water availability, low temperature is the most important environmental factor limiting the productivity and geographical distribution of plants across the world. To cope with cold stress, plant species have evolved several physiological and molecular adaptations to maximize cold tolerance by adjusting their metabolism. The regulation of some gene products represents an additional mechanism of cold tolerance. A consequence of these mechanisms is that plants are able to survive exposure to low temperature via a process known as cold acclimation. In this review, we briefly summarize recent progress in research and hypotheses on how sensitive plants perceive cold. We also explore how this perception is translated into changes within plants following exposure to low temperatures. Particular emphasis is placed on physiological parameters as well as transcriptional, post-transcriptional and post-translational regulation of cold-induced gene products that occur after exposure to low temperatures, leading to cold acclimation.
Low temperatures damage many temperate crops, including grapevine, which, when exposed to chilling, can be affected by symptoms ranging from reduced yield up to complete infertility. We have previously demonstrated that Burkholderia phytofirmans PsJN, a plant growth-promoting rhizobacteria (PGPR) that colonizes grapevine, is able to reduce chilling-induced damage. We hypothesized that the induced tolerance may be explained at least partly by the impact of bacteria on grapevine photosynthesis or carbohydrate metabolism during cold acclimation. To investigate this hypothesis, we monitored herein the fluctuations of photosynthesis parameters (net photosynthesis [P(n)], intercellular CO(2) concentration, stomatal conductances, ΦPSII, and total chlorophyll concentration), starch, soluble sugars (glucose, fructose, saccharose, mannose, raffinose, and maltose), and their precursors during 5 days of chilling exposure (4°C) on grapevine plantlets. Bacterization affects photosynthesis in a non-stomatal dependent pattern and reduced long-term impact of chilling on P(n). Furthermore, all studied carbohydrates known to be involved in cold stress tolerance accumulate in non-chilled bacterized plantlets, although some of them remained more concentrated in the latter after chilling exposure. Overall, our results suggest that modification of carbohydrate metabolism in bacterized grapevine plantlets may be one of the major effects by which this PGPR reduces chilling-induced damage.
The distribution of carbon (C) into whole grapevine fruiting cuttings was investigated during flower development to determine
the relative contribution of inflorescence and leaf photoassimilates in the total C balance and to investigate their partitioning
towards other plant organs. A 13C labelling procedure was used to label C photoassimilates by leaves and inflorescences in grapevine. Investigations were
carried out at various stages of flower/berry development, from separated cluster to fruit set, using grapevine fruiting cuttings
with four leaves (Vitis vinifera L. cv. Chardonnay). This is the first study reporting that, during its development, (i) the carbon needs of the inflorescence
were met by both leaf and inflorescence photosynthesis, and (ii) the inflorescence amazingly participated significantly to
the total C balance of grapevine cuttings by redistributing an important part of its own assimilates to other plant organs.
With regard to flowering, 29% of C assimilated by the inflorescence remained in the inflorescence, while partitioning towards
the stem reached 42% and, as a lower proportion, 15% in leaves, and 14% in roots.
Our previous study indicated that PSII is more sensitive to chilling and light stress than PSI in tropical trees, and Erythrophleum guineense is more sensitive to chilling stress than Dalbergia odorifera and Khaya ivorensis, but the underlying physiological mechanisms are unclear. Although recent studies have reported that cyclic electron flow (CEF) plays an important role in photoprotection, the role of CEF in protecting PSI and PSII of tropical tree species remains unclear. We investigated the effect of temporal chilling temperature on energy distribution in PSII, the redox state of P700 and CEF in the above-mentioned tropical evergreen tree species grown in an open field. Our results indicated that the overclosure of PSII reaction centers at chilling temperature led to excess excitation pressure in PSII. At the temporal chilling temperature under low light, PSI acceptor side limitation [Y(NA)] was lower than those at 25°C for all species. Although the effective quantum yield of CEF [Y(CEF)] was not significantly stimulated in E. guineense and K. ivorensis under temporal chilling at low light levels, the ratio of Y(CEF) to the effective quantum yield of PSII [Y(II)] significantly increased. Under chilling conditions Y(CEF)/Y(II) was stimulated much more in K. ivorensis and D. odorifera compared with that in the chilling-sensitive E. guineense. These results suggested that stimulation of Y(CEF)/Y(II) plays an important role in protecting PSI and PSII from photoinhibition caused by chilling stress.
Trehalose 6-phosphate (T6P), the precursor of trehalose, is a signaling molecule in plants with strong effects on metabolism, growth and development. We recently showed that in growing tissues T6P is an inhibitor of SnRK1 of the SNF1-related group of protein kinases. SnRK1 acts as transcriptional integrator in response to carbon and energy supply. In microarray experiments on seedlings of transgenic Arabidopsis with elevated T6P content we found that expression of SnRK1 marker genes was affected in a manner to be predicted by inhibition of SnRK1 by T6P in vivo. A large number of genes involved in reactions that utilize carbon, e.g., UDP-glucose dehydrogenase genes involved in cell wall synthesis, were upregulated. T6P was also found to affect developmental signaling pathways, probably in a SnRK1-independent manner. This includes upregulation of genes encoding UDP-glycosyltransferases that are involved in the glycosylation of hormones. In addition, genes involved in auxin response and light signaling were affected. Many of these genes belong to pathways that link the circadian clock to plant growth and development. The overall pattern of changes in gene expression supports a role for T6P in coordinating carbon supply with biosynthetic process involved in growth and development.
In recent years, several regulatory systems that link carbon nutrient status to plant growth and development have emerged. In this paper, we discuss the growth promoting functions of the hexokinase (HXK) glucose sensor, the trehalose 6-phosphate (T6P) signal and the Target of Rapamycin (TOR) kinase pathway, and the growth inhibitory function of the SNF1-related Protein Kinase1 (SnRK1) and the C/S1 bZIP transcription factor network. It is crucial that these systems interact closely in regulating growth and in several cases crosstalk has been demonstrated. Importantly, these nutrient controlled systems must interact with other growth regulatory pathways.
While the possible importance of the tricarboxylic acid (TCA) cycle reactions for leaf photosynthesis operation has been recognized, many uncertainties remain on whether TCA cycle biochemistry is similar in the light compared with the dark. It is widely accepted that leaf day respiration and the metabolic commitment to TCA decarboxylation are down-regulated in illuminated leaves. However, the metabolic basis (i.e. the limiting steps involved in such a down-regulation) is not well known. Here, we investigated the in vivo metabolic fluxes of individual reactions of the TCA cycle by developing two isotopic methods, (13)C tracing and fluxomics and the use of H/D isotope effects, with Xanthium strumarium leaves. We provide evidence that the TCA "cycle" does not work in the forward direction like a proper cycle but, rather, operates in both the reverse and forward directions to produce fumarate and glutamate, respectively. Such a functional division of the cycle plausibly reflects the compromise between two contrasted forces: (1) the feedback inhibition by NADH and ATP on TCA enzymes in the light, and (2) the need to provide pH-buffering organic acids and carbon skeletons for nitrate absorption and assimilation.
Cyclic electron flow (CEF) around photosystem I has a role in avoiding photoinhibition of photosystem II (PSII), which occurs under conditions in which the rate of photodamage to PSII exceeds the rate of its repair. However, the molecular mechanism underlying how CEF contributes to photoprotection is not yet well understood. We examined the effect of impairment of CEF and thermal energy dissipation (qE) on photoinhibition using CEF (pgr5) and qE (npq1 and npq4) mutants of Arabidopsis (Arabidopsis thaliana) exposed to strong light. Impairment of CEF by mutation of pgr5 suppressed qE and accelerated photoinhibition. We found that impairment of qE, by mutations of pgr5, npq1, and npq4, caused inhibition of the repair of photodamaged PSII at the step of the de novo synthesis of the D1 protein. In the presence of the chloroplast protein synthesis inhibitor chloramphenicol, impairment of CEF, but not impairment of qE, accelerated photoinhibition, and a similar effect was obtained when leaves were infiltrated with the protonophore nigericin. These results suggest that CEF-dependent generation of DeltapH across the thylakoid membrane helps to alleviate photoinhibition by at least two different photoprotection mechanisms: one is linked to qE generation and prevents the inhibition of the repair of photodamaged PSII at the step of protein synthesis, and the other is independent of qE and suppresses photodamage to PSII.
We found similarities between the effects of low night temperatures (5 degrees C-10 degrees C) and slowly imposed water stress on photosynthesis in grapevine (Vitis vinifera L.) leaves. Exposure of plants growing outdoors to successive chilling nights caused light- and CO(2)-saturated photosynthetic O(2) evolution to decline to zero within 5 d. Plants recovered after four warm nights. These photosynthetic responses were confirmed in potted plants, even when roots were heated. The inhibitory effects of chilling were greater after a period of illumination, probably because transpiration induced higher water deficit. Stomatal closure only accounted for part of the inhibition of photosynthesis. Fluorescence measurements showed no evidence of photoinhibition, but nonphotochemical quenching increased in stressed plants. The most characteristic response to both stresses was an increase in the ratio of electron transport to net O(2) evolution, even at high external CO(2) concentrations. Oxygen isotope exchange revealed that this imbalance was due to increased O(2) uptake, which probably has two components: photorespiration and the Mehler reaction. Chilling- and drought-induced water stress enhanced both O(2) uptake processes, and both processes maintained relatively high rates of electron flow as CO(2) exchange approached zero in stressed leaves. Presumably, high electron transport associated with O(2) uptake processes also maintained a high DeltapH, thus affording photoprotection.
Changes in gas exchange with leaf age and fruit growth were determined in lychee trees (Litchi chinensis Sonn.) growing in subtropical Queensland (27° S). Leaves expanded in a sigmoid pattern over 50 days during spring, with net CO2 assimilation (A) increasing from –4.1 ± 0.9 to 8.3 ± 0.5 μmol m⁻² s⁻¹ as the leaves changed from soft and red, to soft and light green, to hard and dark green. Over the same period, dark respiration (Rd) decreased from 5.0 ± 0.8 to 2.0 ± 0.1 μmol CO2 m⁻² s⁻¹. Net CO2 assimilation was above zero about 30 days after leaf emergence or when the leaves were half fully expanded. Chlorophyll concentrations increased from 0.7 ± 0.2 mg g⁻¹ in young red leaves to 10.3 ± 0.7 mg g⁻¹ in dark green leaves, along with stomatal conductance (gs, from 0.16 ± 0.09 to 0.47 ± 0.17 mol H2O m⁻² s⁻¹).
Fruit growth was sigmoidal, with maximum values of fresh mass (29 g), dry mass (6 g) and fruit surface area (39 cm²) occurring 97 to 115 days after fruit set. Fruit CO2 exchange in the light (Rl) and dark (Rd) decreased from fruit set to fruit maturity, whether expressed on a surface area (10 to 3 μmol CO2 m⁻² s⁻¹ and 20 to 3 μmol CO2 m⁻² s⁻¹, respectively) or on a dry mass basis (24 to 2 nmol CO2 g⁻¹ s⁻¹ and 33 to 2 nmol CO2 g⁻¹ s⁻¹, respectively). Photosynthesis never exceeded respiration, however, the difference between Rl and Rd was greatest in young green fruit (4 to 8 μmol CO2 m⁻² s⁻¹). About 90% of the carbon required for fruit growth was accounted for in the dry matter of the fruit, with the remainder required for respiration. Fruit photosynthesis contributed about 3% of the total carbon requirement of the fruit over the season. Fruit growth was mainly dependent on CO2 assimilation in recently expanded dark green leaves.
Microarrays have emerged as the
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Protoplasts were tested to determine whether the freezing sensitivity of the sfr4 (sensitive to freezing) mutant of Arabidopsis was due to the mutant's deficiency in soluble sugars after cold acclimation. When grown under nonacclimated conditions, sfr4 protoplasts possessed freezing tolerance similar to that of wild type, with the temperature at which 50% of protoplasts are injured (LT(50)) of -4.5 degrees C. In both wild-type and sfr4 protoplasts, expansion-induced lysis was the predominant lesion between -2 degrees C and -4 degrees C, but its incidence was low (approximately 10%); below -5 degrees C, loss of osmotic responsiveness (LOR) was the predominant lesion. After cold acclimation, the LT(50) was decreased to only -5.6 degrees C for sfr4 protoplasts, compared with -9.1 degrees C for wild-type protoplasts. Although expansion-induced lysis was precluded in both types of protoplasts, the sfr4 protoplasts remained susceptible to LOR. After incubation of seedlings in Suc solution in the dark at 2 degrees C, freezing tolerance and the incidence of freeze-induced lesions in sfr4 protoplasts were examined. The freezing tolerance of isolated protoplasts (LT(50) of -9 degrees C) and the incidence of LOR were now similar for wild type and sfr4. These results indicate that the freezing sensitivity of cold-acclimated sfr4 is due to its continued susceptibility to LOR (associated with lyotropic formation of the hexagonal II phase) and associated with the low sugar content of its cells.
Carbohydrate metabolism is important in plant sexual reproduction since sugar contents are determining factors for both flower initiation and floral organ development. In woody plants, flowering represents the most energy-consuming step crucial to reproductive success. Nevertheless, in these species, the photosynthesis performed by flowers supplies the carbon required for reproduction. In grapevine (Vitis vinifera), the inflorescence has a specific status since this organ imports carbohydrates at the same time as it exports photoassimilates. In this study, fluctuations in carbohydrate metabolism were monitored by analyzing gas exchanges, photosynthetic electron transport capacity, carbohydrate contents and some activities of carbohydrate metabolism enzymes, in the inflorescences of Pinot noir and Gewurztraminer, two cultivars with a different sensitivity to coulure phenomenon. Our results showed that photosynthetic activity and carbohydrate metabolism are clearly different and differently regulated during the floral development in the two cultivars. Indeed, the regulation of the linear electron flow and the cyclic electrons flow are not similar. Moreover, the regulation of PSII activity, with a higher Y(NPQ)/Y(NO) ratio in Gewurztraminer, can be correlated with the higher protection of the photosynthetic chain and consequently with the higher yield under optimal conditions of this cultivar. At least, our results showed a higher photosynthetic activity and a better protection of PSI in Pinot noir during the floral development.
We found similarities between the effects of low night temperatures (5°C–10°C) and slowly imposed water stress on photosynthesis in grapevine (Vitis vinifera L.) leaves. Exposure of plants growing outdoors to successive chilling nights caused light- and CO2-saturated photosynthetic O2 evolution to decline to zero within 5 d. Plants recovered after four warm nights. These photosynthetic responses were confirmed in potted plants, even when roots were heated. The inhibitory effects of chilling were greater after a period of illumination, probably because transpiration induced higher water deficit. Stomatal closure only accounted for part of the inhibition of photosynthesis. Fluorescence measurements showed no evidence of photoinhibition, but nonphotochemical quenching increased in stressed plants. The most characteristic response to both stresses was an increase in the ratio of electron transport to net O2evolution, even at high external CO2 concentrations. Oxygen isotope exchange revealed that this imbalance was due to increased O2 uptake, which probably has two components: photorespiration and the Mehler reaction. Chilling- and drought-induced water stress enhanced both O2 uptake processes, and both processes maintained relatively high rates of electron flow as CO2 exchange approached zero in stressed leaves. Presumably, high electron transport associated with O2uptake processes also maintained a high ΔpH, thus affording photoprotection.
Photoinhibition was defined originally as the decrease in photosynthetic activity that occurs upon excess illumination. The
site of photoinhibition has generally been considered to be located in PSII. However, a novel type of photoinhibition has
recently been characterized in chillingsensitive plants. This photoinhibition occurs under relatively weak illumination at
chilling temperatures and the main site of damage is in PSI. The photoinhibition of PSI is initiated by the inactivation of
the acceptor side, with the subsequent destruction of the reaction center and the degradation of the product of the psaB gene, which is one of the two major subunit polypeptides of the PSI reaction center complex. Chilling and oxidative stress
(the presence of reactive species of oxygen) are characteristic requirements for the photoinhibition of PSI in vivo.
A series of experiments is presented investigating short term and long term changes of the nature of the response of rate of CO2 assimilation to intercellular p(CO2). The relationships between CO2 assimilation rate and biochemical components of leaf photosynthesis, such as ribulose-bisphosphate (RuP2) carboxylase-oxygenase activity and electron transport capacity are examined and related to current theory of CO2 assimilation in leaves of C3 species. It was found that the response of the rate of CO2 assimilation to irradiance, partial pressure of O2, p(O2), and temperature was different at low and high intercellular p(CO2), suggesting that CO2 assimilation rate is governed by different processes at low and high intercellular p(CO2). In longer term changes in CO2 assimilation rate, induced by different growth conditions, the initial slope of the response of CO2 assimilation rate to intercellular p(CO2) could be correlated to in vitro measurements of RuP2 carboxylase activity. Also, CO2 assimilation rate at high p(CO2) could be correlated to in vitro measurements of electron transport rate. These results are consistent with the hypothesis that CO2 assimilation rate is limited by the RuP2 saturated rate of the RuP2 carboxylase-oxygenase at low intercellular p(CO2) and by the rate allowed by RuP2 regeneration capacity at high intercellular p(CO2).
Sucrose accumulation and its related enzyme activities in the juice sacs were compared between the fruit from conventional crop load (CCL, leaf–fruit ratio is about 25) and low crop load (LCL, leaf–fruit ratio is more than 50) trees in satsuma mandarin (Citrus unshiu Mark). Sucrose in the juice began to increase in September and the increase continued to harvest (late November) in fruit on trees with both crop loads, but the rate of increase was significantly higher in CCL fruit. Synthetic sucrose synthase (SS, EC 2.4.1.13) increased greatly as sucrose accumulated in the juice sacs, whereas its cleavage activity decreased. In spite of lower accumulation of sucrose in LCL fruit, synthetic SS activity was higher than in CCL fruit. Acid invertase (EC 3.2.1.26) activity, which decreased with fruit development, was significantly higher in LCL fruit than in CCL ones until late October. Thus, sucrose synthesized by SS may be broken through this higher activity of acid invertase in LCL fruit, resulting in repression of sucrose accumulation. When diurnal changes in SS activity in juice sacs were measured under orchard conditions in mid-November, sucrose increase was estimated to be 0.4% per fruit per day. This activity is enough to accumulate sucrose to harvest level (7%) within 20 days. In satsuma mandarin fruit, therefore, sucrose concentration in the juice may be regulated by both synthetic SS and acid invertase activities in the juice sacs, and crop load of the tree may greatly affect sucrose accumulation by controlling these enzyme activities.
As a part of a project aimed at elucidating the causal relationship between reserve mobilisation and the extent of shedding in Vitis vinifera L., we compared storage and fate of carbon (C) and nitrogen (N) reserves in two varieties differing in their susceptibility to fruitlet abscission. Merlot (susceptible) and Pinot Noir (P. Noir, not susceptible) vines were grown in trenches under semi-controlled conditions over a 3-y period after planting. Mobilisation of stored C and N, distribution of reserve materials within the vines and 15N uptake were followed particularly during the spring growth flush and floral development in the third year. At dormancy, starch levels in the perennial tissues (roots, trunk, canes) were higher in Merlot than in P. Noir. During the spring growth flush, starch level decreased markedly in the roots of both cultivars until early bloom. At that time, starch started to accumulate in P. Noir but not in Merlot. Similar variations were found with total N. Accordingly, 15N analysis showed that translocation of storage N to the annual tissues was nearly achieved at early bloom in P. Noir while it continued until pea berry size in Merlot. In parallel, N uptake increased during the spring growth flush, and it was higher in P. Noir than in Merlot. These results indicate that transition between heterotrophic (root) and autotrophic (leaf) mode of nutrient allocation towards the developing inflorescences occurs earlier in P. Noir. Possible consequences are discussed in relation to the susceptibility of each cultivar to shedding.
Plastic mini-chambers were used as tiny ‘glass houses’ to increase bud temperature in the vineyard. Open containers, with holes cut in them for ventilation, were used as controls, and inflorescences produced in those chambers were compared with inflorescences from modified chambers where either shade cloth or reflective foil were used to alter internal levels of photosynthetically-active radiation (PAR) as well as temperature. Buds were treated for either14 days prior to budburst or for 13 days subsequent to budburst. Temperature and PAR were monitored immediately adjacent to the buds. Applied prior to budburst, the closed mini-chambers increased bud temperatures and reduced flower numbers per inflorescence. Both ‘clear’ and ‘reflective foil’ treatments resulted in similar flower numbers. However, the shading treatment increased flower numbers by approximately 13%. Prior to budburst, there was a significant but weak correlation between average temperature and flower number per inflorescence for both the basal and apical inflorescence. Average PAR was not significantly correlated with flower number on either inflorescence, and did not improve the correlation when included with temperature in a multiple linear regression. Subsequent to budburst, flower numbers per inflorescence were decreased by the closed container but were unaffected by either the shading or foil treatment. The correlation between temperature and flower number on the apical inflorescence was maintained but the correlation between temperature and flower number on the basal inflorescence was no longer apparent. These results suggest that temperatures encountered in a vineyard during budburst can influence the number of flowers per inflorescence to the extent of a 15 to 25% variation in flower number. PAR, apart from influencing bud temperature, does not appear to influence flower number. The effect of temperature on flower differentiation diminishes as budburst advances.
To study ovule development, small Chardonnay and Shiraz vines were grown in pots under controlled conditions at 25°/20°C day/night temperature until flowering or were transferred to 12°/9°C two days before flowering and then returned to 25°/20°C after one week. The ovules of flowers at three positions within the inflorescence were excised on the day after they had opened. The ovules exposed to the lower temperatures tended to be smaller and less advanced in development, even when otherwise normal, especially on Chardonnay, and the ovules of king flowers tended to be larger than those of the other flowers. Over half of the ovules of Chardonnay exposed to the low temperatures were abnormal (with abnormal or no embryo sacs, and some also having a degenerated nucellus) while 35% of the Shiraz ovules were without normal embryo sacs. To study pollination and pollen tube growth, vines similar to those of the ovule study were exposed to 12°/9°C two days before or on the day of flowering. In the pistils exposed to low temperature, a reduction occurred in the number of pollen tubes present in the lower ovary on days 2 and 4 after flowering, from about four to almost nil in Chardonnay, but a smaller reduction occurred in Shiraz. In the absence of cool temperatures, pollen tubes penetrated on average less than one ovule per ovary in Chardonnay and about one ovule per ovary in Shiraz. It is concluded that temperature sensitivity to fruit-set is a varietal characteristic, expressing itself in quantitative differences in the damage imparted to the structure of the ovules and the function of the pollen.
Small Chardonnay and Shiraz vines were grown under controlled conditions at 25°/20°C day/night temperatures but transferred for one week to 17°/14°C or 12°/9°C at four stages of growth between budburst and flowering. In Chardonnay, half the vines were given supplementary pollination with pollen produced under favourable conditions. Per cent fruit-set and berry number per bunch of Chardonnay were reduced by about one third to one half by 12°/9°C, applied just before and at the early stage of flowering. At these stages, and with this low-temperature treatment, supplementary pollination had significant positive effects. Total seed number per berry was not affected by treatment, but the proportion of hollow seeds increased in the treatments which suffered reduced set. This resulted in a positive relation between the number of functional seeds per berry and the number of berries per bunch. In Shiraz, the differences between the various treatment means were small and mostly not significant. It is concluded that the reduction in fruit-set of Chardonnay due to cool temperatures near flowering is due to detrimental effects on both pollen and ovules.
Respiratory activity of plants in the light, measured as carbon dioxide release from the tricarboxylic acid (TCA) cycle or oxygen consumption by the respiratory chain, is generally reported to lie between 25 and 100% of that in the dark. While this has been interpreted as evidence for an inhibition of respiration during photosynthesis, an increasing body of evidence indicates that mitochondrial respiration plays an important role in photosynthetic tissues. Historically, the view from experiments using specific respiratory inhibitors has been that oxidative phosphorylation in the mitochondria provides the cytosol with adenosine triphosphate even in the light. However, functioning of TCA cycle reactions is also required for the export of carbon skeletons necessary for nitrate reduction in the cytosol. In addition, export of TCA cycle-derived reducing equivalents may also be necessary for photorespiration (for hydroxypyruvate reduction in the peroxisomes). The work with respiratory inhibitors has recently been complemented by a range of transgenic experiments that provide direct evidence for the importance of the TCA cycle in the illuminated leaves. These transgenesis experiments hint at an important role for ascorbate in coordinating the major pathways of energy metabolism within the leaf and are in keeping with current thinking that redox signals emanating from the mitochondria are important in setting the cellular machinery to maintain overall redox balance. In this review we intend to synthesize recent experimental data to postulate a model of the function of the TCA cycle in the illuminated leaf.
The Gewurztraminer (GW) and the Pinot noir (PN) cultivars of grapevine differ in their sensitivity to environmental factors that can cause flower abscission, cv. GW being the most sensitive. In order to further define the mechanisms leading to abscission, and owing to the importance of sugars in the achievement of sexual organ ontogenesis, we attempted to correlate the chronology of flower ontogenesis with the variations of carbohydrates in the inflorescence. In the vineyard, under optimal climatic conditions, fruit set of cv. GW and cv. PN was 82% and 65%, respectively. The sugar distribution was different in their inflorescences during the entire duration of flower development. Between stages 15 and 17, flowers of GW and PN reached the crucial meiosis stage. At that time, the inflorescences of cv. GW exhibited higher concentrations of starch and sucrose, whereas those of PN presented higher levels of glucose and fructose. Despite higher starch concentrations in GW inflorescences, starch reserves were present in the ovules and anthers of PN but not in those of GW. These results suggest that the higher content of reserve and transport carbohydrates in the inflorescences of GW favour flower development and fruit set under optimal environmental conditions. Furthermore, since meiosis represents a key step of female development, the different sugar concentrations in the inflorescences of the two cultivars at stages 15 and 17 could be related to the sensitivity to flower abscission under climatic stress. In particular, the presence of starch granules in PN ovules and anthers might explain the higher resistance of this cultivar to flower abscission.
Jatropha curcas, one of the most important energy plant resources, is vulnerable to chilling. To evaluate the effects of chilling on photosynthesis of J. curcas and intraspecific differences in chilling tolerance, seedlings of twelve populations were treated with the temperature of 4–6°C for five consecutive nights with normal environmental temperature during the day. Night chilling treatment decreased light-saturated photosynthetic rate (P
max) significantly for all populations. Stomatal limitation could not explain the decreased P
max because intracellular CO2 concentration was not significantly reduced by night chilling in all populations (with only one exception). The decreased soluble-protein content, which may be related to the increased malondialdehyde (MDA) content, contributed to the decreased P
max. The increased MDA content indicated that oxidative stress occurred after night chilling, which was associated with the larger decrease in P
max compared with the decrease in actual photochemical efficiency of photosystem II, and the slight increase in thermal dissipation of excessive energy. After five-day recovery, MDA (with two exceptions) and P
max still did not recover to the levels as those before night chilling treatment for all populations, indicating that J. curcas was vulnerable to chilling. Chilling tolerance was significantly different among populations. Populations originating from high elevations had greater chilling-tolerant abilities than populations originating from low elevations, showing a local adaptation to environmental temperatures of origins. Our study shed light on the possibility to find or breed chilling-tolerant genotypes of J. curcas.
Photosynthetic irradiance response of vegetative and reproductive structures of the green-flowered deciduous perennial green hellebore was studied by the comparative use of chlorophyll (Chl) fluorescence techniques and gas exchange measurements. All the Chl-containing organs (leaves, sepals, stalks, and fruits) examined were photosynthetically active showing high intrinsic efficiencies of photosystem 2 (Fv/Fm: 0.75–0.79) after dark adaptation. Even in the smaller fertile and sterile parts of the flower (nectaries and anthers) a remarkable photosynthetic competence was detected. With increasing photon flux densities (PFD) electron transport rates, actual quantum yields, and photochemical quenching coefficients of the main photosynthetic organs decreased in the order: leaf>sepal>fruit>stalk. At moderate to high PFDs the sepals achieved maximum electron transport rates corresponding to about 80 % of concomitant mature leaves. In contrast, maximum net photosynthetic rate of the sepals [2.3 mol(CO2) m–2 s–1] were less than one fourth of the leaves [10.6 mol(CO2) m–2 s–1]. This difference is explained by a 70–80 % lower stomatal density of sepals in comparison to leaves. As the basal leaves emerge late during fruit development, the photosynthetically active sepals are a major source of assimilates, contributing more than 60 % of whole-plant CO2 gain in early spring. The ripening dehiscent fruits are characterized by an effective internal re-fixation of the respirational carbon loss and thus additionally improve the overall carbon budget.
Gas chromatography coupled to time-of-flight mass spectrometry (GC-TOF-MS) has become a promising technique for simultaneous
and rapid analysis of small metabolites in complex mixtures. The aim of this work was to establish the quantitative nature
of the information generated by amino acid analysis of crude leaf extracts using GC-TOF-MS. Dried aliquots of methanol/water
extracts of Arabidopsis leaves were analysed in parallel by GC-TOF-MS following trimethylsilylation or high performance liquid chromatography and
fluorescence detection of o-phthaldialdehyde derivatives (OPA-HPLC). Twenty amino acids could be routinely detected in leaf extracts by both methods.
Because of instability of some trimethylsilylated derivatives, all GC-TOF-MS analyses were performed within a window of 2h
30min following derivatization. Repeatability studies showed that relative standard deviations for multiple injections of
a single extract were below 20% for both techniques, though significantly smaller for OPA-HPLC. Similar between-extract variability
and condition-independent biological variation were detected by OPA-HPLC and GC-TOF-MS, and both techniques detected similar
environmentally induced changes in four major amino acids. Recovery of standard compounds through the extraction procedure
was between 80% and 120% for OPA-HPLC but more variable when analysed by GC-TOF-MS. When quantified on the basis of tissue
fresh weight according to response factors of mixed standards, the two techniques gave consistent values for a number of amino
acids but divergent values for others. Taken together, the results suggest GC-TOF-MS analysis of Arabidopsis leaves with the present protocol can be used for absolute quantification of 4–7 amino acids, accurate relative quantification
of 8–11 amino acids, and more limited quantification for five compounds of this class.
SummaryIn addition to the green leaves, commonly considered as the primary sources of photosynthate production, higher plants can potentially use almost all vegetative and reproductive structures to perform photosynthetic CO2 assimilation. Green leaves, stems and green sterile flower organs, optimized for light harvesting and photosynthetic performance, are characterized by net photosynthetic assimilation utilizing mainly the atmospheric carbon dioxide. In contrast, chlorophyll-containing bark and wood tissue, most fruit, root and fertile flower organs are principally sub-ordinated to non-photosynthetic functions, but typically perform an effective internal CO2 recycling using the respiratory released CO2. Non-foliar photosynthesis, either manifested as net photosynthesis or internal CO2 refixation is regarded as an important strategy of additional carbon-acquisition. While chlorophyllous stems or aerial roots even can serve as primary photosynthetic organs, reproductive structures could derive up to 60%; of their total carbon requirement from own CO2 fixation. In the review, the main strategies of additional carbon acquisition by non-foliar photosynthetic organs are illustrated, presenting an extensive compilation of published data completed with relevant own studies.
Confronted to changes in temperatures, plants readjust their biochemical makeup to adapt and survive. The fact that temperature changes can induce cellular responses indicates that temperature is sensed and that the temperature signal is transduced into the cell. While the signalling pathways triggered temperature changes are well described, the way plants sense temperature is often considered as elusive. This review is focused on the mechanisms by which plants sense temperature. We show that plants have no internal thermometer as such, but that the very alterations in cellular equilibria triggered by temperature changes act as networked thermostats to sense heat and cold. Amongst these temperature-sensitive devices, we identified membrane fluidity, protein conformation, cytoskeleton depolymerization, and metabolic reactions. Besides, other molecular switches are proposed. A model of the temperature sensing “machinery” is proposed. Finally, we discuss the specificities of temperature sensing, showing that signalling events can feed-back perception steps.
Measurements of the quantum yields of chlorophyll fluorescence and CO2 assimilation for a number of plant species exposed to changing light intensity and atmospheric CO2 concentrations and during induction of photosynthesis are used to examine the relationship between fluorescence quenching parameters and the quantum yield of non-cyclic electron transport. Over a wide range of physiological conditions the quantum yield of non-cyclic electron transport was found to be directly proportional to the product of the photochemical fluorescence quenching (qQ) and the efficiency of excitation capture by open Photosystem II (PS II) reaction centres (Fv/Fm). A simple fluorescence parameter, ΔφF/φFm, which is defined by the difference in fluorescence yield at maximal φFm, and steady-state φFs, divided by φFm, can be used routinely to estimate changes in the quantum yield of non-cyclic electron transport. It is demonstrated that both the concentration of open PS II reaction centres and the efficiency of excitation capture by these centres will determine the quantum yield of non-cyclic electron transport in vivo and that deactivation of excitation within PS II complexes by non-photochemical processes must influence the quantum yield of non-cyclic electron transport.
During the last decade, there has been growing interest in the role of trehalose metabolism in tolerance to abiotic stress in higher plants, especially cold stress. So far, this metabolism has not yet been studied in Vitis vinifera L., despite the economic importance of this crop. The goal of this paper was to investigate the involvement of trehalose metabolism in the response of grapevine to chilling stress, and to compare the response in plants bacterised with Burkholderia phytofirmans strain PsJN, a plant growth-promoting rhizobacterium that confers grapevine chilling tolerance, with mock-inoculated plants. In silico analysis revealed that the V. vinifera L. genome contains genes encoding the enzymes responsible for trehalose synthesis and degradation. Transcript analysis showed that these genes were differentially expressed in various plant organs, and we also characterised their response to chilling. Both trehalose and trehalose 6-phosphate (T6P) were present in grapevine tissues and showed a distinct pattern of accumulation upon chilling. Our results suggest a role for T6P as the main active molecule in the metabolism upon chilling, with a possible link with sucrose metabolism. Furthermore, plants colonised by B. phytofirmans and cultivated at 26°C accumulated T6P and trehalose in stems and leaves at concentrations similar to non-bacterised plants exposed to chilling temperatures for 1 day. Overall, our data suggest that T6P and trehalose accumulate upon chilling stress in grapevine and might participate in the resistance to chilling stress conferred by B. phytofirmans.
The most photosynthetically active leaves of rice seedlings were severely damaged when shoots but not roots were chilled (10°C/25°C, respectively), but no such injury was observed when the whole seedling was chilled (10°C/10°C). To elucidate the mechanisms, we compared the photosynthetic characteristics of the seedlings during the dark chilling treatments. Simultaneous analyses of Chl fluorescence and the change in absorbance of P700 showed that electron transport almost disappeared in both PSII and PSI in the 10°C/25°C leaves, whereas the electron transport rate in PSI in the 10°C/10°C leaves was similar to or higher than that in non-chilled control leaves. Light-induced non-photochemical quenching in PSII was inhibited in the 10°C/25°C leaves, occurring at only half the level in the 10°C/10°C leaves, whereas non-light-induced non-photochemical quenching remained high in the 10°C/25°C leaves. The light induction of Chl a fluorescence (OJIP curves) in the 10°C/25°C leaves was similar to that in leaves treated with DCMU. The fluorescence decay after a single turnover saturating flash in the 10°C/25°C leaves was much slower than in the 10°C/10°C leaves. In vivo analyses of the 550-515 nm difference signal indicated decreased formation of a proton gradient across the thylakoid membrane and decreased zeaxanthin formation in the 10°C/25°C leaves. Our results suggest that electron transport was blocked between Q(A) and Q(B) in the dark 10°C/25°C leaves, but without irreversible damage to the components of this system. The consequent light-dependent losses of electron transport, proton gradient formation across the thylakoids and thermal dissipation may therefore be responsible for the visible injury.
The disaccharide trehalose is involved in stress response in many organisms. However, in plants, its precise role remains unclear, although some data indicate that trehalose has a protective role during abiotic stresses. By contrast, some trehalose metabolism mutants exhibit growth aberrations, revealing potential negative effects on plant physiology. Contradictory effects also appear under biotic stress conditions. Specifically, trehalose is essential for the infectivity of several pathogens but at the same time elicits plant defense. Here, we argue that trehalose should not be regarded only as a protective sugar but rather like a double-faced molecule and that further investigation is required to elucidate its exact role in stress tolerance in plants.
The natural (13)C/(12)C isotope composition (delta(13)C) of plants and organic compounds within plant organs is a powerful tool to understand carbon allocation patterns and the regulation of photosynthetic or respiratory metabolism. However, many enzymatic fractionations are currently unknown, thus impeding our understanding of carbon trafficking pathways within plant cells. One of them is the (12)C/(13)C isotope effect associated with invertases (EC 3.2.1.26) that are cornerstone enzymes for Suc metabolism and translocation in plants. Another conundrum of isotopic plant biology is the need to measure accurately the specific delta(13)C of individual carbohydrates. Here, we examined two complementary methods for measuring the delta(13)C value of sucrose, glucose and fructose, that is, off-line high-performance liquid chromatography (HPLC) purification followed by elemental analysis and isotope ratio mass spectrometry (EA-IRMS) analysis, and gas chromatography-combustion (GC-C)-IRMS. We also used these methods to determine the in vitro (12)C/(13)C isotope effect associated with the yeast invertase. Our results show that, although providing more variable values than HPLC approximately EA-IRMS, and being sensitive to derivatization conditions, the GC-C-IRMS method gives reliable results. When applied to the invertase reaction, both methods indicate that the (12)C/(13)C isotope effect is rather small and it is not affected by the use of heavy water (D(2)O).
The problem of multiple comparisons is discussed in the context of medical research. The need for more powerful procedures than classical multiple comparison procedures is indicated. To this end some new, general and simple procedures are discussed and demonstrated by two examples from the medical literature: the neuropsychologic effects of unidentified childhood exposure to lead, and the sleep patterns of sober chronic alcoholics.
During photosynthesis, plants must control the utilization of light energy in order to avoid photoinhibition. We isolated an Arabidopsis mutant, pgr5 (proton gradient regulation), in which downregulation of photosystem II photochemistry in response to intense light was impaired. PGR5 encodes a novel thylakoid membrane protein that is involved in the transfer of electrons from ferredoxin to plastoquinone. This alternative electron transfer pathway, whose molecular identity has long been unclear, is known to function in vivo in cyclic electron flow around photosystem I. We propose that the PGR5 pathway contributes to the generation of a Delta(pH) that induces thermal dissipation when Calvin cycle activity is reduced. Under these conditions, the PGR5 pathway also functions to limit the overreduction of the acceptor side of photosystem I, thus preventing photosystem I photoinhibition.