Single-Cell Ribonucleic Acid Sequencing Clarifies Cold Tolerance Mechanisms in the Pacific White Shrimp (Litopenaeus Vannamei)

To characterize the cold tolerance mechanism of the Pacific white shrimp (Litopenaeus vannamei), we performed single-cell RNA sequencing (scRNA-seq) of ∼5185 hepatopancreas cells from cold-tolerant (Lv-T) and common (Lv-C) L. vannamei at preferred and low temperatures (28°C and 10°C, respectively). The cells fell into 10 clusters and 4 cell types: embryonic, resorptive, blister-like, and fibrillar. We identified differentially expressed genes between Lv-T and Lv-C, which were mainly associated with the terms “immune system,” “cytoskeleton,” “antioxidant system,” “digestive enzyme,” and “detoxification,” as well as the pathways “metabolic pathways of oxidative phosphorylation,” “metabolism of xenobiotics by cytochrome P450,” “chemical carcinogenesis,” “drug metabolism-cytochrome P450,” and “fatty acid metabolism.” Reconstruction of fibrillar cell trajectories showed that, under low temperature stress, hepatopancreas cells had two distinct fates, cell fate 1 and cell fate 2. Cell fate 1 was mainly involved in signal transduction and sensory organ development. Cell fate 2 was mainly involved in metabolic processes. This study preliminarily clarifies the molecular mechanisms underlying cold tolerance in L. vannamei, which will be useful for the breeding of shrimp with greater cold tolerance.

Localization of QTL for cold tolerance at the germination stage of rice (Oryza sativa L.) using recombinant inbred lines

The cold tolerance of germinating direct-sown rice (Oryza sativa L.) has an increased rate of emergence, which ensures vigorous seedling growth. Research on QTL localization for cold tolerance at the germination stage can assist in molecular marker-assisted selection and enhance breeding efficiency. In this study, 94 populations of recombinant self-incompatible lines from Heigu and Ha 9366 were selected to investigate germination rates at low temperatures. It was found that two QTL loci (qLTG-3 and qLTG-12) were located at different germination times on chromosomes 3 and 12, respectively. The two QTLs at three different germination times, located using QTL, accounted for 21.3–25.9% of the phenotypic variation. Moreover, a reciprocal effect was detected between the two QTLs. The double QTLs increased the germination rate by 22–27% in this population. Additionally, qLTG-12 improved cold tolerance at the seedling stage. The results of this study might provide the materials and molecular markers for future molecular marker-assisted breeding for cold tolerance at the germination stage.

Additional Blue LED during Cultivation Induces Cold Tolerance in Tomato Fruit but Only to an Optimum

Tomato is a chilling-sensitive fruit. The aim of this study is to examine the role of preharvest blue LED lighting (BL) to induce cold tolerance in ‘Foundation’ tomatoes. Blue and red supplemental LED light was applied to achieve either 0, 12 or 24% additional BL (0B, 12B and 24B). Mature green (MG) or red (R) tomatoes were harvested and cold stored at 4 °C for 0, 5, 10, 15 and 20 d, and then stored for 20 d at 20 °C (shelf life). Chilling injury (CI) indices, color and firmness, hydrogen peroxide, malondialdehyde, ascorbic acid and catalase activity were characterized. At harvest, R tomatoes cultivated at 12B were firmer and showed less coloration compared to fruit of other treatments. These fruits also showed higher loss of red color during cold storage and lower CI symptoms during shelf-life. MG tomatoes cultivated at 12B showed delayed coloring (non-chilled) and decreased weight loss (long cold stored) during shelf life compared to fruit in the other treatments. No effects of light treatments, both for MG and R tomatoes, were observed for the selected antioxidant capacity indicators. Improved cold tolerance for R tomatoes cultivated at 12B points to lycopene having higher scavenging activity at lower concentrations to mitigate chilling injury.

Jasmonate and Melatonin Act Synergistically to Potentiate Cold Tolerance in Tomato Plants

Both jasmonic acid (JA) and melatonin (MT) have been demonstrated to play positive roles in cold tolerance, however, whether and how they crosstalk in the cold responses in plants remain elusive. Here, we report that JA and MT act synergistically in the cold tolerance in tomato plants (Solanum lycopersicum). It was found that JA and MT were both substantially accumulated in response to cold stress and foliar applications of methyl jasmonate (MeJA) and MT promoted cold tolerance as evidenced by increased Fv/Fm, decreased relative electrolyte leakage (EL) and declined H2O2 accumulation in tomato plants. Inhibition of MT biosynthesis attenuated MeJA-induced cold tolerance, while inhibition of JA biosynthesis reduced MT accumulation in tomato plants under cold conditions. Furthermore, qRT-PCR analysis showed that the expressions of two MT biosynthetic genes, SlSNAT and SlAMST, were strongly induced by MeJA, whereas suppression of SlMYC2, a master JA signaling regulator, abated the expressions of SlSNAT and SlAMST under cold stress. Additionally, suppression of SlMYC2 reduced MT accumulation, decreased Fv/Fm and increased EL in cold-stressed tomato plants. Interestingly, exogenous MT promoted JA accumulation, while inhibition of MT biosynthesis significantly reduced JA accumulation in tomato plants under the cold condition. Taken together, these results suggest that JA and MT act cooperatively in cold tolerance and form a positive feedback loop, amplifying the cold responses of tomato plants. Our findings might be translated into the development of cold-resistant tomato cultivars by genetically manipulating JA and MT pathways.

ERF Transcription Factor OsBIERF3 Positively Contributes to Immunity against Fungal and Bacterial Diseases but Negatively Regulates Cold Tolerance in Rice

We previously showed that overexpression of the rice ERF transcription factor gene OsBIERF3 in tobacco increased resistance against different pathogens. Here, we report the function of OsBIERF3 in rice immunity and abiotic stress tolerance. Expression of OsBIERF3 was induced by Xanthomonas oryzae pv. oryzae, hormones (e.g., salicylic acid, methyl jasmonate, 1-aminocyclopropane-1-carboxylic acid, and abscisic acid), and abiotic stress (e.g., drought, salt and cold stress). OsBIERF3 has transcriptional activation activity that depends on its C-terminal region. The OsBIERF3-overexpressing (OsBIERF3-OE) plants exhibited increased resistance while OsBIERF3-suppressed (OsBIERF3-Ri) plants displayed decreased resistance to Magnaporthe oryzae and X. oryzae pv. oryzae. A set of genes including those for PRs and MAPK kinases were up-regulated in OsBIERF3-OE plants. Cell wall biosynthetic enzyme genes were up-regulated in OsBIERF3-OE plants but down-regulated in OsBIERF3-Ri plants; accordingly, cell walls became thicker in OsBIERF3-OE plants but thinner in OsBIERF3-Ri plants than WT plants. The OsBIERF3-OE plants attenuated while OsBIERF3-Ri plants enhanced cold tolerance, accompanied by altered expression of cold-responsive genes and proline accumulation. Exogenous abscisic acid and 1-aminocyclopropane-1-carboxylic acid, a precursor of ethylene biosynthesis, restored the attenuated cold tolerance in OsBIERF3-OE plants while exogenous AgNO3, an inhibitor of ethylene action, significantly suppressed the enhanced cold tolerance in OsBIERF3-Ri plants. These data demonstrate that OsBIERF3 positively contributes to immunity against M. oryzae and X. oryzae pv. oryzae but negatively regulates cold stress tolerance in rice.

Functional Identification of ICE Transcription Factors in Rubber Tree

ICE (inducer of CBF expression) is a positive regulator of cold signaling pathway in plants. Identification of ICE transcription factors is important for the sustainable development of the natural rubber planting industry in nontraditional regions where sudden cold waves often occur. In this study, five ICE genes were isolated from genome of rubber tree (Hevea brasiliensis Muell. Arg.) for analysing tolerance to cold stress. They shared an ICE-specific region in the highly conserved bHLH-ZIP domain and were localized in the nucleus. The HbICEs were different in transcript abundance and expression patterns in response to cold and drought stresses and among different rubber tree clones. Generally, the expression level of HbICEs was significantly higher in the cold-tolerant rubber tree clones than that in the cold-sensitive rubber tree clones. Overexpression of HbICE1, HbICE2, and HbICE4 significantly enhanced the cold tolerance of transgenic Arabidopsis and tobacco, which showed a significant increase in chlorophyll content and decrease in relative water content and conductivity at the early stage of cold stress in comparison with wild-type plants. Furthermore, overexpression of HbICE2 and HbICE4, but also HbICE1 enhanced drought tolerance in transgenic Arabidopsis. The cold tolerance of rubber tree clones is positively controlled by the expression level of HbICE1, HbICE2, and HbICE4.

Characterization of transcription factor genes related to cold tolerance in Brassica napus

Brassica napus is the third most important oilseed crop in the world; however, in Korea, it is greatly affected by cold stress, limiting seed growth and production. Plants have developed specific stress responses that are generally divided into three categories: cold-stress signaling, transcriptional/post-transcriptional regulation, and stress-response mechanisms. Large numbers of functional and regulatory proteins are involved in these processes when triggered by cold stress. Here, our objective was to investigate the different genetic factors involved in the cold-stress responses of B. napus. Consequently, we treated the Korean B. napus cultivar Naehan at the 4-week stage in cold chambers under different conditions, and RNA and cDNA were obtained. An in silico analysis included 80 cold-responsive genes downloaded from the National Center for Biotechnology Information (NCBI) database. Expression levels were assessed by reverse transcription polymerase chain reaction, and 14 cold-triggered genes were identified under cold-stress conditions. The most significant genes encoded zinc-finger proteins (33.7%), followed by MYB transcription factors (7.5%). In the future, we will select genes appropriate for improving the cold tolerance of B. napus.

Transcriptomic Analysis of Camellia Sinensis (L.) Kuntze Var. Niaowangensis Q. H. Chen Leaves Under Cold Stress

Fine varieties of the Yunwu Tribute Tea (Camellia Sinensis (L.) Kuntze var. niaowangensis Q. H. Chen) are distributed on the Yunwu Mountain, Guiding County, Guizhou province, China. Cold stress usually occurs in winter and is one of the most significant environmental factors restricting the growth of this plant as well as its geographical distribution. However, only few systematic studies have examined the molecular mechanism of cold tolerance in the Yunwu Tribute Tea. Hence, in this study, Illumina HiSeq technology was applied to investigate the cold-tolerance mechanism and for this purpose, cDNA libraries were obtained from two groups of samples namely, the cold-treated group (DW) and the control group (CK). A total of 185,973 unigenes were produced from 511,987 assembled transcripts and among these, 16,020 differentially expressed genes (DEGs) (corrected p-value <0.01, |log2(fold change)| >3), including 9,606 upregulated and 6,414 downregulated genes, were obtained. Moreover, the antioxidant enzyme system, plant hormone signal transduction, proline metabolism, tyrosine metabolism pathway, and transcription factors were analyzed and based on the results, a series of candidate genes related to cold stress were screened out and discussed. The physiological indexes related to the low temperature response were tested, along with five DEGs which were validated by quantitative real-time PCR. For this study, it is expected that the results of the transcriptome sequence of Yunwu Tribute Tea will provide valuable clues for genetic studies while helping to screen candidate genes for cold-resistance breeding in tea plants.