The quantification of tomato microRNAs response to viral infection by stem-loop real-time RT-PCR

Feng, Jia; Wang, Kai; Liu, Xin; Chen, Shaoning; Chen, Jishuang

doi:10.1016/j.gene.2009.01.017

Cited by 110 publications

(76 citation statements)

References 32 publications

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“…The integrity of the total RNAs was determined by electrophoresis on 1% (w/v) agarose gel. Isolated RNA was dissolved in 50 µl of RNase free H 2 O and stored at −80 °C (Feng et al, 2009). …”

Section: Extraction Of Total Rnasmentioning

confidence: 99%

Responses of phospholipase D and lipoxygenase to mechanical wounding in postharvest cucumber fruits

Zhao

Qian

Chen

et al. 2010

J. Zhejiang Univ. Sci. B

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Abstract:This study was to investigate the responses of phospholipase D (PLD) and lipoxygenase (LOX) to mechanical wounding in postharvest cucumber (Cucumis sativus L. cv. Biyu-2) fruits. Membrane-associated Ca 2+ content, activities and gene expression of PLD and LOX, and contents of phosphatidylcholine (PC), phosphatidylinositol (PI), and phosphatidic acid (PA) were determined in cucumber fruits following mechanical wounding. Results show that PLD and LOX activities increased with the PLD and LOX mRNAs which are upregulated upon wounding, while membrane-associated Ca 2+ content decreased. Accompanying with the increase of PLD and LOX activities, accumulation of PA and losses of PC and PI were observed in all fruits, but there were differences of degrees between wounded and control fruits. Results suggest that PLD and LOX might be the main hydrolytic enzymes of phospholipids in postharvest cucumber fruits participating in the mechanical wounding injury. The activation of PLD and LOX might be the result of gene expression, which could be stimulated by the Ca 2+ flowing from the membrane to the cytoplasm upon receiving the wounding signals.

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Section: Extraction Of Total Rnasmentioning

confidence: 99%

Responses of phospholipase D and lipoxygenase to mechanical wounding in postharvest cucumber fruits

Zhao

Qian

Chen

et al. 2010

J. Zhejiang Univ. Sci. B

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“…Although miRNAs are initially discovered using genetic screening technology (Lee et al, 1993;Wightman et al, 1993), this method is limited as it is expensive, time consuming and dominated by chance (Zhang et al, 2006b). Direct cloning of miRNAs, followed by construction and sequencing of a small RNA library, has been successfully employed to identify miRNAs in plants (Feng et al, 2009;Jian et al, 2010;Sunkar et al, 2005;Yao et al, 2007). However, only abundant miRNA genes can be easily detected by cloning or Northern blot.…”

Section: Introductionmentioning

confidence: 99%

Identification and Validation of Potential Conserved microRNAs and Their Targets in Peach (Prunus persica)

Gao

Luo

Shi

et al. 2012

Molecules and Cells

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1MicroRNAs are a class of small, endogenous, non-coding RNA molecules that negatively regulate gene expression at the transcriptional or the post-transcriptional level. Although a large number of miRNAs have been identified in many plant species, especially from model plants and crops, they remain largely unknown in peach. In this study, 110 potential miRNAs belonging to 37 families were identified using computational methods. A total of 43 potential targets were found for 21 families based on near-perfect or perfect complementarity between the plant miRNA and the target sequences. A majority of the targets were transcription factors which play important roles in peach development. qRT-PCR analysis of RNA samples prepared from different peach tissues for 25 miRNA families revealed that miRNAs were differentially expressed in different tissues. Furthermore, two target genes were experimentally verified by detection of the miRNA-mediated mRNA cleavage sites in peach using RNA ligase-mediated 5' rapid amplification of cDNA ends (RLM-RACE). Finally, we studied the expression pattern of the two target genes in three different tissues of peach to further understand the mechanism of the interaction between miRNAs and their target genes.

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“…, where ∆∆C T = (C T target -C T 18S rRNA) sample X -(C T target -C T 18S rRNA) sample 1 (Feng et al, 2009). In this study, sample 1 of the candidate gene acted as the mock infection, whereas sample X was the PCR product of the gene at 2-day intervals from 1 to 7 dpi.…”

Section: Real-time Pcrmentioning

confidence: 96%

Molecular characterization and functional analysis of the Nep1-like protein-encoding gene from Phytophthora capsici

Feng

Li²

2013

Genet. Mol. Res.

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ABSTRACT. Phytophthora capsici is an aggressive plant pathogen that affects solanaceous and cucurbitaceous hosts. Nep1-like proteins (NLPs) are a group of effectors found particularly in oomycetes and considered important virulence factors. We identified an NLP gene (phcnlp1) from the highly virulent P. capsici strain Phyc12 that had an encoded polypeptide of 476-amino acid residues and a predicted molecular mass of 51.75 kDa. We performed quantitative reverse transcription-polymerase chain reaction to detect the expression pattern of phcnlp1 during various phases of interaction with the host plant, and the results showed that phcnlp1 was increasingly expressed during symptom development after P. capsici infection of pepper leaves. We also confirmed that phcnlp1 caused significant necrosis on tobacco plants when expressed based on potato virus agroinfection. All results indicated that phcnlp1 belongs to the NLP gene family and is important for the pathogenesis of P. capsici in its hosts.

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The quantification of tomato microRNAs response to viral infection by stem-loop real-time RT-PCR

Cited by 110 publications

References 32 publications

Responses of phospholipase D and lipoxygenase to mechanical wounding in postharvest cucumber fruits

Responses of phospholipase D and lipoxygenase to mechanical wounding in postharvest cucumber fruits

Identification and Validation of Potential Conserved microRNAs and Their Targets in Peach (Prunus persica)

Molecular characterization and functional analysis of the Nep1-like protein-encoding gene from Phytophthora capsici

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