Cold Acclimation Process Research Articles

Physiological and molecular responses of 'Hamlin' sweet orange trees expressing the VvmybA1 gene under cold stress conditions.

Overexpression of VvmybA1 transcription factor in 'Hamlin' citrus enhances cold tolerance by increasing anthocyanin accumulation. This results in improved ROS scavenging, altered gene expression, and stomatal regulation, highlighting anthocyanins' essential role in citrus cold acclimation. Cold stress is a significant threat to citrus cultivation, impacting tree health and productivity. Anthocyanins are known for their role as pigments and have emerged as key mediators of plant defense mechanisms against environmental stressors. This study investigated the potential of anthocyanin overexpression regulated by grape (Vitis vinifera) VvmybA1 transcription factor to enhance cold stress tolerance in citrus trees. Transgenic 'Hamlin' citrus trees overexpressing VvmybA1 were exposed to a 30-day cold stress period at 4°C along with the control wild-type trees. Our findings reveal that anthocyanin accumulation significantly influences chlorophyll content and their fluorescence parameters, affecting leaf responses to cold stress. Additionally, we recorded enhanced ROS scavenging capacity and distinct expression patterns of key transcription factors and antioxidant-related genes inthe transgenic leaves. Furthermore, VvmybA1 overexpression affected stomatal aperture regulation by moderating ABA biosynthesis, resulting in differential responses in a stomatal opening between transgenic and wild-type trees under cold stress. Transgenic trees exhibited reduced hydrogen peroxide levels, enhanced flavonoids, radical scavenging activity, and altered phytohormonal profiles. These findings highlighted the role of VvmybA1-mediated anthocyanin accumulation in enhancing cold tolerance. The current study alsounderlines the potential of anthocyanin overexpression as a critical regulator of the cold acclimation process by scavenging ROS in plant tissues.

Does deacclimation reverse the changes in structural/physicochemical properties of the chloroplast membranes that are induced by cold acclimation in oilseed rape?

Winter crops acquire frost tolerance during the process of cold acclimation when plants are exposed to low but non-freezing temperatures that is connected to specific metabolic adjustments. Warm breaks during/after cold acclimation disturb the natural process of acclimation, thereby decreasing frost tolerance and can even result in a resumption of growth. This phenomenon is called deacclimation. In the last few years, studies that are devoted to deacclimation have become more important (due to climate changes) and necessary to be able to understand the mechanisms that occur during this phenomenon. In the acclimation of plants to low temperatures, the importance of plant membranes is indisputable; that is why the main aim of our studies was to answer the question of whether (and to what extent) deacclimation alters the physicochemical properties of the plant membranes. The studies were focused on chloroplast membranes from non-acclimated, cold-acclimated and deacclimated cultivars of winter oilseed rape. The analysis of the membranes (formed from chloroplast lipid fractions) using the Langmuir technique revealed that cold acclimation increased membrane fluidity (expressed as the Alim values), while deacclimation generally decreased the values that were induced by cold. Moreover, because the chloroplast membranes were penetrated by lipophilic molecules such as carotenoids or tocopherols, the relationships between the structure of the lipids and the content of these antioxidants in the chloroplast membranes during the process of the cold acclimation and deacclimation of oilseed rape are discussed.

Changes in protein expression in Acacia mearnsii De Wild (black wattle) as a result of exposure to frost-inducing temperatures

Acacia mearnsii De Wild (black wattle) is one of the most important forestry crops in South Africa. The bark of this species contains high-quality tannins, while its wood is used for wood chips, charcoal and mining timber. One of the main problems faced by black wattle farmers is frost damage, thus there is an urgent need to develop frost-tolerant black wattle varieties. A quick method for the screening of frost tolerance variation would be beneficial for black wattle breeders and would result in the need for fewer families to undergo field trials. Frost tolerance in plants is directly related to their ability to undergo cold acclimation in response to changing weather conditions. During the cold acclimation process, many physiological changes occur, including modifications in protein expression. In this study, two-dimensional gel electrophoresis was used to investigate changes in protein expression among 40 black wattle families with varying levels of frost tolerance before and after a cold-stress treatment. The proteomic data generated from this was analysed using multivariate analysis. Six proteins were observed to be upregulated in frosttolerant black wattle families. The proteins were identified and have been previously shown to contribute to the protection of cellular membranes, maintenance of photosynthetic processes and the prevention of protein misfolding and aggregation.

Cold Acclimation and Deacclimation of Winter Oilseed Rape, with Special Attention Being Paid to the Role of Brassinosteroids.

Winter plants acclimate to frost mainly during the autumn months, through the process of cold acclimation. Global climate change is causing changes in weather patterns such as the occurrence of warmer periods during late autumn or in winter. An increase in temperature after cold acclimation can decrease frost tolerance, which is particularly dangerous for winter crops. The aim of this study was to investigate the role of brassinosteroids (BRs) and BR analogues as protective agents against the negative results of deacclimation. Plants were cold-acclimated (3 weeks, 4 °C) and deacclimated (1 week, 16/9 °C d/n). Deacclimation generally reversed the cold-induced changes in the level of the putative brassinosteroid receptor protein (BRI1), the expression of BR-induced COR, and the expression of SERK1, which is involved in BR signal transduction. The deacclimation-induced decrease in frost tolerance in oilseed rape could to some extent be limited by applying steroid regulators. The deacclimation in plants could be detected using non-invasive measurements such as leaf reflectance, chlorophyll a fluorescence, and gas exchange monitoring.

Cold Acclimation and Deacclimation Processes in Grapevine Buds: Insights into the Regulation of Cod Hardiness and ICE-CBF-COR Gene Expression

Cold Acclimation and Deacclimation Processes in Grapevine Buds: Insights into the Regulation of Cod Hardiness and ICE-CBF-COR Gene Expression

Physiological and Biochemical Background of Deacclimation in Plants, with Special Attention Being Paid to Crops: A Minireview

Global climate change, which is connected to global warming and changes in weather patterns, affects various parts of the environment, including the growth/development of plants. Generally, a number of plant species are capable of acquiring tolerance to frost after exposure to cold (in the cold-acclimation/cold-hardening process). In the last few decades, there have been more and more frequent periods of higher temperatures—warm periods that, e.g., break down the process of cold acclimation. This generates deacclimation, which could stimulate growth and lower frost tolerance in plants. Generally, deacclimation causes the reversal of changes induced by cold acclimation (i.e., in concentration of sugars, accumulation of protective proteins, or hormonal homeostasis). Unlike cold acclimation, the phenomenon of deacclimation has been less studied. The aim of this article was (1) to briefly describe the problem of deacclimation, with more attention being paid to its significance for economically important winter crop species, (2) to review and characterize the physiological-biochemical changes that are induced in plants by deacclimation, and (3) to discuss the possibilities of detecting deacclimation earlier in order to counteract its effects on crops.

Transcriptomic analysis of Asparagus officinalis cultivars with varying levels of freezing tolerance over fall acclimation and spring deacclimation periods.

Asparagus (Asparagus officinalis L.) is an important vegetable crop in southern Ontario, Canada, where winter air and soil temperatures below 0°C are common. Consequently, cultivars growing in this area must possess winterhardiness and freezing tolerance for survival. Asparagus acquires freezing tolerance in the fall through cold acclimation and loses freezing tolerance in the spring through deacclimation. To understand the molecular bases of these processes, transcriptomic analysis (RNA-Seq) was conducted on two cultivars, one adapted, 'Guelph Millennium' (GM), and one unadapted, 'UC157' (UC), to the winter conditions of southern Ontario. RNA extracted from bud and rhizome tissues, sampled on three dates during early spring and late fall, was subjected to sequencing. In the fall, the numbers of differentially expressed (DE) genes at the second and third harvests increased, relative to the first harvest, in dormant buds and rhizomes as freezing tolerance of cultivars increased, and the majority of DE genes were downregulated. In spring, freezing tolerance decreased as plants deacclimated and most genes DE at second and third harvests were upregulated in both cultivars. GM had lower LT50 (lethal temperature at which 50% of plants die) values and hence higher freezing tolerance than UC on specific sampling dates during both spring and fall, and expression patterns of specific genes were correlated with LT50 differences. Functional analysis revealed that these genes were involved in carbohydrate metabolic process, plant hormone signal transduction (auxin and gibberellin), proline metabolism, biosynthesis of secondary metabolites, circadian rhythm, and late embryogenesis abundant proteins and could be associated with cold acclimation and deacclimation processes. These findings will help researchers understand the molecular mechanisms of freezing tolerance in asparagus, leading to breeding and genetic strategies to improve the trait.

Screening Cold Tolerance in St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) for USDA Hardiness Zone 7

Among warm-season turfgrasses, St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] generally has poor cold tolerance yet excellent shade tolerance. As mostly hot summers follow cold winters in USDA Hardiness Zone 7, severely damaging tall fescue [Festuca arundineacea Schreb.] and centipedegrass [Eremochloa ophiuroides (Munro) Hack.], a St. Augustinegrass cultivar cold tolerant enough to be grown for shady lawns would greatly benefit both home owners and sod growers in this and potentially other regions. Eight St. Augustinegrass samples were selected from an initial collection of 30, including industry standards ‘Raleigh’ and ‘Palmetto’, for further testing from an established germplasm collection of material collected from lawns grown in USDA Hardiness Zone 7. Percent regrowth was calculated 4 weeks after samples underwent a 2 week cold acclimation process, followed by freezing at 0°C, -4°C, and -6°C separately for 3 h. The germplasm samples designated at ‘A’, ‘G’, and ‘H’ had greater clipping regrowth percentages compared to the industry cold tolerant standard, ‘Raleigh’ at either -4°C and -6°C indicating possible increased cold tolerance for several selections compared to ‘Raleigh’. An increased cold tolerant St. Augustinegrass would provide a wider selection of desirable turfgrasses in the transition zone growing region of the world, especially shady sites.

Mechanisms of cold-induced immunity in plants.

Overwintering plants acquire substantial levels of freezing tolerance through cold acclimation or winter hardening. This process is essential for the plants survival to harsh winter conditions. In the areas where persistent snow cover lasts several months, plants are protected from freezing but are, however, exposed to other harsh conditions such as dark, cold, and high humidity. These conditions facilitate infection of psychrophilic pathogens, which are termed 'snow molds'. To fight against infection of snow molds, overwintering plants develop disease resistance via the process of cold acclimation. Compared with pathogen-induced disease resistance, the molecular mechanisms of cold-induced disease resistance have yet to be fully elucidated. In this review, we outline the recent progress in our understanding of disease resistance acquired through cold acclimation. This article is protected by copyright. All rights reserved.

The Effect of White Light Spectrum Modifications by Excess of Blue Light on the Frost Tolerance, Lipid- and Hormone Composition of Barley in the Early Pre-Hardening Phase.

It is well established that cold acclimation processes are highly influenced, apart from cold ambient temperatures, by light-dependent environmental factors. In this study we investigated whether an extra blue (B) light supplementation would be able to further improve the well-documented freezing tolerance enhancing effect of far-red (FR) enriched white (W) light. The impact of B and FR light supplementation to white light (WFRB) on hormone levels and lipid contents were determined in winter barley at moderate (15 °C) and low (5 °C) temperatures. Low R:FR ratio effectively induced frost tolerance in barley plantlets, but additional B light further enhanced frost hardiness at both temperatures. Supplementation of WFR (white light enriched with FR light) with B had a strong positive effect on abscisic acid accumulation while the suppression of salicylic acid and jasmonic acid levels were observed at low temperature which resembles the shade avoidance syndrome. We also observed clear lipidomic differences between the individual light and temperature treatments. WFRB light changed the total lipid content negatively, but monogalactosyldiacylglycerol (MGDG) content was increased, nonetheless. Our results prove that WFRB light can greatly influence phytohormone dynamics and lipid contents, which eventually leads to more efficient pre-hardening to avoid frost damage.

Cold Hardiness of Prunus mume ‘Xiang Ruibai’ and Its Parents Based on Biological Indexes and Physical Parameters

Low temperature is a primary factor limiting the distribution of Prunus mume. In order to produce a variety that has both cold tolerance and the characteristic fragrance of true mume, previous researchers crossbred a strong-tolerance variety apricot mei, P. mume ‘DF’ (‘Dan Fenghou’) and the weak-tolerance variety of true mume, P. mume ‘BY’ (‘Beijing Yudie’). They gained an offspring variety named P. mume ‘XR’ (‘Xiang Ruibai’), but its cold tolerance is unknown at this point. Here, three varieties (XR, BY, and DF) were selected as the materials, and different low-temperature treatments were used, with temperature as the only variable. Conventional biological methods, such as ion leakage rate, different tissues, and plant viability statistics, were used, as well as an innovative use of infrared engineering and moisture monitoring for dynamic observation of the water-to-ice process in tissues. The results were as follows: DF cold tolerance was the highest, followed by XR and then BY. The LT50 of XR was increased by 6 °C after five days of cold priming at 4 °C, which indicated a stronger cold acclimation ability than the parent varieties. The XR variety enhanced the antioxidant capacity by increasing SOD and POD enzyme activities during low temperature treatment, thus enhancing the cold tolerance. The antioxidant enzyme genes PmSOD3, PmPOD2, PmPOD19, and PmPOD22 had important regulatory roles in XR’s cold acclimation process.

Isolation and characterization of wheat ice recrystallisation inhibition gene promoter involved in low temperature and methyl jasmonateresponses.

It is well known that plant growth, development, survival and geographical distribution are constrained by extreme climatic conditions, especially extreme low temperature. Under cold stress, cold-inducible promoters were identified as important molecular switches to transcriptionally regulate the initiation of genes associated with cold acclimation processes and enhance the adaptability of plants to cold stimulation. Wheat (Triticum aestivum L.) is one of the most dominating food crops in the world, andwheat crops are generally overwintering with strong cold resistance. Our previous study already proved that heterologous expression of wheat ice recrystallization inhibition (IRI) genes enhanced freezing tolerance in tobacco. However, the upstream regulatory mechanisms of TaIRI are ambiguous. In this study, the space-time specific expression of TaIRI genes in wheat was analyzed by quantitative real-time PCR (qRT-PCR), and results showed that the expression of TaIRI in all tissues was cold-induced and accelerate by exogenous methyl jasmonate (MeJA). Three promoters of TaIRI genes were isolated from wheat genome, and various 5'-deletion fragments of TaIRIp were integrated into β-glucuronidase (GUS) within vector pCAMBIA1301. The promoteractivity of TaIRI geneswas determined through transient expression system of tobacco and stable expression of Arabidopsis thaliana. Results revealed that the GUS activity were significantly strengthened by cold and MeJA treatments. This study will provide insights into elucidating the transcription-regulatory mechanism of IRI proteins responding to low temperature.

Time-series RNA-Seq transcriptome profiling reveals novel insights about cold acclimation and de-acclimation processes in an evergreen shrub of high altitude

The high-altitude alpine regions are characterized by highly variable and harsh environmental conditions. However, relatively little is known about the diverse mechanisms adopted by alpine plants to adapt to these stressful conditions. Here, we studied variation in transcriptome and physiological adjustments occurring across the year at high elevation environments in the leaf tissue of Rhododendron anthopogon, an evergreen shrub of Himalaya. The samples were collected at 12 different time-points, from August until snowfall in November 2017, and then from June to September 2018. It was observed that with a drop in both ambient air temperature and photoperiod towards onset of winter, the freezing resistance of plants increased, resulting in ‘cold acclimation’. Further, ‘de-acclimation’ was associated with a decrease in freezing resistance and increase in photosynthetic efficiency of leaves during spring. A considerable amount of variation was observed in the transcriptome in a time-dependent sequential manner, with a total of 9,881 differentially expressed genes. Based on gene expression profiles, the time-points could be segregated into four clusters directly correlating with the distinct phases of acclimation: non-acclimation (22-August-2017, 14-August-2018, 31-August-2018), early cold acclimation (12-September-2017, 29-September-2017), late cold acclimation (11-October-2017, 23-October-2017, 04-November-2017, 18-September-2018) and de-acclimation (15-June-2018, 28-June-2018, 14-July-2018). Cold acclimation was a gradual process, as indicated by presence of an intermediate stage (early acclimation). However, the plants can by-pass this stage when sudden decrease in temperature is encountered. The maximum variation in expression levels of genes occurred during the transition to de-acclimation, hence was ‘transcriptionally’ the most active phase. The similar or higher expression levels of genes during de-acclimation in comparison to non-acclimation suggested that molecular functionality is re-initiated after passing through the harsh winter conditions.

Cold adaptation strategies in plants-An emerging role of epigenetics and antifreeze proteins to engineer cold resilient plants.

Cold stress adversely affects plant growth, development, and yield. Also, the spatial and geographical distribution of plant species is influenced by low temperatures. Cold stress includes chilling and/or freezing temperatures, which trigger entirely different plant responses. Freezing tolerance is acquired via the cold acclimation process, which involves prior exposure to non-lethal low temperatures followed by profound alterations in cell membrane rigidity, transcriptome, compatible solutes, pigments and cold-responsive proteins such as antifreeze proteins. Moreover, epigenetic mechanisms such as DNA methylation, histone modifications, chromatin dynamics and small non-coding RNAs play a crucial role in cold stress adaptation. Here, we provide a recent update on cold-induced signaling and regulatory mechanisms. Emphasis is given to the role of epigenetic mechanisms and antifreeze proteins in imparting cold stress tolerance in plants. Lastly, we discuss genetic manipulation strategies to improve cold tolerance and develop cold-resistant plants.

Light Quality Modulates Plant Cold Response and Freezing Tolerance.

The cold acclimation process is regulated by many factors like ambient temperature, day length, light intensity, or hormonal status. Experiments with plants grown under different light quality conditions indicate that the plant response to cold is also a light-quality-dependent process. Here, the role of light quality in the cold response was studied in 1-month-old Arabidopsis thaliana (Col-0) plants exposed for 1 week to 4°C at short-day conditions under white (100 and 20 μmol m−2s−1), blue, or red (20 μmol m−2s−1) light conditions. An upregulated expression of CBF1, inhibition of photosynthesis, and an increase in membrane damage showed that blue light enhanced the effect of low temperature. Interestingly, cold-treated plants under blue and red light showed only limited freezing tolerance compared to white light cold-treated plants. Next, the specificity of the light quality signal in cold response was evaluated in Arabidopsis accessions originating from different and contrasting latitudes. In all but one Arabidopsis accession, blue light increased the effect of cold on photosynthetic parameters and electrolyte leakage. This effect was not found for Ws-0, which lacks functional CRY2 protein, indicating its role in the cold response. Proteomics data confirmed significant differences between red and blue light-treated plants at low temperatures and showed that the cold response is highly accession-specific. In general, blue light increased mainly the cold-stress-related proteins and red light-induced higher expression of chloroplast-related proteins, which correlated with higher photosynthetic parameters in red light cold-treated plants. Altogether, our data suggest that light modulates two distinct mechanisms during the cold treatment - red light-driven cell function maintaining program and blue light-activated specific cold response. The importance of mutual complementarity of these mechanisms was demonstrated by significantly higher freezing tolerance of cold-treated plants under white light.

Dietary salt supplementation adversely affects thermal acclimation responses of flight ability in Drosophila melanogaster

Cold acclimation may enhance low temperature flight ability, and salt loading can alter an insects’ cold tolerance by affecting their ability to maintain ion balance in the cold. Presently however, it remains unclear if dietary salt impacts thermal acclimation of flight ability in insects. Here, we examined the effect of a combination of dietary salt loading (either NaCl or KCl) and low temperature exposure on the flight ability of Drosophila melanogaster at low (15 °C) and benign (optimal, 22 °C) temperatures. Additionally, we determined whether dietary salt supplementation translates into increased K+ and Na+ levels in the bodies of D. melanogaster. Lastly, we determined whether salt supplementation impacts body mass and wing morphology, to ascertain whether any changes in flight ability were potentially driven by flight-related morphometric variation. In control flies, we find that cold acclimation enhances low temperature flight ability over non-acclimated flies confirming the beneficial acclimation hypothesis. By contrast, flies supplemented with KCl that were cold acclimated and tested at a cold temperature had the lowest flight ability, suggesting that excess dietary KCl during development negates the beneficial cold acclimation process that would have otherwise taken place. Overall, the NaCl-supplemented flies and the control group had the greatest flight ability, whilst those fed a KCl-supplemented diet had the lowest. Dietary salt supplementation translated into increased Na+ and K+ concentration in the body tissues of flies, confirming that dietary shifts are reflected in changes in body composition and are not simply regulated out of the body by homeostasis over the course of development. Flies fed with a KCl-supplemented diet tended to be larger with larger wings, whilst those reared on the control or NaCl-supplemented diet were smaller with smaller wings. Additionally, the flies with greater flight ability tended to be smaller and have lower wing loading. In conclusion, dietary salts affected wing morphology as well as ion balance, and dietary KCl seemed to have a detrimental effect on cold acclimation responses of flight ability in D. melanogaster.

Genome-wide identification of CBF genes and their responses to cold acclimation in Taraxacum kok-saghyz.

C-repeat binding factors (CBFs) are transcription factors that are known to play important roles in plant cold acclimation. They are highly conserved in most higher plants. Taraxacum kok-saghyz (TKS) is an herb native to China and Kazakhstan and is well-known for its production of rubber silk with industrial and economic value. To understand cold acclimation mechanisms, we conducted a genome-wide discovery of the CBF family genes in TKS and revealed ten CBF genes. A bioinformatic analysis of the CBF genes was carried out to analyze the phylogenetic relationship, protein conservative motifs, protein physicochemical properties, gene structure, promoter cis-acting elements, and the gene expression patterns under cold acclimation and control conditions. It was found that most of these genes were highly responsive at the late stage of cold acclimation, indicating that they play important roles in the cold acclimation processes of TKS. This study provides a theoretical basis for the study of the molecular functions of the CBF gene family in TKS, and a useful guidance for the genetic improvement of the cold tolerance traits of TKS and other plants, including crops.

Post-translational activation of CBF for inducing freezing tolerance.

Plants can acquire increased freezing tolerance through cold-acclimation involving the ICE1-CBF-COR pathway. Recently, Lee et al. investigated a potential link between the functional activation of CBF and cellular redox state. We propose that redox-mediated CBF activation could be a hub of low temperature as well as light signaling in the cold-acclimation process.

Plant Low-Temperature Stress: Signaling and Response

Cold stress has always been a significant limitation for plant development and causes substantial decreases in crop yield. Some temperate plants, such as Arabidopsis, have the ability to carry out internal adjustment, which maintains and checks the metabolic machinery during cold temperatures. This cold acclimation process requires prior exposure to low, chilling temperatures to prevent damage during subsequent freezing stress and maintain the overall wellbeing of the plant despite the low-temperature conditions. In comparison, plants of tropical and subtropical origins, such as rice, are sensitive to chilling stress and respond differently to low-temperature stress. Plants have evolved various physiological, biochemical, and molecular mechanisms to sense and respond to low-temperature stress, including membrane modifications and cytoskeletal rearrangement. Moreover, the transient increase in cytosolic calcium level leads to the activation of many calcium-binding proteins and calcium-dependent protein kinases during low-temperature stress. Recently, mitogen-activated protein kinases have been found to regulate low-temperature signaling through ICE1. Besides, epigenetic control plays a crucial role during the cold stress response. This review primarily focuses on low-temperature stress experienced by plants and their strategies to overcome it. We have also reviewed recent progress and previous knowledge for a better understanding of plant cold stress response.

The protective effect of cold acclimation on the low temperature stress of the lotus (Nelumbo nucifera)

This study compared the protective effect of cold acclimation on the cold tolerance in the lotus (Nelumbo nucifera). The cold acclimation increased the sprouting rate and leaf expansion rate of the lotus by about 36% at 0 °C, and the cold acclimation could enhance the levels of the stress related osmolytes including higher proline, soluble protein, and soluble sugar contents. The electrolyte leakage and lipid peroxidation level of the control samples increased significantly, but these indices did not change significantly in the cold acclimation group during low temperature stress. Furthermore, the cold acclimated rhizomes had higher antioxidant enzyme activities and a more stable ROS homeostasis response to the low temperature stress. Some stress-related genes were significantly up-regulated after the cold acclimation, especially the antioxidase related genes (CAT1, GPX, APX and MSD) were up-regulated nearly five times higher than that of the control group at the 0 °C condition. Additionally, the ICE1-CBF-COR pathway was involved in the lotus cold acclimation process. These results suggested that cold acclimation can obviously improve the stress tolerance of the lotus by the stable ROS homeostasis, enhance the antioxidant enzyme activity, regulate the stress-related gene expression and alleviate the stress damage.