Cold Acclimation in Brachypodium Is Accompanied by Changes in Above-Ground Bacterial and Fungal Communities

Shifts in microbiota undoubtedly support host plants faced with abiotic stress, including low temperatures. Cold-resistant perennials prepare for freeze stress during a period of cold acclimation that can be mimicked by transfer from growing conditions to a reduced photoperiod and a temperature of 4 °C for 2–6 days. After cold acclimation, the model cereal, Brachypodium distachyon, was characterized using metagenomics supplemented with amplicon sequencing (16S ribosomal RNA gene fragments and an internal transcribed spacer region). The bacterial and fungal rhizosphere remained largely unchanged from that of non-acclimated plants. However, leaf samples representing bacterial and fungal communities of the endo- and phyllospheres significantly changed. For example, a plant-beneficial bacterium, Streptomyces sp. M2, increased more than 200-fold in relative abundance in cold-acclimated leaves, and this increase correlated with a striking decrease in the abundance of Pseudomonas syringae (from 8% to zero). This change is of consequence to the host, since P. syringae is a ubiquitous ice-nucleating phytopathogen responsible for devastating frost events in crops. We posit that a responsive above-ground bacterial and fungal community interacts with Brachypodium’s low temperature and anti-pathogen signalling networks to help ensure survival in subsequent freeze events, underscoring the importance of inter-kingdom partnerships in the response to cold stress.

Cold Acclimation Affects Physiological and Biochemical Characteristics of Betula platyphylla S. under Freezing Stress

Cold and freezing stress is one of the most harmful environmental stresses, especially in temperate and subtropical areas, that adversely affects plant growth, development, and yield production. Betula platyphylla Sukaczev, also known as white birch, is one of the most valuable, important, and widely distributed tree species in East Asia. This study explored the effects of cold acclimation (CA) in reducing the destructive effect of freezing stress in B. platyphylla seedlings. We measured the physiological and biochemical characteristics of B. platyphylla seedlings, such as chlorophyll content, electrolyte leakage (EL), malondialdehyde (MDA), antioxidant enzymes (such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)), and proline content before and after freezing stress to observe the contribution of CA in reducing the detrimental effects of freezing stress. The results showed that CA increased physiological and biochemical characteristics of B. platyphylla seedlings before and after freezing stress, except for chlorophyll content. Antioxidant enzymes were significantly positively correlated with proline, MDA, and EL content, and negatively correlated with chlorophyll content. Moreover, histochemical detection (H2O2 and O2−) and cell death were revealed to be induced by cold stress in B. platyphylla seedlings. Furthermore, it was revealed that increased time and decreased temperature of the CA process significantly influenced the physiological and biochemical parameters. Overall, the CA process significantly reduced the detrimental effects of freezing stress compared to the control treatment in B. platyphylla seedlings. Taken together, these findings provide beneficial information toward understanding the mechanism of CA and freezing stress in B. platyphylla. Furthermore, the substantial activity of physiological and biochemical results could be used as selection criteria for screening time and temperature points of cold/freezing stress in further omics analyses. In addition, the combination of current study results, further omics analyses, and genetic engineering techniques directly contribute to sustainable forest management systems, tree plantations, and conservation of tree species, especially non-cold/non-freezing tolerant tree species.

Acclimations to Cold and Warm Conditions Differently Affect the Energy Metabolism of Diapausing Larvae of the European Corn Borer Ostrinia nubilalis (Hbn.)

The European corn borer Ostrinia nubilalis is a pest species, whose fifth instar larvae gradually develop cold hardiness during diapause. The physiological changes underlying diapause progression and cold hardiness development are still insufficiently understood in insects. Here, we follow a complex of changes related to energy metabolism during cold acclimation (5°C) of diapausing larvae and compare this to warm-acclimated (22°C) and non-diapause controls. Capillary electrophoresis of nucleotides and coenzymes has shown that in gradually cold-acclimated groups concentrations of ATP/ADP and, consequently, energy charge slowly decrease during diapause, while the concentration of AMP increases, especially in the first months of diapause. Also, the activity of cytochrome c oxidase (COX), as well as the concentrations of NAD+ and GMP, decline in cold-acclimated groups, until the latter part of diapause, when they recover. Relative expression of NADH dehydrogenase (nd1), coenzyme Q-cytochrome c reductase (uqcr), COX, ATP synthase (atp), ADP/ATP translocase (ant), and prohibitin 2 (phb2) is supressed in cold-acclimated larvae during the first months of diapause and gradually increases toward the termination of diapause. Contrary to this, NADP+ and UMP levels significantly increased in the first few months of diapause, after gradual cold acclimation, which is in connection with the biosynthesis of cryoprotective molecules, as well as regeneration of small antioxidants. Our findings evidence the existence of a cold-induced energy-saving program that facilitates long-term maintenance of larval diapause, as well as gradual development of cold hardiness. In contrast, warm acclimation induced faster depletion of ATP, ADP, UMP, NAD+, and NADP+, as well as higher activity of COX and generally higher expression of all energy-related genes in comparison to cold-acclimated larvae. Moreover, such unusually high metabolic activity, driven by high temperatures, lead to premature mortality in the warm-acclimated group after 2 months of diapause. Thus, our findings strongly support the importance of low temperature exposure in early diapause for gradual cold hardiness acquisition, successful maintenance of the resting state and return to active development. Moreover, they demonstrate potentially adverse effects of global climate changes and subsequent increase in winter temperatures on cold-adapted terrestrial organisms in temperate and subpolar regions.