scholarly journals Identification of copper (Cu) stress-responsive grapevine microRNAs and their target genes by high-throughput sequencing

2019 ◽  
Vol 6 (1) ◽  
pp. 180735 ◽  
Author(s):  
Songtao Jiu ◽  
Xiangpeng Leng ◽  
Muhammad Salman Haider ◽  
Tianyu Dong ◽  
Le Guan ◽  
...  
Keyword(s):  
Plant Growth ◽  
High Throughput ◽  
Stress Responses ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Expression Patterns ◽  
Qrt Pcr ◽  
Cu Stress ◽  
Gene Information

MicroRNAs (miRNAs) are a class of single-stranded non-coding small RNAs (sRNAs) that are 20–24 nucleotides (nt) in length. Extensive studies have indicated that miRNAs play important roles in plant growth, development and stress responses. With more copper (Cu) and copper containing compounds used as bactericides and fungicides in plants, Cu stress has become one of the serious environmental problems that affect plant growth and development. In order to uncover the hidden response mechanisms of Cu stress, two small RNA libraries were constructed from Cu-treated and water-treated (Control) leaves of ‘Summer Black’ grapevine. Following high-throughput sequencing and filtering, a total of 158 known and 98 putative novel miRNAs were identified in the two libraries. Among these, 100 known and 47 novel miRNAs were identified as differentially expressed under Cu stress. Subsequently, the expression patterns of nine Cu-responsive miRNAs were validated by quantitative real-time PCR (qRT-PCR). There existed some consistency in expression levels of Cu-responsive miRNAs between high throughput sequencing and qRT-PCR assays. The targets prediction of miRNAs indicates that miRNA may regulate some transcription factors, including AP2, SBP, NAC, MYB and ARF during Cu stress. The target genes for two known and two novel miRNAs showed specific cleavage sites at the 10th and/or 11th nucleotide from the 5′-end of the miRNA corresponding to their miRNA complementary sequences. The findings will lay the foundation for exploring the role of the regulation of miRNAs in response to Cu stress and provide valuable gene information for breeding some Cu-tolerant grapevine cultivars.

scholarly journals Identification of Drought-Responsive microRNAs from Roots and Leaves of Alfalfa by High-Throughput Sequencing

2017 ◽  
Author(s):  
Yue Li ◽  
Liqiang Wan ◽  
Shuyi Bi ◽  
Xiufu Wan ◽  
Zhenyi Li ◽  
...  
Keyword(s):  
Drought Stress ◽  
High Throughput ◽  
Stress Responses ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Plant Stress ◽  
Online Tool ◽  
Bioenergy Plant ◽  
Plant Stress Responses

Alfalfa, an important legume forage, is an ideal crop for sustainable agriculture and a potential bioenergy plant. Drought, one of the most common environmental stresses, substantially affects plants’ growth, development and productivity. MicroRNAs (miRNAs) are newly discovered gene expression regulators that have been linked to several plant stress responses. To elucidate the role of miRNAs in drought stress regulation of alfalfa, a high-throughput sequencing approach was used to analyze 12 small RNA libraries comprising of 4 samples, each with 3 biological replicates. We identified 348 known miRNAs, belonging to 80 miRNA families, from the 12 libraries and 281 novel miRNAs using Mireap software. 18 known miRNAs in roots and 12 known miRNAs in leaves were screened out as drought-responsive miRNAs. Except for miR319d and miR157a which were upregulated under drought stress, the expression pattern of drought-responsive miRNAs were different between roots and leaves in alfalfa. This is the first study discovering miR157a, miR1507, miR3512, miR3630, miR5213, miR5294, miR5368 and miR6173 are drought-responsive miRNAs. Target transcripts of drought-responsive miRNAs were computationally predicted. All 447 target genes for the known miRNAs were predicted using an online tool. This study provides a significant insight on understanding drought-responsive mechanisms of alfalfa.

scholarly journals High-throughput sequencing analysis identified microRNAs associated with egg production in ducks ovaries

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8440
Author(s):  
Mohan Qiu ◽  
Zengrong Zhang ◽  
Xia Xiong ◽  
Huarui Du ◽  
Qingyun Li ◽  
...  
Keyword(s):  
High Throughput ◽  
Egg Production ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Expression Patterns ◽  
Egg Laying ◽  
Sequencing Analysis ◽  
Pathway Network ◽  
Qrt Pcr ◽  
Meat Type

Background MicroRNAs (miRNAs) exist widely and are involved in multiple biological processes in ducks, whereas the regulatory mechanism of miRNAs in egg laying of ducks has remained unclear. This study aims to reveal key miRNAs involved in the regulation of egg production in duck ovaries. Methods High-throughput sequencing was performed on four egg-type duck ovaries and four egg-meat-type duck ovaries at the start of the egg-laying stage. Quantitative reverse transcription PCR (qRT-PCR) validation was performed on differentially expressed miRNAs (DE miRNAs). Gene network of DEmiRNA-mRNA-pathway was constructed by Cytoscape. Results A total of 251 know miRNAs and 1,972 novel miRNAs were obtained from whole clean reads. Among the known miRNAs, we identified 21 DEmiRNAs, including eight down-regulated and 13 up-regulated miRNAs in egg-type ducks compared with egg-meat-type ducks. Among the novel miRNAs, we identified 70 DEmiRNAs, including 58 down-regulated and 12 up-regulated in egg-type ducks compared with egg-meat-type ducks. The expression patterns of four miRNAs were verified by qRT-PCR. The DEmiRNAs were involved in the function of response to folic acid and the pathway of valine, leucine and isoleucine degradation. Specific target genes of DEmiRNAs enrichment was found in some egg-laying regulation pathways, such as dopaminergic synapse, ovarian steroidogenesis and oocyte meiosis. The DEmiRNA-mRNA-pathway network including three DEmiRNAs, nine mRNAs and 11 pathways. apl-miR-194-5p and apl-miR-215-5p may be potential key miRNAs in regulating egg laying. Conclusions This study provided miRNAs profiles in ducks about egg laying and establish a theoretical basis for subsequent selection or modification of duck phenotypes at the molecular level.

scholarly journals The Effect of Different Pollination on the Expression of Dangshan Su Pear MicroRNA

2017 ◽  
Vol 2017 ◽  
pp. 1-18 ◽  
Author(s):  
Xi Cheng ◽  
Chongchong Yan ◽  
Jinyun Zhang ◽  
Chenhui Ma ◽  
Shumei Li ◽  
...  
Keyword(s):  
High Throughput ◽  
Fruit Quality ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Cell Content ◽  
Qrt Pcr ◽  
Stone Cell ◽  
Negative Effect

The high-throughput sequencing of pear “Dangshan Su” × “Yali” (whose fruits lignin and stone cell content are high and quality is poor) and pear “Dangshan Su” × “Wonhwang” (whose fruits with low content of lignin and stone cell and the quality are better ) found that the expressions of these two miRNAs (pyr-1809 and pyr-novel-miR-144-3p) were significantly different; their corresponding target genes encode two kinds of laccase (Pbr018935.1 and Pbr003857.1). qRT-PCR results showed that these two enzymes are involved in the formation of lignin and stone cells and the existence of these two miRNAs has a negative effect on them. It was concluded that the effect of pollination on the development of stone cells may affect the synthesis of lignin, through the regulation of laccase controlled by miRNAs, and ultimately affect the formation of stone cell and fruit quality.

miR166h Directed Cleavage of Target Upon PGPR Inoculation Under Drought Stress and Tissue-Specific Expression Analysis Under Abiotic Stresses in Chickpea.

2021 ◽  
Author(s):  
Ankita Yadav ◽  
Sanoj Kumar ◽  
Rita Verma ◽  
Shashi Pandey Rai ◽  
Charu Lata ◽  
...  
Keyword(s):  
Abiotic Stress ◽  
Plant Growth ◽  
Stress Responses ◽  
Expression Patterns ◽  
Crop Improvement ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Growth Promoting

Abstract Legumes are an indispensable food after cereals with extensive production across the world. The legume production is imposed with limitations and has been augmented by various environmental stresses. The symbiotic relations between legumes and rhizobacteria have been an intriguing topic of research in view of their roles in plant growth, development and various stress responses. Recent advances on gene networks involving plethora of evolutionarily conserved miRNAs have been investigated pertaining to their roles in plant stress responses. The interaction between plant growth promoting rhizobacteria (PGPR) strain Pseudomonas putida RA, MTCC5279 and abiotic stress responsive miRNAs have previously been studied with roles in abiotic stress mitigation by modulating stress responsive miRNAs and their target genes. The present studyis an investigation involving the role of RA in abiotic stress responsive miR166h for drought mitigation in tolerant desi chickpea genotype. miRNA166 directed cleavage of its target, ATHB15 has been drifted of drought treated plantlets upon RA inoculation using 5´RLM-RACE analysis. Drought stressed chickpea plants when inoculated with growth promoting rhizobacteria, RA, the inverse correlation in expression patterns were noticed in miR166h and its validated target, ATHB15. Tissue-specific expression patterns in 15 days old chickpea seedlings including leaves, shoot and roots when exposed to salinity, drought and abscisic acid at different time points indicated the role of miR166 in different abiotic stress response. In view of the results, validation and functional characterization of such interactions involving stress responsive miRNAs along with microbial stress management techniques could be an important technique for crop improvement.

scholarly journals Overexpression of Medicago sativa cv. Pianguan miR397-5p gene enhances drought tolerance in K326 tobacco

2020 ◽  
Author(s):  
Huisen Zhu ◽  
Rong Jia ◽  
Jiali Tao ◽  
Junbing Jiang ◽  
Kuanhu Dong
Keyword(s):  
Drought Stress ◽  
Plant Growth ◽  
Medicago Sativa ◽  
Stress Responses ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Abiotic Factors ◽  
Differential Expression Analysis ◽  
Plasmid Vector ◽  
Sequencing Analysis

Abstract Background Alfalfa (Medicago sativa cv. Pianguan) is one of the most widely cultivated perennial leguminous forage. Drought is one of the major abiotic factors that affect alfalfa productivity. MicroRNAs (miRNAs) have been proved playing important roles in plant growth, development and stress response. Results In this study, High-throughput sequencing (HTSeq) was used to identify the miRNAs and their target related to drought stress from polyethylene glycol treated and control alfalfa samples, differential expression analysis showed that a large number of miRNAs were down-regulated or up-regulated as a result of PEG-6000 stress. qRT-PCR and sequencing analysis results showed that the relative expression of 9 miRNAs and their target genes related to drought stress, 3 miRNAs were considered potential drought-responsive miRNAs, including miR159b, miR397-5p and unconservative_6_4438. The recombinant plasmid vector pBWA(V)KS-miR397 was constructed and introduced into tobacco successfully. 31 positive seedlings were obtained through the process of infection, co-culture, differentiation and rooting. The physiologieal variation of transgenic tobacco under drought stress was researched. Finally, a new tobacco variety with high drought resistance was obtained. Conclusion These finding although need further studies to confirm, these data provide a useful evidence for the possible involvement of miRNAs in the process of drought response in the alfalfa plant which could help explain the drought stress responses and to alleviate the adverse effects of drought stress on plant growth and development.

scholarly journals snoDB: an interactive database of human snoRNA sequences, abundance and interactions

2019 ◽  
Vol 48 (D1) ◽  
pp. D220-D225 ◽  
Author(s):  
Philia Bouchard-Bourelle ◽  
Clément Desjardins-Henri ◽  
Darren Mathurin-St-Pierre ◽  
Gabrielle Deschamps-Francoeur ◽  
Étienne Fafard-Couture ◽  
...  
Keyword(s):  
High Throughput ◽  
Ribosome Biogenesis ◽  
High Throughput Sequencing ◽  
Expression Patterns ◽  
Sequencing Data ◽  
Ribonucleoprotein Complexes ◽  
High Throughput Sequencing Data ◽  
Non Coding Rna ◽  
Rna Interaction

Abstract Small nucleolar RNAs (snoRNAs) are an abundant type of non-coding RNA with conserved functions in all known eukaryotes. Classified into two main families, the box C/D and H/ACA snoRNAs, they enact their most well characterized role of guiding site specific modifications in ribosomal RNA, through the formation of specific ribonucleoprotein complexes, with fundamental implications in ribosome biogenesis. However, it is becoming increasingly clear that the landscape of snoRNA cellular functionality is much broader than it once seemed with novel members, non-uniform expression patterns, new and diverse targets as well as several emerging non-canonical functions ranging from the modulation of alternative splicing to the regulation of chromatin architecture. In order to facilitate the further characterization of human snoRNAs in a holistic manner, we introduce an online interactive database tool: snoDB. Its purpose is to consolidate information on human snoRNAs from different sources such as sequence databases, target information, both canonical and non-canonical from the literature and from high-throughput RNA–RNA interaction datasets, as well as high-throughput sequencing data that can be visualized interactively.

scholarly journals Identification and Characterization of Known and Novel MicroRNAs in Five Tissues of Wax Gourd (Benincasa hispida) Based on High-Throughput Sequencing

2021 ◽  
Vol 11 (21) ◽  
pp. 10068
Author(s):  
Jinqiang Yan ◽  
Min Wang ◽  
Wenrui Liu ◽  
Dasen Xie ◽  
Xiaoming He ◽  
...  
Keyword(s):  
High Throughput ◽  
Small Rna ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Expression Patterns ◽  
Small Rna Sequencing ◽  
Specific Expression ◽  
Nac Transcription Factors ◽  
Benincasa Hispida ◽  
Wax Gourd

MicroRNAs (miRNAs) are endogenous single-stranded non-coding small RNAs of 20-24 nucleotides and play important roles in many plant biological and metabolic processes. Wax gourd is an important vegetable of Cucurbitacea family, with great economic and medicinal value. Although miRNAs have been extensively studied in model plant species, less is known in wax gourd (Benincasa hispida). In this study, in order to identify miRNAs in wax groud, five independent small RNA libraries were constructed using leaf, root, stem, flower, and fruit of B227. Based on high-throughput Illumina deep sequencing. In total, 422 known and 409 novel miRNAs were identified from five libraries. Comparative analysis revealed that many miRNAs were differentially expressed among different tissues, indicating tissue-specific expression of some miRNAs. qRT-PCR verified the reliability of small RNA sequencing results. Furthermore, miRNAs with similar expression patterns among five tissues were clustered into the same profile, among which many miRNAs were found with relatively high expression in the fruit of wax gourd. MiR164-x had the highest expression in fruit than in other tissues and many NAC transcription factors were predicted as its target genes. We propose that miR164 might regulate fruit development by forming miR164-NAC module in wax gourd. Taken together, this study provides the first global miRNAs profiling of wax gourd, and lays the foundation for understanding the regulatory roles of miRNAs in the growth and development processes of wax gourd.

Bioinformatics Analysis and Validation of Differentially Expressed MicroRNAs with Their Target Genes Involved in GLP-1RA Facilitated Osteogenesis

2020 ◽  
Vol 15 ◽  
Author(s):  
Na Wang ◽  
Yukun Li ◽  
Sijing Liu ◽  
Liu Gao ◽  
Chang Liu ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Target Genes ◽  
Bioinformatics Analysis ◽  
Differentially Expressed ◽  
Functional Study ◽  
Regulation Network ◽  
Glucagon Like Peptide 1 ◽  
G Protein Coupled ◽  
Lower Expression

Background: Recent studies revealed that the hypoglycemic hormone, glucagon-like peptide-1 (GLP-1), acted as an important modulator in osteogenesis of bone marrow derived mesenchymal stem cells (BMSCs). Objectives: The aim of this study was to identify the specific microRNA (miRNA) using bioinformatics analysis and validate the presence of differentially expressed microRNAs with their target genes after GLP-1 receptor agonist (GLP-1RA) administration involved in ostogenesis of BMSCs. Methods: MiRNAs were extracted from BMSCs after 5 days’ treatment and sent for high-throughput sequencing for differentially expressed (DE) miRNAs analyses. Then the expression of the DE miRNAs verified by the real-time RT-PCR analyses. Target genes were predicted, and highly enriched GOs and KEGG pathway analysis were conducted using bioinformatics analysis. For the functional study, two of the target genes, SRY (sex determining region Y)-box 5 (SOX5) and G protein-coupled receptor 84 (GPR84), were identified. Results: A total of 5 miRNAs (miRNA-509-5p, miRNA-547-3p, miRNA-201-3p, miRNA-201-5p, and miRNA-novel-272-mature) were identified differentially expressed among groups. The expression of miRNA-novel-272-mature were decreased during the osteogenic differentiation of BMSCs, and GLP-1RA further decreased its expression. MiRNA-novel-272-mature might interact with its target mRNAs to enhance osteogenesis. The lower expression of miRNA-novel-272-mature led to an increase in SOX5 and a decrease in GPR84 mRNA expression, respectively. Conclusions: Taken together, these results provide further insights to the pharmacological properties of GLP-1RA and expand our knowledge on the role of miRNAs-mRNAs regulation network in BMSCs’ differentiation.

scholarly journals Discovery of MicroRNAs and Their Target Genes Related to Drought in Paulownia “Yuza 1” by High-Throughput Sequencing

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Minjie Deng ◽  
Yabing Cao ◽  
Zhenli Zhao ◽  
Lu Yang ◽  
Yanfang Zhang ◽  
...  
Keyword(s):  
Drought Stress ◽  
Molecular Mechanisms ◽  
High Throughput Sequencing ◽  
Plant Hormone ◽  
Target Genes ◽  
Reference Data ◽  
Relevant Information ◽  
Putative Target

Understanding the role of miRNAs in regulating the molecular mechanisms responsive to drought stress was studied in Paulownia “yuza 1.” Two small RNA libraries and two degradome libraries were, respectively, constructed and sequenced in order to detect miRNAs and their target genes associated with drought stress. A total of 107 miRNAs and 42 putative target genes were identified in this study. Among them, 77 miRNAs were differentially expressed between drought-treated Paulownia “yuza 1” and the control (60 downregulated and 17 upregulated). The predicted target genes were annotated using the GO, KEGG, and Nr databases. According to the functional classification of the target genes, Paulownia “yuza 1” may respond to drought stress via plant hormone signal transduction, photosynthesis, and osmotic adjustment. Furthermore, the expression levels of seven miRNAs (ptf-miR157b, ptf-miR159b, ptf-miR398a, ptf-miR9726a, ptf-M2153, ptf-M2218, and ptf-M24a) and their corresponding target genes were validated by quantitative real-time PCR. The results provide relevant information for understanding the molecular mechanism of Paulownia resistance to drought and reference data for researching drought resistance of other trees.

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