Shedding Light on Penetration of Cereal Host Stomata by Wheat Stem Rust Using Improved Methodology

Solanki, Shyam; Ameen, Gazala; Borowicz, Paweł; Brueggeman, Robert

doi:10.1038/s41598-019-44280-6

Cited by 28 publications

(16 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The stomatal closure by ABA cannot be a permanent strategy to prevent microbial entry, as pathogens, such as Puccinia, can enter leaves through places other than stomata ( Mendgen et al, 1996 ; Solanki et al, 2019 ). Therefore, we do not mean to overemphasize the role of ABA-induced stomatal closure as the sole mode of adaptation.…”

Section: Countermeasures By Pathogensmentioning

confidence: 99%

Abscisic Acid-Induced Stomatal Closure: An Important Component of Plant Defense Against Abiotic and Biotic Stress

2021

View full text Add to dashboard Cite

Abscisic acid (ABA) is a stress hormone that accumulates under different abiotic and biotic stresses. A typical effect of ABA on leaves is to reduce transpirational water loss by closing stomata and parallelly defend against microbes by restricting their entry through stomatal pores. ABA can also promote the accumulation of polyamines, sphingolipids, and even proline. Stomatal closure by compounds other than ABA also helps plant defense against both abiotic and biotic stress factors. Further, ABA can interact with other hormones, such as methyl jasmonate (MJ) and salicylic acid (SA). Such cross-talk can be an additional factor in plant adaptations against environmental stresses and microbial pathogens. The present review highlights the recent progress in understanding ABA’s multifaceted role under stress conditions, particularly stomatal closure. We point out the importance of reactive oxygen species (ROS), reactive carbonyl species (RCS), nitric oxide (NO), and Ca2+ in guard cells as key signaling components during the ABA-mediated short-term plant defense reactions. The rise in ROS, RCS, NO, and intracellular Ca2+ triggered by ABA can promote additional events involved in long-term adaptive measures, including gene expression, accumulation of compatible solutes to protect the cell, hypersensitive response (HR), and programmed cell death (PCD). Several pathogens can counteract and try to reopen stomata. Similarly, pathogens attempt to trigger PCD of host tissue to their benefit. Yet, ABA-induced effects independent of stomatal closure can delay the pathogen spread and infection within leaves. Stomatal closure and other ABA influences can be among the early steps of defense and a crucial component of plants’ innate immunity response. Stomatal guard cells are quite sensitive to environmental stress and are considered good model systems for signal transduction studies. Further research on the ABA-induced stomatal closure mechanism can help us design strategies for plant/crop adaptations to stress.

show abstract

Section: Countermeasures By Pathogensmentioning

confidence: 99%

Abscisic Acid-Induced Stomatal Closure: An Important Component of Plant Defense Against Abiotic and Biotic Stress

2021

View full text Add to dashboard Cite

show abstract

“…It is hypothesized that P. graminis f. sp. tritici hijacks the stomatal aperture regulation of the cereal hosts in order to enter through the natural openings at night without being detected (64,65). The nec3-γ1 mutant in Bowman background contains the stem rust resistance gene Rpg1 (66,67), which elicits an effective race specific immunity response yet did not elicit the nec3 phenotype.…”

Section: Discussionmentioning

confidence: 99%

“…The nec3-γ1 mutant in Bowman background contains the stem rust resistance gene Rpg1 (66,67), which elicits an effective race specific immunity response yet did not elicit the nec3 phenotype. Recent research has reported that Rpg1 -mediated defenses are non-HR responses (64,68), thus suggesting that the nec3 phenotype may still be elicited by HR-mediated resistance responses.…”

Section: Discussionmentioning

confidence: 99%

Mutations in a barley cytochrome P450 gene enhances pathogen induced programmed cell death and cutin layer instability

Ameen

Solanki

Sager-Bittara

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Disease lesion mimic mutants (DLMMs) are characterized by spontaneous development of necrotic spots with various phenotypes designated as necrotic ( nec ) mutants in barley. The nec mutants were traditionally considered to have aberrant regulation of programmed cell death (PCD) pathways, which have roles in plant immunity and development. Most barley nec3 mutants express cream to orange necrotic lesions contrasting them from typical spontaneous DLMMs that develop dark pigmented lesions indicative of serotonin/phenolics deposition. Also, barley nec3 mutants grown under sterile conditions did not exhibit necrotic phenotypes until inoculated with adapted pathogens suggesting that they are not typical DLMMs. The F 2 progeny of a cross between nec3- γ1 and variety Quest segregated as a single recessive gene post inoculation with Bipolaris sorokiniana, the causal agent of the disease spot blotch . Nec3 was genetically delimited to 0.14 cM representing 16.5 megabases of physical sequence containing 149 annotated high confidence genes. RNAseq and comparative analysis of wild type and five independent nec3 mutants identified a single candidate cytochrome P450 gene (HORVU.MOREX.r2.6HG0460850) that was validated as nec3 by independent mutations that result in predicted nonfunctional proteins. Histology studies determined that nec3 mutants had an unstable cutin layer that disrupted normal Bipolaris sorokiniana germ tube development.

show abstract

“…T-PMT was assigned for differential interface contrast (DIC) imaging. For multilayer visualization Z-stack images were taken and computation from Z stack was performed in Imaris 9.0.1 software (Bitplane, South Windsor, CT) as described previously [18]. During image capture the negative control was assessed first to set the strength of LASER and exposer time.…”

Section: Image Capturingmentioning

confidence: 99%

Visualization of spatial gene expression in plants by modified RNAscope fluorescent in situ hybridization

et al. 2020

Self Cite

View full text Add to dashboard Cite

Background: In situ analysis of biomarkers such as DNA, RNA and proteins are important for research and diagnostic purposes. At the RNA level, plant gene expression studies rely on qPCR, RNAseq and probe-based in situ hybridization (ISH). However, for ISH experiments poor stability of RNA and RNA based probes commonly results in poor detection or poor reproducibility. Recently, the development and availability of the RNAscope RNA-ISH method addressed these problems by novel signal amplification and background suppression. This method is capable of simultaneous detection of multiple target RNAs down to the single molecule level in individual cells, allowing researchers to study spatiotemporal patterning of gene expression. However, this method has not been optimized thus poorly utilized for plant specific gene expression studies which would allow for fluorescent multiplex detection. Here we provide a step-bystep method for sample collection and pretreatment optimization to perform the RNAscope assay in the leaf tissues of model monocot plant barley. We have shown the spatial distribution pattern of HvGAPDH and the low expressed disease resistance gene Rpg1 in leaf tissue sections of barley and discuss precautions that should be followed during image analysis. Results: We have shown the ubiquitous HvGAPH and predominantly stomatal guard cell associated subsidiary cell expressed Rpg1 expression pattern in barley leaf sections and described the improve RNAscope methodology suitable for plant tissues using confocal laser microscope. By addressing the problems in the sample collection and incorporating additional sample backing steps we have significantly reduced the section detachment and experiment failure problems. Further, by reducing the time of protease treatment, we minimized the sample disintegration due to over digestion of barley tissues. Conclusions: RNAscope multiplex fluorescent RNA-ISH detection is well described and adapted for animal tissue samples, however due to morphological and structural differences in the plant tissues the standard protocol is deficient and required optimization. Utilizing barley specific HvGAPDH and Rpg1 RNA probes we report an optimized method which can be used for RNAscope detection to determine the spatial expression and semi-quantification of target RNAs. This optimized method will be immensely useful in other plant species such as the widely utilized Arabidopsis.

show abstract

Shedding Light on Penetration of Cereal Host Stomata by Wheat Stem Rust Using Improved Methodology

Cited by 28 publications

References 31 publications

Abscisic Acid-Induced Stomatal Closure: An Important Component of Plant Defense Against Abiotic and Biotic Stress

Abscisic Acid-Induced Stomatal Closure: An Important Component of Plant Defense Against Abiotic and Biotic Stress

Mutations in a barley cytochrome P450 gene enhances pathogen induced programmed cell death and cutin layer instability

Visualization of spatial gene expression in plants by modified RNAscope fluorescent in situ hybridization

Contact Info

Product

Resources

About