scholarly journals Transcriptomic profiling of germinating seeds under cold stress and characterization of the cold-tolerant gene LTG5 in rice

2020 ◽  
Author(s):  
yinghua Pan ◽  
Haifu Liang ◽  
Lijun Gao ◽  
Gaoxing Dai ◽  
Weiwei Chen ◽  
...  

Abstract Background: Wild rice ( Oryza rufipogon Griff.) is an important germplasm resource for rice improvement. It has superior tolerance for many abiotic stresses including cold stress, but little is known about the mechanism underlying its resistance to cold. Low temperature is one of the most prevalent factors that limit rice productivity and geographical distribution.Results: This study aimed to elucidate the molecular genetic mechanisms of wild rice in tolerating low temperature. Comprehensive transcriptome profiles of two rice genotypes (cold-sensitive ce 253 and cold-tolerant Y12-4) at the germinating stage under cold stress were comparatively analyzed. A total of 42.44–68.71 million readings were obtained, resulting in the alignment of 29128 and 30131 genes in genotypes 253 and Y12-4, respectively. Many common and differentially expressed genes (DEGs) were analyzed in cold-sensitive and cold-tolerant genotypes. Results showed more upregulated DEGs in cold-tolerant genotypes than in cold-sensitive genotypes at four stages under cold stress. Gene ontology enrichment analyses indicated more upregulated genes than downregulated ones in cold-tolerant genotypes based on cellular process, metabolic process, response stimulus, membrane part, and catalytic activity. To confirm the RNA Sequencing (RNA-seq) data, Quantitative real time polymerase chain reaction (qRT-PCR) was performed on seven randomly selected DEGs. These genes showed similar expression patterns corresponding with the RNA-Seq method. We also explored a gene for cold tolerance LTG5 , encoding a UDP-glucosyltransferase. The overexpression of LTG5 gene conferred cold tolerance to indica rice.Conclusion: Overall, our results suggested that gene resources related to cold stress from wild rice can be valuable for improving the cold tolerance of crop plants.

2020 ◽  
Author(s):  
Yinghua Pan ◽  
Haifu Liang ◽  
Lijun Gao ◽  
Gaoxing Dai ◽  
Weiwei Chen ◽  
...  

Abstract Background: Low temperature is a limiting factor of rice productivity and geographical distribution. Wild rice (Oryza rufipogon Griff.) is an important germplasm resource for rice improvement. It has superior tolerance to many abiotic stresses, including cold stress, but little is known about the mechanism underlying its resistance to cold. Results: This study elucidated the molecular genetic mechanisms of wild rice in tolerating low temperature. Comprehensive transcriptome profiles of two rice genotypes (cold-sensitive ce 253 and cold-tolerant Y12-4) at the germinating stage under cold stress were comparatively analyzed. A total of 42.44–68.71 million readings were obtained, resulting in the alignment of 29 128 and 30 131 genes in genotypes 253 and Y12-4, respectively. Many common and differentially expressed genes (DEGs) were analyzed in the cold-sensitive and cold-tolerant genotypes. Results showed more upregulated DEGs in the cold-tolerant genotype than in the cold-sensitive genotype at four stages under cold stress. Gene ontology enrichment analyses based on cellular process, metabolic process, response stimulus, membrane part, and catalytic activity indicated more upregulated genes than downregulated ones in the cold-tolerant genotype than in the cold-sensitive genotype. Quantitative real-time polymerase chain reaction was performed on seven randomly selected DEGs to confirm the RNA Sequencing (RNA-seq) data. These genes showed similar expression patterns corresponding with the RNA-Seq method. Weighted gene co-expression network analysis(WGCNA)revealed Y12-4 showed more positive genes than 253 under cold stress. We also explored the cold tolerance gene LTG5 (Low Temperature Growth 5) encoding a UDP-glucosyltransferase. The overexpression of the LTG5 gene conferred cold tolerance to indica rice. Conclusion: Gene resources related to cold stress from wild rice can be valuable for improving the cold tolerance of crops.


2020 ◽  
Author(s):  
Changbing Huang ◽  
Chun Jiang ◽  
limin Jin ◽  
Huanchao Zhang

Abstract Background:Hemerocallis fulva is a perennial herb belonging to Hemerocallis of Hemerocallis. Because of the large and bright colors, it is often used as a garden ornamental plant. But most varieties of H. fulva on the market will wither in winter, which will affect their beauty. It is very important to study the effect of low temperature stress on the physiological indexes of H. fulva and understand the cold tolerance of different H. fulva. MiRNA is a kind of endogenous non coding small molecular RNA with length of 21-24nt. It mainly inhibits protein translation by cutting target genes, and plays an important role in the development of organisms, gene expression and biological stress. Low temperature is the main abiotic stress affecting the production of H. fulva in China, which hinders the growth and development of plants. A comprehensive understanding of the expression pattern of microRNA in H. fulva under low temperature stress can improve our understanding of microRNA mediated stress response. Although there are many studies on miRNAs of various plants under cold stress at home and abroad, there are few studies on miRNAs related to cold stress of H. fulva. It is of great significance to explore the cold stress resistant gene resources of H. fulva, especially the identification and functional research of miRNA closely related to cold stress, for the breeding of excellent H. fulva.Results A total of 5619 cold-responsive miRNAs, 315 putative novel and 5 304 conserved miRNAs, were identified from the leaves and roots of two different varieties ‘Jinyan’ (cold-tolerant) and ‘Lucretius ’ (cold-sensitive), which were stressed under -4 oC for 24 h. Twelve conserved and three novel miRNAs (novel-miR10, novel-miR19 and novel-miR48) were differentially expressed in leaves of ‘Jinyan’ under cold stress. Novel-miR19, novel-miR29 and novel-miR30 were up-regulated in roots of ‘Jinyan’ under cold stress. Thirteen and two conserved miRNAs were deferentially expressed in leaves and roots of ‘Lucretius’ after cold stress. The deferentially expressed miRNAs between two cultivars under cold stress include novel miRNAs and the members of the miR156, miR166 and miR319 families. A total of 6 598 target genes for 6 516 known miRNAs and 82 novel miRNAs were predicted by bioinformatic analysis, mainly involved in metabolic processes and stress responses. Ten differentially expressed miRNAs and predicted target genes were confirmed by quantitative reverse transcription PCR(q-PCR), and the expressional changes of target genes were negatively correlated to differentially expressed miRNAs. Our data indicated that some candidate miRNAs (e.g., miR156a-3-p, miR319a, and novel-miR19) may play important roles in plant response to cold stress.Conclusions Our study indicates that some putative target genes and miRNA mediated metabolic processes and stress responses are significant to cold tolerance in H. fulva.


Forests ◽  
2019 ◽  
Vol 10 (11) ◽  
pp. 1039
Author(s):  
Qianqian Wang ◽  
Jing Tao ◽  
Yurong Li ◽  
Yabei Xu ◽  
Xinhai Liu ◽  
...  

Eogystia hippophaecola Hua, Chou, Fang et Chen (Lepidoptera: Cossidae) is an important borer pest of the sea buckthorn forest (Hippophae rhamnoides L.) in China. Its larvae, which are highly cold tolerant, mainly overwinter in sea buckthorn roots. Heat shock proteins (Hsps) are important molecular chaperones that have been linked to cold tolerance in insects. In this study, we cloned the open reading frames (ORFs) of two Hsp90 genes from E. hippophaecola, EhHsp90-1 and EhHsp90-2, and analyzed their expression under cold stress by qRT-PCR. EhHsp90-1 and EhHsp90-2 are 2154 and 2346 bp in length, respectively, encoding 717 and 781 amino acids. The deduced amino acid sequences contain the conserved signature sequences of the Hsp90 family and the C-terminus characteristic sequence of cytoplasmic or endoplasmic reticulum Hsp90 protein. Phylogenetic analysis revealed the amino acid sequences of EhHsp90-1 and EhHsp90-2 were very similar to the corresponding proteins from Lepidoptera. Under various low-temperature treatments lasting 2 h, EhHsp90-1 and EhHsp90-2 exhibited similar expression patterns, increasing first and then decreasing. At −5 °C, EhHsp90-1 was significantly up-regulated after 12 h, whereas EhHsp90-2 was up-regulated after just 1 h and reached its highest level at 2 h; however, the overall degree of upregulation was greater for EhHsp90-1. Subsequently, the expression level of EhHsp90-2 fluctuated with time. Our results suggest that the two Hsp90s play important roles in E. hippophaecola larvae response to cold stress, but that their response times and the magnitudes of their responses to low-temperature stress differed significantly, providing a theoretical basis for further studying the molecular mechanism of cold tolerance in E. hippophaecola larvae.


Author(s):  
Maciej Jończyk ◽  
Alicja Sobkowiak ◽  
Joanna Trzcinska-Danielewicz ◽  
Paweł Sowiński

Abstract Maize is a subtropical, cold-sensitive species. However, some varieties of this species have the potential to withstand long-term low temperatures, even at the seedling stage. The molecular basis of this phenomenon has not been determined. In a chromatin-level study, we compared the cold-stress reaction of seedlings of two maize inbred lines showing contrasting levels of cold sensitivity. The cold-tolerant line was selected based on field data and previous physiological and transcriptomic level studies. The first condition of gene expression—chromatin accessibility—was assessed by formaldehyde-aided isolation of regulatory elements method and DNA sequencing. Potentially expressed genes and cis-regulatory sequences open for interaction with transcription factors have been defined. The results of this study suggest that during cold stress, the tolerant maize line shifted resources from growth to defense. This shift was shown by potential hormone-level events—degradation of growth-promoting gibberellins and synthesis of jasmonic and abscisic acids. This finding is congruent with the xeromorphic morphology of seedlings of the cold-tolerant line and their ability to regrow when stress ceases. It is a common reaction of cold-tolerant maize lines. Moreover, in the cold-tolerant line, several genes from the low-temperature signaling pathways were potentially expressed. Additionally, numerous stress-response AP2/EREBP-binding cis-motifs were accessible in the cold-tolerant line. Differently in the cold-sensitive B73 line, MADS-binding cis-motifs were the most abundant. Development of the photosynthetic apparatus is crucial for the survival of maize seedlings at low temperature. Our results suggest efficient photosynthesis in seedlings of the cold-tolerant line, as was described earlier in physiological-level analyses.


2019 ◽  
Vol 20 (20) ◽  
pp. 5089 ◽  
Author(s):  
Hui Guo ◽  
Tingkai Wu ◽  
Shuxing Li ◽  
Qiang He ◽  
Zhanlie Yang ◽  
...  

Chilling stress is considered the major abiotic stress affecting the growth, development, and yield of rice. To understand the transcriptomic responses and methylation regulation of rice in response to chilling stress, we analyzed a cold-tolerant variety of rice (Oryza sativa L. cv. P427). The physiological properties, transcriptome, and methylation of cold-tolerant P427 seedlings under low-temperature stress (2–3 °C) were investigated. We found that P427 exhibited enhanced tolerance to low temperature, likely via increasing antioxidant enzyme activity and promoting the accumulation of abscisic acid (ABA). The Methylated DNA Immunoprecipitation Sequencing (MeDIP-seq) data showed that the number of methylation-altered genes was highest in P427 (5496) and slightly lower in Nipponbare (Nip) and 9311 (4528 and 3341, respectively), and only 2.7% (292) of methylation genes were detected as common differentially methylated genes (DMGs) related to cold tolerance in the three varieties. Transcriptome analyses revealed that 1654 genes had specifically altered expression in P427 under cold stress. These genes mainly belonged to transcription factor families, such as Myeloblastosis (MYB), APETALA2/ethylene-responsive element binding proteins (AP2-EREBP), NAM-ATAF-CUC (NAC) and WRKY. Fifty-one genes showed simultaneous methylation and expression level changes. Quantitative RT-PCR (qRT-PCR) results showed that genes involved in the ICE (inducer of CBF expression)-CBF (C-repeat binding factor)—COR (cold-regulated) pathway were highly expressed under cold stress, including the WRKY genes. The homologous gene Os03g0610900 of the open stomatal 1 (OST1) in rice was obtained by evolutionary tree analysis. Methylation in Os03g0610900 gene promoter region decreased, and the expression level of Os03g0610900 increased, suggesting that cold stress may lead to demethylation and increased gene expression of Os03g0610900. The ICE-CBF-COR pathway plays a vital role in the cold tolerance of the rice cultivar P427. Overall, this study demonstrates the differences in methylation and gene expression levels of P427 in response to low-temperature stress, providing a foundation for further investigations of the relationship between environmental stress, DNA methylation, and gene expression in rice.


2020 ◽  
Author(s):  
Yan Jia ◽  
Hualong Liu ◽  
Zhaojun Qu ◽  
Jin Wang ◽  
Xinpeng Wang ◽  
...  

Abstract Background: Rice ( Oryza sativa L.), one of the most important crops cultivated in both tropical and temperate regions, has a high sensitivity to cold stress. Chilling stress limits N uptake and nitrogen metabolism in rice. To identify the genes and pathways involved in cold tolerance, specifically within the nitrogen metabolism pathway, we compared gene and protein expression differences between a cold-tolerant cultivar, Dongnong428 (DN), and a cold-sensitive cultivar, Songjing10 (SJ). Results: Using isobaric tags for relative or absolute quantification (iTRAQ) with high-throughput mRNA sequencing (RNA-seq) techniques, we identified 5,549 genes and 450 proteins in DN and 6,145 genes and 790 proteins in SJ, that were differentially expressed during low water temperature (T w ) treatment. There were 354 transcription factor (TF) genes (212 down, 142 up), and 366 TF genes (220 down, 146 up), including 47 gene families, differentially expressed in the DN under control (CKDN) vs. DN under low-T w (D15DN) and CKSJ vs. D15SJ, respectively. Genes related to rice cold-related biosynthesis pathways, particularly the mitogen-activated protein kinase (MAPK) signaling pathway, zeatin biosynthesis, and plant hormone signal transduction pathways, were significantly differentially expressed in both rice cultivars. Differentially expressed proteins (DEPs) related to rice cold-related biosynthesis pathways and particularly glutathione metabolism were significantly differentially expressed in both rice cultivars. Transcriptome and proteome analysis of the nitrogen metabolism pathways showed that major genes and proteins that participated in γ-aminobutyric acid (GABA) and glutamine synthesis were downregulated. Conclusion: Under cold stress conditions during reproductive growth, genes and proteins related to the biosynthesis pathways of cold stress were significantly differentially expressed in DN and SJ. The present study confirmed the known cold stress-associated genes and identified a number of putative new cold-responsive genes. We also found that translational regulation under cold stress plays an important role in cold-tolerant DN. Low-T w treatments affected N uptake and N metabolism in rice, and promoted Glu metabolism, and the synthesis of ornithine and proline in cold-sensitive SJ.


2020 ◽  
Author(s):  
Yinghua Pan ◽  
Lei Chen ◽  
Xinghai Yang ◽  
Lijun Gao ◽  
Baoxuan Nong ◽  
...  

Abstract Low-temperature stress significantly affects rice growth and causes serious loss of yield in temperate and high-altitude areas of the world. Rice lacks cold tolerance (CT) at germination stage, which reduces seedling vigor, hinders crop establishment and crop growth, and even affect direct-seeded rice. A chromosome segment substitution line (CSSL) population, including 271 lines, was derived from cold-tolerant donor wild rice Y11 (Oryza rufipogon Griff.) crossed with cold-sensitive rice variety GH998 to explore new genetic resource with CT and further investigate quantitative trait loci (QTLs) responsible for germination properties under low temperature. The germination rates of Y11 and GH998 were 95% and 73.33%, respectively, under cold stress. The range of variance of the germination rate in the CSSLs was between 0% and 98.33%. In addition, the CSSLs and parents were sequenced via whole genome resequencing. Results showed 33.46, 33.36, and 475.65 Gbp of clean data of GH998, Y11, and CSSLs, respectively. In 12 linkage groups, the general map distance was 941.49 cM, while the average map distance was 0.63 cM. On thebasis of 1484 bins, a high-density linkage map of the CSSLs was developed. The average distance of the linkage map ranged from 0.42 cM to 1.39 cM. The phenotype of CSSLs under low temperature and 615,466 single nucleotide polymorphisms (SNPs) between Y11 and GH998 were used for QTL analysis. Two QTLs, namely, low-temperature germination 8 (qLTG8) and qLTG11, were responsible for the germination ability under low temperature in rice. qLTG8 was mapped on chromosome 8, and it explained 14.55% of the total phenotypic variation explained (PVE) during the germination stage. qLTG8 was in 195.5 kb, and 32 genes were predicted based on the Rice Genome Annotation Project. qLTG11 was located on chromosome 11, and it explained 14.31% of the total PVE during the germination stage. qLTG11 was mapped at a narrow distance in 78.83 kb, and only 12 genes fall within this range according to the Rice Genome Annotation Project. The expression patterns of these 32 genes in the qLTG8 region demonstrated that LOC_Os08g01120, LOC_Os08g01140, LOC_Os08g01390, LOC_Os08g01170, and LOC_Os08g01380 were highly induced by cold stress in Y11 compared with GH998. The expression patterns of 12 genes in the qLTG11 region suggested that LOC_Os11g32880 and LOC_Os11g32940 were highly induced by cold stress in Y11 compared with GH998. This study provides an effective method, i.e., constructing CSSLs of wild rice, to explore excellent genes of wild rice and create new genetic resources. The results also provide a basis for identifying the genes underlying qLTG8 and qLTG11, indicating that QTL could be used for genetic improvement of CT in cultivar rice.


2001 ◽  
Vol 126 (2) ◽  
pp. 216-220 ◽  
Author(s):  
M.R. Foolad ◽  
G.Y. Lin

The genetic relationship between cold tolerance (CT) during seed germination and vegetative growth in tomato (Lycopersicon esculentum Mill.) was determined. An F2 population of a cross between accession PI120256 (cold tolerant during both seed germination and vegetative growth) and UCT5 (cold sensitive during both stages) was evaluated for germination under cold stress and the most cold tolerant progeny (the first 5% germinated) were selected. Selected progeny were grown to maturity and self-fertilized to produce F3 families (referred to as the selected F3 population). The selected F3 population was evaluated for CT separately during seed germination and vegetative growth and its performance was compared with that of a nonselected F3 population of the same cross. Results indicated that selection for CT during seed germination significantly improved CT of the progeny during germination; a realized heritability of 0.75 was obtained for CT during seed germination. However, selection for CT during germination did not affect plant CT during vegetative growth; there was no significant difference between the selected and nonselected F3 populations in either absolute CT [defined as shoot fresh weight (FW) under cold stress] or relative CT (defined as shoot FW under cold as a percentage of control). Results indicated that, in PI120256, CT during seed germination was genetically independent of CT during vegetative growth. Thus, to develop tomato cultivars with improved CT during different developmental stages, selection protocols that include all critical stages are necessary.


Horticulturae ◽  
2021 ◽  
Vol 7 (12) ◽  
pp. 572
Author(s):  
Faisal Hayat ◽  
Chengdong Ma ◽  
Shahid Iqbal ◽  
Xiao Huang ◽  
Ouma Kenneth Omondi ◽  
...  

Japanese apricot (Prunus mume) is remarkably valuable for its high ornamental and economic importance due to its distinctive features. Low temperature is a serious environmental constraint for this species, restricting its cultivation and dispersal in the north of China. To address this issue, breeding requires an understanding of the molecular mechanisms underlying responses to cold stress. We examined the leaf physiological and transcriptome profile by RNA sequencing in ‘Bungo’ scion cultivar grafted onto Prunus mume (cold-sensitive) and Prunus armeniaca (cold-tolerant) rootstocks at 4 °C for 0, 6, and 24 h. Our results revealed that the increased MDA concentration in the leaves of P. mume cultivar (cold-sensitive) suggests that cold stress might cause oxidative damage and increased sensitivity. Moreover, the cold-tolerant cultivar (P. armeniaca) considerably enhances the enzyme activities (i.e., SOD, POD, and CAT), as well as osmo-protectants (soluble sugars and proline) compared with sensitive cultivar, which helps plants to withstand oxidative damage caused by cold stress. Additionally, differentially expressed genes were shown to be enriched in plant hormone signal transduction, ribosome, MAPK signaling, and circadian rhythm pathway. After 24 h of cold stress, genes related to PYL4, histidine kinase 1, SAUR36, bHLH130, bHLH123, TIFY 6B-like, WRKY 40, WRKY 57, and 60S acidic ribosomal protein P1 were differentially expressed, implying that these DEGs involved in multiple pathways are involved in cold tolerance in Japanese apricot. This study improved our current understanding of the mechanism of cold tolerance in Japanese apricot, and the findings could be utilized for other related fruit species.


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