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

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

Frontiers in Plant Science ◽

10.3389/fpls.2021.763284 ◽

2022 ◽

Vol 12 ◽

Author(s):

Fei Ding ◽

Liming Ren ◽

Fang Xie ◽

Meiling Wang ◽

Shuoxin Zhang

Keyword(s):

Cold Stress ◽

Cold Tolerance ◽

Feedback Loop ◽

Pcr Analysis ◽

Tomato Plants ◽

Qrt Pcr ◽

Cold Condition ◽

Tomato Cultivars ◽

Cold Responses ◽

Ja Signaling

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

10.21203/rs.3.rs-1172962/v1 ◽

2021 ◽

Author(s):

Ying Wang ◽

Cheng Wan ◽

Leijia Li ◽

Zhun Xiang ◽

Jihong Wang ◽

...

Keyword(s):

Cold Stress ◽

Candidate Genes ◽

Cold Tolerance ◽

Camellia Sinensis ◽

Resistance Breeding ◽

Guizhou Province ◽

Cdna Libraries ◽

Control Group ◽

P Value ◽

Illumina Hiseq

Abstract 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.

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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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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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Genome-Wide Association Study Reveals the Genetic Basis of Cold Tolerance in Rice at the Seedling Stage

Agriculture ◽

10.3390/agriculture11040318 ◽

2021 ◽

Vol 11 (4) ◽

pp. 318

Author(s):

Tae-Ho Ham ◽

Yebin Kwon ◽

Yoonjung Lee ◽

Jisu Choi ◽

Joohyun Lee

Keyword(s):

Association Study ◽

Candidate Genes ◽

Cold Tolerance ◽

Genome Wide Association Study ◽

Seedling Stage ◽

Genome Wide Association ◽

Matrix Analysis ◽

Rice Seedlings ◽

Genome Wide ◽

A Genome

We conducted a genome-wide association study (GWAS) of cold tolerance in a collection of 127 rice accessions, including 57 Korean landraces at the seedling stage. Cold tolerance of rice seedlings was evaluated in a growth chamber under controlled conditions and scored on a 0–9 scale, based on their low-temperature response and subsequent recovery. GWAS, together with principal component analysis (PCA) and kinship matrix analysis, revealed four quantitative trait loci (QTLs) on chromosomes 1, 4, and 5 that explained 16.5% to 18.5% of the variance in cold tolerance. The genomic region underlying the QTL on chromosome four overlapped with a previously reported QTL associated with cold tolerance in rice seedlings. Similarly, one of the QTLs identified on chromosome five overlapped with a previously reported QTL associated with seedling vigor. Subsequent bioinformatic and haplotype analyses revealed three candidate genes affecting cold tolerance within the linkage disequilibrium (LD) block of these QTLs: Os01g0357800, encoding a pentatricopeptide repeat (PPR) domain-containing protein; Os05g0171300, encoding a plastidial ADP-glucose transporter; and Os05g0400200, encoding a retrotransposon protein, Ty1-copia subclass. The detected QTLs and further evaluation of these candidate genes in the future will provide strategies for developing cold-tolerant rice in breeding programs.

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Comparative Transcriptome Analysis Reveals Potential Gene Modules Associated with Cold Tolerance in Sugarcane (Saccharum officinarum L.)

Journal of Plant Growth Regulation ◽

10.1007/s00344-021-10437-9 ◽

2021 ◽

Author(s):

Xing Huang ◽

Yongsheng Liang ◽

Baoqing Zhang ◽

Xiupeng Song ◽

Yangrui Li ◽

...

Keyword(s):

Cold Stress ◽

Cold Tolerance ◽

Cold Resistance ◽

Soluble Sugar ◽

Resistance Mechanism ◽

Saccharum Officinarum ◽

Calcium Signal ◽

Protein Kinase Activity ◽

Comparative Transcriptome ◽

Climate Conditions

AbstractSugarcane is an important crop worldwide, and most sugar is derived directly from sugarcane. Due to its thermophilic nature, the yield of sugarcane is largely influenced by extreme climate conditions, especially cold stress. Therefore, the development of sugarcane with improved cold tolerance is an important goal. However, little is known about the multiple mechanisms underlying cold acclimation at the bud stage in sugarcane. In this study, we emphasized that sensitivity to cold stress was higher for the sugarcane variety ROC22 than for GT42, as determined by physical signs, including bud growth capacity, relative conductivity, malonaldehyde contents, and soluble sugar contents. To understand the factors contributing to the difference in cold tolerance between ROC22 and GT42, comparative transcriptome analyses were performed. We found that genes involved in the regulation of the stability of the membrane system were the relative determinants of difference in cold tolerance. Additionally, genes related to protein kinase activity, starch metabolism, and calcium signal transduction were associated with cold tolerance. Finally, 25 candidate genes, including 23 variety-specific and 2 common genes, and 7 transcription factors were screened out for understanding the possible cold resistance mechanism. The findings of this study provide candidate gene resources for cold resistance and will improve our understanding of the regulation of cold tolerance at the bud stage in sugarcane.

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Physiological and transcriptome changes induced by exogenous putrescine in anthurium under chilling stress

Botanical Studies ◽

10.1186/s40529-020-00305-2 ◽

2020 ◽

Vol 61 (1) ◽

Author(s):

Xiangli Sun ◽

Zebin Yuan ◽

Bo Wang ◽

Liping Zheng ◽

Jianzhong Tan ◽

...

Keyword(s):

Antioxidant Activities ◽

Chilling Stress ◽

Chilling Tolerance ◽

Membrane Lipid ◽

Control Group ◽

Qrt Pcr ◽

Differential Gene ◽

Anthurium Andraeanum ◽

Chilling Response

Abstract Background Chilling stress is the major factor limiting plant productivity and quality in most regions of the world. In the present study, we aimed to evaluate the effects of putrescine (Put) and polyamine inhibitor d-arginine (d-arg) on the chilling tolerance of anthurium (Anthurium andraeanum). Results Anthurium seedlings were pretreated with five different concentrations of Put solution or d-arg solution. Subsequently, the seedlings were subjected to chilling stress at 6 °C for 3 days, followed by a recovery at 25 °C for 1 day. Relative permeability of the plasma membrane, as well as physiological and morphologic parameters was assessed during the experiments. Additionally, transcriptome sequencing and patterns of differential gene expression related to chilling response were analyzed by qRT-PCR in 1.0 mM Put-treated and untreated anthurium seedlings. Results indicated that the supplementation of exogenous Put decreased the extent of membrane lipid peroxidation and the accumulation of malondialdehyde (MDA), promoted the antioxidant activities and proline content and maintained the morphologic performances compared with the control group. This finding indicated that the application of exogenous Put could effectively decrease the injury and maintain the quality of anthurium under chilling conditions. In contrast, the treatment of d-arg exhibited the opposite effects, which confirmed the effects of Put. Conclusions This research provided a possible approach to enhance the chilling tolerance of anthurium and reduce the energy consumption used in anthurium production.

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GRAS-domain transcription factor PAT1 regulates jasmonic acid biosynthesis in grape cold stress response

Plant Physiology ◽

10.1093/plphys/kiab142 ◽

2021 ◽

Author(s):

Zemin Wang ◽

Darren Chern Jan Wong ◽

Yi Wang ◽

Guangzhao Xu ◽

Chong Ren ◽

...

Keyword(s):

Stress Response ◽

Cold Stress ◽

Cold Tolerance ◽

Cold Treatment ◽

Ectopic Expression ◽

Bimolecular Fluorescence Complementation ◽

Luciferase Reporter ◽

Vitis Amurensis ◽

Mobility Shift

Abstract Cultivated grapevine (Vitis) is a highly valued horticultural crop, and cold stress affects its growth and productivity. Wild Amur grape (Vitis amurensis) PAT1 (Phytochrome A signal transduction 1, VaPAT1) is induced by low temperature, and ectopic expression of VaPAT1 enhances cold tolerance in Arabidopsis (Arabidopsis thaliana). However, little is known about the molecular mechanism of VaPAT1 during the cold stress response in grapevine. Here, we confirmed the overexpression of VaPAT1 in transformed grape calli enhanced cold tolerance. Yeast two-hybrid and bimolecular fluorescence complementation assays highlighted an interaction between VaPAT1 with INDETERMINATE-DOMAIN 3 (VaIDD3). A role of VaIDD3 in cold tolerance was also indicated. Transcriptome analysis revealed VaPAT1 and VaIDD3 overexpression and cold treatment coordinately modulate the expression of stress-related genes including lipoxygenase 3 (LOX3), a gene encoding a key jasmonate biosynthesis enzyme. Co-expression network analysis indicated LOX3 might be a downstream target of VaPAT1. Both electrophoretic mobility shift and dual luciferase reporter assays showed the VaPAT1-IDD3 complex binds to the IDD-box (AGACAAA) in the VaLOX3 promoter to activate its expression. Overexpression of both VaPAT1 and VaIDD3 increased the transcription of VaLOX3 and JA levels in transgenic grape calli. Conversely, VaPAT1-SRDX (dominant repression) and CRISPR/Cas9-mediated mutagenesis of PAT1-ED causing the loss of the C-terminus in grape calli dramatically prohibited the accumulation of VaLOX3 and JA levels during cold treatment. Together, these findings point to a pivotal role of VaPAT1 in the cold stress response in grape by regulating JA biosynthesis.

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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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Gene Expression in First Trimester Preeclampsia Placenta

Biological Research For Nursing ◽

10.1177/1099800410385448 ◽

2010 ◽

Vol 13 (2) ◽

pp. 134-139 ◽

Cited By ~ 21

Author(s):

Sandra A. Founds ◽

Lauren A. Terhorst ◽

Kirk P. Conrad ◽

W. Allen Hogge ◽

Arun Jeyabalan ◽

...

Keyword(s):

Candidate Genes ◽

Chorionic Villus ◽

First Trimester ◽

Chorionic Villus Sampling ◽

Qrt Pcr ◽

Individualized Care ◽

Additional Control ◽

Polymerase Chain ◽

The Difference

Background. The goal of this study was to further validate eight candidate genes identified in a microarray analysis of first trimester placentas in preeclampsia. Material and method. Surplus chorionic villus sampling (CVS) specimens of 4 women subsequently diagnosed with preeclampsia (PE) and 8 control women (C) without preeclampsia analyzed previously by microarray and 24 independent additional control samples (AS) were submitted for confirmatory studies by quantitative real-time polymerase chain reaction (qRT-PCR). Results. Downregulation was significant in FSTL3 in PE as compared to C and AS (p = .04). PAEP was downregulated, but the difference was only significant between C and AS (p = .002) rather than between PE and either of the control groups. Expression levels for CFH, EPAS1, IGFBP1, MMP12, and SEMA3C were not statistically different among groups, but trends were consistent with microarray results; there was no anti-correlation. S100A8 was not measurable in all samples, probably because different probes and primers were needed. Conclusions. This study corroborates reduced FSTL3 expression in the first trimester of preeclampsia. Nonsignificant trends in the other genes may require follow-up in studies powered for medium or medium/large effect sizes. qRT-PCR verification of the prior microarray of CVS may support the placental origins of preeclampsia hypothesis. Replication is needed for the candidate genes as potential biomarkers of susceptibility, early detection, and/or individualized care of maternal—infant preeclampsia.

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