A small <scp>RNA</scp> targets pokeweed antiviral protein transcript

Klenov, Alexander; Neller, Kira C. M.; Burns, Lydia A.; Krivdova, Gabriela; Hudak, Katalin A.

doi:10.1111/ppl.12393

Cited by 2 publications

(4 citation statements)

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“…As herbivores are often viral vectors, the JA pathway also has important implications for virus resistance. We showed recently that PAP mRNA and protein levels increase in the presence of JA ( Klenov et al, 2015 ). A link between jasmonate and other plant RIPs has previously been established.…”

Section: Introductionmentioning

confidence: 99%

The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid

2016

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The American pokeweed plant, Phytolacca americana, is recognized for synthesizing pokeweed antiviral protein (PAP), a ribosome inactivating protein (RIP) that inhibits the replication of several plant and animal viruses. The plant is also a heavy metal accumulator with applications in soil remediation. However, little is known about pokeweed stress responses, as large-scale sequencing projects have not been performed for this species. Here, we sequenced the mRNA transcriptome of pokeweed in the presence and absence of jasmonic acid (JA), a hormone mediating plant defense. Trinity-based de novo assembly of mRNA from leaf tissue and BLASTx homology searches against public sequence databases resulted in the annotation of 59 096 transcripts. Differential expression analysis identified JA-responsive genes that may be involved in defense against pathogen infection and herbivory. We confirmed the existence of several PAP isoforms and cloned a potentially novel isoform of PAP. Expression analysis indicated that PAP isoforms are differentially responsive to JA, perhaps indicating specialized roles within the plant. Finally, we identified 52 305 natural antisense transcript pairs, four of which comprised PAP isoforms, suggesting a novel form of RIP gene regulation. This transcriptome-wide study of a Phytolaccaceae family member provides a source of new genes that may be involved in stress tolerance in this plant. The sequences generated in our study have been deposited in the SRA database under project # SRP069141.

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Section: Introductionmentioning

confidence: 99%

The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid

2016

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“…Pokeweed plants at the 4-leaf stage of growth were sprayed with 5 mM JA suspended in 0.5% ethanol, or 0.5% ethanol alone (control), and leaf tissue was harvested 24 h post treatment (Neller et al, 2016 ). Through time-course analysis, we established previously that PAP is expressed maximally 24 h following JA treatment (Klenov et al, 2016 ). This time point was chosen for both the mRNA transcriptome-wide analysis of pokeweed (Neller et al, 2016 ) and the current paired miRNA analysis, as we were interested in identifying defense genes co-expressed with PAP.…”

Section: Methodsmentioning

confidence: 99%

“…Total RNA was extracted from leaf tissue of pokeweed plants as reported in Neller et al ( 2016 ) and the small RNA fraction was sequenced according to Klenov et al ( 2016 ). Briefly, the 15–30 nt size RNA portion was gel purified and ligated to directional 5p and 3p primers.…”

Section: Methodsmentioning

confidence: 99%

“…Each replicate consisted of an equal amount of small RNA pooled from three independent plants. The raw sequencing reads were processed as described in Klenov et al ( 2016 ). Non-coding RNA (rRNA, tRNA, snRNA, and snoRNA) and mature miRNA sequences were annotated by mapping reads to the sugar beet genome (Dohm et al, 2014 ) and the plant miRNA database (Zhang et al, 2010 ; http://bioinformatics.cau.edu.cn/PMRD/ ).…”

Section: Methodsmentioning

confidence: 99%

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Integration of the Pokeweed miRNA and mRNA Transcriptomes Reveals Targeting of Jasmonic Acid-Responsive Genes

et al. 2018

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The American pokeweed plant, Phytolacca americana, displays broad-spectrum resistance to plant viruses and is a heavy metal hyperaccumulator. However, little is known about the regulation of biotic and abiotic stress responses in this non-model plant. To investigate the control of miRNAs in gene expression, we sequenced the small RNA transcriptome of pokeweed treated with jasmonic acid (JA), a hormone that mediates pathogen defense and stress tolerance. We predicted 145 miRNAs responsive to JA, most of which were unique to pokeweed. These miRNAs were low in abundance and condition-specific, with discrete expression change. Integration of paired mRNA-Seq expression data enabled us to identify correlated, novel JA-responsive targets that mediate hormone biosynthesis, signal transduction, and pathogen defense. The expression of approximately half the pairs was positively correlated, an uncommon finding that we functionally validated by mRNA cleavage. Importantly, we report that a pokeweed-specific miRNA targets the transcript of OPR3, novel evidence that a miRNA regulates a JA biosynthesis enzyme. This first large-scale small RNA study of a Phytolaccaceae family member shows that miRNA-mediated control is a significant component of the JA response, associated with widespread changes in expression of genes required for stress adaptation.

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A small RNA targets pokeweed antiviral protein transcript

Cited by 2 publications

References 40 publications

The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid

The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid

Integration of the Pokeweed miRNA and mRNA Transcriptomes Reveals Targeting of Jasmonic Acid-Responsive Genes

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