Closure of plasmodesmata in maize (Zea mays) at low temperature: a new mechanism for inhibition of photosynthesis

Background and Aims

Photosynthesis is one of the processes most susceptible to low-temperature inhibition in maize, a tropical C4 crop not yet fully adapted to a temperate climate. C4 photosynthesis relies on symplasmic exchange of large amounts of photosynthetic intermediates between Kranz mesophyll (KMS) and bundle sheath (BS) cells. The aim of this study was to test the hypothesis that the slowing of maize photosynthesis at low temperature is related to ultrastructural changes in the plasmodesmata between KM and BS as well as BS and vascular parenchyma (VP) cells.

Methods

Chilling-tolerant (CT) KW 1074 and chilling-sensitive (CS) CM 109 maize (Zea mays) lines were studied. The effect of moderate chilling (14 °C) on the rate of photosynthesis, photosynthate transport kinetics, and the ultrastructure of plasmodesmata linking the KMS, BS and VP cells were analysed. Additionally, the accumulation of callose and calreticulin was studied by the immunogold method.

Key Results

Chilling inhibited photosynthesis, photosynthate transfer to the phloem and photosynthate export from leaves in both lines. This inhibition was reversible upon cessation of chilling in the CT line but irreversible in the CS line. Simultaneously to physiological changes, chilling induced swelling of the sphincters of plasmodesmata linking KMS and BS cells and a decreased lumen of the cytoplasmic sleeve of plasmodesmata at the BS/VP interface in the CS line but not in the CT line. Accumulation of calreticulin, which occurred near the neck region of the closed plasmodesmata was observed after just 4 h of chilling and over-accumulation of callose at the KMS/BS and BS/VP interfaces occurred after 28 h of chilling.

Conclusions

Stronger chilling sensitivity of the CM 109 maize line compared with the KW 1074 line, shown by decreased photosynthesis and assimilate export from a leaf, is related to changes in the ultrastructure of leaf plasmodesmata at low temperature. The chain of reactions to chilling is likely to include calreticulin action resulting in rapid and efficient closure of the plasmodesmata at both KMS/BS and BS/VP interfaces. Callose deposition in a leaf was a secondary effect of chilling.     

Abscisic acid is involved in brassinosteroids-induced chilling tolerance in the suspension cultured cells from Chorispora bungeana

The objective of this study was to investigate whether abscisic acid (ABA), a second messenger in chilling stress responses, is involved in brassinosteroids (BRs)-induced chilling tolerance in suspension cultured cells from Chorispora bungeana. The suspension cells were treated with 24-epibrassinolide (EBR), ABA, ABA biosynthesis inhibitor fluridone (Flu) and EBR in combination with Flu. Their effects on chilling tolerance, reactive oxygen species (ROS) levels and antioxidant defense system were analyzed. The results showed that EBR treatment markedly alleviated the decrease of cell viability and the increases of ion leakage and lipid peroxidation induced by chilling stress, suggesting that application of EBR could improve the chilling tolerance of C. bungeana suspension cultures. In addition, similar results were observed when exogenous ABA was applied. Treatment with Flu alone and in combination with EBR significantly suppressed cell viability and increased ion leakage and lipid peroxidation under low temperature conditions, indicating that the inhibition of ABA biosynthesis could decrease the chilling tolerance of C. bungeana suspension cultures and the EBR-enhanced chilling tolerance. Further analyses showed that EBR and ABA enhanced antioxidant defense and slowed down the accumulation of ROS caused by chilling. However, Flu application differentially blocked these protective effects of EBR. Moreover, EBR was able to mimic the effect of ABA by markedly increasing ABA content in the suspension cells under chilling conditions, whereas the EBR-induced ABA accumulation was inhibited by the addition of Flu. Taken together, these results demonstrate that EBR may confer chilling tolerance to C. bungeana suspension cultured cells by enhancing the antioxidant defense system, which is partially mediated by ABA, resulting in preventing the overproduction of ROS to alleviate oxidative injury induced by chilling.     

Understanding chilling responses in Arabidopsis seeds and their contribution to life history

Winter chilling is of central importance in the phenology of temperate annual and perennial plants. Chilling accelerates flowering through the process of vernalization and breaks both bud and seed dormancy, permitting the onset of growth in the spring. The quantitative effects of chilling in floral promotion in winter annual Arabidopsis accessions are well-documented, but very little is known about the basic physiology underlying summer annual responses to winter chilling, which acts on seeds within the soil seed bank. Here, we analyse the response of wild accessions to extended chilling in seeds, and explore the interaction between seed-maturation temperature and chilling responses. We show that two weeks of chilling induces secondary dormancy, and that this time period is not dependent on seed-maturation temperature. In addition, we found that seeds for most accessions set under simulated summer conditions in the laboratory are unable to overwinter in the soil seed bank, as they germinate without light during extended chilling treatments. This shows that these seeds are committed to re-establishment in the same growing season. Understanding how winter chilling affects the timing of Arabidopsis phenology will enable us to explore the genetics behind adaptation to changing climates, and inform rational approaches to breeding crops with improved performance under new climate scenarios and develop a systems ecology of Arabidopsis.     

From observations to experiments in phenology research: investigating climate change impacts on trees and shrubs using dormant twigs

Background and Aims Climate change is advancing the leaf-out times of many plant species and mostly extending the growing season in temperate ecosystems. Laboratory experiments using twig cuttings from woody plant species present an affordable, easily replicated approach to investigate the relative importance of factors such as winter chilling, photoperiod, spring warming and frost tolerance on the leafing-out times of plant communities. This Viewpoint article demonstrates how the results of these experiments deepen our understanding beyond what is possible via analyses of remote sensing and field observation data, and can be used to improve climate change forecasts of shifts in phenology, ecosystem processes and ecological interactions.Scope The twig method involves cutting dormant twigs from trees, shrubs and vines on a single date or at intervals over the course of the winter and early spring, placing them in containers of water in controlled environments, and regularly recording leaf-out, flowering or other phenomena. Prior to or following leaf-out or flowering, twigs may be assigned to treatment groups for experiments involving temperature, photoperiod, frost, humidity and more. Recent studies using these methods have shown that winter chilling requirements and spring warming strongly affect leaf-out and flowering times of temperate trees and shrubs, whereas photoperiod requirements are less important than previously thought for most species. Invasive plant species have weaker winter chilling requirements than native species in temperate ecosystems, and species that leaf-out early in the season have greater frost tolerance than later leafing species.Conclusions This methodology could be extended to investigate additional drivers of leaf-out phenology, leaf senescence in the autumn, and other phenomena, and could be a useful tool for education and outreach. Additional ecosystems, such as boreal, southern hemisphere and sub-tropical forests, could also be investigated using dormant twigs to determine the drivers of leaf-out times and how these ecosystems will be affected by climate change.     

A novel aspartic acid protease gene from pineapple fruit (Ananas comosus): Cloning,characterization and relation to postharvest chilling stress resistance

A full-length cDNA encoding a putative aspartic acid protease (AcAP1) was isolated for the first time from the flesh of pineapple (Ananas comosus) fruit. The deduced sequence of AcAP1 showed all the common features of a typical plant aspartic protease phytepsin precursor. Analysis of AcAP1 gene expression under postharvest chilling treatment in two pineapple varieties differing in their resistance to blackheart development revealed opposite trends. The resistant variety showed an up-regulation of AcAP1 precursor gene expression whereas the susceptible showed a down-regulation in response to postharvest chilling treatment. The same trend was observed regarding specific AP enzyme activity in both varieties. Taken together our results support the involvement of AcAP1 in postharvest chilling stress resistance in pineapple fruits.     

Arrhenius plots and the involvement of thermotropic phase transitions of the thylakoid membrane in chilling impairment of photosynthesis in thermophilic higher plants

Abstract Breaks and discontinuities in Arrhenius plots of physiological and physical properties of thylakoids are not diagnostic of thermotropic lipid phase transitions of the membrane. Bulk lipid transitions, as first inferred by the membrane phase transition hypothesis, do not occur in any higher plant at chilling temperatures. Solidification of some varying, but always minor, fraction of the total membrane lipid does take place. However, the presence of minor domains of solid thylakoid membrane lipid at chilling temperatures is not unique to chilling sensitive plants but is also found in tolerant species. Minor solidification may in some plants, or groups of plants, be controlled by the specific molecular species of phosphatidylglycerol only recently investigated. In plants containing little, or no, phosphatidylglycerol with this positional distribution of fatty acids, other yet unknown constituents of the membrane must fill a similar function, since DSC thermograms indicate minor solidification also in isolated, unperturbed thylakoids from chilling tolerant species. However, chilling induced phase transitions, or other perturbations, of the thylakoid membrane are not the reason for the chilling lability of net photosynthesis in the intact plant. This conclusion follows from detailed comparison between photosynthetic membranes isolated from prechilled plants and the effects of chilling exposure on CO₂ fixation of the whole plant. Damage at the level of the thylakoid membrane does occur, although not to the extent where it can account for the proportionally much larger damage to CO₂ fixation.     

Chromium in plants

Chromium is an essential trace element and is associated with some biological pathways, especially with glucose tolerance. For these reasons, we decided to determine the concentration of chromium in two sets of Brazilian medicinal plants. The first group consisted of plants that are considered as antidiabetic, whereas the second included plants that do not have this therapeutic property. The concentration of chromium was determined by flameless atomic absorption. All the plants analyzed contain chromium in the normal range for this element, but the hypoglycemic plants contain more chromium than the others (1–4 μg/g compared to 0.5–1.5 μg/g).     

Chilling injury: a plea for uniform terminology

Abstract This article proposes definitions for the terms now commonly applied to studies of both the molecular and cellular aspects of chilling injury. They are proposed with the intent of increasing the precision and clarity of communications on the subject and arc based on the present understanding and current hypotheses regarding the molecular events underlying the development of the visible symptoms of chilling injury.     

Antioxidant compound responses to chilling stress in differentially sensitive inbred maize lines

Chilling temperatures increase the amounts of potentially lethal toxic oxygen compounds present within plants. These toxic oxygen compounds can be scavenged by antioxidant compounds such as ascorbate and β-carotene. Three developmental stages (first, third and fifth leaf) of four inbred lines of maize ( Zea mays L.) exhibiting differential sensitivity to chilling were examined in order to determine if the chilling-sensitive line had lower concentrations of antioxidant compounds than did the tolerant lines. Plants were exposed to one of three treatments: (1) control (25°C constant), (2) control treatment plus a short-term chilling exposure of 11°C one day prior to harvesting, and (3) long-term (11°C constant) chilling exposure. Total ascorbate, total glutathione, β-carotene, α-tocopherol and chlorophyll contents were quantified, and ratios of dehydroascorbate/ascorbate and reduced/oxidized glutathione were determined. Lower concentrations of β-carotene were found in the chilling-sensitive relative to those in the chilling-tolerant lines for the first-leaf stage under both short- and long-term chilling treatments. Concentrations of total ascorbate and glutathione and β-carotene in the chilling-sensitive line increased as the chilling treatment progressed and as the plants developed until they ultimately became either significantly higher or no different relative to the tolerant lines. Results suggest that this sensitive line became less sensitive to chilling-induced oxidative stress with development.     

Antioxidants and antioxidative enzymes in wild-type and transgenic Lycopersicon genotypes of different chilling tolerance

The Mehler–Ascorbate–Peroxidase cycle is a protection system against reactive oxygen species (ROS) occurring during over-excitation of the photosynthetic apparatus. In the cultivated tomato, Lycopersicon esculentum, long-term chilling under moderate light leads to oxidation of the Calvin cycle key enzyme, ribulose-1,5-bisphosphate carboxylase (rubisco), presumably by generation of ROS. In contrast, high-altitude lines of the wild tomato species L. peruvianum were tolerant against the same chilling stress. In the present study, we analysed leaf contents of antioxidants (ascorbate, glutathione) and activities of enzymes of the Mehler–Ascorbate–Peroxidase cycle in the two Lycopersicon species. While antioxidant levels and activities of chloroplast superoxide dismutase (SOD) and ascorbate peroxidase (APX), both inducible by chilling stress, were similar in chilling-tolerant and chilling-sensitive genotypes, chilled L. esculentum showed lower glutathione reductase (GR) activities than high-altitude L. peruvianum. We constructed transgenic plants overexpressing an Escherichia coli GR in the chloroplast (approximately 60-fold of the wild-type (WT) activity). However, these plants resembled identical chilling sensitivity of the photosynthetic apparatus as WT plants as measured after a photoinhibition treatment and by the effect of long-term chilling on rubisco activity. We conclude that the Mehler–Ascorbate–Peroxidase cycle is not the limiting factor for the sensitivity of the photosynthetic apparatus of L. esculentum towards long-term chilling under moderate light. We suggest that a possible cause for the higher chilling tolerance of L. peruvianum is prevention of ROS formation by better conversion of light energy to photochemistry at suboptimal temperatures.     

Initiation and Development of Chilling Injury in Leaves of Chilling-Sensitive Plants

Based on author’s own and literature data, possible mechanisms of initiation, development, and reparation of cell damages in chilling-sensitive plants during and after chilling are reviewed. A conception of initiation and development of chilling injury, based on a key role of oxidative stress, is put forward. Possible mechanisms of structural and functional changes in cells of chilling-sensitive plants subjected to chilling stress are discussed.__________Translated from Fiziologiya Rastenii, Vol. 52, No. 4, 2005, pp. 608–613.Original Russian Text Copyright © 2005 by Lukatkin.     

低温弱光胁迫对野生大豆和大豆栽培种光系统功能的影响

以野生大豆和栽培大豆为材料,通过同时测定大豆叶片的叶绿素荧光快速诱导动力学曲线和对820nm光的吸收曲线,以及测定超氧化物歧化酶（SOD）和抗坏血酸过氧化物酶（APX）的活性,分析了低温弱光胁迫及常温弱光恢复下这2种大豆光系统Ⅱ（PSⅡ）和光系统Ⅰ（PSI）功能的变化。结果表明,低温弱光胁迫对这2种大豆的PSI和PSⅡ的功能都造成伤害;在低温弱光胁迫下,维持较高的SOD和APX活性和维持PSI和PSⅡ的协调性是野生大豆比栽培大豆耐低温的一个重要原因。     

Environmental and Genetic Control of Dormancy in Picea abies

The effects of temperature, photoperiod and chilling on the leafing-out of Norway spruce, Picea abies (L.) Karst. were studied. High temperature promotes breakage of post-dormancy, long photoperiods having no such effect. Photoperiod and chilling cause the breakage of true dormancy. However, under field conditions, photoperiod will have no effect on leafing-out date in the spring. By use of clonal material it was possible to show substantial genetic differences between individuals in response to temperature and photoperiod. When the effect of clones was accounted for, treatments could be compared more precisely. Differences between clones were apparent in heat-sum required for leafing-out, in rapidity of response to favorable post chilling conditions, and in chilling requirement This latter quantity was given a new definition, applicable when both chilling and post chilling temperatures are controlled and specified. This is that period beyond which a further 10 days of chilling accelerates leafing-out by less than one day, i.e. the point at which the slope of the line relating days till leafing-out to chilling period, is equal to minus 0.10. Differences in leafing-out date were shown between provenances taken from throughout the range of Picea abies. These differences were related to latitude with provenances of high latitude leafing-out first.     

Differences in photosynthetic characteristics among northern and southern C4 plants

Both responses to short-term changes of temperature and to chilling under high light were analyzed in populations of Echinochloa crus-galli var. crus-galli (L.) Beauv. from Québec. North Carolina and Mississippi to improve the understanding of C₄ photosynthesis at low temperature. Comparison also included plants of Eleusine indica (L.) Gaertn. from Mississippi to provide for differences among species and populations. Plants were grown at two thermoperiods (28/22°C, 21/15°C). After transfer from cool (21/15°C) to warm (28/22°C) growth conditions, Echinochloa from Mississippi achieved the highest photosynthetic rates. Plants from Québec maintained the highest rates of CO₂ uptake upon transfer to cool conditions. Exposure to 7°C for 3 days at a photon fluence rate of 1000 μmol m−²s−¹ resulted in a reduction in the growth rates of all populations. This reduction was paralleled by a decrease in net photosynthesis and in stomatal conductance. Following chilling under hight light, the reduction in growth parameters was less important for plants from Québec than for the other populations. It suggests that, among other characteristics, northern plants had developed a certain tolerance to chilling under light.     

An heuristic hypothesis of chilling injury in plants: a role for calcium as the primary physiological transducer of injury

Abstract. It is suggested that increased levels of free cytosolic calcium ([Ca²⁺]_cyt) may serve as the primary physiological transducer of chilling injury in plants. Numerous similarities between the effects of [Ca²⁺]_cyt-raising treatments on plants and the effects of chilling temperatures on chilling-sensitive (CS) plants are noted. It is proposed that chilling temperatures may lead to increases in [Ca²⁺]_cyt in CS plant cells by reducing the rate at which they exclude Ca²⁺ from their cytosol and that rapid cooling (coldshock) may cause rapid increases in [Ca²⁺]_cyt due to the activation of voltage-dependent cation channels. Chill-induced increases in [Ca²⁺]_cyt in the cells of CS plants may reflect either an inherent inability of such plants to maintain homeostatic levels of Ca²⁺ at low temperatures or a stress-induced reaction which has evolved to enable such cells to cope more effectively with the short-term hardships imposed by cold. Previous proposals concerning the physiological transduction of chilling injury are also discussed. It is argued that there is little evidence to suggest that the immediate effects of low temperatures on CS cells include either decreases in ATP levels, general increases in the passive permeability of membranes, or increased rates of fermentation.