Improved Cold Tolerance Research Articles

PpERF17 alleviates peach fruit postharvest chilling injury under elevated CO2 by activating jasmonic acid and γ-aminobutyric acid biosynthesis

Abstract Internal browning (IB) is a common chilling injury (CI) feature in peach fruit after prolonged cold storage. Our previous study demonstrated that low O2 and elevated CO2 (eCO2) condition of modified atmosphere (MA) storage alleviated CI by facilitating the accumulation of jasmonic acids (JAs) and γ-aminobutyric acid (GABA) in ‘Zhonghuashoutao’ (‘ZHST’) peach fruit. Here we show that 10 % CO2 alone can improve cold tolerance, with ethylene response factor 17 (PpERF17) identified as a pivotal transcription factor (TF) that promotes biosynthesis of JAs and GABA. Stable transformation of PpERF17 in tobacco resulted in reduced cold damage, attributed to decreased levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA), as well as enhanced accumulation of JAs and GABA. Moreover, under eCO2, PpMYC2.1, the master regulator of JA signaling, was found to activate transcription of 13S-lipoxygenase (Pp13S-LOX), allene oxide synthase (PpAOS), 12-oxophytodienoate reductase 3 (PpOPR3) and glutamate decarboxylase (PpGAD), while also inducing the expression of the upstream TF PpERF17, thereby establishing positive feedback loops upregulating JA and GABA biosynthesis. Finally, application of methyl jasmonate (MeJA) to fruit before shelf transfer from cold storage alleviated chilling injury development, due to increased accumulation of JAs and GABA as a result of raised expression of related biosynthetic genes. Collectively, our results suggest that eCO2-induced PpERF17 enhances JAs and GABA accumulation while activating the JA signaling pathway. This contributes to a positive feedback loop mediated by PpMYC2.1, ultimately alleviating CI of peach fruit through the sustained accumulation of JAs and GABA.

Comprehensive analysis of amino acid/auxin permease family genes reveal the positive role of GhAAAP128 in cotton tolerance to cold stress.

Amino acid/auxin permeases (AAAPs) play crucial roles in plant development and response to environmental stimuli. They have been characterized at genome-wide levels in several plant species. However, little is known about the AAAP genes in Gossypium. Here, we identified 149, 141, 73, and 70 AAAPs from four Gossypium species G. hirsutum, G. barbadense, G. arboreum, and G. raimondii, respectively. All the AAAPs were categorized into eight subfamilies (Groups I-VIII). Moreover, we found that 182 and 179 AAAP paralogous gene pairs existed within G. hirsutum and G. barbadense genomes, respectively, and whole genome duplication (WGD) or segmental duplication contributed to the expansion of these AAAPs during evolution. Additionally, many cis-elements related to abiotic stress responses were detected in the promoter regions of GhAAAPs and GbAAAPs. Consistently, the expression of multiple AAAPs was significantly induced by NaCl, polyethylene glycol 6000, and especially cold stress. Among these GhAAAPs, GhAAAP128 had clearly positive roles in cotton and Arabidopsis seedling tolerance to cold stress. GhAAAP128 may also improve cold tolerance by up-regulating the expression of some anthocyanin synthesis genes rather than CBF (C-repeat binding factor) signaling genes. Our findings provide important information for further functional analysis of GhAAAPs in response to stressful cues, particularly cold stress in cotton plants.

ERD14 regulation by the HY5- or HY5-MED2 module mediates the cold signal transduction of asparagus bean.

Cold stress affects the growth, development, and yield of asparagus bean (Vigna unguiculata subsp. sesquipedalis). Mediator (MED) complex subunits regulate the cold tolerance of asparagus bean, but the underlying regulatory mechanisms remain unclear. Here, VunMED2 positively responds to cold stress of asparagus beans. Under cold acclimation and freezing treatment, the survival rate, ROS scavenging activity, and expression levels of VunMED2 were increased in VunMED2 transgenic plants. Natural variation in the promoter of VunMED2 in two different cold-tolerant asparagus beans was observed. Under cold stress, the expression of the GUS reporter gene was higher in cold-tolerant plants than in cold-sensitive plants, and the expression of the GUS reporter gene was tissue-specific. VunHY5 positively influenced the expression of VunMED2 by binding to the E-box motif, and the transcriptional activation of the promoter was stronger in the cold-tolerant variety than in cold-sensitive plants. VunHY5 overexpression improved plant freezing resistance by increasing the antioxidant capacity and expression of dehydrin genes. VunHY5 and VunMED2 play a synergistic role in binding to the G-box/ABRE motif and transcriptionally activating the expression of VunERD14. VunERD14 complemented the med2 mutant, which could positively respond to plant freezing resistance by reducing membrane lipid peroxidation and improving the antioxidant capacity. Therefore, the VunHY5-VunERD14 module and the VunHY5-VunMED2-VunERD14 positive cascade effect are involved in the cold signal transduction in asparagus bean. Our findings have implications for the breeding of asparagus bean varieties with improved cold tolerance.

Cold Intolerance Following Digital Nerve Injury: A Multicenter Prospective Randomized Comparison of Decellularized Nerve Allograft Versus Nerve Conduits.

Cold intolerance following digital nerve injury burdens patients significantly. To better understand how cold intolerance evolves in the setting of digital nerve injuries, a sub-analysis of a trial comparing conduit-based (CONDUIT) and processed nerve allograft (PNA) repairs was conducted. It was hypothesized that PNA repairs would alleviate cold intolerance more effectively, especially for longer nerve gaps. A multicenter trial across 20 US-based medical centers was undertaken of patients 18- to 69-year-old presenting with 5 to 25 mm digital nerve gaps within 24 weeks of injury. Patients were randomized (1:1) to PNA or collagen CONDUIT repairs. Cold Intolerance Symptom Severity (CISS) scores and sensory function testers were assessed at first patient visit (FPV), 1-, 3-, 6-, 9-, and 12-months post-surgery, with patients and assessors blinded to treatment. In total, 220 patients were enrolled, with 183 patients included in final analysis with ≥6 months follow-up. At the last evaluable visit (LEV), mean CISS score decreased from FPV for both PNA (from 31.2 ± 27 to 20.8 ± 19) and CONDUIT (from 31.2 ± 30 to 25.9 ± 24). On sub-analysis, more patients converted from severe/extremely severe cold intolerance to mild cold intolerance for PNA compared with CONDUIT at 1 month and LEV (P < 0.05). The CISS scores correlated significantly with sensory function testing. Although no correlation was demonstrated with nerve gap size, digital nerve gap repaired with PNA had significantly improved cold tolerance outcomes for patients with more severe cold intolerance at FPV relative to nerves repaired with CONDUIT.

Multifunctional Transcription Factor YABBY6 Regulates Morphogenesis, Drought and Cold Stress Responses in Rice

The roles of plant-specific transcription factor family YABBY may vary among different members. OsYABBY6 is a rice YABBY gene, whose function is not well elucidated so far. In this paper, we show that OsYABBY6 is a nucleus-localized protein with transcriptional activation activity. OsYABBY6 is predominantly expressed in the palea and lemma, as well as in the sheath, culm and node. OsYABBY6 RNA interference (RNAi) plants exhibited altered plant height and larger grain size. Under cold treatment, OsYABBY6 overexpression (OE) plants had up-regulated expression of cold responsive genes, and accumulated less reactive oxygen species but more proline compared to wild type, resulting in improved cold tolerance. On the other hand, RNAi plants showed enhanced drought tolerance compared to the wild type by slower water loss, less reactive oxygen species but more proline and soluble sugar accumulation. In addition, endogenous abscisic acid (ABA) level was reduced in OsYABBY6 RNAi plants, and RNAi and OE plants were more and less sensitive to ABA treatment, respectively. Accordingly, we deduce that OsYABBY6 positively regulates cold response but negatively regulates drought response through different pathways. Our study reveals the crucial roles of OsYABBY6 in plant architecture and grain development, as well as in abiotic stress response, providing new insights into the functions of YABBYs in rice.

Further enhancement of cold tolerance in rice seedlings by Piriformospora indica collaborating with plant growth-promoting bacteria: Evidence from the antioxidant defense, osmoregulation, photosynthesis, and related genes

As the global population continues to grow, so does the demand for food. However, cold stress due to global climate change greatly threatens the safe production of rice. Piriformospora indica plays a pivotal role as a functional microbiota in improving cold tolerance in rice. However, the addition of P. indica alone has a limited effect. In this study, an attempt was made to select plant growth-promoting bacteria (Agrobacterium rhizogenes and Bacillus subtilis) as helper bacteria to help P. indica function better and further enhance rice cold tolerance. Under cold stress, the co-addition of the three beneficial microorganisms significantly increased the biomass, photosynthetic performance indicators, osmoregulatory substance contents, and antioxidant enzyme activities of rice. P. indica and helper bacteria significantly reduced the cold stress-induced increases in malondialdehyde content and electrolyte leakage rate in rice leaves and triggered a substantial upregulation of cold-tolerant genes in rice. This suggests that P. indica and helper bacteria can synergistically improve the cold resistance of rice. The findings of the research offer a conceptual foundation and practical assistance for enhancing rice's resilience to cold temperatures.

Molecular regulation by H2S of antioxidant and glucose metabolism in cold-sensitive Capsicum

BackgroundCold is an important environmental limiting factor affecting plant yield and quality. Capsicum (chili pepper), a tropical and subtropical vegetable crop, is extremely sensitive to cold. Although H2S is an important signaling regulator in the responses of plant growth and development to abiotic stress, few studies have examined its effects on cold-sensitive capsicum varieties. Through biotechnology methods to enhance the cold resistance of peppers, to provide some reference for pepper breeding, investigated molecular regulation by H2S of responses to cold stress in cold-sensitive capsicum plants, via physiological and transcriptomic analyses.ResultsIn capsicum seedlings, exogenous H2S enhanced relative electrical conductivity (REC) and levels of malondialdehyde (MDA) under cold stress, maintained membrane integrity, increased the activity of enzymatic and non-enzymatic antioxidants, balanced reactive oxygen species levels (O2·− and H2O2), and improved photosynthesis, mitigating the damage caused by cold. In addition, 416 differentially expressed genes (DEGs) were involved in the response to cold stress after H2S treatment. These DEGs were mainly enriched in the ascorbate–glutathione and starch–sucrose metabolic pathways and plant hormone signal-transduction pathways. Exogenous H2S altered the expression of key enzyme-encoding genes such as GST, APX, and MDHAR in the ascorbate–glutathione metabolism pathway, as well as that of regulatory genes for stimulatory hormones (auxin, cytokinins, and gibberellins) and inhibitory hormones (including jasmonate and salicylic acid) in the plant hormone signal-transduction pathway, helping to maintain the energy supply and intracellular metabolic stability under cold stress.ConclusionsThese findings reveal that exogenous H2S improves cold tolerance in cold-sensitive capsicum plants, elucidating the molecular mechanisms underlying its responses to cold stress. This study provides a theoretical basis for exploring and improving cold tolerance in capsicum plants.

Muti-omics revealed the mechanisms of MT-conferred tolerance of Elymus nutans Griseb. to low temperature at XiZang

BackgroundLow temperature seriously limited the development of grass and crops in plateau. Thus, it is urgent to develop an effective strategy for improving the plant cold tolerance and elucidate the underlying mechanisms.ResultsWe found that MT alleviated the effects of cold stress on suppressing ENG growth, then improved cold tolerance of ENG. Integration of transcriptome and metabolome profiles showed that both cold exposure (TW) and MT reprogrammed the transcription pattern of galactose and flavonoids biosynthesis, leading to changes in compositions of soluble sugar and flavonoids in ENG. Additionally, TW inhibited the photosynthesis, and destroyed the antioxidant system of ENG, leading to accumulation of oxidant radicals represented by MDA. By contrast, MT promoted activities of antioxidant enzymes and flavonoid accumulation in ENG under cold condition, then reduced the MDA content and maintained normal expression of photosynthesis-related genes in ENG even under TW. Importantly, MT mainly enhanced cold tolerance of ENG via activating zeatin synthesis to regulate flavonoid biosynthesis in vivo. Typically, WRKY11 was identified to regulate MT-associated zeatin synthesis in ENG via directly binding on zeatin3 promoter.ConclusionsMT could enhance ENG tolerance to cold stress via strengthening antioxidant system and especially zeatin synthesis to promote accumulation of flavonoids in ENG. Thus, our research gain insight into the global mechanisms of MT in promoting cold tolerance of ENG, then provided guidance for protecting plant from cold stress in plateau.

High lipolytic capacity improves cold tolerance in red swamp crayfish (Procambarus clarkii)

Temperature is one of the most prevalent environmental factors that affects the growth and development of organisms. The survival of aquatic animals is closely related to the changes in temperature, low temperature could cause physiological damage even death. In this study, we revealed the regulatory mechanisms of low-temperature stimulation from the transcriptomics in the hepatopancreas of the red swamp crayfish, Procambarus clarkii. Transcriptome sequencing revealed that lipid metabolism pathways were significantly enriched at low temperature. Crayfish improved their cold tolerance by increasing expression levels of thermogenic gene prdm16 and uncoupling protein genes ucp4 and ucp5. Furthermore, lipid and triglyceride content increased significantly in cold-sensitive crayfish, which in turn increased the expression levels of proinflammatory factors (cox2 and tgfβr) and the amount of hepatopancreas bacteria. In addition, low-temperature stimulation could cause more severe damage to cold-sensitive crayfish spermatophores, accompanied by downregulated of spermatozoa viability-related genes such as alp and ldh. Estradiol 17β-dehydrogenase 8-like (17βde8) may contain a critical SNP site at exon 544, the bases of this site were “G” in cold-tolerant crayfish and “A” in cold-sensitive crayfish. Above all, our results reveal that lipid metabolism is the main factor affecting cold tolerance in crayfish and reveal the key loci for cold tolerance, providing a reference for cold tolerance breeding.

The seed endophytic microbe Microbacterium testaceum M15 enhances the cold tolerance and growth of rice (Oryza sativa L.)

The potential of seed endophytic microbes to enhance plant growth and resilience is well recognized, yet their role in alleviating cold stress in rice remains underexplored due to the complexity of these microbial communities. In this study, we investigated the diversity of seed endophytic microbes in two rice varieties, the cold-sensitive CB9 and the cold-tolerant JG117. Our results revealed significant differences in the abundance of Microbacteriaceae, with JG117 exhibiting a higher abundance under both cold stress and room temperature conditions compared to CB9. Further analysis led to the identification of a specific cold-tolerant microbe, Microbacterium testaceum M15, in JG117 seeds. M15-inoculated CB9 plants showed enhanced growth and cold tolerance, with a germination rate increase from 40 % to 56.67 % at 14℃ and a survival rate under cold stress (4℃) doubling from 22.67 % to 66.67 %. Additionally, M15 significantly boosted chlorophyll content by over 30 %, increased total protein by 16.31 %, reduced malondialdehyde (MDA) levels by 37.76 %, and increased catalase activity by 26.15 %. Overall, our study highlights the potential of beneficial endophytic microbes like M. testaceum M15 in improving cold tolerance in rice, which could have implications for sustainable agricultural practices and increased crop productivity in cold-prone regions.

Enhancement of Thermal Tolerance and Growth Performances of Asian Seabass (Lates calcarifer) Fed with Grape Extract Supplemented Feed.

Cold snaps during the winter present a critical challenge for Asian seabass (Lates calcarifer) in Taiwan, as sudden temperature drops significantly affect their growth and survival. This study explores the effects of dietary grape extract (GE) from Vitis vinifera on the growth performance, oxidative stress regulation, and thermal tolerance of this commercially valuable fish. Over a 60-day feeding trial, four dietary groups were tested: a control diet without GE and three diets supplemented with GE at 2% (GE20), 3% (GE30), and 4% (GE40) with commercial feed. The results demonstrated that GE supplementation positively influenced growth, with the GE20 group achieving the best weight gain and feed conversion ratio among all groups. The upregulation of the growth-related gene igf-1 in the liver of the GE20 group further supported its superior growth performance. Additionally, GE-fed groups showed increased expression of antioxidant-related genes sod1 and sod2 in the liver, while gpx1 exhibited a significant increase only in the GE20 group, indicating enhanced antioxidant defenses. Cat gene expression remained unchanged, and higher GE doses reduced the expression of gpx1, cat, and igf-1. Furthermore, GE supplementation improved cold tolerance in all treated groups compared to the control. These findings suggest that dietary GE at 20 g/kg is particularly effective in enhancing growth performance and cold tolerance in Asian seabass, offering a promising strategy for boosting fish health and adaptability in aquaculture.

Dietary potassium and cold acclimation additively increase cold tolerance in Drosophila melanogaster

In the cold, chill susceptible insects lose the ability to regulate ionic and osmotic gradients. This leads to hemolymph hyperkalemia that drives a debilitating loss of cell membrane polarization, triggering cell death pathways and causing organismal injury. Biotic and abiotic factors can modulate insect cold tolerance by impacting the ability to mitigate or prevent this cascade of events. In the present study, we test the combined and isolated effects of dietary manipulations and thermal acclimation on cold tolerance in fruit flies. Specifically, we acclimated adult Drosophila melanogaster to 15 or 25 °C and fed them either a K+-loaded diet or a control diet. We then tested the ability of these flies to recover from and survive a cold exposure, as well as their capacity to protect transmembrane K+ gradients, and intracellular Na+ concentration. As predicted, cold-exposed flies experienced hemolymph hyperkalemia and cold-acclimated flies had improved cold tolerance due to an improved maintenance of the hemolymph K+ concentration at low temperature. Feeding on a high-K+ diet improved cold tolerance additively, but paradoxically reduced the ability to maintain extracellular K+ concentrations. Cold-acclimation and K+-feeding additively increased the intracellular K+ concentration, aiding in maintenance of the transmembrane K+ gradient during cold exposure despite cold-induced hemolymph hyperkalemia. There was no effect of acclimation or diet on intracellular Na+ concentration. These findings suggest intracellular K+ loading and reduced muscle membrane K+ sensitivity as mechanisms through which cold-acclimated and K+-fed flies are able to tolerate hemolymph hyperkalemia.

Transcriptome analysis reveals the potential mechanism of methyl jasmonate alleviated ripening disorder in mango fruit at low temperature

High susceptibility of mangoes to low temperature leads to ripening failure that restricts the marketability of products. This study investigated the effect of methyl jasmonate (MeJA) on ripening disorder and mechanism involved in mangoes during refrigeration. Results showed that 50 μM MeJA ameliorated ripening disorder, as indicated by accelerated advancement of ripening-related parameters. Transcriptome analysis revealed that 17,414 significantly differentially expressed genes were mainly enriched in ethylene synthesis, cell wall degradation, starch degradation and sugar transport. Moreover, 8 AP2/ERF transcription factors and 12 ripening-related genes were characterized via qRT-PCR. Afterwards, through the analysis of transcription factor binding sites and cis-acting elements, a regulatory network of ERFs mediated alleviation of ripening disorder conferred by MeJA was constructed. Finally, the interactions between MiERFs and the promoters of target genes were verified by yeast one-hybrid assay. Our findings provide a theoretical basis for improving cold tolerance via counteracting ripening disorder in mangoes.

Integration of transcriptional responses in cold contrasting sorghum genotypes under low temperatures

AbstractSorghum is a crop that is susceptible to low temperatures. However, due to the extreme global temperatures, new genotypes have been bred to adapt better to cold climates. Therefore, to better understand the metabolic and molecular differences that may contribute to the improved cold tolerance of these genotypes, it is essential to investigate whether specific molecules play a key role in determining the degree of sorghum tolerance. Furthermore, identifying genes associated with cold tolerance may assist in the development of more robust and productive sorghum cultivars. Here, we analysed the changes in selected elements of the antioxidant metabolism, genes, and miRNAs in two sorghum varieties with different degrees of tolerance to cold stress at different times of exposure to cold stress. The high expression of SbCBF6 in response to cold treatment in early time in the tolerant variety suggests its involvement in the cold response, thereby facilitating a better adaptation of the plant to cold conditions. MiR398 and miR394 showed contrasting behaviour in both varieties in low temperatures, resulting in interesting miRNAs in the degree of sorghum tolerance. MiR319, a marker for the selection of cold‐tolerant cultivars, presents a decrease, suggesting that the better capacity to repress the expression of several members of TCP family of transcription factor genes might be involved in better tolerance to cold stress. This study shows the changes of molecules of interest in response to cold stress in sorghum, marking the difference between cold‐susceptible and cold‐tolerant sorghum plants, which can contribute to selecting tolerant genotypes, allowing new strategies to enhance crop productivity and food security, in regions where low temperatures significantly constrain crop growth and yield.

CIPK11 phosphorylates GSTU23 to promote cold tolerance in Camellia sinensis.

Cold stress negatively impacts the growth, development, and quality of Camellia sinensis (Cs, tea) plants. CBL-interacting protein kinases (CIPK) comprise a pivotal protein family involved in plant development and response to multiple environmental stimuli. However, their roles and regulatory mechanisms in tea plants (Camellia sinensis (L.) O. Kuntze) remain unknown. Here we show that CsCBL-interacting protein kinase 11 (CsCIPK11), whose transcript abundance was significantly induced at low temperatures, interacts and phosphorylates tau class glutathione S-transferase 23 (CsGSTU23). CsGSTU23 was also a cold-inducible gene and has significantly higher transcript abundance in cold-resistant accessions than in cold-susceptible accessions. CsCIPK11 phosphorylated CsGSTU23 at Ser37, enhancing its stability and enzymatic activity. Overexpression of CsCIPK11 in Arabidopsis thaliana resulted in enhanced cold tolerance under freezing conditions, while transient knockdown of CsCIPK11 expression in tea plants had the opposite effect, resulting in decreased cold tolerance and suppression of the C-repeat-binding transcription factor (CBF) transcriptional pathway under freezing stress. Furthermore, the transient overexpression of CsGSTU23 in tea plants increased cold tolerance. These findings demonstrate that CsCIPK11 plays a central role in the signaling pathway to cold signals and modulates antioxidant capacity by phosphorylating CsGSTU23, leading to improved cold tolerance in tea plants.

FveDREB1B improves cold tolerance of woodland strawberry by positively regulating FveSCL23 and FveCHS.

Cold stress has seriously inhibited the growth and development of strawberry during production. CBF/DREB1 is a key central transcription factor regulating plant cold tolerance, but its regulatory mechanisms are varied in different plants. Especially in strawberry, the molecular mechanism of CBF/DREB1 regulating cold tolerance is still unclear. In this study, we found that FveDREB1B was most significantly induced by cold stress in CBF/DREB1 family of diploid woodland strawberry. FveDREB1B was localized to the nucleus, and DREB1B sequences were highly conserved in diploid and octoploid strawberry, and even similar in Rosaceae. And FveDREB1B overexpressed strawberry plants showed delayed flowering and increased cold tolerance, while FveDREB1B silenced plants showed early flowering and decreased cold tolerance. Under cold stress, FveDREB1B activated FveSCL23 expression by directly binding to its promoter. Meanwhile, FveDREB1B and FveSCL23 interacted with FveDELLA, respectively. In addition, we also found that FveDREB1B promoted anthocyanin accumulation in strawberry leaves by directly activating FveCHS expression after cold treatment and recovery to 25°C. DREB1B genes were also detected to be highly expressed in cold-tolerant strawberry resources 'Fragaria mandschurica' and 'Fragaria nipponica'. In conclusion, our study reveals the molecular mechanism of FveDREB1B-FveSCL23-FveDELLA module and FveDREB1B-FveCHS module to enhance the cold tolerance of woodland strawberry. It provides a new idea for improving the cold tolerance of cultivated strawberry and evaluating the cold tolerance of strawberry germplasm resources.

Integrated Transcriptomic and Proteomic Analysis Reveals Molecular Mechanisms of the Cold Stress Response during the Overwintering Period in Blueberries (Vaccinium spp.).

In China, the Liaodong Peninsula is an important growing area for blueberries because of the high organic matter content in the soil, the abundance of light, and the large temperature difference between day and night. However, the low temperature and relative humidity of the air during the winter and early spring in the Liaodong Peninsula are the main reasons for the damage to blueberry plants. Here, we documented the transcriptome and proteome dynamics in response to cold stress in three blueberry cultivars ('Northland', 'Bluecrop', and 'Berkeley'). Functional enrichment analysis indicated that many differentially expressed genes (DEGs) and differentially abundant proteins (DAPs) were mainly involved in the pathways of protein processing in the endoplasmic reticulum, the glutathione metabolism pathway, and ribosomes. We identified 12,747 transcription factors (TFs) distributed in 20 families. Based on our findings, we speculated that cold tolerance development was caused by the expression of calcium-related genes (CDPKs and CMLs), glutathione proteins, and TFs (NAC, WRKY, and ERF). Our investigation found that three cultivars experienced cold damage when exposed to temperatures between -9 °C and -15 °C in the field. Therefore, the cold resistance of blueberries during overwintering should not only resist the influence of low temperatures but also complex environmental factors such as strong winds and low relative humidity in the air. The order of cold resistance strength in the three blueberry cultivars was 'Berkeley', 'Bluecrop', and 'Northland'. These results provide a comprehensive profile of the response to cold stress, which has the potential to be used as a selection marker for programs to improve cold tolerance in blueberries.

Identification of candidate genes controlling cold tolerance at the early seedling stage from Dongxiang wild rice by QTL mapping, BSA-Seq and RNA-Seq

BackgroundThe cold tolerance of rice is closely related to its production and geographic distribution. The identification of cold tolerance-related genes is of important significance for developing cold-tolerant rice. Dongxiang wild rice (Oryza rufipogon Griff.) (DXWR) is well-adapted to the cold climate of northernmost-latitude habitats ever found in the world, and is one of the most valuable rice germplasms for cold tolerance improvement.ResultsTranscriptome analysis revealed genes differentially expressed between Xieqingzao B (XB; a cold sensitive variety) and 19H19 (derived from an interspecific cross between DXWR and XB) in the room temperature (RT), low temperature (LT), and recovery treatments. The results demonstrated that chloroplast genes might be involved in the regulation of cold tolerance in rice. A high-resolution SNP genetic map was constructed using 120 BC5F2 lines derived from a cross between 19H19 and XB based on the genotyping-by-sequencing (GBS) technique. Two quantitative trait loci (QTLs) for cold tolerance at the early seedling stage (CTS), qCTS12 and qCTS8, were detected. Moreover, a total of 112 candidate genes associated with cold tolerance were identified based on bulked segregant analysis sequencing (BSA-seq). These candidate genes were divided into eight functional categories, and the expression trend of candidate genes related to ‘oxidation-reduction process’ and ‘response to stress’ differed between XB and 19H19 in the RT, LT and recovery treatments. Among these candidate genes, the expression level of LOC_Os12g18729 in 19H19 (related to ‘response to stress’) decreased in the LT treatment but restored and enhanced during the recovery treatment whereas the expression level of LOC_Os12g18729 in XB declined during recovery treatment. Additionally, XB contained a 42-bp deletion in the third exon of LOC_Os12g18729, and the genotype of BC5F2 individuals with a survival percentage (SP) lower than 15% was consistent with that of XB. Weighted gene coexpression network analysis (WGCNA) and modular regulatory network learning with per gene information (MERLIN) algorithm revealed a gene interaction/coexpression network regulating cold tolerance in rice. In the network, differentially expressed genes (DEGs) related to ‘oxidation-reduction process’, ‘response to stress’ and ‘protein phosphorylation’ interacted with LOC_Os12g18729. Moreover, the knockout mutant of LOC_Os12g18729 decreased cold tolerance in early rice seedling stage signifcantly compared with that of wild type.ConclusionsIn general, study of the genetic basis of cold tolerance of rice is important for the development of cold-tolerant rice varieties. In the present study, QTL mapping, BSA-seq and RNA-seq were integrated to identify two CTS QTLs qCTS8 and qCTS12. Furthermore, qRT-PCR, genotype sequencing and knockout analysis indicated that LOC_Os12g18729 could be the candidate gene of qCTS12. These results are expected to further exploration of the genetic mechanism of CTS in rice and improve cold tolerance of cultivated rice by introducing the cold tolerant genes from DXWR through marker-assisted selection.

Exploring cotton SFR2’s conundrum in response to cold stress

ABSTRACT Cotton is an important agricultural crop to many regions across the globe but is sensitive to low-temperature exposure. The activity of the enzyme SENSITIVE TO FREEZING 2 (SFR2) improves cold tolerance of plants and produces trigalactosylsyldiacylglycerol (TGDG), but its role in cold sensitive plants, such as cotton remains unknown. Recently, it was reported that cotton SFR2 produced very little TGDG under normal and cold conditions. Here, we investigate cotton SFR2 activation and TGDG production. Using multiple approaches in the native system and transformation into Arabidopsis thaliana, as well as heterologous yeast expression, we provide evidence that cotton SFR2 activates differently than previously found among other plant species. We conclude with the hypothesis that SFR2 in cotton is not activated in a similar manner regarding acidification or freezing like Arabidopsis and that other regions of SFR2 protein are critical for activation of the enzyme than previously reported.

Bi-environmental evaluation of oat (Avena sativa L.) genotypes for yield and nutritional traits under cold stress conditions using multivariate analysis

AbstractOat being a rabi/winter crop in Kashmir, experiences extremely low temperatures which has detrimental effects on its growth and development. Therefore, this study was designed to evaluate a set of 130 oat genotypes in multi-location trials across temperate conditions of Kashmir valley from 2018 to 2022. From the preliminary data of 56 genotypes, including five checks, were selected and evaluated for nutritional and yield attributing traits under cold stress conditions at two locations. The results demonstrated significant genetic variation and high heritability for majority of traits, except for days to 50% flowering, days to maturity and dry fodder. Positive correlations were observed between green fodder yield and other traits, indicating their potential for enhancing yield. Principal component analysis identified four principal components that accounted for 69.87% of the total variation. Cluster analysis categorized the genotypes into two main clusters and six sub-clusters. Frost damage assessment was conducted at tillering stage after the snow melted in late January 2021 and 2022 using cold tolerance rating scale and subsequently tested for chilling injury through an electrolyte leakage test. From field and lab data analysis, five most promising cold tolerant, nutritious and high-yielding genotypes were identified. These genotypes have significant potential for utilization in future breeding programmes to improve cold tolerance in cultivated oats within the Kashmir valley thus promoting agricultural productivity and sustainability. The outcomes also provide valuable insights into the genetic variation, heritability, genotype-by-environment interactions, correlations and cold tolerance of oat genotypes in Kashmir.