To detect potential pathogens, plants perceive the fungal polysaccharide chitin through receptor complexes containing lysin motif receptor-like kinases (LysM-RLKs). To investigate the ligand-induced ...spatial dynamics of chitin receptor components, we studied the subcellular behaviour of two Arabidopsis thaliana LysM-RLKs involved in chitin signalling, CHITIN ELICITOR RECEPTOR KINASE1 (CERK1) and LYSIN MOTIF-CONTAINING RECEPTOR-LIKE KINASE5.
We performed standard and quantitative confocal laser scanning microscopy on stably transformed A. thaliana plants expressing fluorescently tagged CERK1 and LYK5 from their native promoters. Microscopy approaches were complemented by biochemical analyses in plants and in vitro.
Both CERK1 and LYK5 localized to the plasma membrane and showed constitutive endomembrane trafficking. After chitin treatment, however, CERK1 remained at the plasma membrane while LYK5 relocalized into mobile intracellular vesicles. Detailed analyses revealed that chitin perception transiently induced the internalization of LYK5 into late endocytic compartments. Plants that lacked CERK1 or expressed an enzymatically inactive CERK1 variant did not exhibit chitin-induced endocytosis of LYK5. CERK1 could phosphorylate LYK5 in vitro and chitin treatment induced CERK1-dependent phosphorylation of LYK5 in planta.
Our results suggest that chitin-induced phosphorylation by CERK1 triggers LYK5 internalization. Thus, our work identifies phosphorylation as a key regulatory step in endocytosis of plant RLKs and also provides evidence for receptor complex dissociation after ligand perception.
Sporisorium reilianum
f. sp.
zeae
(SRZ) is a biotrophic fungus causing head smut in maize. Maize infection with SRZ leads to very little cell death suggesting the presence of cell-death suppressinpg ...effectors. Several hundred effector proteins have been predicted based on genome annotation, genome comparison, and bioinformatic analysis. For only very few of these effectors, an involvement in virulence has been shown. In this work, we started to test a considerable subset of these predicted effector proteins for a possible function in suppressing cell death. We generated an expression library of 62 proteins of SRZ under the control of a strong constitutive plant promoter for delivery into plant cells
via Agrobacterium tumefaciens
-mediated transient transformation. Potential apoplastic effectors with high cysteine content were cloned with signal peptide while potential intracellular effectors were also cloned without signal peptide to ensure proper localization after expression in plant cells. After infiltration of
Nicotiana benthamiana
leaves, infiltration sites were evaluated for apparent signs of hypersensitive cell death in absence or presence of the elicitin INF1 of
Phytophthora infestans
. None of the tested candidates was able to induce cell death, and most were unable to suppress INF1-induced cell death. However, the screen revealed one predicted cytoplasmic effector (sr16441) of SRZ that was able to reliably suppress INF1-induced cell death when transiently expressed in
N. benthamiana
lacking its predicted secretion signal peptide. This way, we discovered a putative function for one new effector of SRZ.
Biotrophic pathogens, such as the related maize pathogenic fungi Ustilago maydis and Sporisorium reilianum, establish an intimate relationship with their hosts by secreting protein effectors. Because ...secreted effectors interacting with plant proteins should rapidly evolve, we identified variable genomic regions by sequencing the genome of S. reilianum and comparing it with the U. maydis genome. We detected 43 regions of low sequence conservation in otherwise well-conserved syntenic genomes. These regions primarily encode secreted effectors and include previously identified virulence clusters. By deletion analysis in U. maydis, we demonstrate a role in virulence for four previously unknown diversity regions. This highlights the power of comparative genomics of closely related species for identification of virulence determinants.
Introduction Gestational hypertension is a prevalent condition, occurring in 10% of pregnancies. Physical exercise has a valuable effect on lowering blood pressure. Device-guided breathing is a ...beneficial strategy for treating high blood pressure. The purpose of this study was to compare aerobic exercise vs. device-guided breathing in gestational hypertension. Methods The study included 90 singleton pregnant women (at the 21st week of gestation) diagnosed with gestational hypertension. They were randomly divided into 3 groups. The aerobic exercise group (n = 30) received aerobic exercise until the 36th week of gestation. The device-guided breathing group (n = 30) received device-guided breathing until the 36th week of gestation. The third group was a control group. All groups received the same antihypertensive medications. They were assessed before and after treatment with a mercury column sphygmomanometer to measure systolic and diastolic blood pressure. Results All groups showed a significant reduction in their systolic and diastolic blood pressure values after the end of the training program. However, the comparison with the corresponding values measured before treatment revealed that the participants in the device-guided breathing group exhibited a greater reduction in the systolic and diastolic blood pressure values than those in the aerobic exercise and control groups. Conclusions Device-guided breathing is more effective than aerobic exercise in decreasing systolic and diastolic blood pressure in gestational hypertensive women.
Silicon induced resistance (SiIR) in tomato against bacterial wilt caused by
Ralstonia solanacearum. The mechanism of SiIR is ambiguous. Therefore we tracked the expression of twelve defense marker ...genes over time using qRT-PCR. Besides up-regulated expression of the jasmonic acid/ethylene marker genes
JERF3,
TSRF1and
ACCO during SiIR, the expression of the oxidative stress markers
FD-I and
POD and the basal defense marker
AGP-1g were also up-regulated. The expression of defense marker genes was generally induced upon challenging the silicon-treated plants with
R. solanacearum and reached its highest levels at 72
h post inoculation. A global transcriptome profiling was performed to uncover further changes at this time point using the TOM2 microarray. Significant regulation of additional 16 genes was revealed after silicon treatment in plants challenged with
R. solanacearum. Twelve genes, involved in defense, signal transduction, response to stresses, transcription, ubiquitinylation and metabolism, were up-regulated. A
JAZ1-similar gene showed the highest level of up-regulation indicating prior induction and fine tuning of jasmonic acid signaling. Thus, we conclude that silicon primed plants, thereby alleviating biotic stress imposed by the pathogen. The primed state could be mediated via ethylene, jasmonic acid and/or reactive oxygen species signaling pathways. We also suggest that the protective role of silicon can not only be explained with its mechanical properties.
► This paper reveals for the first time gene expression underlying silicon-induced resistance (SiIR) in a silicon non-accumulator plant, tomato, in response to challenge with a bacterial pathogen, and that SiIR is expressed in form of priming. ► The paper presents a follow-up on transcript level of our biochemical and immunohistochemical work on silicon induced resistance, and confirms our histochemical observations of induction of basal resistance (e.g. PMPP 64: 233–243, 68: 41–50, 70: 120–129). ► From the transcription profile we could develop a hypothetical model of the SiIR, in which JA/ET and/ROS signaling pathways could be involved. ► We also suggest that the protective role of silicon can not only be explained with its mechanical properties.
The phytohormones salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) are central regulators of biotic and abiotic stress responses in
. Here, we generated modular fluorescent protein-based ...reporter lines termed COLORFUL-PR1pro, -VSP2pro, and -PDF1.2apro. These feature hormone-controlled nucleus-targeted transcriptional output sensors and the simultaneous constitutive expression of spectrally separated nuclear reference and plasma membrane-localized reporters. This set-up allowed the study of cell-type specific hormone activities, cellular viability and microbial invasion. Moreover, we developed a software-supported high-throughput confocal microscopy imaging protocol for output quantification to resolve the spatio-temporal dynamics of respective hormonal signaling activities at single-cell resolution. Proof-of-principle analyses in
leaves revealed distinguished hormone sensitivities in mesophyll, epidermal pavement and stomatal guard cells, suggesting cell type-specific regulatory protein activities. In plant-microbe interaction studies, we found that virulent and avirulent
(
) isolates exhibit different invasion dynamics and induce spatio-temporally distinct hormonal activity signatures. On the cellular level, these hormone-controlled reporter signatures demarcate the nascent sites of
entry and progression, and highlight initiation, transduction and local containment of immune signals.
Background
There are many physiological changes occur during pregnancy that affect oxygenation of the pregnant women. It was considered that aerobic and breathing exercises are safe for the mother ...and the fetus especially moderate intensity exercise during pregnancy.
Objective
This study was designed to investigate the blood oxygenation response to aerobic exercise combined with breathing exercises in pregnant women.
Methodology
Forty pregnant women were selected from obstetrics and gynecology outpatient clinic at Kasr El Einy Hospital, Egypt, confidentiality was assured. They were ranged from 25 to 30 years old, with body mass index (BMI) less than 30 kg/m
2
and at the beginning of 3
rd
trimester, this study was conducted from September 2019 to April 2020. They were assigned into two groups: group A performed aerobic exercise in a form of walking for 20 min. On the treadmill at 60–75% of the maximum heart rate (MHR) of each woman, three times per week in addition to deep breathing exercises in form of diaphragmatic and lateral costal breathing; group B who performed deep breathing exercises only in form of diaphragmatic and lateral costal breathing. The program continued for three months, three times per week. The oxygen saturation (SaO2) was measured twice time, firstly, at the beginning of 3
rd
trimester of pregnancy then after three months of treatment program.
Results
The results of this study revealed a statistically significant difference in the oxygen saturation in group A than in group B.
Conclusion
It could be concluded that there was significant statistical effect of aerobic exercise combined with breathing exercises on blood oxygenation in pregnant women. It improved the oxygen saturation in pregnant women.
Subdiffraction super‐resolution fluorescence microscopy, or nanoscopy, has seen remarkable developments in the last two decades. Yet, for the visualization of plant cells, nanoscopy is still rarely ...used. In this study, we established RESOLFT nanoscopy on living green plant tissue. Live‐cell RESOLFT nanoscopy requires and utilizes comparatively low light doses and intensities to overcome the diffraction barrier. We generated a transgenic Arabidopsis thaliana plant line expressing the reversibly switchable fluorescent protein rsEGFP2 fused to the mammalian microtubule‐associated protein 4 (MAP4) in order to ubiquitously label the microtubule cytoskeleton. We demonstrate the use of RESOLFT nanoscopy for extended time‐lapse imaging of cortical microtubules in Arabidopsis leaf discs. By combining our approach with fluorescence lifetime gating, we were able to acquire live‐cell RESOLFT images even close to chloroplasts, which exhibit very strong autofluorescence. The data demonstrate the feasibility of subdiffraction resolution imaging in transgenic plant material with minimal requirements for sample preparation.
Genome comparison between the maize pathogens
and
revealed a large diversity region (19-1) containing nearly 30 effector gene candidates, whose deletion severely hampers virulence of both fungi. ...Dissection of the
gene cluster resulted in the identification of one major contributor to virulence,
(
;
). Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) experiments revealed high expression of
during biotrophic growth of
. Using the yeast secretion trap assay, we confirmed the existence of a functional signal peptide allowing protein secretion via the conventional secretory pathway. We identified the cytoplasmic maize chorismate mutase ZmCM2 by yeast two-hybrid screening as a possible interaction partner of Vag2. Interaction of the two proteins in planta was confirmed by bimolecular fluorescence complementation. qRT-PCR experiments revealed
-dependent downregulation of salicylic acid (SA)-induced genes, which correlated with higher SA levels in plant tissues colonized by Δ
deletion strains relative to
wildtype strains. Metabolite analysis suggested rewiring of pathogen-induced SA biosynthesis by preferential conversion of the SA precursor chorismate into the aromatic amino acid precursor prephenate by ZmCM2 in the presence of Vag2. Possibly, the binding of Vag2 to ZmCM2 inhibits the back reaction of the ZmCM2-catalyzed interconversion of chorismate and prephenate, thus contributing to fungal virulence by lowering the plant SA-induced defenses.
Background
The most common vasomotor symptoms (VMS) are hot flashes and night sweats, which occur in as many as 68.5% of women as a result of menopause. Symptoms caused by fluctuating levels of ...estrogen may be alleviated by hormone therapy (HT), but a marked global decline in its use has resulted from concerns about the risks and benefits of HT. Consequently, many women in the postmenopausal period search for alternative natural treatment options to manage menopause. As large numbers of women are choosing not to take HT, it is increasingly important to identify evidence-based foot reflexology with paced respiration that has the potential to reduce vasomotor menopausal symptoms.
Aim
To examine the combined effect of foot reflexology with paced respiration versus paced respiration on VMS in symptomatic menopausal women.
Patients and methods
A total of 50 women of menopausal age, 45–55 years were studied; their BMI was less than 30 kg/m
2
. They were allocated randomly to two groups with equal numbers of participants (A and B). The participants in group A received foot reflexology in addition to paced respiration, whereas the participants in group B received the paced respiration training only. The treatment program was conducted three times per week for 8 weeks. Assessment of all participants in both groups (A and B) was carried out before and after the treatment program throughout by determining blood cortisol level in addition to the use of the menopause rating scale (MRS).
Results
Both groups (A and B) showed a significant reduction in their blood cortisol and MRS values after the end of the 8 weeks of the training program. The mean values of blood cortisol after treatment were 15.40±1.47, 16.32±1.70 in both groups A and B, respectively. The mean values of total MRS were 8.12±2.19, 12.56±2.96 in both groups A and B, respectively.
However, the participants who received foot reflexology plus paced respiration, group A, showed a greater reduction in the blood cortisol value and total MRS (
P
<0.001 and <0.001), respectively.
Conclusion
It could be concluded that foot reflexology in addition to paced respiration are more effective than paced respiration only in decreasing blood cortisol level as well as VMS in symptomatic menopausal women.
PDF
