Differentiation Zone Research Articles

Root cap cell corpse clearance limits microbial colonization in Arabidopsis thaliana.

Programmed cell death occurring during plant development (dPCD) is a fundamental process integral for plant growth and reproduction. Here, we investigate the connection between developmentally controlled PCD and fungal accommodation in Arabidopsis thaliana roots, focusing on the root cap-specific transcription factor ANAC033/SOMBRERO (SMB) and the senescence-associated nuclease BFN1. Mutations of both dPCD regulators increase colonization by the beneficial fungus Serendipita indica, primarily in the differentiation zone. smb-3 mutants additionally exhibit hypercolonization around the meristematic zone and a delay of S. indica-induced root-growth promotion. This demonstrates that root cap dPCD and rapid post-mortem clearance of cellular corpses represent a physical defense mechanism restricting microbial invasion of the root. Additionally, reporter lines and transcriptional analysis revealed that BFN1 expression is downregulated during S. indica colonization in mature root epidermal cells, suggesting a transcriptional control mechanism that facilitates the accommodation of beneficial microbes in the roots.

Root cap cell corpse clearance limits microbial colonization in Arabidopsis thaliana

Programmed cell death occurring during plant development (dPCD) is a fundamental process integral for plant growth and reproduction. Here, we investigate the connection between developmentally controlled PCD and fungal accommodation in Arabidopsis thaliana roots, focusing on the root cap-specific transcription factor ANAC033/SOMBRERO (SMB) and the senescence-associated nuclease BFN1. Mutations of both dPCD regulators increase colonization by the beneficial fungus Serendipita indica, primarily in the differentiation zone. smb-3 mutants additionally exhibit hypercolonization around the meristematic zone and a delay of S. indica-induced root-growth promotion. This demonstrates that root cap dPCD and rapid post-mortem clearance of cellular corpses represent a physical defense mechanism restricting microbial invasion of the root. Additionally, reporter lines and transcriptional analysis revealed that BFN1 expression is downregulated during S. indica colonization in mature root epidermal cells, suggesting a transcriptional control mechanism that facilitates the accommodation of beneficial microbes in the roots.

Tomato roots exhibit distinct, development-specific responses to bacterial-derived peptides.

Plants possess cell-surface recognition receptors that detect molecular patterns from microbial invaders and initiate an immune response. Understanding the conservation of pattern-triggered immunity within different plant organs and across species is crucial to its sustainable and effective use in plant disease management but is currently unclear. We examined the activation and immune response patterns of three pattern recognition receptors (PRRs: Sl FLS2, Sl FLS3, and Sl CORE) in different developmental regions of roots and in leaves of multiple accessions of domesticated and wild tomato ( Solanum lycopersicum and S. pimpinellifolium ) using biochemical and genetic assays. Roots from different tomato accessions differed in the amplitude and dynamics of their immune response, but all exhibited developmental-specific PTI responses in which the root early differentiation zone was the most sensitive to molecular patterns. PRR signaling pathways also showed distinct but occasionally overlapping responses downstream of each immune receptor in tomato roots.These results reveal that each PRR initiates a unique PTI pathway and suggest that the specificity and complexity of tomato root immunity are tightly linked to the developmental stage, emphasizing the importance of spatial and temporal regulation in PTI.

MyFeedback: an online writing feedback service with students as partners

MyFeedback is an online feedback service run by the academic tutoring team at the University of East London (UEL). The service provides a formative type of feedback on the academic writing of students based on drafts they upload to a digital inbox. The service is a differentiating support mechanism that complements the one-to-one guidance provided by the academic writing tutors. This is possible by training and supervising students (online feedback initiators) to provide reviews of assignment drafts. In this ALDCon24 workshop, we discussed how the development and operation of the service has thrived through a partnership between learning developers and students, and thus establishing a community of practice (Wenger, 1998) of shared learning and vision. We provided insights into the working model and the ways in which students become partners and occasionally co-creators. We highlighted the opportunities and benefits that are shared among all those involved and the roles they assume. Student users of the service engage in a self-regulated learning experience (Nicol and Macfarlane-Dick, 2006) enacted by their repeated exposure to peer feedback interactions that is underpinned by pedagogical principles, such as the zone of proximal development (Vygotsky, 1978) and differentiation, for example in Tomlinson (2001). Tutors and student partners collaborate to consolidate their feedback mechanisms by bringing together teaching expertise, fresh enthusiasm, first-hand student experiences, and a sense of belonging, inclusivity and collaboration. At the same time, tutors direct their capacity efficiently to students who can benefit more from a live dialogue based on their assessment at hand.

An auxin regulated Universal stress protein (TaUSP_3B-1) interacts with TaGolS and provides tolerance under drought stress and ER stress.

A previously identified wheat drought stress responsive Universal stress protein, TaUSP_3B-1 has been found to work in an auxin dependent manner in the plant root tissues in the differentiation zone. We also found a novel interacting partner, TaGolS, which physically interacts with TaUSP_3B-1 and colocalizes in the endoplasmic reticulum. TaGolS is a key enzyme in the RFO (Raffinose oligosaccharides) biosynthesis which is well reported to provide tolerance under water deficit conditions. TaUSP_3B-1 overexpression lines showed an early flowering phenotype under drought stress which might be attributed to the increased levels of AtTPPB and AtTPS transcripts under drought stress. Moreover, at the cellular levels ER stress induced TaUSP_3B-1 transcription and provides tolerance in both adaptive and acute ER stress via less ROS accumulation in the overexpression lines. TaUSP_3B-1 overexpression plants had increased silique numbers and a denser root architecture as compared to the WT plants under drought stress.

The Morphological Parameters and Cytosolic pH of Cells of Root Zones in Tobacco Plants (Nicotiana tabacum L.): Nonlinear Effects of NaCl Concentrations

Salinity impacts important processes in plants, reducing their yield. The effect of salinity on the cytosolic pH (pHcyt) has been little studied. In this research, we employed transgenic tobacco plants expressing the pH sensor Pt-GFP to investigate the alterations in pHcyt in cells across various root zones. Furthermore, we examined a wide spectrum of NaCl concentrations (ranging from 0 to 150 mM) and assessed morphological parameters and plant development. Our findings revealed a pattern of cytosolic acidification in cells across all root zones at lower NaCl concentrations (50, 100 mM). Interestingly, at 150 mM NaCl, pHcyt levels either increased or returned to normal, indicating a nonlinear effect of salinity on pHcyt. Most studied parameters related to development and morphology exhibited an inhibitory influence in response to NaCl. Notably, a nonlinear relationship was observed in the cell length within the elongation and differentiation zones. While cell elongation occurred at 50 and 100 mM NaCl, it was not evident at 150 mM NaCl. This suggests a complex interplay between stimulating and inhibitory effects of salinity, contributing to the nonlinear relationship observed between pHcyt, cell length, and NaCl concentration.

A double-negative feedback loop between miR319c and JAW-TCPs establishes growth pattern in incipient leaf primordia in Arabidopsis thaliana.

The microRNA miR319 and its target JAW-TCP transcription factors regulate the proliferation-to-differentiation transition of leaf pavement cells in diverse plant species. In young Arabidopsis leaf primordia, JAW-TCPs are detected towards the distal region whereas the major mRNA319-encoding gene MIR319C, is expressed at the base. Little is known about how this complementary expression pattern of MIR319C and JAW-TCPs is generated. Here, we show that MIR319C is initially expressed uniformly throughout the incipient primordia and is later abruptly down-regulated at the distal region, with concomitant distal appearance of JAW-TCPs, when leaves grow to ~100 μm long. Loss of JAW-TCPs causes distal extension of the MIR319C expression domain, whereas ectopic TCP activity restricts MIR319C more proximally. JAW-TCPs are recruited to and are capable of depositing histone H3K27me3 repressive marks on the MIR319C chromatin. JAW-TCPs fail to repress MIR319C in transgenic seedlings where the TCP-binding cis-elements on MIR319C are mutated, causing miR319 gain-of-function-like phenotype in the embryonic leaves. Based on these results, we propose a model for growth patterning in leaf primordia wherein MIR319C and JAW-TCPs repress each other and divide the uniformly growing primordia into distal differentiation zone and proximal proliferation domain.

Exploring the Gene Pool of Silver Fir in Southern Germany on the Search for Climate-Smart Seed Sources

Central European populations of silver fir (Abies alba Mill.) grow under a relatively wide amplitude of environmental conditions. Assuming that forest tree stands are locally adapted, the use of forest reproductive material from sites with limited water availability is expected to increase drought tolerance in future forests. At the same time, genetic diversity is of utmost importance as the basis of adaptation to a changing environment. Currently, a selection of potential sources for climate-smart reproductive material of silver fir is under way in Southern Germany. It is based on a multidisciplinary approach elucidating the genetic variation, site conditions, as well as tree resilience based on a dendroecological approach. The analysis encompasses a large number of stands representing the whole range of the species’ ecological requirements. The population genetic analysis based on molecular markers (nuclear microsatellites) provided important information concerning the gene pool of the species in Southern Germany. On one hand, it revealed genetic differentiation and transition zones between western and eastern clusters. On the other hand, the results indicated gradients and regional variation of genetic diversity. These patterns can be explained by post glacial recolonization and the peripheral character of the species at the northern limit of its distribution. Together with the outcomes of the site condition and dendroecological approaches, the results of the genetic analysis will be used to inform future provenance recommendations.

A spatially concerted epidermal auxin signaling framework steers the root hair foraging response under low nitrogen

As a major determinant of the nutrient-acquiring root surface, root hairs (RHs) provide a low-input strategy to enhance nutrient uptake. Although primary and lateral roots exhibit elongation responses under mild nitrogen (N)deficiency, the foraging response of RHs and underlying regulatory mechanisms remain elusive. Employing transcriptomics and functional studies revealed a framework of molecular components composing a cascade of auxin synthesis, transport, and signaling that triggers RH elongation for N acquisition. Through upregulation of Tryptophan Aminotransferase of Arabidopsis 1 (TAA1) and YUCCA8, low N increases auxin accumulation in the root apex. Auxin is then directed to the RH differentiation zone via the auxin transport machinery, AUXIN TRANSPORTER PROTEIN 1 (AUX1) and PIN-FORMED 2 (PIN2). Upon arrival to the RH zone, auxin activates the transcription factors AUXIN RESPONSE FACTOR 6 and 8 (ARF6/8) to promote the epidermal and auxin-inducible transcriptional module ROOT HAIR DEFECTIVE 6 (RHD6)-LOTUS JAPONICA ROOT HAIRLESS-LIKE 3 (LRL3) to steer RH elongation in response to low N. Our study uncovers a spatially defined regulatory signaling cascade for N foraging by RHs, expanding the mechanistic framework of hormone-regulated nutrient sensing in plant roots.

Advanced imaging and quantification of the cambium and developing xylem in eucalypts using X-ray micro- and nano-computed tomography

Summary In recent years, the popularity of X-ray computed tomography (CT), as a non-destructive imaging technique, has continued to expand in various research domains. In wood research, X-ray CT has proven to be useful for three-dimensional (3D) structural studies investigating the complex tissues of trees. Wood formation (i.e., xylogenesis) initiates in the cambium and a narrow zone of subsequent differentiation, both of which play key roles in plant growth and development. However, the dynamics of xylogenesis in eucalypts remain relatively poorly understood, in large part due to challenges in sampling, imaging, and characterizing the cambium. Therefore, the aim of this study was to present a workflow to evaluate the feasibility of using X-ray CT to characterize and quantify the structural properties of the cambium in eucalypts. The growth responses of Corymbia hybrid seedlings, exposed to either irrigated or droughted conditions, was investigated by monitoring the structural development of the cambium. To track microstructural changes in the cambium, the same seedlings were imaged with X-ray micro-CT (μCT) one day before the treatments and again six days after the respective treatments. After the last X-ray μCT scan, X-ray nano-CT was also applied. Using image analysis techniques, the morphological characteristics of the cambium could be determined. X-ray μCT displayed a larger, thicker cambial zone in irrigated plants, while a much thinner cambium was visible in droughted seedlings. X-ray nano-CT revealed that droughted plants were associated with a significantly () smaller cambium volume containing smaller cells, compared to the cambium of irrigated plants. Light microscopy was used to validate the CT results and demonstrated no significant () difference in the cambium width and cell diameter obtained from the two respective CT techniques. The findings of this study proved X-ray CT to be a valuable tool for examining the effect of changing environmental conditions on the complex cambium structure of Corymbia hybrid seedlings.

Genetic Interaction between Arabidopsis SUR2/CYP83B1 andGNOM Indicates the Importance ofStabilizing Local Auxin Accumulation inLateral Root Initiation.

Lateral root (LR) formation is an important developmental event for the establishment of the root system in most vascular plants. In Arabidopsis thaliana, the fewer roots (fwr) mutation in the GNOM gene, encoding a guanine nucleotide exchange factor of ADP ribosylation factor that regulates vesicle trafficking, severely inhibits LR formation. Local accumulation of auxin response for LR initiation is severely affected in fwr. To better understand how local accumulation of auxin response for LR initiation is regulated, we identified a mutation, fewer roots suppressor1 (fsp1), that partially restores LR formation in fwr. The gene responsible for fsp1 was identified as SUPERROOT2 (SUR2), encoding CYP83B1 that positions at the metabolic branch point in the biosynthesis of auxin/indole-3-acetic acid (IAA) and indole glucosinolate. The fsp1 mutation increases both endogenous IAA levels and the number of the sites where auxin response locally accumulates prior to LR formation in fwr. SUR2 is expressed in the pericycle of the differentiation zone and in the apical meristem in roots. Time-lapse imaging of the auxin response revealed that local accumulation of auxin response is more stable in fsp1. These results suggest that SUR2/CYP83B1 affects LR founder cell formation at the xylem pole pericycle cells where auxin accumulates. Analysis of the genetic interaction between SUR2 and GNOM indicates the importance of stabilization of local auxin accumulation sites for LR initiation.

Multi-objective optimization methodology for green-gray coupled runoff control infrastructure adapting spatial heterogeneity of natural endowment and urban development

Cost-effective runoff control scheme drafting involves localization, multi-sector coordination, and configuration of multifunctional infrastructures. Numerous independent variables, parameters, weights, and objectives make runoff control optimization quantitatively arduous. This study innovatively proposed a multi-objective optimization methodology for green-gray coupled runoff control infrastructure adapting spatial heterogeneity of natural endowment and urban development. The quantitative methods of multi-objective evaluation, hydrological feature partition, and pressure-adapted multi-objective weight assignment were proposed. Remote sensing inversion of water quality, hydrological model simulation (using SWAT and SWMM software), landscape pattern index calculation, life cycle cost (LCC), life cycle assessment (LCA) on ecological impact, and NSGA-II optimization algorithm were applied. Wuhan, the most water-sensitive city in China, was studied as a case. Runoff control function (RCF), capital investment (CI), and ecological return on investment (EROI) served as optimized objectives. High, medium, and low built-up regions in Wuhan urban development planning district were extracted by topographic factors and landscape patterns, which comprised 28, 34, and 38% of the case area, respectively. Three corresponding hydrological models were then built to illustrate distinct runoff control cost-efficiency in each region. Pressure distributions on runoff control, economic constraints, and ecological resource scarcity were quantitatively evaluated. And four pressure zones were clustered, which occupied 36, 29, 16, and 19% of the case area, respectively. Then the zonal weighted optimization decision-making matrix (with 3 hydrological models and 5 wt) was established by overlaying the pressure zone and built-up zone. In high, medium, and low built-up regions, optimized solutions reduced annual runoff volume by 86, 82%, and 77%The average runoff investments per square meter of impervious underlying surface in high, medium, and low built-up regions were 34.2, 18.7, and 7.9 RMB yuan, respectively. Medium and low built-up regions may only need 55 and 23% of the high built-up region for the unitary impervious underlying surface to balance runoff control and ecological benefits. Runoff control and financial utilization efficiency enhance with hydrological differentiation zones. Thus, the optimization solutions are zonal adaptive, refined, comparable, replicable, and implementable.

A dual-flow RootChip enables quantification of bi-directional calcium signaling in primary roots.

One sentence summary: Bi-directional-dual-flow-RootChip to track calcium signatures in Arabidopsis primary roots responding to osmotic stress. Plant growth and survival is fundamentally linked with the ability to detect and respond to abiotic and biotic factors. Cytosolic free calcium (Ca2+) is a key messenger in signal transduction pathways associated with a variety of stresses, including mechanical, osmotic stress and the plants' innate immune system. These stresses trigger an increase in cytosolic Ca2+ and thus initiate a signal transduction cascade, contributing to plant stress adaptation. Here we combine fluorescent G-CaMP3 Arabidopsis thaliana sensor lines to visualise Ca2+ signals in the primary root of 9-day old plants with an optimised dual-flow RootChip (dfRC). The enhanced polydimethylsiloxane (PDMS) bi-directional-dual-flow-RootChip (bi-dfRC) reported here adds two adjacent inlet channels at the base of the observation chamber, allowing independent or asymmetric chemical stimulation at either the root differentiation zone or tip. Observations confirm distinct early spatio-temporal patterns of salinity (sodium chloride, NaCl) and drought (polyethylene glycol, PEG)-induced Ca2+ signals throughout different cell types dependent on the first contact site. Furthermore, we show that the primary signal always dissociates away from initially stimulated cells. The observed early signaling events induced by NaCl and PEG are surprisingly complex and differ from long-term changes in cytosolic Ca2+ reported in roots. Bi-dfRC microfluidic devices will provide a novel approach to challenge plant roots with different conditions simultaneously, while observing bi-directionality of signals. Future applications include combining the bi-dfRC with H2O2 and redox sensor lines to test root systemic signaling responses to biotic and abiotic factors.

Stepping up to genome scan allows stock differentiation in the worldwide distributed blue shark Prionace glauca.

The blue shark Prionace glauca is a top predator with one of the widest geographical distributions of any shark species. It is classified as Critically Endangered in the Mediterranean Sea, and Near Threatened globally. Previous genetic studies did not reject the null hypothesis of a single global population. The blue shark was proposed as a possible archetype of the "grey zone of population differentiation," coined to designate cases where population structure may be too recent or too faint to be detected using a limited set of markers. Here, blue shark samples collected throughout its global range were sequenced using a specific RAD method (DArTseq), which recovered 37,655 genome-wide single nucleotide polymorphisms (SNPs). Two main groups emerged, with Mediterranean Sea and northern Atlantic samples (Northern population) differentiated significantly from the Indo-west Pacific samples (Southern population). Significant pairwise FST values indicated further genetic differentiation within the Atlantic Ocean, and between the Atlantic Ocean and the Mediterranean Sea. Reconstruction of recent demographic history suggested divergence between Northern and Southern populations occurred about 500 generations ago and revealed a drastic reduction in effective population size from a large ancestral population. Our results illustrate the power of genome scans to detect population structure and reconstruct demographic history in highly migratory marine species. Given that the management plans of the blue shark (targeted or bycatch) fisheries currently assume panmictic regional stocks, we strongly recommend that the results presented here be considered in future stock assessments and conservation strategies.

Histomorphological variants of oral squamous cell carcinoma and their relation with prognosis of the disease

The key to wealth is health but in present days cancer is going to exhaust mental, physical and economic conditions. Oral Cancer is the 6 most common cancer worldwide. Approximately 630,000 new patients diagnosed annually resulting in more than 350,000 deaths every year. Squamous cell carcinoma is by far the most important and the most common malignant mucosal neoplasm of the head and neck accounting for over 90% of all malignancies.Histomorphological features of oral squamous cell carcinoma (OSCC) and their relation with the pathological prognostic factors i.e. histological stage and grade of malignancy is the key feature, which help early and easy prognosis of OSCC. :To study the histomorphological features of oral squamous cell carcinoma and their relation with the pathological prognostic factors i.e. histological stage and grade of malignancy. Recent reports suggest an increase in oral squamous cell carcinoma (OSCC) frequency. To improve programs in public health, it is necessary to understand the epidemiological conditions. A retrospective review of all OSCC cases diagnosed by the Pathology Department was performed. Demographic data, in addition to anatomic zone and histological degree of differentiation were obtained. Central tendency, dispersion and prevalence rate per 100,000 individuals were determined. A total of 150 patients were diagnosed with OSCC and majority of them were men 118(79%) were men. OSCC were predominantly observed in the age of 40-60 years. The predominant anatomic zone was the Buccal mucosa, GBS, Angle of Mouth 99(66%), followed by the tongue and lip 42(28%) and Retromolar Triagone 09(06%).The most frequent histological degree was well differentiated in 110 cases (73.3%). The rates of OSCC prevalence showed similar patterns in terms across time.

Cell-Type-Specific Length and Cytosolic pH Response of Superficial Cells of Arabidopsis Root to Chronic Salinity.

Soil salinity negatively affects the growth, development and yield of plants. Acidification of the cytosol in cells of glycophytes was reported under salinity, while various types of plant cells can have a specific reaction under the same conditions. Transgenic Arabidopsis plants expressing the pH sensor Pt-GFP in the cytosol were used in this work for determination of morphometric changes and cytosolic pH changes in the superficial cells of Arabidopsis roots under chronic salinity in vitro. We did not find changes in the length of the root cap cells, while there was a decrease in the length of the differentiation zone under 50, 75 mM NaCl and the size of the epidermal cells of the differentiation zone under 75 mM NaCl. The most significant changes of cytosolic pH to chronic salinity was noted in columella (decrease by 1 pH unit at 75 mM NaCl) and epidermal cells of the differentiation zone (decrease by 0.6 and 0.4 pH units at 50 and 75 mM NaCl, respectively). In developed lateral root cap cells, acidification of cytosol by 0.4 units occurred only under 75 mM NaCl in the medium. In poorly differentiated lateral cells of the root cap, there were no changes in pH under chronic salinity.

Maize lateral rootless 1 encodes a homolog of the DCAF protein subunit of the CUL4-based E3 ubiquitin ligase complex.

In maize (Zea mays L.), lateral roots are formed in the differentiation zone of all root types in a multi-step process. The maize mutant lateral rootless 1 (lrt1) is defective in lateral root formation in primary and seminal roots but not in shoot-borne roots. We cloned the lrt1 gene by mapping in combination with BSA-seq and subsequent validation via CRISPR/Cas9. The lrt1 gene encodes a 209 kDa homolog of the DDB1-CUL4-ASSOCIATED FACTOR (DCAF) subunit of the CUL4-based E3 ubiquitin ligase (CRL4) complex localized in the nucleus. DDB1-CUL4-ASSOCIATED FACTOR proteins are encoded by an evolutionary old gene family already present in nonseed plants. They are adaptors that bind substrate proteins and promote their ubiquitylation, thus typically marking them for subsequent degradation in the 26S proteasome. Gene expression studies demonstrated that lrt1 transcripts are expressed preferentially in the meristematic zone of all root types of maize. Downregulation of the rum1 gene in lrt1 mutants suggests that lrt1 acts upstream of the lateral root regulator rum1. Our results demonstrate that DCAF proteins play a key role in root-type-specific lateral root formation in maize. Together with its role in nitrogen acquisition in nitrogen-poor soil, lrt1 could be a promising target for maize improvement.

Associations of dietary, lifestyle, and other participant characteristics with APC, β-catenin, E-cadherin, and MSH2 expression in the normal mucosa of sporadic colorectal adenoma patients

Abnormal expression of Wnt pathway and DNA mismatch repair proteins is common during colorectal carcinogenesis. To investigate cross-sectional associations of lifestyle, dietary, and other participant characteristics with the expression of such proteins, we measured APC, β-catenin, E-cadherin, and MSH2 colorectal crypt expression in biopsies of normal-appearing colorectal mucosa from 104 sporadic colorectal adenoma patients using automated immunohistochemistry and quantitative image analysis. We used multivariable general linear models to compare adjusted mean biomarker expression across categories of participant characteristics. Example findings include that among women relative to men, mean APC expression in whole crypts, the upper 40% of crypts (differentiation zone), and the lower 60% of crypts (proliferation zone) was 322.9% higher (p<0.01), 296.7% higher (p<0.01), and 399.1% higher (p<0.01), respectively. Among participants with higher alcohol consumption, APC expression in the crypt differentiation zone was estimated to be 15.9% lower (p=0.08). Among those with higher total meat consumption, β-catenin expression in whole crypts and the crypt proliferation zone was estimated to be 20.5% higher (p=0.07) and 19.6% higher (p=0.06), respectively, and MSH2 expression in the crypt differentiation zone was estimated to be 64.4% lower (p=0.10). Among those with a higher body mass index, MSH2 expression in the crypt differentiation zone was estimated to be 87.5% lower (p=0.15). These pilot study findings suggest that being male, higher adiposity, and higher alcohol and meat consumption may be unfavorably associated with biomarkers of colorectal carcinogenesis pathway proteins in the normal-appearing colorectal mucosa of sporadic colorectal adenoma patients and support further investigation in larger studies.

Trifluoperazine (TFP)-mediated fluorescence imaging approach reveals a probable calmodulin (CaM)-independent calcium signaling accompanying differential protein phosphorylation in NaCl-stressed sunflower seedlings (Helianthus annuus L. var. KBSH44)

The impact of calcium (Ca2+) under abiotic stress is manifested by both calmodulin (CaM)-dependent and calmodulin-independent pathways of signaling mechanisms in plants. Present work demonstrates the impact of NaCl (120 mM) stress on Ca2+-calmodulin activity and its spatial distribution in roots of 2d old sunflower seedlings (Helianthus annuus). NaCl stress significantly increases [Ca2+]cyt in roots (69%) which corresponds with 8% increase in activated calmodulin (Ca2+-CaM) levels. Thus, NaCl stress induced Ca2+ signaling in roots is likely to majorly operate through CaM-independent routes. Trifluoperazine-mediated fluorescence imaging reveals differential distribution of Ca2+-CaM complex in the elongation and differentiation zone of seedling roots which indicate spatial regulation of Ca2+-CaM signaling operative in the presence of NaCl stress. NaCl stress-induced CaM-independent Ca2+ signaling is further accompanied by differential protein phoshorylation of cytosolic proteins in seedling roots and cotyledons. Phosphoproteins exhibiting qualitative differences in roots in response to salt-stress were of both low and high molecular weight range (76–17.5 kDa) unlike in cotyledons where only high molecular weight (>76 kDa) phosphoproteins exhibited major differential expressions in response to 120 mM NaCl stress. Among the various polypeptides in the fraction of OB membrane proteins two polypeptides of 48 and >76 kDa exhibited increased protein phosphorylation in the presence of NaCl stress. NaCl stress for 48 h induces differences in the expression of phosphoproteins in roots and cotyledons, suggesting the long distance signaling of Na+ stress from roots to cotyledons. Roots seem to exhibit much stringent regulation of phosphoproteins in comparison with cotyledons which have lesser qualitative differences. Thus, present work provides evidence on the modulation of Ca2+-CaM signaling accompanying differential protein phosphorylation in sunflower seedling roots and cotyledons under NaCl stress.

Genome-Wide Identification of Potential mRNAs in Drought Response in Wheat (Triticum aestivum L.).

Plant cell metabolism inevitably forms an important drought-responsive mechanism, which halts crop productivity. Globally, more than 30% of the total harvested area was affected by dehydration. RNA-seq technology has enabled biologists to identify stress-responsive genes in relatively quick times. However, one shortcoming of this technology is the inconsistent data generation compared to other parts of the world. So, we have tried, here, to generate a consensus by analyzing meta-transcriptomic data available in the public microarray database GEO NCBI. In this way, the aim was set, here, to identify stress genes commonly identified as differentially expressed (p < 0.05) then followed by downstream analyses. The search term “Drought in wheat” resulted in 233 microarray experiments from the GEO NCBI database. After discarding empty datasets containing no expression data, the large-scale meta-transcriptome analytics and one sample proportional test were carried out (Bonferroni adjusted p < 0.05) to reveal a set of 11 drought-responsive genes on a global scale. The annotation of these genes revealed that the transcription factor activity of RNA polymerase II and sequence-specific DNA-binding mechanism had a significant role during the drought response in wheat. Similarly, the primary root differentiation zone annotations, controlled by TraesCS5A02G456300 and TraesCS7B02G243600 genes, were found as top-enriched terms (p < 0.05 and Q < 0.05). The resultant standard drought genes, glycosyltransferase; Arabidopsis thaliana KNOTTED-like; bHLH family protein; Probable helicase MAGATAMA 3; SBP family protein; Cytochrome c oxidase subunit 2; Trihelix family protein; Mic1 domain-containing protein; ERF family protein; HD-ZIP I protein; and ERF family protein, are important in terms of their worldwide proved link with stress. From a future perspective, this study could be important in a breeding program contributing to increased crop yield. Moreover, the wheat varieties could be identified as drought-resistant/sensitive based on the nature of gene expression levels.