Transcriptomic Analysis of Stropharia Rugosoannulata Reveals Potential Carbohydrate Metabolism and Cold Resistance Mechanisms Under Low-Temperature Stress

2021 ◽  
Author(s):  
Haibo Hao ◽  
Jinjing Zhang ◽  
Shengdong Wu ◽  
Jing Bai ◽  
Xinyi Zhuo ◽  
...  
Keyword(s):  
Heat Shock ◽  
Low Temperature ◽  
Heat Shock Protein ◽  
Carbohydrate Metabolism ◽  
Temperature Stress ◽  
Molecular Mechanisms ◽  
Low Temperature Stress ◽  
Rna Seq ◽  
Oxidoreductase Activity ◽  
Heat Shock Protein Genes

Abstract Low temperature is an important environmental factor that restricts the growth of Stropharia rugosoannulata; however, the molecular mechanisms underlying S. rugosoannulata responses to low-temperature stress are largely unknown. In this study, we performed a transcriptome analysis of a high-sensitivity strain (DQ-1) and low-sensitivity strain (DQ-3) under low-temperature stress. The liquid hyphae of S. rugosoannulata treated at 25°C and 10°C were analyzed by RNA-Seq, and a total of 9499 differentially expressed genes (DEGs) were identified. GO and KEGG enrichment analyses showed that these genes were enriched in “xenobiotic biodegradation and metabolism”, “carbohydrate metabolism”, “lipid metabolism” and “oxidoreductase activity”. Further research found that carbohydrate enzyme (AA, GH, CE, and GT) genes were downregulated more significantly in DQ-1 than DQ-3 and several cellulase activities were also reduced to a greater extent. Moreover, the CAT1, CAT2, GR, and POD genes and more heat shock protein genes (HSP20, HSP78 and sHSP) were upregulated in the two strains after low-temperature stress, and the GPX gene and more heat shock protein genes were upregulated in DQ-3. In addition, the enzyme activity and qRT–PCR results showed trends similar to those of the RNA-Seq results. This result indicates that low-temperature stress reduces the expression of different AA, GH, CE, and GT enzyme genes and reduces the secretion of cellulase, thereby reducing the carbohydrate metabolism process and mycelial growth of S. rugosoannulata. Moreover, the expression levels of different types of antioxidant enzymes and heat shock proteins are also crucial for S. rugosoannulata to resist low-temperature stress. In short, this study will provide a basis for further research on important signaling pathways, gene functions and variety breeding of S. rugosoannulata related to low-temperature stress.

scholarly journals Full-length transcriptome sequencing reveals the low-temperature-tolerance mechanism of Medicago falcata roots

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Guowen Cui ◽  
Hua Chai ◽  
Hang Yin ◽  
Mei Yang ◽  
Guofu Hu ◽  
...  
Keyword(s):  
Low Temperature ◽  
Temperature Stress ◽  
Electrolyte Leakage ◽  
Temperature Tolerance ◽  
Low Temperature Stress ◽  
Full Length ◽  
Rna Seq ◽  
Smrt Sequencing ◽  
Tolerance Mechanism ◽  
Low Temperature Tolerance

Abstract Background Low temperature is one of the main environmental factors that limits crop growth, development, and production. Medicago falcata is an important leguminous herb that is widely distributed worldwide. M. falcata is related to alfalfa but is more tolerant to low temperature than alfalfa. Understanding the low temperature tolerance mechanism of M. falcata is important for the genetic improvement of alfalfa. Results In this study, we explored the transcriptomic changes in the roots of low-temperature-treated M. falcata plants by combining SMRT sequencing and NGS technologies. A total of 115,153 nonredundant sequences were obtained, and 8849 AS events, 73,149 SSRs, and 4189 lncRNAs were predicted. A total of 111,587 genes from SMRT sequencing were annotated, and 11,369 DEGs involved in plant hormone signal transduction, protein processing in endoplasmic reticulum, carbon metabolism, glycolysis/gluconeogenesis, starch and sucrose metabolism, and endocytosis pathways were identified. We characterized 1538 TF genes into 45 TF gene families, and the most abundant TF family was the WRKY family, followed by the ERF, MYB, bHLH and NAC families. A total of 134 genes, including 101 whose expression was upregulated and 33 whose expression was downregulated, were differentially coexpressed at all five temperature points. PB40804, PB75011, PB110405 and PB108808 were found to play crucial roles in the tolerance of M. falcata to low temperature. WGCNA revealed that the MEbrown module was significantly correlated with low-temperature stress in M. falcata. Electrolyte leakage was correlated with most genetic modules and verified that electrolyte leakage can be used as a direct stress marker in physiological assays to indicate cell membrane damage from low-temperature stress. The consistency between the qRT-PCR results and RNA-seq analyses confirmed the validity of the RNA-seq data and the analysis of the regulatory mechanism of low-temperature stress on the basis of the transcriptome. Conclusions The full-length transcripts generated in this study provide a full characterization of the transcriptome of M. falcata and may be useful for mining new low-temperature stress-related genes specific to M. falcata. These new findings could facilitate the understanding of the low-temperature-tolerance mechanism of M. falcata.

scholarly journals Different Genes Express Analysis of root crown of alfalfa under low temperature stress

2019 ◽  
Author(s):  
Xiaolong Wang ◽  
Huiqing Jin ◽  
Kai Meng ◽  
Zhenyu Jia ◽  
Shiyuan Yan ◽  
...  
Keyword(s):  
Survival Rate ◽  
Low Temperature ◽  
Temperature Stress ◽  
Expression Patterns ◽  
Northern China ◽  
Winter Hardiness ◽  
Low Temperature Stress ◽  
Winter Survival ◽  
Rna Seq ◽  
Alfalfa Varieties

Abstract Abstract Background: Alfalfa ( Medicago sativa ) is a perennial forage crop widely cultivated in northern China. The root crown of alfalfa is an important storage organ in the process of wintering, and it is closely related to the winter hardiness of alfalfa. At present, the specific molecular mechanism of response to winter hardiness in alfalfa root crown is unclear. The transcriptome database created by RNA sequencing (RNA-seq) is widely used to identify the critical genes related to winter hardiness. Results: The transcriptomes of alfalfa varieties, such as “Lomgmu 806” (with high winter survival rate) and “Sardi” (with low winter survival rate) have been sequenced in the study. Among the identified 57,712 unigenes, 2,299 differentially expressed genes (DEGs) were up-regulated, and 2,143 unigenes were down-regulated in the Lomgmu 806 vs Sardi root crown. The KEGG pathway annotations showed that 1,159 unigenes were mainly annotated to 116 pathways. Seven DEGs belonging to “plant hormone signaling transduction”, “peroxidase” pathway and transcription factors family (MYB, B3, AP2/ERF, WRKY) genes involved in alfalfa winter hardiness. Among them, the expression patterns of seven DEGs were verified by real-time quantitative PCR (RT-qPCR) analyses, which verified the reliable results of transcriptome sequencing analyses. Conclusions: RNA-Seq was used to discover genes associated with the wintering differences between alfalfa varieties. The transcriptome data showed that the gene regulation response of alfalfa to low temperature stress, which provides a valuable resource for further identification and functional analysis of candidate genes for winter hardiness of alfalfa. In addition, these data provide references for future study of genetic breeding and winter hardiness in alfalfa.

scholarly journals Transcriptomic, proteomic, and physiological comparative analyses of flooding mitigation of the damage induced by low-temperature stress in direct seeded early indica rice at the seedling stage

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Wenxia Wang ◽  
Jie Du ◽  
Liming Chen ◽  
Yongjun Zeng ◽  
Xueming Tan ◽  
...  
Keyword(s):  
Low Temperature ◽  
Temperature Stress ◽  
Molecular Mechanisms ◽  
Tricarboxylic Acid Cycle ◽  
Indica Rice ◽  
Low Temperature Stress ◽  
Seedling Stage ◽  
Differentially Expressed ◽  
Rice Seedlings ◽  
Direct Seeded

Abstract Background Low temperature (LT) often occurs at the seedling stage in the early rice-growing season, especially for direct seeded early-season indica rice, and using flooding irrigation can mitigate LT damage in rice seedlings. The molecular mechanism by which flooding mitigates the damage induced by LT stress has not been fully elucidated. Thus, LT stress at 8 °C, LT accompanied by flooding (LTF) and CK (control) treatments were established for 3 days to determine the transcriptomic, proteomic and physiological response in direct seeded rice seedlings at the seedling stage. Results LT damaged chloroplasts, and thylakoid lamellae, and increased osmiophilic bodies and starch grains compared to CK, but LTF alleviated the damage to chloroplast structure caused by LT. The physiological characteristics of treated plants showed that compared with LT, LTF significantly increased the contents of rubisco, chlorophyll, PEPCK, ATP and GA3 but significantly decreased soluble protein, MDA and ABA contents. 4D-label-free quantitative proteomic profiling showed that photosynthesis-responsive proteins, such as phytochrome, as well as chlorophyll and the tricarboxylic acid cycle were significantly downregulated in LT/CK and LTF/CK comparison groups. However, compared with LT, phytochrome, chlorophyllide oxygenase activity and the glucan branching enzyme in LTF were significantly upregulated in rice leaves. Transcriptomic and proteomic studies identified 72,818 transcripts and 5639 proteins, and 4983 genes that were identified at both the transcriptome and proteome levels. Differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were significantly enriched in glycine, serine and threonine metabolism, biosynthesis of secondary metabolites, glycolysis/gluconeogenesis and metabolic pathways. Conclusion Through transcriptomic, proteomic and physiological analyses, we determined that a variety of metabolic pathway changes were induced by LT and LTF. GO and KEGG enrichment analyses demonstrated that DEGs and DEPs were associated with photosynthesis pathways, antioxidant enzymes and energy metabolism pathway-related proteins. Our study provided new insights for efforts to reduce the damage to direct seeded rice caused by low-temperature stress and provided a breeding target for low temperature flooding-resistant cultivars. Further analysis of translational regulation and metabolites may help to elucidate the molecular mechanisms by which flooding mitigates low-temperature stress in direct seeded early indica rice at the seedling stage.

scholarly journals Differential expression analysis of genes in the root crown of alfalfa under low-temperature stress

2019 ◽  
Author(s):  
Xiaolong Wang ◽  
Huiqing Jin ◽  
Kai Meng ◽  
Zhenyu Jia ◽  
Shiyuan Yan ◽  
...  
Keyword(s):  
Low Temperature ◽  
Temperature Stress ◽  
Expression Patterns ◽  
Differential Expression Analysis ◽  
Northern China ◽  
Winter Hardiness ◽  
Low Temperature Stress ◽  
Rna Seq ◽  
Real Time Quantitative Pcr ◽  
Alfalfa Varieties

Abstract Background: Alfalfa ( Medicago sativa ) is a perennial forage crop widely cultivated in northern China. The root crown is an important storage organ of alfalfa, especially in the wintering process, as it is closely related to winter hardiness. However, the molecular mechanism underlying the winter hardiness of the alfalfa root crown remains unclear. To investigate these gaps in knowledge, the RNA sequencing (RNA-Seq) technology was used to identify critical genes related to winter hardiness. Results: In this study, the winter survival rate of the Lomgmu 806 variety was approximately 3.68-fold higher than that of the Sardi variety. We sequenced the transcriptomes of the root crown of the two alfalfa varieties. Among the 57,712 unigenes identified, 2,299 differentially expressed genes (DEGs) were upregulated, and 2,143 DEGs were downregulated. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotations showed that 1,159 unigenes were mainly annotated in 116 pathways. Seven DEGs belonging to the “plant hormone signalling transduction” pathway, the “peroxisome” pathway and transcription factor family (MYB, B3, AP2/ERF, and WRKY) and involved in alfalfa winter hardiness were identified. As a result, the expression patterns of seven DEGs were verified by real-time quantitative PCR (RT-qPCR) analyses, which verified the reliability of the RNA-Seq analyses. Conclusions: The RNA-Seq data revealed the gene regulation response of alfalfa to low-temperature stress, which provides a valuable resource for the further identification and functional analysis of candidate genes related to winter hardiness in alfalfa. Furthermore, these data provide references for future in-depth studies of winter hardiness mechanisms in alfalfa.

scholarly journals Transcriptome Analysis of Photosynthetic Characteristics was Induced by Low Temperature Stress in Brassica napus L.

2019 ◽  
Author(s):  
Jin Jiaojiao ◽  
Liu Zigang ◽  
Mi Wenbo ◽  
Sun Wancang ◽  
Wu Junyan ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Low Temperature ◽  
Temperature Stress ◽  
Photosynthetic Characteristics ◽  
Low Temperature Stress ◽  
Rna Seq ◽  
Brassica Napus L ◽  
Regulated Expression ◽  
Tolerant Variety ◽  
Cold Tolerant

Abstract Background RNA Sequencing (RNA-Seq) technique could be utilized to compare the transcription groups of two different cold-resistant rapeseed leaves responding to low temperature at the seedling stage, analyze the photosynthetic characteristics of rapeseed subjected to low temperature stress, and identify the related genes for low temperature induction in rapeseed leaves. Results Using cold-tolerant variety 17NS and sensitive variety NF24 as experimental materials, carrying out RNA-Seq analysis by photosynthetic parameter determination and Illumina HiSeqTM platform. and screen out the KEGG significant enrichment pathway related to photosynthetic characteristics under low temperature stress. Differential Expressed Genes (DEGs) were used for real-time PCR to verify the reliability of RNA-Seq results. The results showed that the response of Brassica napus L. to low temperature stress mainly was achieved by inhibiting photosynthesis, the cold-tolerant variety 17NS had a strong ability to maintain membrane system stability and structural integrity after 24 h of low temperature stress, while the sensitive variety NF24 photosynthesis was significantly inhibited. Two pathways of Photosynthesis and Photosynthesis-antennas, which were significantly correlated with photosynthetic characteristics and low temperature stress were screened by KEGG enrichment. The results of DEGs indicated that 64 differentially expressed genes in these two pathways were induced by low temperature stress, and 8 of them were up-regulated expression and 56 of them were down-regulated expression. The expression pattern of DEGs was consistent with the results of RNA-Seq analysis by qRT-PCR detection and confirmed the reliability of RNA-Seq results. Conclusion Our study analysis and identified 17 low-temperature-induced photosynthetic-related candidate genes in Brassica napus L., and the GO and KEGG metabolic pathways clarified the molecular function of differentially expressed genes.

scholarly journals Full-length transcriptome sequencing reveals a low-temperature-tolerance mechanism in Medicago falcata roots

2019 ◽  
Author(s):  
Guowen Cui ◽  
Hua Chai ◽  
Hang Yin ◽  
Mei Yang ◽  
Guofu Hu ◽  
...  
Keyword(s):  
Low Temperature ◽  
Temperature Stress ◽  
Electrolyte Leakage ◽  
Temperature Tolerance ◽  
Low Temperature Stress ◽  
Full Length ◽  
Rna Seq ◽  
Tolerance Mechanism ◽  
Medicago Falcata ◽  
Low Temperature Tolerance

Abstract Background Low temperature is one of the main environmental factors that limits crop growth, development and production. Medicago falcata is an economically and ecologically important legume that is closely related to alfalfa and exhibits better tolerance to low temperature than alfalfa. Understanding the low-temperature-tolerance mechanism of M. falcata is important for the genetic improvement of alfalfa. Results In this study, we explored the transcriptomic changes in low-temperature-treated M. falcata roots by combining SMRT and NGS technologies. A total of 115,153 nonredundant sequences were obtained, and 8,849 AS events, 73,149 SSRs and 4,189 LncRNAs were predicted. A total of 111,587 genes from SMRT were annotated, and 11,369 DEGs were identified in this paper that are involved in plant hormone signal transduction, protein processing in endoplasmic reticulum, carbon metabolism, glycolysis/gluconeogenesis, starch and sucrose metabolism, and endocytosis pathways. We characterized 1,538 TF genes into 45 TF gene families, and the most abundant TF family was WRKY, followed by ERF, MYB, bHLH and NAC. A total of 134 genes were differentially coexpressed at all five temperature points, including 101 upregulated genes and 33 downregulated genes. PB40804, PB75011, PB110405 and PB108808 were found to play crucial roles in the tolerance of M. falcata to low temperature. The WGCNA results showed that the MEbrown module was significantly correlated with low-temperature stress in M. falcata. Electrolyte leakage was correlated with most genetic modules and corroborated that electrolyte leakage can be used as direct stress markers to reflect cell membrane damage from low-temperature stress in physiological assays. The consistency between the qRT-PCR results and RNA-Seq analyses confirm the validity of the RNA-Seq data and the analysis of the regulation of low-temperature stress in the transcriptome. Conclusions The full-length transcripts generated in this study provided a full characterization of the gene transcription of M. falcata and are useful for mining new low-temperature stress-related genes specific to M. falcata. These new findings facilitate the understanding of low-temperature-tolerance mechanisms in M. falcata.

scholarly journals Full-length transcriptome sequencing reveals the low-temperature-tolerance mechanism of Medicago falcata roots

2019 ◽  
Author(s):  
Guowen Cui ◽  
Hua Chai ◽  
Hang Yin ◽  
Mei Yang ◽  
Guofu Hu ◽  
...  
Keyword(s):  
Low Temperature ◽  
Temperature Stress ◽  
Electrolyte Leakage ◽  
Temperature Tolerance ◽  
Low Temperature Stress ◽  
Full Length ◽  
Rna Seq ◽  
Smrt Sequencing ◽  
Tolerance Mechanism ◽  
Low Temperature Tolerance

Abstract Background: Low temperature is one of the main environmental factors that limits crop growth, development, and production. Medicago falcata is an important leguminous herb that is widely distributed worldwide. M. falcata is related to alfalfa but is more tolerant to low temperature than alfalfa. Understanding the low temperature tolerance mechanism of M. falcata is important for the genetic improvement of alfalfa. Results: In this study, we explored the transcriptomic changes in the roots of low-temperature-treated M. falcata plants by combining SMRT sequencing and NGS technologies. A total of 115,153 nonredundant sequences were obtained, and 8,849 AS events, 73,149 SSRs, and 4,189 lncRNAs were predicted. A total of 111,587 genes from SMRT sequencing were annotated, and 11,369 DEGs involved in plant hormone signal transduction, protein processing in endoplasmic reticulum, carbon metabolism, glycolysis/gluconeogenesis, starch and sucrose metabolism, and endocytosis pathways were identified. We characterized 1,538 TF genes into 45 TF gene families, and the most abundant TF family was the WRKY family, followed by the ERF, MYB, bHLH and NAC families. A total of 134 genes, including 101 whose expression was upregulated and 33 whose expression was downregulated, were differentially coexpressed at all five temperature points. PB40804, PB75011, PB110405 and PB108808 were found to play crucial roles in the tolerance of M. falcata to low temperature. WGCNA revealed that the MEbrown module was significantly correlated with low-temperature stress in M. falcata. Electrolyte leakage was correlated with most genetic modules and verified that electrolyte leakage can be used as a direct stress marker in physiological assays to indicate cell membrane damage from low-temperature stress. The consistency between the qRT-PCR results and RNA-seq analyses confirmed the validity of the RNA-seq data and the analysis of the regulatory mechanism of low-temperature stress on the basis of the transcriptome. Conclusions: The full-length transcripts generated in this study provide a full characterization of the transcriptome of M. falcata and may be useful for mining new low-temperature stress-related genes specific to M. falcata. These new findings could facilitate the understanding of the low-temperature-tolerance mechanism of M. falcata.

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