HbSnRK2.6 Functions in ABA-Regulated Cold Stress Response by Promoting HbICE2 Transcriptional Activity in Hevea brasiliensis

Low temperature remarkably limits rubber tree (Hevea brasiliensis Muell. Arg.) growth, latex production, and geographical distribution, but the underlying mechanisms of Hevea brasiliensis cold stress response remain elusive. Here, we identified HbSnRK2.6 as a key component in ABA signaling functions in phytohormone abscisic acid (ABA)-regulated cold stress response in Hevea brasiliensis. Exogenous application of ABA enhances Hevea brasiliensis cold tolerance. Cold-regulated (COR) genes in the CBF pathway are upregulated by ABA. Transcript levels of all five HbSnRK2.6 members are significantly induced by cold, while HbSnRK2.6A, HbSnRK2.6B, and HbSnRK2.6C can be further activated by ABA under cold conditions. Additionally, HbSnRK2.6s are localized in the cytoplasm and nucleus, and can physically interact with HbICE2, a crucial positive regulator in the cold signaling pathway. Overexpression of HbSnRK2.6A or HbSnRK2.6B in Arabidopsis extensively enhances plant responses to ABA and expression of COR genes, leading to increased cold stress tolerance. Furthermore, HbSnRK2.6A and HbSnRK2.6B can promote transcriptional activity of HbICE2, thus, increasing the expression of HbCBF1. Taken together, we demonstrate that HbSnRK2.6s are involved in ABA-regulated cold stress response in Hevea brasiliensis by regulating transcriptional activity of HbICE2.

Demyristoylation of the Cytoplasmic Redox Protein Trx-h2 Is Critical for Inducing a Rapid Cold Stress Response in Plants

In Arabidopsis, the cytosolic redox protein thioredoxin h2 (Trx-h2) is anchored to the cytoplasmic endomembrane through the myristoylated second glycine residue (Gly2). However, under cold stress, the cytosolic Trx-h2 is rapidly translocated to the nucleus, where it interacts with and reduces the cold-responsive C-repeat-binding factors (CBFs), thus activating cold-responsive (COR) genes. In this study, we investigated the significance of fatty acid modification of Trx-h2 under cold conditions by generating transgenic Arabidopsis lines in the trx-h2 mutant background, overexpressing Trx-h2 (Trx-h2OE/trx-h2) and its point mutation variant Trx-h2(G/A) [Trx-h2(G/A)OE/trx-h2], in which the Gly2 was replaced by alanine (Ala). Due to the lack of Gly2, Trx-h2(G/A) was incapable of myristoylation, and a part of Trx-h2(G/A) localized to the nucleus even under warm temperature. As no time is spent on the demyristoylation and subsequent nuclear translocation of Trx-h2(G/A) under a cold snap, the ability of Trx-h2(G/A) to protect plants from cold stress was greater than that of Trx-h2. Additionally, COR genes were up-regulated earlier in Trx-h2(G/A)2OE/trx-h2 plants than in Trx-h2OE/trx-h2 plants under cold stress. Consequently, Trx-h2(G/A)2OE/trx-h2 plants showed greater cold tolerance than Col-0 (wild type) and Trx-h2OE/trx-h2 plants. Overall, our results clearly demonstrate the significance of the demyristoylation of Trx-h2 in enhancing plant cold/freezing tolerance.

Exogenous Methyl Jasmonate Improves Cold Tolerance with Parallel Induction of Two Cold-Regulated (COR) Genes Expression in Triticum aestivum L.

Methyl jasmonate (MJ) is an important plant growth regulator that plays a key role in tolerance to biotic and abiotic stresses. In this research, the effects of exogenous MJ on cold tolerance, photosynthesis, activity and gene expression of antioxidant enzymes, proline accumulation, and expression of cold-regulated (COR) genes in wheat seedlings under low temperature (4 °C) were investigated. Exogenous MJ treatment (1 µM) promoted wheat cold tolerance before and during cold exposure. Low temperature significantly decreased photosynthetic parameters, whereas MJ application led to their partial recovery under cold exposure. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels increased in response to low temperature, and this was counteracted by MJ application. Exogenous MJ significantly enhanced the activities of antioxidant enzymes and upregulated the expression of MnSOD and CAT during cold exposure. MJ application also led to enhanced proline content before 4 °C exposure, whereas the P5CS gene expression was upregulated by MJ’s presence at both normal (22 °C) and low (4 °C) temperatures. It was also shown that MJ tended to upregulate the expression of the COR genes WCS19 and WCS120 genes. We conclude that exogenous MJ can alleviate the negative effect of cold stress thus increasing wheat cold tolerance.

Physiological and Molecular Mechanism Involved in Cold Stress Tolerance in Plants

Previous studies have reported that low temperature (LT) constrains plant growth and restricts productivity in temperate regions. However, the underlying mechanisms are complex and not well understood. Over the past ten years, research on the process of adaptation and tolerance of plants during cold stress has been carried out. In molecular terms, researchers prioritize research into the field of the ICE-CBF-COR signaling pathway which is believed to be the important key to the cold acclimation process. Inducer of CBF Expression (ICE) is a pioneer of cold acclimation and plays a central role in C-repeat binding (CBF) cold induction. CBFs activate the expression of COR genes via binding to cis-elements in the promoter of COR genes. An ICE-CBF-COR signaling pathway activates the appropriate expression of downstream genes, which encodes osmoregulation substances. In this review, we summarize the recent progress of cold stress tolerance in plants from molecular and physiological perspectives and other factors, such as hormones, light, and circadian clock. Understanding the process of cold stress tolerance and the genes involved in the signaling network for cold stress is essential for improving plants, especially crops.

Endogenous accumulation of glycine betaine confers improved low temperature resistance on transplastomic potato plants

Glycine betaine (GB) plays a crucial role in plant response to abiotic stress, and its accumulation in chloroplasts is more effective than in the cytosol in improving the resistance of transgenic plants. Here, we report that the codA gene from Arthrobacter globiformis, which encodes a choline oxidase catalysing the conversion of choline to GB, was successfully introduced into the plastid genome of potato (Solanum tuberosum L.). Transgenic plants with plastid expression of codA showed increased tolerance to low temperature stress compared with the wild type (WT). Further studies revealed that under low temperature stress condition, transgenic plants presented a significantly higher photosynthetic performance by regulating the electron transport and energy distribution in PSII, and higher antioxidant enzyme activities and lower O2– and H2O2 accumulation than did the WT plants. A higher expression of the COR genes was also observed in transgenic plants. Our results suggest that chloroplast biosynthesis of GB could be an effective strategy for the engineering of plants with increased resistance to low temperature stress.

Identification and Characterization of a Well-Defined Series of Coronatine Biosynthetic Mutants of Pseudomonas syringae pv. tomato DC3000

To identify Pseudomonas syringae pv. tomato genes involved in pathogenesis, we carried out a screen for Tn5 mutants of P. syringae pv. tomato DC3000 with reduced virulence on Arabidopsis thaliana. Several mutants defining both known and novel virulence loci were identified. Six mutants contained insertions in biosynthetic genes for the phytotoxin coronatine (COR). The P. syringae pv. tomato DC3000 COR genes are chromosomally encoded and are arranged in two separate clusters, which encode enzymes responsible for the synthesis of coronafacic acid (CFA) or coronamic acid (CMA), the two defined intermediates in COR biosynthesis. High-performance liquid chromatography fractionation and exogenous feeding studies confirmed that Tn5 insertions in the cfa and cma genes disrupt CFA and CMA biosynthesis, respectively. All six COR biosynthetic mutants were significantly impaired in their ability to multiply to high levels and to elicit disease symptoms on A. thaliana plants. To assess the relative contributions of CFA, CMA, and COR in virulence, we constructed and characterized cfa6 cmaA double mutant strains. These exhibited virulence phenotypes on A. thalliana identical to those observed for the cmaA or cfa6 single mutants, suggesting that reduced virulence of these mutants on A. thaliana is caused by the absence of the intact COR toxin. This is the first study to use biochemically and genetically defined COR mutants to address the role of COR in pathogenesis.

Genetic analysis of the expression of the cold-regulated gene cor14b: a way toward the identification of components of the cold response signal transduction in Triticeae

Extensive molecular biological studies have led to the cloning of many cold-regulated genes (cor) whose expression is up-regulated in winter cereals during exposure to cold. Among them is cor14b, a nuclear gene coding for a chloroplast-localized protein whose expression level has been associated with frost resistance in Triticeae. Although frost tolerance is recognised as a complex quantitative character, members of homologous group 5 in Triticeae are known to carry major loci determining frost tolerance. In this work we summarize a number of recent experiments where the cold-regulated gene cor14b was used as a tool (i) to understand the genetic relationship between the expression of cor genes and the frost resistance loci and (ii) to identify barley mutants revealing a chloroplast role in the signal transduction pathway leading to the molecular cold response.Key words: Triticeae, cold hardening, frost resistance, cor genes.