New Insights into the Roles of Osmanthus Fragrans Heat-Shock Transcription Factors in Cold and Other Stress Responses

Sweet osmanthus (Osmanthus fragrans) is an evergreen woody plant that emits a floral aroma and is widely used in the landscape and fragrance industries. However, its application and cultivation regions are limited by cold stress. Heat-shock transcription factor (HSF) family members are widely present in plants and participate in, and regulate, the defense processes of plants under various abiotic stress conditions, but now, the role of this family in the responses of O. fragrans to cold stress is still not clear. Here, 46 OfHSF members were identified in the O. fragrans genome and divided into three subfamilies on the basis of a phylogenetic analysis. The promoter regions of most OfHSFs contained many cis-acting elements involved in multiple hormonal and abiotic stresses. RNA-seq data revealed that most of OfHSF genes were differentially expressed in various tissues, and some OfHSF members were induced by cold stress. The qRT-PCR analysis identified four OfHSFs that were induced by both cold and heat stresses, in which OfHSF11 and OfHSF43 had contrary expression trends under cold stress conditions and their expression patterns both showed recovery tendencies after the cold stress. OfHSF11 and OfHSF43 localized to the nuclei and their expression patterns were also induced under multiple abiotic stresses and hormonal treatments, indicating that they play critical roles in responses to multiple stresses. Furthermore, after a cold treatment, transient expression revealed that the malondialdehyde (MDA) content of OfHSF11-transformed tobacco significantly increased, and the expression levels of cold-response regulatory gene NbDREB3, cold response gene NbLEA5 and ROS detoxification gene NbCAT were significantly inhibited, implying that OfHSF11 is a negative regulator of cold responses in O. fragrans. Our study contributes to the further functional characterization of OfHSFs and will be useful in developing improved cold-tolerant cultivars of O. fragrans.

Seed Germination and Seedling Growth of Robinia pseudoacacia Depending on the Origin of Different Geographic Provenances

Black locust (Robinia pseudoacacia) is recognised as a forest species of interest due to its multiple uses. The management of forest genetic resources and their efficient conservation suffer from variations in traits and start with seed germination. The aim of the current study was to investigate the germination of seeds obtained from plus trees selected in eight Romanian provenances, as well as to investigate the influence of the origin upon plants’ growth and development. Two experiments were undertaken to test seed germination: one treatment involved water-soaked seeds and heat/cold treatment, while the other treatment was based on sulphuric acid, at different concentrations (50, 70, 90%). The results were correlated with the morphological analysis of the seeds. Satu-Mare had the lowest germination rate within both treatments. Sulphuric acid did not improve seed germination as much as the heat treatment. The highest germination rate occurred for the water and temperature treatment on seeds from Bihor provenance (68.2%). The most distant provenance was Bihor, in inverse correlation with Bistrița Năsăud and grouped separately within the hierarchical dendrogram of cluster analysis based on the analysed parameters of the provenances investigated. The results demonstrated that the genotypes and environmental heterogeneity of the seed origin within the provenances may finally result in different performances.

GhGLK1 a Key Candidate Gene From GARP Family Enhances Cold and Drought Stress Tolerance in Cotton

Drought and low-temperature stresses are the most prominent abiotic stresses affecting cotton. Wild cotton being exposed to harsh environments has more potential to cope with both biotic and abiotic stresses. Exploiting wild cotton material to induce resistant germplasm would be of greater interest. The candidate gene was identified in the BC2F2 population among Gossypium tomentosum and Gossypium hirsutum as wild male donor parent noted for its drought tolerance and the recurrent parent and a high yielding but drought susceptible species by genotyping by sequencing (GBS) mapping. Golden2-like (GLK) gene, which belongs to the GARP family, is a kind of plant-specific transcription factor (TF) that was silenced by virus-induced gene silencing (VIGS). Silencing of GhGLK1 in cotton results in more damage to plants under drought and cold stress as compared with wild type (WT). The overexpression of GhGLK1 in Arabidopsis thaliana showed that the overexpressing plants showed more adaptability than the WT after drought and cold treatments. The results of trypan blue and 3,3′-diaminobenzidine (DAB) staining showed that after drought and cold treatment, the leaf damage in GhGLK1 overexpressed plants was less as compared with the WT, and the ion permeability was also lower. This study suggested that the GhGLK1 gene may be involved in the regulation of drought and cold stress response in cotton. Our current research findings add significantly to the existing knowledge of cold and drought stress tolerance in cotton.

Leptin Gene Protects Against Cold Stress in Antarctic Toothfish

Leptin is a cytokine-like peptide, predominantly biosynthesized in adipose tissue, which plays an important role in regulating food intake, energy balance and reproduction in mammals. However, how it may have been modified to enable life in the chronic cold is unclear. Here, we identified a leptin-a gene (lepa) in the cold-adapted and neutrally buoyant Antarctic toothfish Dissostichus mawsoni that encodes a polypeptide carrying four α-helices and two cysteine residues forming in-chain disulfide bonds, structures shared by most vertebrate leptins. Quantitative RT-PCR confirmed that mRNA levels of the leptin-a gene of D. mawsoni (DM-lepa) were highest in muscle, followed by kidney and liver; detection levels were low in the gill, brain, intestine, and ovary tissues. Compared with leptin-a genes of fishes living in warmer waters, DM-lepa underwent rapid evolution and was subjected to positive selection. Over-expression of DM-lepa in the zebrafish cell line ZFL resulted in signal accumulation in the cytoplasm and significantly increased cell proliferation both at the normal culture temperature and under cold treatment. DM-lepa over-expression also reduced apoptosis under low-temperature stress and activated the STAT3 signaling pathway, in turn upregulating the anti-apoptotic proteins bcl2l1, bcl2a, myca and mdm2 while downregulating the pro-apoptotic baxa, p53 and caspase-3. These results demonstrate that DM-lepa, through STAT3 signaling, plays a protective role in cold stress by preventing apoptotic damage. Our study reveals a new role of lepa in polar fish.

Genome-wide Identification and Expression Analysis of the Cucumber PP2C Genes Family

Background: Type 2C protein phosphatase (PP2Cs) is a negative regulator of ABA signaling pathway, which play important roles in stress signal transduction in plants. However, cucumber (Cucumis sativus L.), as an important economic vegetable, has little research on its PP2C genes family. Results: This study conducted a genome-wide investigation of CsPP2C gene family. Through bioinformatics analysis, 56 CsPP2C genes were identified in cucumber. Based on phylogenetic analysis, the PP2C genes of cucumber and Arabidopsis were divided into 13 groups. Gene structure and conserved motif analysis showed that CsPP2C genes in the same group had similar gene structure and conserved domains. Collinearity analysis showed that segmental duplication events played a key role in the expansion of cucumber PP2C genes family. In addition, the expression of CsPP2Cs under different abiotic treatments was analyzed by qRT-PCR. The results showed that CsPP2C family genes showed different expression patterns under ABA, drought, salt and cold treatment, and a significantly responsive gene CsPP2Cs was obtained (CsPP2C3). By predicting the cis-elements in the promoter, we found that all CsPP2C members contained ABA response elements (ABRE) and drought response elements (MYC). Additionally, the expression patterns of CsPP2C genes were specific in different tissues. Conclusions: The results of this study provide a reference for the genome-wide identification of PP2C gene family in other species, and provide a basis for future studies on the function of PP2C gene in cucumber.

Comparative Evaluation of Methods for Stabilizing Alcoholic Beverages Based on Cornel Distillate

Обработки для обеспечения розливостойкости спиртных напитков на основе фруктовых дистиллятов - плодовых водок являются завершающим этапом в технологии производства. Настоящая статья посвящена выбору способа и определению режимных параметров обработки кизиловой водки. С этой целью в работе была проведена сравнительная оценка двух способов обработки - холодом с выдержкой и последующей холодной фильтрацией и ступенчатая фильтрация через мембраны с разным рейтингом пор. Объектами исследования в работе служили контрольный образец спиртного напитка крепостью 45%об., приготовленный из кизилового дистиллята с использованием дистиллированной воды и опытные образцы, подвергнутые стабилизации. На 1-м этапе работы было изучено влияние температуры и длительности обработки холодом на розливостойкость спиртного напитка. Установлено, что при снижении температуры продолжительность обработки до достижения требуемой розливостойкости сокращается. Сравнительная оценка влияния способа обработки (холодом или фильтрацией) показала неэффективность последовательной фильтрации в аспекте стойкости напитка на основе кизилового дистиллята к помутнениям физико-химического характера. Показано, что применение способа, основанного на обработке напитка с использованием низких температур (от -3 до -10 °С), влияет на концентрацию отдельных летучих компонентов. В зависимости от температуры и времени воздействия отмечено снижение суммарной концентрации основных высших спиртов. В наибольшей степени уменьшалась концентрация изобутанола - от 4,6 до 16,7%. Максимальное снижение отмечено в образце, обработанном при -10 °С. В образцах, обработанных холодом, также снижалась концентрация ацетальдегида и метанола. По результатам работы рекомендован способ, основанный на обработке холодом, режимы обработки: -10 °С в течение 2 ч или при температуре -5 °С в течение 6 ч. Processing to ensure the bottling resistance of alcoholic beverages based on fruit distillates-fruit vodkas is the final stage in the production technology. This article is devoted to the choice of the processing method and the determination of the operating parameters of the processing of Cornel vodka. For this purpose, a comparative evaluation of two processing methods was carried out - cold with exposure and subsequent cold filtration and step filtration through membranes with different pore ratings. The objects of research in the work were a control sample of an alcoholic beverage with a strength of 45%vol., prepared from Cornel distillate using distilled water and experimental samples subjected to stabilization. At the first stage of the work, the influence of temperature and duration of cold treatment on the bottling resistance of an alcoholic beverage was studied. It is established that when the processing temperature decreases, the processing time until the required filling resistance is achieved is reduced. A comparative assessment of the effect of the processing method (cold or filtration) showed the ineffectiveness of sequential filtration in terms of the resistance of a drink based on dogwood distillate to turbidity of a physico-chemical nature. It is shown that the use of a method based on the processing of a drink using low temperatures (from -3 °C to -10 °C) affects the concentration of individual volatile components. Depending on the temperature and time of exposure, a decrease in the total concentration of the main higher alcohols was noted. The concentration of isobutanol decreased to the greatest extent - from 4.6 to 16.7%. The maximum decrease was observed in the sample treated at -10 °C. The concentration of acetaldehyde and methanol also decreased in the samples treated with cold. According to the results of the work, a method based on cold treatment is recommended, processing modes: -10 °C for 2 hours or at a temperature of -5 °C for 6 hours.

Macrophages expressing uncoupling protein 1 increase in adipose tissue in response to cold in humans

Acute cold induces beige adipocyte protein marker expression in human subcutaneous white adipose tissue (SC WAT) from both the cold treated and contralateral leg, and the immune system regulates SC WAT beiging in mice. Cold treatment significantly increased the gene expression of the macrophage markers CD68 and 86 in SC WAT. Therefore, we comprehensively investigated the involvement of macrophages in SC WAT beiging in lean and obese humans by immunohistochemistry. Cold treatment significantly increased CD163/CD68 macrophages in SC WAT from the cold treated and contralateral legs of lean and obese subjects, and had similar effects on CD206/CD68 macrophages, whereas the effects on CD86/CD68 macrophages were inconsistent between lean and obese. However, linear regression analysis did not find significant relationships between the change in macrophage numbers and the change in UCP1 protein abundance. A high percentage of CD163 macrophages in SC WAT expressed UCP1, and these UCP1 expressing CD163 macrophages were significantly increased by cold treatment in SC WAT of lean subjects. In conclusion, our results suggest that CD163 macrophages are involved in some aspect of the tissue remodeling that occurs during SC WAT beiging in humans after cold treatment, but they are likely not direct mediators of the beiging process.

The cold - resistance mechanism of a mutagenic Volvariella volvacea strain VH3 with outstanding traits revealed by transcriptome profiling

Background The straw mushroom (Volvariella volvacea) is one of the important vegetables that is popular for its delicious taste. However, the straw mushroom is sensitive to low temperature, resulting in economic loss during transportation and storage. We obtained a novel straw mushroom strain, named VH3, via ultraviolet mutagenesis. Results Our study revealed that VH3 exhibited high cold resistance compared to an ordinary straw mushroom cultivar, V23. We found that the electrolyte leakages of VH3 were always significantly lower than that of V23 treated with 4 °C for 0 h, 2 h,4 h, 8 h, 16 h, and 24 h. Before cold treatment (0 h), there were no difference of MDA contents, SOD activities, and CAT activities between VH3 and V23. At the late stage (8 h, 26 h, and 24 h) of cold treatment, the MDA contents of VH3 were lower while both the SOD and CAT activities were higher than those of V23. To investigate the potential mechanisms of VH3 cold resistance, we performed transcriptome sequencing to detect the transcriptome profiling of VH3 and V23 after 0 h and 4 h cold treatment. Transcriptome sequencing revealed that 111 differentially expressed genes (DEG) between V23 (0 h) and VH3 (0 h) (V23–0_vs_VH3–0), consisting 50 up-regulated and 61 down-regulated DEGs. A total of 117 DEGs were obtained between V23 (4 h) and VH3(4 h) (V23–4_vs_VH3–4), containing 94 up-regulated and 23 down-regulated DEGs. Among these DEGs, VVO_00021 and VVO_00017 were up-regulated while VVO_00003, VVO_00004, VVO_00010, and VVO_00030 were down-regulated in V23–0_vs_VH3–0 and VH3–4_vs_V23–4. KEGG and GO analysis revealed that the 6 DEGs were annotated to pathways related to cold stress. Besides, the GA3 content was also decreased in VH3. Conclusions Collectively, our study first revealed that the increased cold resistance of VH3 might be caused by the expression change of VVO_00003, VVO_00004, VVO_00017, VVO_00021, and VVO_00030, and decreased GA3.