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Samuel W Wilkinson,1 Emilie L Ungricht,1,2 William B West,1,2 Jacob T Harris,1,2 Brian Zaugg,1 Randall J Olson,1 Jeff H Pettey1 1Department of Ophthalmology and Visual Sciences, John A. Moran Eye Center, University of Utah, Salt Lake City, UT, USA; 2University of Utah School of Medicine, Salt Lake City, UT, USACorrespondence: Jeff H Pettey, John A. Moran Eye Center, University of Utah, 65 Mario Capecchi Drive, Salt Lake City, UT, 84132, USA, Tel +1 801-581-2352, Fax +1 801-581-3357, Email jeff.pettey@hsc.utah.eduPurpose: To determine the difference, if any, in grooving efficiency at various settings on the Whitestar Signature Pro phacoemulsification (phaco) system.Methods: Cataractous lenses were simulated by exposing porcine lenses to formalin for 2 hours. A total of 120 lenses were analyzed at various power settings on both longitudinal and transversal handpieces. Twenty trials each were performed with power set to 25%, 50%, and 75% on both handpieces. A Whitestar Phaco Handpiece System was used to provide longitudinal power, and a Whitestar Signature Ellipsis Handpiece provided transversal power. Lenses were placed within a plastic chamber and grooved by an investigator blinded to settings. A second investigator recorded times and adjusted settings. The Whitestar Signature Pro phaco system was used for grooving.Results: There was no significant difference in grooving times between the longitudinal and transverse handpieces at any power setting (P > 0.05). There was a significant decrease in grooving times when comparing the 25% power setting with the 75% power setting for the transversal handpiece (P=0.021).Conclusion: Both longitudinal and transversal handpieces on the Whitestar Signature Pro phaco system produce similar results to one another at each power setting. There is a general trend toward shorter grooving times, reflecting greater efficiency, at higher power settings. Grooving efficiency on the transversal handpiece may be more affected by changes in the power settings as compared with the longitudinal settings.Keywords: phacoemulsification, cataract, phacoemulsification energy, transversal ultrasound, phacoemulsification efficiency

To compare the protection of the corneal endothelium provided by a cohesive and a dispersive ophthalmic viscosurgical device (OVD) against damage from torsional ultrasound and simulated lenticular debris during phacoemulsification.University setting, Salt Lake City, Utah.Experimental study.15 New Zealand rabbits were included. After incision, each eye received cohesive or dispersive OVD (ProVisc or Viscoat). 10 1.0 mm beads were injected to simulate lenticular debris. The Intrepid Balanced tip (Centurion Ozil handpiece) was inserted into the anterior chamber, and 60% torsional ultrasound with 50 mL/min flow, 70 mm Hg intraocular pressure, and 600 mm Hg vacuum was applied (20 seconds). Cumulative dissipated energy (CDE) was noted. After euthanasia and enucleation, corneas were removed, stained with trypan blue/alizarin red, and photographed (5 ×400 magnification photos and 1 overview photograph from each cornea). ImageJ was used to evaluate cell damage and loss.CDE was 4.70 ± 0.26 and 4.64 ± 0.10 in the cohesive and dispersive OVD groups, respectively ( P = .8647). The analysis of the ×400 photographs showed that the percentage of intact cells was statistically higher in the dispersive OVD group ( P = .0002), whereas the percentages of damaged and lost cells were statistically higher in the cohesive OVD group ( P = .0002 and .0059, respectively). Overview photographs revealed the presence of residual OVD on the endothelium, especially in the dispersive group.In this experimental study using a rabbit model, the corneal endothelium protection provided by the dispersive OVD was superior to that provided by the cohesive OVD.

To evaluate mechanical damage of simulated lenticular debris on corneal endothelial cells during phacoemulsification.University setting, Salt Lake City, Utah.Experimental study.12 New Zealand rabbits underwent bilateral surgery. After incision, 10 1.0 mm beads were injected into the anterior chamber of test eyes to simulate lenticular debris. A balanced salt solution was injected into the contralateral eyes with the same injector. The Intrepid Balanced Tip of the Centurion Ozil handpiece was inserted into the anterior chamber, and 60% torsional ultrasound was applied for 20 seconds with 50 mL/min flow, 70 mm Hg intraocular pressure, and 600 mm Hg vacuum. Cumulative dissipated energy (CDE) was noted. After euthanasia and enucleation, the corneas were removed, stained with trypan blue/alizarin red, and photographed in a standardized manner (×400 photographs from 5 specific areas and 1 overview photograph from each corneal button). The ImageJ program was used to evaluate cell damage and loss in the photographs.CDE was 4.64 ± 0.19 for test eyes with beads and 4.66 ± 0.17 for control eyes without beads ( P = .58). The percentage of intact cells was statistically higher in the control group ( P = .0005), whereas the percentages of damaged and lost cells were statistically higher in the test group ( P = .0010 and .0005, respectively). This was observed in the analysis of the ×400 photographs and overview photographs.This study suggests that lenticular debris swirling around the anterior chamber may cause significant damage to the corneal endothelium, providing further insight on mechanisms of damage during phacoemulsification.

Norway spruce (Picea abies) is an economically and ecologically important tree species that grows across northern and central Europe. Treating Norway spruce with jasmonate has long-lasting beneficial effects on tree resistance to damaging pests, such as the European spruce bark beetle Ips typographus and its fungal associates. The (epi)genetic mechanisms involved in such long-lasting jasmonate induced resistance (IR) have gained much recent interest but remain largely unknown. In this study, we treated 2-year-old spruce seedlings with methyl jasmonate (MeJA) and challenged them with the I. typographus vectored necrotrophic fungus Grosmannia penicillata. MeJA treatment reduced the extent of necrotic lesions in the bark 8 weeks after infection and thus elicited long-term IR against the fungus. The transcriptional response of spruce bark to MeJA treatment was analysed over a 4-week time course using mRNA-seq. This analysis provided evidence that MeJA treatment induced a transient upregulation of jasmonic acid, salicylic acid and ethylene biosynthesis genes and downstream signalling genes. Our data also suggests that defence-related genes are induced while genes related to growth are repressed by methyl jasmonate treatment. These results provide new clues about the potential underpinning mechanisms and costs associated with long-term MeJA-IR in Norway spruce.

The plant immune system protects against pests and diseases. The recognition of stress-related molecular patterns triggers localised immune responses, which are often followed by longer-lasting systemic priming and/or up-regulation of defences. In some cases, this induced resistance (IR) can be transmitted to following generations. Such transgenerational IR is gradually reversed in the absence of stress at a rate that is proportional to the severity of disease experienced in previous generations. This review outlines the mechanisms by which epigenetic responses to pathogen infection shape the plant immune system across expanding time scales. We review the cis- and trans-acting mechanisms by which stress-inducible epigenetic changes at transposable elements (TEs) regulate genome-wide defence gene expression and draw particular attention to one regulatory model that is supported by recent evidence about the function of AGO1 and H2A.Z in transcriptional control of defence genes. Additionally, we explore how stress-induced mobilisation of epigenetically controlled TEs acts as a catalyst of Darwinian evolution by generating (epi)genetic diversity at environmentally responsive genes. This raises questions about the long-term evolutionary consequences of stress-induced diversification of the plant immune system in relation to the long-held dichotomy between Darwinian and Lamarckian evolution.

In response to various stimuli, plants acquire resistance against pests and/or pathogens. Such acquired or induced resistance allows plants to rapidly adapt to their environment. Spraying the bark of mature Norway spruce (Picea abies) trees with the phytohormone methyl jasmonate (MeJA) enhances resistance to tree‐killing bark beetles and their associated phytopathogenic fungi. Analysis of spruce chemical defenses and beetle colonization success suggests that MeJA treatment both directly induces immune responses and primes inducible defenses for a faster and stronger response to subsequent beetle attack. We used metabolite and transcriptome profiling to explore the mechanisms underlying MeJA‐induced resistance in Norway spruce. We demonstrated that MeJA treatment caused substantial changes in the bark transcriptional response to a triggering stress (mechanical wounding). Profiling of mRNA expression showed a suite of spruce inducible defenses are primed following MeJA treatment. Although monoterpenes and diterpene resin acids increased more rapidly after wounding in MeJA‐treated than control bark, expression of their biosynthesis genes did not. We suggest that priming of inducible defenses is part of a complex mixture of defense responses that underpins the increased resistance against bark beetle colonization observed in Norway spruce. This study provides the most detailed insights yet into the mechanisms underlying induced resistance in a long‐lived gymnosperm.

Methyl jasmonate (MeJA) treatment elicits induced resistance (IR) against pests and diseases in Norway spruce (Picea abies). We recently demonstrated using mRNA-seq that this MeJA-IR is associated with both a prolonged upregulation of inducible defenses and defense priming. Gene expression can be regulated at both a transcriptional and post-transcriptional level by small RNAs, including microRNAs (miRNAs). Here we explore the effects of MeJA treatment and subsequent challenge by wounding on the Norway spruce miRNA transcriptome. We found clusters of prolonged down- or upregulated miRNAs as well as miRNAs whose expression was primed after MeJA treatment and subsequent wounding challenge. Differentially expressed miRNAs included miR160, miR167, miR172, miR319, and the miR482/2118 superfamily. The most prominent mRNA targets predicted to be differentially expressed by miRNA activity belonged to the nucleotide-binding site leucine-rich repeat (NBS-LRR) family. Among other predicted miRNA targets were genes regulating jasmonic acid biosynthesis. Our results indicate that miRNAs have an important role in the regulation of MeJA-IR in Norway spruce.

Minimising losses to pests and diseases is essential for producing sufficient food to feed our rapidly growing population. The necrotrophic fungus Botrytis cinerea triggers devastating pre- and post-harvest yield losses in tomato (Solanum lycopersicum). Current control methods are based on the pre-harvest use of fungicides, which are limited by strict legislation. Here, we have tested whether induction of resistance by β-aminobutyric acid (BABA) at different developmental stages, provides an alternative strategy to protect tomato fruit post-harvest against B. cinerea. Soil-drenching plants with BABA once fruit had already formed, had no impact on tomatoes susceptibility to B. cinerea. Whereas BABA application to seedlings was found to significantly reduce the post-harvest infection of fruit. This resistance response was not associated with a yield reduction, however there was a delay in fruit ripening. Untargeted metabolomics unravelled differences between fruit from water and BABA-treated plants, demonstrating that BABA triggered a defence-associated metabolomics profile that was long-lasting. Targeted analysis of defence hormones suggested a role of abscisic acid (ABA) in the resistance phenotype. Post-harvest application of ABA to the fruit of water-treated plants induced susceptibility to B. cinerea. This phenotype was absent from the ABA exposed fruit of BABA-treated plants, suggesting a complex role of ABA in the BABA-induced resistance phenotype. A final targeted metabolomic analysis detected trace residues of BABA accumulated in the red fruit. Overall, we have demonstrated that β-aminobutyric acid induces post-harvest resistance in tomato fruit against B. cinerea with no penalties in yield. This article is protected by copyright. All rights reserved.

Exudates released from plant roots can recruit beneficial microorganisms that boost plant growth and immunity. In Arabidopsis thaliana and tomato, active DNA demethylation regulates the production of myo-inositol, a root exudate which recruits a specific plant growth-promoting rhizobacterium.

Stress can alter important plant life-history traits. Here, we report the long-term effects of the stress hormone jasmonic acid (JA) on the defence phenotype, transcriptome and DNA-methylome of Arabidopsis. Three weeks after transient JA signalling activity, 5-week-old plants retained induced resistance (IR) against herbivory but showed enhanced susceptibility to necrotrophic and biotrophic pathogens. Transcriptome analysis of these plants revealed priming and/or up-regulation of JA-dependent defence genes but repression of ethylene- and salicylic acid-dependent genes. Long-term JA-IR against herbivory was associated with shifts in glucosinolate composition and required MYC2/3/4 transcription factors, DNA (de)methylation pathways and the small RNA (sRNA)-binding protein ARGONOUTE1 (AGO1). Although methylome analysis did not reveal consistent changes in DNA methylation near MYC2/3/4-controlled genes, JA-treated plants were specifically enriched with hypomethylated ATREP2 transposable elements (TEs), while ATREP2-derived sRNAs showed increased association with AGO1. Our results indicate that AGO1-associated sRNAs from hypomethylated ATREP2 TEs trans-regulate long-lasting memory of JA-dependent immunity.