Identification of Radish (Raphanus sativus L.) miRNAs and Their Target Genes to Explore miRNA-Mediated Regulatory Networks in Lead (Pb) Stress Responses by High-Throughput Sequencing and Degradome Analysis

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.

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Revealing New Landscape of Turbot (Scophthalmus maximus) Spleen Infected with Aeromonas salmonicida through Immune Related circRNA-miRNA-mRNA Axis

Biology ◽

10.3390/biology10070626 ◽

2021 ◽

Vol 10 (7) ◽

pp. 626

Author(s):

Ting Xue ◽

Yiping Liu ◽

Min Cao ◽

Mengyu Tian ◽

Lu Zhang ◽

...

Keyword(s):

High Throughput ◽

Regulatory Networks ◽

High Throughput Sequencing ◽

Target Genes ◽

Scophthalmus Maximus ◽

Aeromonas Salmonicida ◽

Circular Rnas ◽

Sequencing Data ◽

Turbot Scophthalmus Maximus ◽

Immune Related Genes

Increasing evidence suggests that non-coding RNAs (ncRNA) play an important role in a variety of biological life processes by regulating gene expression at the transcriptional and post-transcriptional levels. Turbot (Scophthalmus maximus) has been threatened by various pathogens. In this study, the expression of circular RNAs (circRNAs), microRNAs (miRNAs), and mRNA in the immune organs spleen of turbot infected with Aeromonas salmonicida was analyzed by high-throughput sequencing, and a circRNA-miRNA-mRNA network was constructed, so as to explore the function of non-coding RNA in the immune system of teleost. Illumina sequencing was performed on the uninfected group and infected group. A total of 119 differential expressed circRNAs (DE-circRNAs), 140 DE-miRNAs, and 510 DE-mRNAs were identified in the four infected groups compared with the uninfected group. Most DE-mRNAs and the target genes of DE-ncRNAs were involved in immune-related pathways. The quantitative real-time PCR (qRT-PCR) results verified the reliability and accuracy of the high-throughput sequencing data. Ninety-six differentially expressed circRNA-miRNA-mRNA regulatory networks were finally constructed. Among them, 15 circRNA-miRNA-mRNA were presented in the form of “up (circRNA)-down (miRNA)-up (mRNA)” or “down-up-down”. Immune-related genes gap junction CX32.2, cell adhesion molecule 3, and CC chemokine were also found in these networks. These results indicate that ncRNA may regulate the expression of immune-related genes through the circRNA-miRNA-mRNA regulatory network and thus participate in the immune response of turbot spleen after pathogen infection.

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

10.20944/preprints201702.0085.v1 ◽

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.

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