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.

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Combining QTL-seq and linkage mapping to fine map a candidate gene in qCTS6 for cold tolerance at the seedling stage in rice

BMC Plant Biology ◽

10.1186/s12870-021-03076-5 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Luomiao Yang ◽

Jingguo Wang ◽

Zhenghong Han ◽

Lei Lei ◽

Hua Long Liu ◽

...

Keyword(s):

Cold Stress ◽

Cold Tolerance ◽

Regulatory Gene ◽

Snp Markers ◽

Seedling Stage ◽

Pcr Analysis ◽

Sequencing Data ◽

Coding Region ◽

Rice Varieties ◽

Major Qtl

Abstract Background Cold stress caused by low temperatures is an important factor restricting rice production. Identification of cold-tolerance genes that can stably express in cold environments is crucial for molecular rice breeding. Results In this study, we employed high-throughput quantitative trait locus sequencing (QTL-seq) analyses in a 460-individual F2:3 mapping population to identify major QTL genomic regions governing cold tolerance at the seedling stage in rice. A novel major QTL (qCTS6) controlling the survival rate (SR) under low-temperature conditions of 9°C/10 days was mapped on the 2.60-Mb interval on chromosome 6. Twenty-seven single-nucleotide polymorphism (SNP) markers were designed for the qCST6 region based on re-sequencing data, and local QTL mapping was conducted using traditional linkage analysis. Eventually, we mapped qCTS6 to a 96.6-kb region containing 13 annotated genes, of which seven predicted genes contained 13 non-synonymous SNP loci. Quantitative reverse transcription PCR analysis revealed that only Os06g0719500, an OsbZIP54 transcription factor, was strongly induced by cold stress. Haplotype analysis confirmed that +376 bp (T>A) in the OsbZIP54 coding region played a key role in regulating cold tolerance in rice. Conclusion We identified OsbZIP54 as a novel regulatory gene associated with rice cold-responsive traits, with its Dongfu-104 allele showing specific cold-induction expression serving as an important molecular variation for rice improvement. This result is expected to further exploration of the genetic mechanism of rice cold tolerance at the seedling stage and improve cold tolerance in rice varieties by marker-assisted selection.

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Identification of miRNAs and Their Response to Cold Stress in Astragalus Membranaceus

Biomolecules ◽

10.3390/biom9050182 ◽

2019 ◽

Vol 9 (5) ◽

pp. 182 ◽

Cited By ~ 6

Author(s):

Merhaba Abla ◽

Huigai Sun ◽

Zhuyun Li ◽

Chunxiang Wei ◽

Fei Gao ◽

...

Keyword(s):

Cold Stress ◽

Redox Homeostasis ◽

Astragalus Membranaceus ◽

Pcr Analysis ◽

Small Rna Sequencing ◽

Factors Affecting ◽

Gene Regulation Network ◽

Qrt Pcr ◽

Yield And Quality ◽

Regulation Network

Astragalus membranaceus is an important medicinal plant widely cultivated in East Asia. MicroRNAs (miRNAs) are endogenous regulatory molecules that play essential roles in plant growth, development, and the response to environmental stresses. Cold is one of the key environmental factors affecting the yield and quality of A. membranaceus, and miRNAs may mediate the gene regulation network under cold stress in A. membranaceus. To identify miRNAs and reveal their functions in cold stress response in A. membranaceus, small RNA sequencing was conducted followed by bioinformatics analysis, and quantitative real time PCR (qRT-PCR) analysis was performed to profile the expression of miRNAs under cold stress. A total of 168 conserved miRNAs belonging to 34 families and 14 putative non-conserved miRNAs were identified. Many miRNA targets were predicted and these targets were involved in diversified regulatory and metabolic pathways. By using qRT-PCR, 27 miRNAs were found to be responsive to cold stress, including 4 cold stress-induced and 17 cold-repressed conserved miRNAs, and 6 cold-induced non-conserved miRNAs. These cold-responsive miRNAs probably mediate the response to cold stress by regulating development, hormone signaling, defense, redox homeostasis, and secondary metabolism in A. membranaceus. These cold-corresponsive miRNAs may be used as the candidate genes in further molecular breeding for improving cold tolerance of A. membranaceus.

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Selection of housekeeping genes for qRT-PCR analysis in potato tubers under cold stress

Molecular Breeding ◽

10.1007/s11032-012-9766-z ◽

2012 ◽

Vol 31 (1) ◽

pp. 39-45 ◽

Cited By ~ 26

Author(s):

Raquel Lopez-Pardo ◽

Jose Ignacio Ruiz de Galarreta ◽

Enrique Ritter

Keyword(s):

Cold Stress ◽

Housekeeping Genes ◽

Pcr Analysis ◽

Potato Tubers ◽

Qrt Pcr ◽

Selection Of

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SiFBA5, a cold-responsive factor from Saussurea involucrate promotes cold resilience and biomass increase in transgenic tomato plants under cold stress

10.21203/rs.3.rs-54573/v1 ◽

2020 ◽

Author(s):

Jianqiang Mu ◽

Yajuan Fu ◽

Bucang Liu ◽

Yao Zhang ◽

Aiying Wang ◽

...

Keyword(s):

Low Temperature ◽

Cold Stress ◽

Cold Tolerance ◽

Stress Responses ◽

Transgenic Tomato ◽

Dihydroxyacetone Phosphate ◽

Tomato Plants ◽

Transgenic Tomato Plants ◽

Photosynthesis Efficiency ◽

Fructose 1,6 Bisphosphate

Abstract Background Saussurea involucrate survives in extreme arctic conditions with strong cold resistant ability. The species occurs in rocky, mountainous habitats at elevations of approximately 2400-4100m with year-round snowfall and freezing temperatures providing a magnificent model and abundant gene pool for dissecting plant stress responses. Fructose-1,6-Bisphosphate Aldolase (FBA) mediates the reverse conversion of fructose 1,6-bisphosphate (FBP) into dihydroxyacetone phosphate (DHAP) and glycerol triphosphate (GAP) during glycolysis or gluconeogenesis, which is the key enzyme in this reaction. Saussurea involucrate can survive in extremely low temperature environment, which shows that it has extremely high photosynthesis efficiency. Significantly, the underlying mechanism of its cold tolerance is yet to be unveiled. Therefore, our work aims to explore potential molecular mechanisms. Results In this study, we identified a cold-responsive gene SiFBA5 that based on a preliminary low-temperature genome-wide transcriptional profiling in S. involucrata. Expression analysis showed that cold temperature rapidly induced transcriptional expression of SiFBA5, suggesting that SiFBA5 participates in initial stress responses. Subcellular localization showed that SiFBA5 is localized to the chloroplast. We then generated transgenic tomato plants overexpressing SiFBA5 derived by a CaMV 35S promoter. Phenotypic observation suggested that the transgenic overexpress plants displayed increased cold tolerance, photosynthesis efficiency, and carbohydrate accumulation in comparison with wild-type plants. Conclusion Collectively, our results demonstrated that SiFBA5 positively regulates plant response to cold stress, making it a promising candidate for improving cold tolerance in crops.

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SiFBA5, a cold-responsive factor from Saussurea involucrata promotes cold resilience and biomass increase in transgenic tomato plants under cold stress

BMC Plant Biology ◽

10.1186/s12870-021-02851-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jianqiang Mu ◽

Yajuan Fu ◽

Bucang Liu ◽

Yao Zhang ◽

Aiying Wang ◽

...

Keyword(s):

Stress Response ◽

Low Temperature ◽

Cold Stress ◽

Cold Tolerance ◽

Crop Yield ◽

Photosynthetic Efficiency ◽

Transgenic Tomato ◽

Saussurea Involucrata ◽

Tomato Plants ◽

Transgenic Tomato Plants

Abstract Background Saussurea involucrata survives in extreme arctic conditions and is very cold-resistant. This species grows in rocky, mountainous areas with elevations of 2400–4100 m, which are snow-covered year-round and are subject to freezing temperatures. S. involucrata’s ability to survive in an extreme low-temperature environment suggests that it has particularly high photosynthetic efficiency, providing a magnificent model, and rich gene pool, for the analysis of plant cold stress response. Fructose-1, 6-bisphosphate aldolase (FBA) is a key enzyme in the photosynthesis process and also mediates the conversion of fructose 1, 6-bisphosphate (FBP) into dihydroxyacetone phosphate (DHAP) and glycerol triphosphate (GAP) during glycolysis and gluconeogenesis. The molecular mechanisms underlying S. involucrata’s cold tolerance are still unclear; therefore, our work aims to investigate the role of FBA in plant cold-stress response. Results In this study, we identified a cold-responsive gene, SiFBA5, based on a preliminary low-temperature, genome-wide transcriptional profiling of S. involucrata. Expression analysis indicated that cold temperatures rapidly induced transcriptional expression of SiFBA5, suggesting that SiFBA5 participates in the initial stress response. Subcellular localization analysis revealed that SiFBA5 is localized to the chloroplast. Transgenic tomato plants that overexpressed SiFBA5 were generated using a CaMV 35S promoter. Phenotypic observation suggested that the transgenic plants displayed increased cold tolerance and photosynthetic efficiency in comparison with wild-type plants. Conclusion Cold stress has a detrimental impact on crop yield. Our results demonstrated that SiFBA5 positively regulates plant response to cold stress, which is of great significance for increasing crop yield under cold stress conditions.

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Selection and Evaluation of Reference Genes for Quantitative Real-Time Reverse-Transcriptase PCR under Cold Stress in Eggplant (Solanum melongena L.) during Fruit Development

10.21203/rs.3.rs-79512/v1 ◽

2020 ◽

Author(s):

Jinkun Yang ◽

Ying Zhang ◽

Yong Lian ◽

Yuhui Chen ◽

Fuzhong Liu

Keyword(s):

Gene Expression ◽

Cold Stress ◽

Cold Tolerance ◽

Fruit Development ◽

Reference Genes ◽

Sequence Data ◽

Expression Patterns ◽

Solanum Melongena ◽

Stable Gene ◽

Qrt Pcr

Abstract Background Eggplant (Solanum melongena L.) is a thermophilic vegetable, and its yield and quality are often affected by cold stress. Therefore, identifying the key genes and mechanisms of cold tolerance has become a significant topic in eggplant. qRT-PCR has been widely used to analyse gene expression patterns, and reliable reference genes are necessary for this technique. Methods To select and evaluate suitable reference genes for qRT-PCR, 18 candidate genes selected from transcriptome sequence data were subjected to analysis of their expression stability under natural cold and normal temperature conditions. Four commonly used programs (geNorm, NormFinder, BestKeeper and RefFinder) were used to determine the stabilities of these genes. Results The results showed that D5, D4 and D1 were the three most stable reference genes among the 18 genes. Then, D5, D4 and D1 were compared with commonly used reference genes. The results showed that D5 was still the most stable gene, followed by APRT, D4, and Actin was the least stable gene. Conclusion D5, APRT and D4 were recommended as a reference gene combination for gene expression normalization under cold stress and at normal temperature during fruit development. Our results provide a molecular foundation for further research on the cold tolerance mechanism of eggplant.

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Genome-Wide Characterization of B-Box Gene Family and Its Roles in Responses to Light Quality and Cold Stress in Tomato

Frontiers in Plant Science ◽

10.3389/fpls.2021.698525 ◽

2021 ◽

Vol 12 ◽

Author(s):

Xin Bu ◽

Xiujie Wang ◽

Jiarong Yan ◽

Ying Zhang ◽

Shunyuan Zhou ◽

...

Keyword(s):

Cold Stress ◽

Cold Tolerance ◽

Genome Mapping ◽

Electron Flow ◽

Regulatory Elements ◽

Photochemical Quenching ◽

Protein Motif ◽

Tomato Plants ◽

High Conservation ◽

Non Photochemical Quenching

Perceiving incoming environmental information is critical for optimizing plant growth and development. Multiple B-box proteins (BBXs) play essential roles in light-dependent developmental processes in plants. However, whether BBXs function as a signal integrator between light and temperature in tomato plants remains elusive. In this study, 31 SlBBX genes were identified from the newly released tomato (Solanum lycopersicum) genome sequences and were clustered into five subgroups. Gene structure and protein motif analyses showed relatively high conservation of closely clustered SlBBX genes within each subgroup; however, genome mapping analysis indicated the uneven distribution of the SlBBX genes on tomato chromosomes. Promoter cis-regulatory elements prediction and gene expression indicated that SlBBX genes were highly responsive to light, hormones, and stress conditions. Reverse genetic approaches revealed that disruption of SlBBX7, SlBBX9, and SlBBX20 largely suppressed the cold tolerance of tomato plants. Furthermore, the impairment of SlBBX7, SlBBX9, and SlBBX20 suppressed the photosynthetic response immediately after cold stress. Due to the impairment of non-photochemical quenching (NPQ), the excess photon energy and electron flow excited by low temperature were not consumed in SlBBX7-, SlBBX9-, and SlBBX20- silenced plants, leading to the over reduction of electron carriers and damage of the photosystem. Our study emphasized the positive roles of light signaling transcription factors SlBBXs in cold tolerance in tomato plants, which may improve the current understanding of how plants integrate light and temperature signals to adapt to adverse environments.

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Selection and Validation of Reference Genes for the qRT-PCR Assays of Populus ussuriensis Gene Expression under Abiotic Stresses and Related ABA Treatment

Forests ◽

10.3390/f11040476 ◽

2020 ◽

Vol 11 (4) ◽

pp. 476

Author(s):

Ming Wei ◽

Yingxi Chen ◽

Mengqiu Zhang ◽

Jingli Yang ◽

Han Lu ◽

...

Keyword(s):

Gene Expression ◽

Cold Stress ◽

Internal Control ◽

Reference Genes ◽

Target Genes ◽

High Salinity ◽

High Yield ◽

Pcr Analysis ◽

Qrt Pcr ◽

Populus Ussuriensis

Populus ussuriensis Kom. is one of the most important tree species for forest renewal in the eastern mountainous areas of Northeast China due to its fast growth, high yield, and significant commercial and ecological value. The selection of optimal reference genes for the normalization of qRT-PCR data is essential for the analysis of relative gene expression. In this study, fourteen genes were selected and assessed for their expression stability during abiotic stress (drought, high salinity, and cold stress) and after the treatment with the drought-related hormone ABA. Three algorithms were used, geNorm, NormFinder, and BestKeeper, and a comprehensive ranking of candidate reference genes was produced based on their output. The most appropriate reference genes were UBQ10 and RPL24 for drought and ABA treatment, UBQ10 and TUB3 for cold stress, and UBQ10 and 60S rRNA for high salinity. Overall, UBQ10 was the most stable reference gene for use as an internal control, whereas PP2A was the least stable. The expression of two target genes (P5CS2 and GI) was used to further verify that the selected reference genes were suitable for gene expression normalization. This work comprehensively assesses the stability of reference genes in Populus ussuriensis and identifies suitable reference genes for normalization during qRT-PCR analysis.

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Methyl jasmonate mediates melatonin-induced cold tolerance of grafted watermelon plants

Horticulture Research ◽

10.1038/s41438-021-00496-0 ◽

2021 ◽

Vol 8 (1) ◽

Author(s):

Hao Li ◽

Yanliang Guo ◽

Zhixiang Lan ◽

Kai Xu ◽

Jingjing Chang ◽

...

Keyword(s):

Cold Stress ◽

Methyl Jasmonate ◽

Cold Tolerance ◽

Feedback Loop ◽

Abiotic Stress Tolerance ◽

Critical Role ◽

Horticultural Crop ◽

Crop Species ◽

Induced Tolerance ◽

Figleaf Gourd

AbstractRoot–shoot communication has a critical role in plant adaptation to environmental stress. Grafting is widely applied to enhance the abiotic stress tolerance of many horticultural crop species; however, the signal transduction mechanism involved in this tolerance remains unknown. Here, we show that pumpkin- or figleaf gourd rootstock-enhanced cold tolerance of watermelon shoots is accompanied by increases in the accumulation of melatonin, methyl jasmonate (MeJA), and hydrogen peroxide (H2O2). Increased melatonin levels in leaves were associated with both increased melatonin in rootstocks and MeJA-induced melatonin biosynthesis in leaves of plants under cold stress. Exogenous melatonin increased the accumulation of MeJA and H2O2 and enhanced cold tolerance, while inhibition of melatonin accumulation attenuated rootstock-induced MeJA and H2O2 accumulation and cold tolerance. MeJA application induced H2O2 accumulation and cold tolerance, but inhibition of JA biosynthesis abolished rootstock- or melatonin-induced H2O2 accumulation and cold tolerance. Additionally, inhibition of H2O2 production attenuated MeJA-induced tolerance to cold stress. Taken together, our results suggest that melatonin is involved in grafting-induced cold tolerance by inducing the accumulation of MeJA and H2O2. MeJA subsequently increases melatonin accumulation, forming a self-amplifying feedback loop that leads to increased H2O2 accumulation and cold tolerance. This study reveals a novel regulatory mechanism of rootstock-induced cold tolerance.

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Transcriptomic Analyses of Camellia oleifera ‘Huaxin’ Leaf Reveal Candidate Genes Related to Long-Term Cold Stress

International Journal of Molecular Sciences ◽

10.3390/ijms21030846 ◽

2020 ◽

Vol 21 (3) ◽

pp. 846 ◽

Cited By ~ 1

Author(s):

Lingli Wu ◽

Jian’an Li ◽

Ze Li ◽

Fanhang Zhang ◽

Xiaofeng Tan

Keyword(s):

Cold Stress ◽

Candidate Genes ◽

Cold Tolerance ◽

Soluble Sugars ◽

Chilling Tolerance ◽

Hunan Province ◽

Camellia Oleifera ◽

Qrt Pcr ◽

Hilly Region

‘Huaxin’ is a new high-yielding timber cultivar of Camellia oleifera of high economic value, and has been widely cultivated in the red soil hilly region of Hunan Province of the People´s Republic of China in recent years. However, its quality and production are severely affected by low temperatures during flowering. To find genes related to cold tolerance and further explore new candidategenes for chilling-tolerance, Illumina NGS (Next Generation Sequencing) technology was used to perform transcriptomic analyses of C. oleifera ‘Huaxin’ leaves under long-term cold stress. Nine cDNA libraries were sequenced, and 58.31 Gb high-quality clean reads were obtained with an average of 5.92 Gb reads for each sample. A total of 191,150 transcripts were obtained after assembly. Among them, 100,703 unigenes were generated, and 44,610 unigenes were annotated. In total, 1564 differentially expressed genes (DEGs) were identified both in the A_B and A_C gene sets. In the current study, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed, andrevealed a group of cold-responsive genes related to hormone regulation, photosynthesis, membrane systems, and osmoregulation; these genes encoded many key proteins in plant biological processes, such as serine/threonine-protein kinase (STPK), transcription factors (TFs), fatty acid desaturase (FAD), lipid-transfer proteins (LTPs), soluble sugars synthetases, and flavonoid biosynthetic enzymes. Some physiological indicators of C. oleifera ‘Huaxin’ were determined under three temperature conditions, and the results were consistent with the molecular sequencing. In addition, the expression levels of 12 DEGs were verified using quantitative real-time polymerase chain reaction (qRT-PCR). In summary, the results of DEGs analysis together with qRT-PCR tests contribute to the understanding of cold tolerance and further exploring new candidate genes for chilling-tolerance in molecular breeding programs of C. oleifera ‘Huaxin’.

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