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预测靶向双生病毒以在辣椒植株中诱导序列特异性基因沉默的候选微小RNA。

Predicting candidate miRNAs for targeting begomovirus to induce sequence-specific gene silencing in chilli plants.

作者信息

Pandey Vineeta, Srivastava Aarshi, Ali Akhtar, Gupta Ramwant, Shahid Muhammad Shafiq, Gaur Rajarshi Kumar

机构信息

Department of Biotechnology, Deen Dayal Upadhyaya Gorakhpur University, Gorakhpur, Uttar Pradesh, India.

Department of Biological Science, The University of Tulsa, Tulsa, OK, United States.

出版信息

Front Plant Sci. 2024 Sep 23;15:1460540. doi: 10.3389/fpls.2024.1460540. eCollection 2024.

Abstract

The begomoviruses are the most economically damaging pathogens that pose a serious risk to India's chilli crop and have been associated with the chilli leaf curl disease (ChiLCD). Chilli cultivars infected with begomovirus have suffered significant decreases in biomass output, negatively impacting their economic characteristics. We used the C-mii tool to predict twenty plant miRNA families from SRA chilli transcriptome data (retrieved from the NCBI and GenBank databases). Five target prediction algorithms, i.e., C-mii, miRanda, psRNATarget, RNAhybrid, and RNA22, were applied to identify and evaluate chilli miRNAs (microRNAs) as potential therapeutic targets against ten begomoviruses that cause ChiLCD. In this study, the top five chilli miRNAs which were identified by all five algorithms were thoroughly examined. Moreover, we also noted strong complementarities between these miRNAs and the AC1 (REP), AC2 (TrAP) and betaC1 genes. Three computational approaches (miRanda, RNA22, and psRNATarget) identified the consensus hybridization site for CA-miR838 at locus 2052. The top predicted targets within ORFs were indicated by CA-miR2673 (a and b). Through Circos algorithm, we identified novel targets and create the miRNA-mRNA interaction network using the R program. Furthermore, free energy calculation of the miRNA-target duplex revealed that thermodynamic stability was optimal for miR838 and miR2673 (a and b). To the best of our knowledge, this was the first instance of miRNA being predicted from chilli transcriptome information that had not been reported in miRbase previously. Consequently, the anticipated biological results substantially assist in developing chilli plants resistant to ChiLCD.

摘要

双生病毒是对印度辣椒作物造成最严重经济损害的病原体,对其构成严重威胁,并与辣椒卷叶病(ChiLCD)相关。感染双生病毒的辣椒品种生物量产量大幅下降,对其经济特性产生负面影响。我们使用C-mii工具从SRA辣椒转录组数据(从NCBI和GenBank数据库检索)中预测了20个植物miRNA家族。应用五种靶标预测算法,即C-mii、miRanda、psRNATarget、RNAhybrid和RNA22,来识别和评估辣椒miRNA(微小RNA)作为针对十种导致ChiLCD的双生病毒的潜在治疗靶标。在本研究中,对所有五种算法鉴定出的前五个辣椒miRNA进行了深入研究。此外,我们还注意到这些miRNA与AC1(REP)、AC2(TrAP)和βC1基因之间有很强的互补性。三种计算方法(miRanda、RNA22和psRNATarget)在2052位点鉴定出CA-miR838的共有杂交位点。CA-miR2673(a和b)表明了开放阅读框内的顶级预测靶标。通过Circos算法,我们鉴定了新的靶标,并使用R程序创建了miRNA-mRNA相互作用网络。此外,miRNA-靶标双链体的自由能计算表明,miR838和miR2673(a和b)的热力学稳定性最佳。据我们所知,这是首次从辣椒转录组信息中预测出miRNA,此前在miRbase中尚未有报道。因此,预期的生物学结果将极大地有助于培育抗ChiLCD的辣椒植株。

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