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The growth of industrialization growth the risk of vanadium (V) contamination. The objective of this study was to examine the impact of 200 µg L - 1 VCI 3 -induced toxicity as well as the potential protective effect of 187.5 mg L - 1 and 375 mg L - 1 Hypericum perforatum (H. perforatum) extracts against this toxicity on the Allium cepa (A. cepa) model organism. For this purpose, a series of investigations were conducted on the growth physiology alterations (germination percentage, root elongation, weight gain), cytogenetic alterations (mitotic index, micronucleus, chromosomal aberrations), biochemical alterations (malondialdehyde, superoxide dismutase, catalase) and defects in meristematic tissue in A. cepa. In addition, the phenolic compound content of H. perforatum extract was determined by the LC/MS-MS method. V application negatively affected all the investigated parameters and caused a serious phytotoxic and genotoxic effect as well as oxidative stress in A. cepa. Conversely, no statistical difference was observed between the parameters of the groups treated with H. perforatum extract and those of the control group. The administration of H. perforatum extract combined with V resulted in a notable enhancement in germination percentage, root elongation, weight gain, mitotic index value, chlorophyll a level and chlorophyll b level. Additionally, it led to a reduction in micronucleus and chromosomal aberrations frequencies, as well as meristematic tissue defects. Furthermore, LC/MS-MS analysis demonstrated that H. perforatum extract contains phenolic compounds, including catechin, epicatechin, quercetin, oleuropein and rutin, which confer antioxidant properties to the extract. The study provided clear evidence that H. perforatum extract attenuates the toxic effects of V in A. cepa, which can be attributed to its high content of bioactive phenols. The findings of the study indicate that H. perforatum extract may serve as a protective natural agent for daily use against heavy metal toxicity., Competing Interests: Declarations Statement regarding experimental research on plants Experimental research and field studies on plants and plant parts, including the collection of plant material, comply with relevant institutional, national, and international guidelines and legislation. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Large scale production of uniform disease-free plants is crucial for Cannabis sativa biotechnology. Existing micropropagation protocols rely heavily on shoot multiplication from existing meristems via direct organogenesis. Such protocols do not allow multiplication of plant material through continuous sub-culturing. Protocols that use indirect regeneration are usually not efficient enough and have very low multiplication rates. In the present study, an efficient protocol that uses a combination of direct organogenesis and callogenesis to induce multiple shoot development cultures is developed. Callogenesis was induced from various explants cultured on the media having various combinations of thidiazuron (TDZ) and naphthaleneacetic acid (NAA); best callogenesis and shoot regeneration was achieved from hypocotyl explants cultured on TDZ 0.4 mg l -1 NAA 0.2 mg l -1 . Hypocotyls with cotyledonary node and shoot apical meristem were significantly better for shoot regeneration than explants without it. Shoots obtained from multiple shoot cultures were successfully rooted and then acclimatized under greenhouse conditions to develop into adult cannabis plants., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Igor Kovalchuk reports financial support was provided by University of Lethbridge. Igor Kovalchuk reports a relationship with University of Lethbridge that includes: employment. n/a If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A randomised open clinical/laboratory study was performed to evaluate the safety and cosmetic efficacy of facial cosmetics for females during the menopausal period. The cosmetics contain active ingredients of meristem cells derived from the medicinal plants Leontopodium alpinum , Buddeleja davidii , Centella asiatica , and Echinacea angustifolia . Recently, the major bioactive molecules of these medicinal plants (leontopodic acid, verbascoside, asiaticoside, and echinacoside, respectively) have been thoroughly evaluated in vitro for molecular pathways and cellular mechanisms and their preventive/curative effects on human skin cells exposed to factors promoting premature skin ageing and cellular senescence. Nevertheless, clinical data on their safety/efficacy to ageing human skin are scarce. This clinical study enrolled 104 Caucasian females in pre-menopause, menopause, or post-menopause periods. They applied cosmetic serums daily for 1 month. Questionnaires and instrumental and biochemical methods were used to assess dermatological/ophthalmological safety and cosmetic efficacy through changes of the skin physiology markers characteristic of ageing/menopause (elasticity, barrier functions, moisture, sebum, ultrasonic properties, and collagen content and structure). Quantitative microbiological tests were carried out for skin microbiota fluctuations. Data showed that the cosmetics were safe, and they shifted the skin physiology parameters to a younger biological age, enhanced collagen synthesis, inhibited lipid peroxidation, and favoured normal microbiota.

Methylisothiazolinone (MIT) is a widely used preservative and biocide to prevent product degradation, yet its potential impact on plant growth remains poorly understood. In this study, we investigated MIT's toxic effects on Arabidopsis thaliana root growth. Exposure to MIT significantly inhibited Arabidopsis root growth, associated with reduced root meristem size and root meristem cell numbers. We explored the polar auxin transport pathway and stem cell regulation as key factors in root meristem function. Our findings demonstrated that MIT suppressed the expression of the auxin efflux carrier PIN1 and major root stem cell regulators (PLT1, PLT2, SHR, and SCR). Additionally, MIT hindered root regeneration by downregulating the quiescent center (QC) marker WOX5. Transcriptome analysis revealed MIT-induced alterations in gene expression related to oxidative stress, with physiological experiments confirming elevated reactive oxygen species (ROS) levels and increased cell death in root tips at concentrations exceeding 50 μM. In summary, this study provides critical insights into MIT's toxicity on plant root development and regeneration, primarily linked to modifications in polar auxin transport and downregulation of genes associated with root stem cell regulation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Australian saltbush (Atriplex spp.) survive in exceptionally saline environments and are often used for pasture in semi-arid areas. To investigate the impact of salinity on saltbush root morphology and root exudates, three Australian native saltbush species (Atriplex nummularia , Atriplex amnicola , and Atriplex vesicaria ) were grown in vitro in optimised sterile, semi-hydroponic systems in media supplemented with different concentrations of salt (NaCl). Histological stains and chromatographic techniques were used to characterise the root apical meristem (RAM) type and root exudate composition of the saltbush seedlings. We report that saltbush species have closed-type RAMs, which release border-like cells (BLCs). Monosaccharide content, including glucose and fructose, in the root mucilage of saltbush was found to be uniquely low, suggesting that saltbush may minimise carbon release in polysaccharides of root exudates. Root mucilage also contained notable levels of salt, plus increasing levels of unidentified compounds at peak salinity. Un-esterified homogalacturonan, xyloglucan, and arabinogalactan proteins between and on the surface of BLCs may aid intercellular adhesion. At the highest salinity levels, root cap morphology was altered but root:shoot ratio remained consistent. While questions remain about the identity of some components in saltbush root mucilage other than the key monosaccharides, this new information about root cap morphology and cell surface polysaccharides provides avenues for future research.

Main Conclusion: Mutation of OsSHR2 adversely impacted root and shoot growth and impaired plant response to N conditions, further reducing the yield per plant. Nitrogen (N) is a crucial factor that regulates the plant architecture. There is still a lack of research on it. In our study, it was observed that the knockout of the SHORTROOT 2 (OsSHR2) which was induced by N deficiency, can significantly affect the regulation of plant architecture response to N in rice. Under N deficiency, the mutation of OsSHR2 significantly reduced root growth, and impaired the sensitivity of the root meristem length to N deficiency. The mutants were found to have approximately a 15% reduction in plant height compared to wild type. But mutants showed a significant increase in tillering at post-heading stage, approximately 26% more than the wild type, particularly in high N conditions. In addition, due to reduced seed setting rate and 1000-grain weight, mutant yield was significantly decreased by approximately 33% under low N fertilizer supply. The mutation also changed the distribution of N between the vegetative and reproductive organs. Our findings suggest that the transcription factor OsSHR2 plays a regulatory role in the response of plant architecture and yield per plant to N in rice., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The conservation of GOLVEN (GLV)/ROOT MERISTEM GROWTH FACTOR (RGF) peptide encoding genes across plant genomes capable of forming roots or root-like structures underscores their potential significance in the terrestrial adaptation of plants. This study investigates the function and role of GOLVEN peptide-coding genes in Medicago truncatula. Five out of fifteen GLV/RGF genes were notably upregulated during nodule organogenesis and were differentially responsive to nitrogen deficiency and auxin treatment. Specifically, the expression of MtGLV9 and MtGLV10 at nodule initiation sites was contingent upon the NODULE INCEPTION transcription factor. Overexpression of these five nodule-induced GLV genes in hairy roots of M. truncatula and application of their synthetic peptide analogues led to a decrease in nodule count by 25-50%. Uniquely, the GOLVEN10 peptide altered the positioning of the first formed lateral root and nodule on the primary root axis, an observation we term 'noduletaxis'; this decreased the length of the lateral organ formation zone on roots. Histological section of roots treated with synthetic GOLVEN10 peptide revealed an increased cell number within the root cortical cell layers without a corresponding increase in cell length, leading to an elongation of the root likely introducing a spatiotemporal delay in organ formation. At the transcription level, the GOLVEN10 peptide suppressed expression of microtubule-related genes and exerted its effects by changing expression of a large subset of Auxin responsive genes. These findings advance our understanding of the molecular mechanisms by which GOLVEN peptides modulate root morphology, nodule ontogeny, and interactions with key transcriptional pathways., (© 2024 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Bleomycin, commonly employed in treating Hodgkin's lymphoma and testicular cancer, is associated with significant pulmonary toxicity. While various studies have assessed the toxic impact of chemotherapeutic agents on aquatic and terrestrial environments, limited data exist on bleomycin's effects, especially concerning higher plants. To address this gap, we utilized the Allium cepa assays, renowned for evaluating chemical and biochemical agents' toxic effects, to investigate bleomycin's impact on the terrestrial ecosystem. Our study aimed to assess bleomycin's cyto-genotoxic effects on A. cepa root tip cells at minimal concentrations (10-40 μg mL -1 ) and varied exposure durations (2, 4, 6, and 24 h). Analysis of nuclear and mitotic abnormalities in bleomycin-treated A. cepa root tip cells, alongside an acridine orange-ethidium bromide double staining assay, illuminated its influence on cell viability. Additionally, agarose gel electrophoresis determined the drug's potential for DNA degradation, unveiling the underlying mechanisms of cyto-genotoxicity. Results also demonstrated a decline in the mitotic index with increased bleomycin concentrations and exposure time, elevated frequencies of various cyto-genotoxic abnormalities, including sticky chromosomes, chromatid breaks, laggards, bridges, polar deviations, nuclear lesions, and hyperchromasia. The study indicated the potential risks of bleomycin even at low concentrations and brief exposures, highlighting its severe adverse effects on genetic material of plant, potentially contributing to cell death. Consequently, this investigation unveils bleomycin's cyto-genotoxic effects on higher plant system, underscoring its threat to terrestrial ecosystems, particularly upon chronic and unmonitored exposure., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Thunbergia coccinea Wall. ex D. Don being a rare, ornamental and medicinal plant of India, is needed to propagate for conserving the germplasm and analyzing its phytochemical compounds in the future. A reliable protocol for direct in vitro propagation using nodal shoot meristem of T. coccinea as explant was standardized. The highest number of shoots per explant (22.17 ± 0.54) with maximum shoot length (2.36 ± 0.28) in cm was obtained in Murashige and Skoog (MS) medium supplemented with 9.70 µM of 6-furfurylaminopurine (Kinetin) and 0.053 µM of α-naphthaleneacetic acid (NAA) combination, among all the different plant growth regulators (PGR's) and concentrations tested. The aforesaid PGR's combination was optimum for axillary shoot bud induction and multiplication in T. coccinea. The best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 0.32) in cm. All the in vitro raised plantlets were acclimatized in sterile sand and soil mixture (1:1) with a survival rate of 70% on earthen pots under greenhouse conditions. PCR-based RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat) molecular markers were employed to determine the genetic homogeneity amongst the plantlets. Twelve (12) RAPD and nine (9) ISSR primers developed a total of 104 and 91 scorable bands, respectively. The band profiles of micropropagated plantlets were monomorphic to the mother, donor in vivo plant, and similarity values varied from 0.9542-1.000. The dendrogram generated through UPGMA (unweighted pair group method with arithmetic mean) showed 99% similarities amongst all tested plants confirming the genetic uniformity of in vitro raised plants., (© 2022. The Author(s).)

Actinomycin D has been reported to selectively inhibit rRNA synthesis and ribosome biogenesis, induce G2 checkpoint of cell cycle arrest in HeLa cells. In Arabidopsis, actinomycin D was also used as agent to preferentially inhibit the ribosome biosynthesis and ribosomal function. However, the function of actinomycin D on Arabidopsis root development remains to be elucidated. In this study, we exposed Arabidopsis seedlings to actinomycin D with the aim of evaluating the effects of ribosome biogenesis on root development. The results demonstrated that actinomycin D inhibited Arabidopsis root growth by reduced meristematic activity in a dose dependent manner. Exposure to actinomycin D decreased the expression of WOX5 and key stem cell niche-defining transcription factors SHR and PLT1, thus the loss function of QC identity and stem cell niche maintenance. In addition, dead cells were observed after actinomycin D treatment in root stele initials and DNA damage response was constitutively activated. Collectively, we propose that ribosome biogenesis plays key role in primary root growth through maintenance of root stem cell niche and DNA damage response in Arabidopsis., Competing Interests: Declaration of competing interest The authors declared that they have no conflicts of interest to this work., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Experiments on Vicia faba root meristem cells exposed to 150 µM cadmium chloride (CdCl 2 ) were undertaken to analyse epigenetic changes, mainly with respect to DNA replication stress. Histone modifications examined by means of immunofluorescence labeling included: (1) acetylation of histone H3 on lysine 56 (H3K56Ac), involved in transcription, S phase, and response to DNA damage during DNA biosynthesis; (2) dimethylation of histone H3 on lysine 79 (H3K79Me2), correlated with the replication initiation; (3) phosphorylation of histone H3 on threonine 45 (H3T45Ph), engaged in DNA synthesis and apoptosis. Moreover, immunostaining using specific antibodies against 5-MetC-modified DNA was used to determine the level of DNA methylation. A significant decrease in the level of H3K79Me2, noted in all phases of the CdCl 2 -treated interphase cell nuclei, was found to correspond with: (1) an increase in the mean number of intranuclear foci of H3K56Ac histones (observed mainly in S-phase), (2) a plethora of nuclear and nucleolar labeling patterns (combined with a general decrease in H3T45Ph), and (3) a decrease in DNA methylation. All these changes correlate well with a general viewpoint that DNA modifications and post-translational histone modifications play an important role in gene expression and plant development under cadmium-induced stress conditions.

After a century of investigations, the function of the obligate betaproteobacterial endosymbionts accommodated in leaf nodules of tropical Rubiaceae remained enigmatic. We report that the α-D-glucose analogue (+)-streptol, systemically supplied by mature Ca. Burkholderia kirkii nodules to their Psychotria hosts, exhibits potent and selective root growth inhibiting activity. We provide compelling evidence that (+)-streptol specifically affects meristematic root cells transitioning to anisotropic elongation by disrupting cell wall organization in a mechanism of action that is distinct from canonical cellulose biosynthesis inhibitors. We observed no inhibitory or cytotoxic effects on organisms other than seed plants, further suggesting (+)-streptol as a bona fide allelochemical. We propose that the suppression of growth of plant competitors is a major driver of the formation and maintenance of the Psychotria-Burkholderia association. In addition to potential agricultural applications as a herbicidal agent, (+)-streptol might also prove useful to dissect plant cell and organ growth processes., (© 2021. The Author(s).)

Signal molecule hydrogen peroxide (H 2 O 2 ) plays critical roles in various processes of plant development. However, H 2 O 2 signaling network, especially the responders that sense and respond to the H 2 O 2 signal remain largely unknown. Here we report two homologous genes H 2 O 2 Response Gene 1 and 2 (HRG1/2) in Arabidopsis that could quickly respond to exogenous or endogenous H 2 O 2 . Knockdown of HRG1/2 facilitated seed germination while overexpression of HRG1/2 greatly retarded seed germination. ROS level in HRG1 overexpression roots was significantly lower than that in HRG1/2 mutants after H 2 O 2 treatment. The expression level of enzymatic antioxidant DHAR3 was upregulated in HRG1 overexpression plants, suggesting that DHAR3 is downstream of HRG1. That the root meristem length and cell number were significantly reduced in hrg1-1 and hrg2-1 plants upon H 2 O 2 treatment compared to that of HRG1 overexpression plants also approves the idea that HRGs function in H 2 O 2 removal. Further evolutionary analysis indicates that this is a dicotyledon-specific pathway responsive to H 2 O 2 . Together, this work reveals HRG1/2 as novel H 2 O 2 responders involved in ROS scavenging to ensure embryonic root meristem activity. These findings provide valuable clues for the of H 2 O 2 signaling and root meristem regulation., (Copyright © 2021 Elsevier B.V. All rights reserved.)

CLAVATA3 (CLV3) is a peptide signal initially identified in the analysis of clv mutants in the model plant Arabidopsis thaliana, as a regulator of meristem homeostasis and floral organ numbers. CLV3 homologs are widely conserved in land plants, collectively called CLV3/ESR-related (CLE) genes. A 12-amino acid CLE peptide with hydroxyproline residues was identified in Zinnia elegans cell culture system, in which cells secrete a CLE peptide called tracheary element differentiation factor (TDIF) into the culture medium. Mature CLV3 peptide is also a post-translationally modified short peptide containing additional triarabinosylation on a hydroxyproline residue. Genetic studies have revealed the involvement of leucin-rich repeat receptor-like kinases (LRR-RLKs) in CLV3 signaling, including CLV1/BAM-CIK, CLV2-CRN and RPK2, although the mechanisms of signal transduction and integration via crosstalk is still largely unknown. Recent studies on bryophyte model species provided a clue to understand evolution and ancestral function of CLV signaling in land plants. Fundamental understanding on CLV signaling provided an opportunity to optimize the crop yield traits using a novel breeding technology with CRISPR/Cas genome editing., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Coumarin is a phytotoxic natural compound able to affect plant growth and development. Previous studies have demonstrated that this molecule at low concentrations (100 µM) can reduce primary root growth and stimulate lateral root formation, suggesting an auxin-like activity. In the present study, we evaluated coumarin's effects (used at lateral root-stimulating concentrations) on the root apical meristem and polar auxin transport to identify its potential mode of action through a confocal microscopy approach. To achieve this goal, we used several Arabidopsis thaliana GFP transgenic lines (for polar auxin transport evaluation), immunolabeling techniques (for imaging cortical microtubules), and GC-MS analysis (for auxin quantification). The results highlighted that coumarin induced cyclin B accumulation, which altered the microtubule cortical array organization and, consequently, the root apical meristem architecture. Such alterations reduced the basipetal transport of auxin to the apical root apical meristem, inducing its accumulation in the maturation zone and stimulating lateral root formation.

Carotenoid-derived regulatory metabolites and hormones are generally known to arise through the oxidative cleavage of a single double bond in the carotenoid backbone, which yields mono-carbonyl products called apocarotenoids. However, the extended conjugated double bond system of these pigments predestines them also to repeated cleavage forming dialdehyde products, diapocarotenoids, which have been less investigated due to their instability and low abundance. Recently, we reported on the short diapocarotenoid anchorene as an endogenous Arabidopsis metabolite and specific signaling molecule that promotes anchor root formation. In this work, we investigated the biological activity of a synthetic isomer of anchorene, iso-anchorene, which can be derived from repeated carotenoid cleavage. We show that iso-anchorene is a growth inhibitor that specifically inhibits primary root growth by reducing cell division rates in the root apical meristem. Using auxin efflux transporter marker lines, we also show that the effect of iso-anchorene on primary root growth involves the modulation of auxin homeostasis. Moreover, by using liquid chromatography-mass spectrometry analysis, we demonstrate that iso-anchorene is a natural Arabidopsis metabolite. Chemical inhibition of carotenoid biosynthesis led to a significant decrease in the iso-anchorene level, indicating that it originates from this metabolic pathway. Taken together, our results reveal a novel carotenoid-derived regulatory metabolite with a specific biological function that affects root growth, manifesting the biological importance of diapocarotenoids., (© 2021 Society for Experimental Biology and John Wiley & Sons Ltd.)

Auxin alone or supplemented with cytokinins and strigolactones were long considered as the main player(s) in the control of apical dominance (AD) and correlative inhibition of the lateral bud outgrowth, the processes that shape the plant phenotype. However, past decade data indicate a more sophisticated pathways of AD regulation, with the involvement of mobile carbohydrates which perform both signal and trophic functions. Here we provide a critical comprehensive overview of the current status of the AD problem. This includes insight into intimate mechanisms regulating directed auxin transport in axillary buds with participation of phytohormones and sugars. Also roles of auxin, cytokinin and sugars in the dormancy or sustained growth of the lateral meristems were assigned. This review not only provides the latest data on implicated phytohormone crosstalk and its relationship with the signaling of sugars and abscisic acid, new AD players, but also focuses on the emerging biochemical mechanisms, at first positive feedback loops involving both sugars and hormones, that ensure the sustained bud growth. Data show that sugars act in concert with cytokinins but antagonistically to strigolactone signaling. A complex bud growth regulating network is demonstrated and unresolved issues regarding the hormone-carbohydrate regulation of AD are highlighted., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Polyamines (PAs) dramatically affect root architecture and development, mainly by unknown mechanisms; however, accumulating evidence points to hormone signaling and reactive oxygen species (ROS) as candidate mechanisms. To test this hypothesis, PA levels were modified by progressively reducing ADC1/2 activity and Put levels, and then changes in root meristematic zone (MZ) size, ROS, and auxin and cytokinin (CK) signaling were investigated. Decreasing putrescine resulted in an interesting inverted-U-trend in primary root growth and a similar trend in MZ size, and differential changes in putrescine (Put), spermidine (Spd), and combined spermine (Spm) plus thermospermine (Tspm) levels. At low Put concentrations, ROS accumulation increased coincidently with decreasing MZ size, and treatment with ROS scavenger KI partially rescued this phenotype. Analysis of double AtrbohD/F loss-of-function mutants indicated that NADPH oxidases were not involved in H 2 O 2 accumulation and that elevated ROS levels were due to changes in PA back-conversion, terminal catabolism, PA ROS scavenging, or another pathway. Decreasing Put resulted in a non-linear trend in auxin signaling, whereas CK signaling decreased, re-balancing auxin and CK signaling. Different levels of Put modulated the expression of PIN1 and PIN2 auxin transporters, indicating changes to auxin distribution. These data strongly suggest that PAs modulate MZ size through both hormone signaling and ROS accumulation in Arabidopsis .

The rampant use of pesticides can cause serious environmental problems. They can be contaminating surface water and groundwater, affecting the surrounding micro and macro biota. In this sense, this work aimed to evaluate the effects of a tebuconazole-based fungicide through endpoints accessed in Lactuca sativa bioassays. Germinated-seeds with roots upon 2 mm were treated with a fungicide containing Tebuconazole (TBZ) as active compound. The final concentration of TBZ in the tested solutions were 0.025 (C1); 0.05 (C2); 0.1 (C3); 0.2 (C4) and 0.4 g/L (C5). L. sativa roots were exposed for 24 h to these solutions and Petri dishes containing the treated seeds were kept in incubation chamber at 24 °C. Two positive controls (PC,) the herbicide trifluralin (0.84 mg/L) and Methanesulfonate (4 ×10 -4 mol/L), were applied. Distilled water was negative control (NC). The following endpoints were analyzed: root growth (RG), cytogenotoxic potential by cell cycle analysis, induction of DNA damage through TUNEL and comet assays. The obtained data were submitted to one-way variance analysis (ANOVA) and then to Tukey or Kruskal Wallis (P < 0.05) tests. The concentrations (C1, C2, C4 and C5) affected negatively the RG of L. sativa, in comparison with the NC. The mitotic index was reduced by 25% from NC to C1 and in the rest of treatments it did not present significant modifications. However, from C3 to C5 great amount of chromosome alterations were observed, in comparison with the NC. TBZ-based fungicide also induced DNA fragmentation as measured by TUNEL and comet assays. Thus, TBZ-based fungicide in some concentrations can have phytotoxic, cytotoxic and genotoxic effects in roots and meristematic cells of L. sativa., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Gravitropism plays a critical role in the growth and development of plants. Previous reports proposed that the disruption of the actin cytoskeleton resulted in enhanced gravitropism; however, the mechanism underlying these phenomena is still unclear. In the present study, real-time observation on the effect of Latrunculin B (Lat B), a depolymerizing agent of microfilament cytoskeleton, on gravitropism of the primary root of Arabidopsis was undertaken using a vertical stage microscope. The results indicated that Lat B treatment prevented the growth of root, and the growth rates of upper and lower flanks of the horizontally placed root were asymmetrically inhibited. The growth of the lower flank was influenced by Lat B more seriously, resulting in an increased differential growth rate between the upper and lower flanks of the root. Further analysis indicated that Lat B affected cell growth mainly in the transition and elongation zones. Briefly, the current data revealed that Lat B treatment inhibited cell elongation, especially the cells in the lower flanks of the transition and elongation zones, which finally manifested as the facilitation of gravitropic curvature of the primary root.

Cross-talk between phytohormones and sugars is intensely involved in plant metabolism, growth and regeneration. We documented alterations in cytokinin (CK) homeostasis in four developmental stages during de novo shoot organogenesis (DNSO) of kohlrabi (Brassica oleracea var. gongylodes cv. Vienna Purple) seedlings induced by exogenous CKs, trans-zeatin (transZ) and thidiazuron (TDZ), added together with elevated sucrose concentration (6% and 9%). Significant impact of CK and sucrose treatment and their interaction was recorded in all investigated stages, including plantlet development before calli formation (T1 and T2), calli formation (T3) and shoot regeneration (T4). Results showed remarkable increase in total CK levels for transZ treatment, particularly with 9% sucrose. This trend was observed for all physiological and structural groups of CKs. Application of TDZ contributed to little or no increase in CK levels regardless of sucrose concentration. Analysis of expression profiles of organogenesis-related genes involved in auxin transport, CK response, shoot apical meristem formation and cell division revealed that higher sugar concentration significantly downregulated the analysed genes, particularly in T3. This continued on TDZ, but transZ induced an opposite effect with 9% sucrose in T4, increasing gene activity. Our results demonstrated that phytohormone metabolism might be triggered by sucrose signalling in kohlrabi DNSO.

Ethnopharmacological Relevance: Aristolochia indica L. (Aristolochiaceae) is a common medicinal plant described in many traditional medicine as well as in Ayurveda used against snakebites. Besides, the plant has also been reported traditionally against fever, rheumatic arthritis, madness, liver ailments, dyspepsia, oedema, leishmaniasis, leprosy, dysmenorrhoea, sexual diseases etc. The plant is known to contain its major bioactive constituent aristolochic acid (AA) known for its anti-snake venom, abortifacient, antimicrobial and antioxidant properties., Materials and Methods: This present work describes a validated, fast and reproducible high performance thin layer chromatography (HPTLC) method to estimate AA from the roots of 20 chemotypes of A. indica procured from 20 diverse geographical locations from the state of West Bengal, India. Further, an evidence-based approach was adopted to investigate the reported anti-venom activity of the aqueous extracts of the A. indica roots by assessing its phospholipase A 2 (PLA 2 ) inhibitory properties since PLA 2 is a major component of many snake-venoms. Finally, the cytotoxicity and genotoxicity of the aqueous root extract of the Purulia (AI 1) chemotype were assessed at various concentrations using Allium cepa root meristematic cells., Results: The highest amount of AA (7643.67 μg/g) was determined in the roots of A. indica chemotype collected from Purulia district followed by the chemotypes collected from Murshidabad, Jalpaiguri and Birbhum districts (7398.34, 7345.09 and 6809.97 μg/g respectively). This study not only determines AA in the plants to select pharmacologically elite chemotypes of A. indica, but it also identifies high AA producing A. indica for further domestication and propagation of the plants for pharmacological and industrial applications. The method was validated via analyzing inter-day and intra-day precision, repeatability, reproducibility, instrumental precision, limit of detection (LOD) and limit of quantification (LOQ) and specificity. Chemotypes with high AA content exhibited superior anti-PLA 2 activity by selectively inhibiting human-group PLA 2 . Moreover, A. indica root extract significantly inhibited mitosis in Allium cepa root tips as a potent clastogen., Conclusions: The present quick, reproducible and validated HPTLC method provides an easy tool to determine AA in natural A. indica plant populations as well as in food and dietary supplements, a potential antivenin at one hand and a possible cause of aristolochic acid nephropathy (AAN) at another. Besides, the cytotoxic and mitotoxic properties of the root extracts should be used with caution especially for oral administration., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Plant cell wall, the first interface or barrier for toxic ions entering into protoplast, suffers from risk. Nitric oxide (NO) plays an important role in plant growth and responses to abiotic stresses, however, it is not clear whether NO is connected with the response of cell wall to aluminum (Al) tolerance in rice (Oryza sativa L.). In this study, we found that the application of 50 µM Al induces nitrate reductase (NR) activity and endogenous NO production, but not nitric oxide synthase (NOS) activity in two rice genotypes. Pretreatment with 100 µM NO donor (sodium nitroprusside, SNP) reduced Al-induced inhibition of root elongation by 32.3% and 91.7%, and Al accumulation in root-tip by 38.4% and 44.3% in Nipponbare and Zhefu802, respectively. The addition of SNP significantly decreased Al-induced accumulation of pectin, hemicellulose 1 and hemicellulose 2 by 43.1%, 13.1% and 19.2% in Zhefu802 and by 16.9%, 13.4% and 14.0% in Nipponbare, compared with roots treated with Al alone, as well as pectin methylesterase (PME) activity. Therefore, the content of Al absorbed in cell walls was decreased, indicating that the Al-induced structure damage to cell walls was alleviated. Furthermore, the activities of peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) treated by Al were all increased by SNP pretreatment, and the lipid peroxidation and damage to plasma membrane of root tips detected with Schiff's reagent and Evans blue reduced. In contrast, the effect was abolished when NO scavenger (cPTIO), and NR inhibitor (NaN 3 ), were added. These results indicated that by regulating the Al-binding capacity to cell walls and lipid peroxidation, the structure of cell walls can be stabilized and that Al toxicity in rice can be alleviated with increased NO., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Soil salinity is a key problem for crop production worldwide. High salt concentration in soil negatively modulates plant growth and development. In roots, salinity affects the growth and development of both primary and lateral roots. The phytohormone auxin regulates various developmental processes during the plant's life cycle, including several aspects of root architecture. Auxin signaling involves the perception by specialized receptors which module several regulatory pathways. Despite their redundancy, previous studies have shown that their functions can also be context-specific depending on tissue, developmental or environmental cues. Here we show that the over-expression of Auxin Signaling F-Box 3 receptor results in an increased resistance to salinity in terms of root architecture and germination. We also studied possible downstream signaling components to further characterize the role of auxin in response to salt stress. We identify the transcription factor SZF1 as a key component in auxin-dependent salt stress response through the regulation of NAC4. These results give lights of an auxin-dependent mechanism that leads to the modulation of root system architecture in response to salt identifying a hormonal cascade important for stress response.

The phytohormone cytokinin regulates diverse aspects of plant growth and development. Our understanding of the metabolism and perception of cytokinin has made great strides in recent years, mostly from studies of the model dicot Arabidopsis Here, we employed a CRISPR/Cas9-based approach to disrupt a subset of cytokinin histidine kinase (HK) receptors in rice ( Oryza sativa ) in order to explore the role of cytokinin in a monocot species. In hk5 and hk6 single mutants, the root growth, leaf width, inflorescence architecture and/or floral development were affected. The double hk5 hk6 mutant showed more substantial defects, including severely reduced root and shoot growth, a smaller shoot apical meristem, and an enlarged root cap. Flowering was delayed in the hk5 hk6 mutant and the panicle was significantly reduced in size and infertile due to multiple defects in floral development. The hk5 hk6 mutant also exhibited a severely reduced cytokinin response, consistent with the developmental phenotypes arising from a defect in cytokinin signaling. These results indicate that HK5 and HK6 act as cytokinin receptors, with overlapping functions to regulate diverse aspects of rice growth and development., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

Background: Hydrogen sulfide (H 2 S) is a gas signal molecule involved in regulating plants tolerance to heavy metals stress. In this study, we investigated the role of H 2 S in cadmium-(Cd-) induced cell death of root tips of cucumber seedlings., Results: The results showed that the application of 200 μM Cd caused cell death, increased the content of reactive oxygen species (ROS), chromatin condensation, the release of Cytochrome c (Cyt c) from mitochondria and activated caspase-3-like protease. Pretreatment of seedlings with 100 μM sodium hydrogen sulfide (NaHS, a H 2 S donor) effectively alleviated the growth inhibition and reduced cell death of root tips caused by Cd stress. Additionally, NaHS + Cd treatment could decrease the ROS level and enhanced antioxidant enzyme activity. Pretreatment with NaHS also inhibited the release of Cyt c from the mitochondria, the opening of the mitochondrial permeability transition pore (MPTP), and the activity of caspase-3-like protease in the root tips of cucumber seedling under Cd stress., Conclusion: H 2 S inhibited Cd-induced cell death in cucumber root tips by reducing ROS accumulation, activating the antioxidant system, inhibiting mitochondrial Cyt c release and reducing the opening of the MPTP. The results suggest that H 2 S is a negative regulator of Cd-induced cell death in the root tips of cucumber seedling.

Plant cells undergo two types of cell cycles-the mitotic cycle in which DNA replication is coupled to mitosis, and the endocycle in which DNA replication occurs in the absence of cell division. To investigate DNA replication programs in these two types of cell cycles, we pulse labeled intact root tips of maize (Zea mays) with 5-ethynyl-2'-deoxyuridine (EdU) and used flow sorting of nuclei to examine DNA replication timing (RT) during the transition from a mitotic cycle to an endocycle. Comparison of the sequence-based RT profiles showed that most regions of the maize genome replicate at the same time during S phase in mitotic and endocycling cells, despite the need to replicate twice as much DNA in the endocycle and the fact that endocycling is typically associated with cell differentiation. However, regions collectively corresponding to 2% of the genome displayed significant changes in timing between the two types of cell cycles. The majority of these regions are small with a median size of 135 kb, shift to a later RT in the endocycle, and are enriched for genes expressed in the root tip. We found larger regions that shifted RT in centromeres of seven of the ten maize chromosomes. These regions covered the majority of the previously defined functional centromere, which ranged between 1 and 2 Mb in size in the reference genome. They replicate mainly during mid S phase in mitotic cells but primarily in late S phase of the endocycle. In contrast, the immediately adjacent pericentromere sequences are primarily late replicating in both cell cycles. Analysis of CENH3 enrichment levels in 8C vs 2C nuclei suggested that there is only a partial replacement of CENH3 nucleosomes after endocycle replication is complete. The shift to later replication of centromeres and possible reduction in CENH3 enrichment after endocycle replication is consistent with a hypothesis that centromeres are inactivated when their function is no longer needed., Competing Interests: The authors have declared that no competing interests exist.

Main Conclusion: Cadmium stress induces WDR5a expression to promote NO accumulation to repress root meristem growth via suppressing auxin transport and synthesis in Arabidopsis. Nitric oxide (NO) synthase (NOS)-like activity plays a vital role in toxic cadmium (Cd)-induced NO production and inhibition of root meristem growth, while factor(s) regulating NOS-like activity and root meristem growth in plant response to Cd has not been identified yet. Here, we report that WD40 repeat 5a (WDR5a) functions in Cd-induced NOS-like activity, NO accumulation and root meristem growth suppression. We found that wdr5a-1 mutant root has increased root meristem growth with lower NOS-like activity and NO accumulation than wild type upon Cd exposure, and exogenous NO donors sodium nitroprusside or nitrosoglutathione can restore its reduced Cd sensitivity. In addition, Cd activates WDR5a expression in roots, and overexpressing WDR5a results in increased NO accumulation and suppressed root meristem growth similar to Cd-stressed wild-type roots, while scavenging NO or inhibiting NOS-like activity significantly reverts these effects of Cd. Furthermore, WDR5a acts in Cd-repressed auxin accumulation through reducing the levels of auxin efflux carriers PIN1/3/7 and biosynthetic enzyme TAA1, and reduced sensitivity of wdr5a-1 root meristem to Cd can be partially reverted by inhibiting TAA1 activity pharmaceutically or mutating TAA1 genetically. This study identified WDR5a as a key factor modulating NO accumulation and root meristem growth in plant response to Cd.

Because zinc sulfate (ZnSO 4 ) is widely used in many fields such as biomedicine, electronics, and chemistry, it is important to evaluate its toxic effects. In this study, the cyto-genotoxic effects of ZnSO 4 on meristematic cells in the root tip of Allium cepa L. were investigated. After calculating the effective concentration (EC 50 = 70 ppm) of ZnSO 4 , A. cepa root tip cells were suspended for 24, 48, 72, and 96 h in solutions of 35 ppm (EC 50 /2), 70 ppm (EC 50 ), and 140 ppm (EC 50 × 2) concentrations. Using the counts of dividing cells, the mitotic index (MI) was calculated. Chromosome aberration index (CAI) was determined from percentages of abnormal cells. When the obtained data were statistically evaluated, it was determined that all application concentrations caused a significant decrease in MI and an increase in CAI compared to the control group (distilled water). It was concluded that increased ZnSO 4 dose concentrations and exposure times caused cytotoxicity and genotoxicity in the root cells of A. cepa L.

Alterations of hydrogen peroxide (H 2 O 2 ) levels have a profound impact on numerous signaling cascades orchestrating plant growth, development, and stress signaling, including programmed cell death. To expand the repertoire of known molecular mechanisms implicated in H 2 O 2 signaling, we performed a forward chemical screen to identify small molecules that could alleviate the photorespiratory-induced cell death phenotype of Arabidopsis thaliana mutants lacking H 2 O 2 -scavenging capacity by peroxisomal catalase2. Here, we report the characterization of pakerine, an m -sulfamoyl benzamide from the sulfonamide family. Pakerine alleviates the cell death phenotype of cat2 mutants exposed to photorespiration-promoting conditions and delays dark-induced senescence in wild-type Arabidopsis leaves. By using a combination of transcriptomics, metabolomics, and affinity purification, we identified abnormal inflorescence meristem 1 (AIM1) as a putative protein target of pakerine. AIM1 is a 3-hydroxyacyl-CoA dehydrogenase involved in fatty acid β-oxidation that contributes to jasmonic acid (JA) and salicylic acid (SA) biosynthesis. Whereas intact JA biosynthesis was not required for pakerine bioactivity, our results point toward a role for β-oxidation-dependent SA production in the execution of H 2 O 2 -mediated cell death.

Ionic lead (Pb) in the environment has accumulated due to anthropogenic activities, causing a potential threat to plants and plant consumers. We conducted this study to reveal the molecular mechanism of Pb stress response in plants. The effects of Pb (5.0 and 15.0 μM) on mitosis, DNA replication, gene expression and proteins in root-tip cells of Allium cepa var. agrogarum L. were addressed. The results indicated that root growth was inhibited dramatically in Pb treatment groups. Chromosomal aberrations were observed and the mitotic index decreased during Pb treatments at different concentrations. The accumulation of reactive oxygen species (ROS) in onion roots was induced by Pb stress. Pb increased DNA damage and suppressed cell cycle progression. The above toxic effects got more serious with increasing Pb concentration and prolonging exposure time. A total of 17 proteins were expressed differentially between control and Pb exposure groups. Under Pb treatment, the decreased expression of Anx D1 indicated decreased defensive response; the decreased expression of SHMT1 indicated decreased respiration; the decreased expression of COMT2 indicated decreased response of other funtions; the increased expression of NDPK indicated increased transcription and protein synthesis; the increased expression of PR1 and CHI1 indicated increased pathogen invasion; the increased expression of ORC5 and MPK5 indicated the reduced DNA replicating activity; the decreased expression of POLD1 indicated the reduced DNA repair activity. Our results provide new insights at the proteomic level into the Pb-induced responses, defensive responses and toxic effects, and provide new molecular markers of the early events of plant responses to Pb toxicity.

The growing tips of plants grow sterile; therefore, disease-free plants can be generated from them. How plants safeguard growing apices from pathogen infection is still a mystery. The shoot apical meristem (SAM) is one of the three stem cells niches that give rise to the above ground plant organs. This is very well explored; however, how signaling networks orchestrate immune responses against pathogen infections in the SAM remains unclear. To reconstruct a transcriptional framework of the differentially expressed genes (DEGs) pertaining to various SAM cellular populations, we acquired large-scale transcriptome datasets from the public repository Gene Expression Omnibus (GEO). We identify here distinct sets of genes for various SAM cellular populations that are enriched in immune functions, such as immune defense, pathogen infection, biotic stress, and response to salicylic acid and jasmonic acid and their biosynthetic pathways in the SAM. We further linked those immune genes to their respective proteins and identify interactions among them by mapping a transcriptome-guided SAM-interactome. Furthermore, we compared stem-cells regulated transcriptome with innate immune responses in plants showing transcriptional separation among their DEGs in Arabidopsis . Besides unleashing a repertoire of immune-related genes in the SAM, our analysis provides a SAM-interactome that will help the community in designing functional experiments to study the specific defense dynamics of the SAM-cellular populations. Moreover, our study promotes the essence of large-scale omics data re-analysis, allowing a fresh look at the SAM-cellular transcriptome repurposing data-sets for new questions.

Poincianella Bracteosa: (Tul.) L.P. Queiroz. (Fabaceae) traditionally is used in Brazilian medicine to treat catarrhal infections, diarrhea, hepatitis and anemia. We investigated the phytochemical profile, and mutagenicity and anti-mutagenicity of aqueous extracts of leaves of P. bracteosa in A. cepa cells and in mice. We investigated four concentrations of extract for the A. cepa bioassay and three doses of extract for administration to mice. For the A. cepa assay, we analyzed 5,000 meristematic cells to determine the mitotic index, mean number of chromosome alterations and percentage of damage reduction. For each animal assay, 2,000 normochromatic erythrocytes were evaluated per mouse to determine the number of micronuclei and the protective effect of the extract. Phytochemical analysis of the extract revealed reducing sugars, tannins and alkaloids, which likely did not interfere with the cell cycle of A. cepa or cause damage to the DNA of A. cepa or mice. The extract exhibited a protective effect in both organisms.

Plants adapt to soil injury and biotic stress via cell regeneration. In Arabidopsis, root tip damage by genotoxic agents, antibiotics, UV light and cutting induces a program that recovers the missing tissues through activation of stem cells and involves ethylene response factor 115 (ERF115), which triggers cell replenishment. Here, we show that mutation of the gene encoding an MED18 subunit of the transcriptional MEDIATOR complex and chromate [Cr(VI)], an environmental pollutant, synergistically trigger a developmental program that enables the splitting of the meristem in vivo to produce twin roots. Expression of the quiescent centre gene marker WOX5, auxin-inducible DR5:GFP reporter and the ERF115 factor traced the changes in cell identity during the conversion of single primary root meristems into twin roots and were induced in an MED18 and chromate-dependent manner during the root twinning events, which also required auxin redistribution and signalling mediated by IAA14/SOLITARY ROOT (SLR1). Splitting of the root meristem allowed dichotomous root branching in Arabidopsis, a poorly understood process in which stem cells may act to enable whole organ regeneration., (© 2020 John Wiley & Sons Ltd.)

Ovule formation is essential for realizing crop yield because it determines seed number. The underlying molecular mechanism, however, remains elusive. Here, we show that cell wall invertase (CWIN) functions as a positive regulator of ovule initiation in Arabidopsis ( Arabidopsis thaliana ). In situ hybridization revealed that CWIN2 and CWIN4 were expressed at the placenta region where ovule primordia initiated. Specific silencing of CWIN2 and CWIN4 using targeted artificial microRNA driven by an ovule-specific SEEDSTICK promoter ( pSTK ) resulted in a substantial reduction of CWIN transcript and activity, which blocked ovule initiation and aggravated ovule abortion. There was no induction of carbon (C) starvation genes in the transgenic lines, and supplementing newly forming floral buds with extra C failed to recover the ovule phenotype. This indicates that suppression of CWIN did not lead to C starvation. A group of hexose transporters was downregulated in the transgenic plants. Among them, two representative ones were spatially coexpressed with CWIN2 and CWIN4 , suggesting a coupling between CWIN and hexose transporters for ovule initiation. RNA-sequencing analysis identified differentially expressed genes encoding putative extracellular receptor-like kinases, MADS-box transcription factors, including STK , and early auxin response genes in response to CWIN-silencing. Our data demonstrate the essential role of CWIN in ovule initiation, which is most likely to occur through sugar signaling instead of C nutrient contribution. We propose that CWIN-mediated sugar signaling may be perceived by, and transmitted through, hexose transporters or receptor-like kinases to regulate ovule formation by modulating downstream auxin signaling and MADS-box transcription factors., (© 2020 American Society of Plant Biologists. All Rights Reserved.)

Glucosinolates (GSLs) are sulfur-containing defense metabolites produced in the Brassicales, including the model plant Arabidopsis ( Arabidopsis thaliana ). Previous work suggests that specific GSLs may function as signals to provide direct feedback regulation within the plant to calibrate defense and growth. These GSLs include allyl-GSL, a defense metabolite that is one of the most widespread GSLs in Brassicaceae and has also been associated with growth inhibition. Here we show that at least three separate potential catabolic products of allyl-GSL or closely related compounds affect growth and development by altering different mechanisms influencing plant development. Two of the catabolites, raphanusamic acid and 3-butenoic acid, differentially affect processes downstream of the auxin signaling cascade. Another catabolite, acrylic acid, affects meristem development by influencing the progression of the cell cycle. These independent signaling events propagated by the different catabolites enable the plant to execute a specific response that is optimal to any given environment., (© 2020 American Society of Plant Biologists. All Rights Reserved.)

On acid soils, the trivalent aluminium ion (Al 3+ ) predominates and is very rhizotoxic to most plant species. For some native plant species adapted to acid soils including tea (Camellia sinensis), Al 3+ has been regarded as a beneficial mineral element. In this study, we discovered that Al 3+ is actually essential for tea root growth and development in all the tested varieties. Aluminum ion promoted new root growth in five representative tea varieties with dose-dependent responses to Al 3+ availability. In the absence of Al 3+ , the tea plants failed to generate new roots, and the root tips were damaged within 1 d of Al deprivation. Structural analysis of root tips demonstrated that Al was required for root meristem development and activity. In situ morin staining of Al 3+ in roots revealed that Al mainly localized to nuclei in root meristem cells, but then gradually moved to the cytosol when Al 3+ was subsequently withdrawn. This movement of Al 3+ from nuclei to cytosols was accompanied by exacerbated DNA damage, which suggests that the nuclear-targeted Al primarily acts to maintain DNA integrity. Taken together, these results provide novel evidence that Al 3+ is essential for root growth in tea plants through maintenance of DNA integrity in meristematic cells., (© 2020 The Authors. Journal of Integrative Plant Biology Published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

Imazalil (IMZ), a fungicide containing imidazole group, is extensively used for the prevention and treatment of fungal diseases in plants. Current study was performed to examine cyto-genotoxic potential of IMZ on Allium cepa roots by following Allium ana-telophase and single cell gel electrophoresis (comet) assays. The concentration which reduced the growth of the root tips of IMZ by 50% compared to the negative control group (EC 50 ) was found to be 1 μg/mL by Allium root growth inhibition test. 0.5, 1, and 2 μg/mL concentrations of IMZ were exposed to Allium roots for intervals of 24, 48, 72, and 96 h. 10 μg/mL of methyl methane sulfonate (MMS) and distilled water were used as control groups, both positive and negative. Statistical analysis was performed by using one-way ANOVA with Duncan's multiple comparison tests at p ≤ 0.05 and Pearson correlation test at p = 0.01. IMZ showed cytotoxic effect by statistically decreasing root growth and mitotic index (MI) and also genotoxic effect by statistically increasing chromosomal aberrations (CAs) and DNA damage compared to the negative control group. With these cyto-genotoxic effects, it should be used carefully and further cyto-genotoxic mechanisms should be investigated along with other toxicity tests.

Root-derived abscisic acid (ABA) is known to regulate shoot physiology, such as stomata closure. Conversely, the basipetal regulatory effect of shoot-derived ABA is poorly understood. Herein, we report that simulation of shoot-ABA accumulation by exogenous application of ABA to shoots basipetally stimulates primary root (PR) growth. ABA applied to shoots accelerates root cell division, as evidenced by the increase in meristem size and cell number and the intensity of CYCB1;1::GFP (a mitosis marker). Root ABA content was not changed following shoot ABA application, although the ABA reporter line RAB18::GFP showed an increase in ABA in the cotyledons. Shoot-ABA application increases basipetal auxin transport by 114 %. Shoot-ABA-promoted PR growth can be abolished by attenuating basipetal auxin flux using 2,3,5-triiodobenzoic acid (TIBA, an auxin transport inhibitor), demonstrating that ABA promotes PR growth by increasing basipetal auxin transport. Root cell elongation, evaluated by the total length of the first 7 cells in the elongation zone (EZ), was increased by 56 % following shoot-ABA application. The cell walls of the root EZ were alkalinized by ABA, as exhibited by 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt staining. Higher pH promotes both PR growth and cell elongation. Thus, shoot-ABA promotes cell elongation by alkalinizing the cell wall. In light of our results, we provide a representative detailed model of the basipetal regulatory effect of ABA on PR growth., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Main Conclusion: In this study, we show that ectopic expression of either HtNHX1 or HtNHX2, from Helianthus tuberosus plant (located at vacuolar and endosome membranes, respectively), in rice plants could enhance its tolerance to aluminum (Al 3+ ) stress and soil acidity. Plant sodium (potassium)/proton (Na + (K + )/H + antiporters of the NHX family have been extensively characterized as they are related to the enhancement of salt tolerance. However, no previous study has reported NHX transporter functions in plant tolerance to Al 3+ toxicity. In this study, we demonstrate their role as a component of the Al 3+ stress tolerance mechanism. We show that the ectopic expression of either HtNHX1 or HtNHX2 , from Helianthus tuberosus plant, in rice (located at vacuole and endosome, respectively) could also enhance rice tolerance to Al 3+ stress and soil acidity. Expression of either HtNHX1 or HtNHX2 reduced the inhibitory effect of Al 3+ on the rice root elongation rate; both genes were reported to be equally effective in improvement of stress conditions. Expression of HtNHX1 enhanced Al 3+ -trigged-secretion of citrate acids, rhizosphere acidification, and also reduced K + efflux from root tissues. In contrast, expression of HtNHX2 prevented Al 3+ -trigged-decrease of H + influx into root tissues. Al 3+ -induced damage of the cell wall extensibility at the root tips was impaired by either HtNHX1 or HtNHX2. Co-expression of HtNHX1 and HtNHX2 further improved rice growth, particularly under the Al 3+ stress conditions. The results demonstrate that HtNHX1 and HtNHX2 improved rice tolerance to Al 3+ via different mechanisms by altering the K + and H + fluxes and the cell wall structure.

Silicon nanoparticles gained a great interest due to its use in biomedical research. It is considered as safe and has been used in nanomedicine. But literature still states its toxicity depending upon the size and dose of silicon nanoparticles. So, current study was aimed to evaluate the cytotoxicity and genotoxicity of silicon dioxide nanoparticles (SiO 2 NPs) by Allium anaphase-telophase and Comet tests. Characterization of SiO 2 NPs showed the particle size as 16.12 ± 3.07 nm. The mean diameter of SiO 2 NPs was having range of 404.66 ± 93.39 nm in solution. Highest total anomalies (18.80 ± 0.45) were observed at 100 µg/mL, whereas least (11.2 ± 0.84) were observed by the 12.5 µg/mL concentration. There was concentration-response association in increased CAs and DNA damage. The highest concentration (100 µg/mL) of SiO 2 NPs induced the significant DNA damage (149.67 ± 1.15), whereas the least was observed by the negative control (2.67 ± 0.58). The current study revealed the cytotoxic and genotoxic effects of SiO 2 NPs on the root meristem cells of A. cepa.

The present experiment was designed to evaluate physico-chemical characteristics and phyto-genotoxicity of arsenic (As) contaminated soil collected from different sites of Lakhimpur, Uttar Pradesh (UP), India by employing Vicia faba L. The analyses included various biochemical as well as cyto-genotoxicity assays. The results showed that soil pH was slightly acidic to neutral in nature. The bulk density (1.18-1.23 gcm -3 ), particle density (2.51-2.59 gcm -3 ) and porosity (44-53%) varied in different places. The level of available nutrients, nitrogen, phosphorus and potassium was found to vary as 124-165 mgkg-1, 173-186 mgkg-1 and 48-98 mgkg-1, respectively. The maximum As levels were found in soil of Fulvareya (27.13 mgkg-1) and Atareya (24.12 mgkg-1), the level of As in water samples of these sites were 0.19 mgl -1 and 0.21 mgl -1 , respectively. Phytotoxicity of the As present in soils was evident through significant increases in stress metabolites, hydrogen peroxide (H 2 O 2 ), malondialdehyde (MDA) and carbonyl groups in root and shoot of V. faba. Cyto-genotoxic effects were also seen through reduced mitotic index (MI) and increased mitotic depression (MD), relative abnormality rate (RAR) as well as other chromosomal abnormalities along with micronuclei in root meristematic cells of V. faba. The phytotoxicity and cyto-genotoxicity assessment suggests the harmful soil properties that might affect biota., (Copyright © 2019. Published by Elsevier Ltd.)

Magnetite nanoparticles (MNPs) are gaining attention because of their biomedical, environmental and industrial applications. However, they have limited uses because of ecotoxicity. On contrast, bacterially synthesized MNPs such as magnetosomes are found to be biocompatible and less toxic due to the lipid bilayer membrane found around magnetite. In this context, this study compares the physio-chemical properties and toxicology effects of MNPs and magnetosomes in different models such as human red blood cells, macrophage cell lines (RAW 264.7), onion root tips ( Allium cepa), Artemia salina (A. salina ) and zebrafish embryo ( Danio rerio ). MNPs showed 38.59% hemolysis whereas the maximum hemolysis induced by magnetosomes was 7.03% for the same concentration (250 μg/ml). The cytotoxicity of MNPs and magnetosomes were 36.01% and 13.4%, respectively, at 250 μg/ml. Onion root tip assay revealed high toxicity when treated with MNPs than magnetosomes. The MNPs were further tested for its toxicity against A. salina and 50% mortality rate was observed. Similarly, notable malformation was seen in zebrafish embryo treated with MNPs. However, magnetosomes did not exhibit any mortality and malformation in A. salina and zebrafish embryo. The study revealed that magnetosomes are safe and do not cause any potential risk to environment compared to synthetic MNPs. Abbreviation: MNPs: Magnetic nanoparticles; ATCC: American Type Culture Collection; MTB: Magnetotactic bacteria; MSR-1: Magnetospirillum gryphiswaldense; DSMZ: Deutsche Sammlung von Mikroorganismen und Zellkulturen; MSGM: Magnetospirillum growth medium; D-PBS: Dulbecco phosphate buffer saline; RBC: Red blood cells; SEM: Scanning electron microscopy; HRTEM: High-resolution transition electron microscope; FTIR: Fourier transform infrared spectroscopy; XRD: X-ray powder diffraction; AFM: Atomic-force microscopy; ZP: Zeta Potential; PSD: Particle Size Distribution; EDX: Energy-dispersive X-ray spectroscopy; PBS: Phosphate buffer saline; DMEM: Dulbecco's modified eagle medium; HEPES: (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid); MTT:3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; DMSO: Dimethyl sulfoxide; ROS: Reactive oxygen species.

The present investigation was designed to monitor the cytotoxic potential of Sodium Hypochlorite using lentil (Lens culinaris) as a bioindicator of toxicity. Sodium hypochlorite (NaClO), is a chemical compound that is used mainly for its disinfectant properties, its effect is widely toxic, which is why it is marketed in low concentrations and it is also a component in various products such as agrochemicals. In the present study the L. culinaris seeds were exposed to different NaClO dose 0, 0.2, 1, 3, 5 and 7 mg L -1 during 24, 48 and 72 h; timeslots in which the root growth was also studied. The cytotoxic potential of NaClO was determined by calculating the mitotic index (MI), calculating cellular anomalies (CA) and observing the longitudinal growth of the roots during the various time periods. The radicular growth was prolonged and it was observed that there was a greater growth at the dose of 1 and 7 mg L -1 in the time of 72 h. The cytotoxic effects could be analyzed in the mitotic index, since the higher the concentration, the lower the mitotic index, as observed in the dose of 7 mg L -1 where a reduction of the mitotic index of the meristematic cells is observed. The results indicate that NaClO has a cytotoxic effect that induces various types of chromosomal abnormalities. This indicates that Sodium Hypochlorite has a cytotoxic effect according to the increase in its dose. Therefore, Lens culinaris turned out to be a kind of appropriate bioindicator to study the cytotoxic effects of various potentially toxic substances., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Glyphosate is a chemical compound used mainly as a broad spectrum herbicide, it is recognized for its proven effectiveness and easy handling. It represents more than 60% of the world market of non-selective herbicides and is used in both agricultural fields and family gardens. The present study was designed to test the cytogenotoxic potential of glyphosate using the Allium cepa test as toxicity bioindicator. Consequently, bulbs of A. cepa were exposed to different concentrations of glyphosate (5, 10, 15, 25 and 30 mgL -1 ) and a control (deionized water), for 72 h; root development was also studied in this period of time. The cytogenotoxic potential of glyphosate was determined by calculating the mitotic index (MI), cellular anomalies (CA) and registering the roots longitudinal growth at 24, 48 and 72 h. Regarding root development, a greater growth was observed in the control treatment in the three measurement times. The mitotic phases analysis, determined that the higher the concentration, the lower the mitotic index, in addition the inhibition of the telophase Mitotic Index (TMI) was observed in any of the concentrations. The results indicate that the exposure to glyphosate of A. cepa meristematic cells induces diverse types of chromosomal anomalies, such as micronuclei (MN), chromosome breaking (CB), nuclear notch (Nn), among others. Therefore, in demonstrates that glyphosate has a highly cytogenotoxic effect for any of the concentrations used., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Visualizing the distribution of hormone signaling activity such as auxin and cytokinins is of key importance for understanding regulation of plant development and physiology. Live imaging and genetically encoded hormone biosensors and reporters allow monitoring the spatial and temporal distribution of these phytohormones. Here, we describe how to cultivate live shoot apical meristems after dissection for observation under the confocal microscope for up to 4 days. The shoot apical meristems are maintained on an appropriate medium allowing them to grow and initiate new organs at a frequency similar to plants grown on soil. Meristems expressing hormone biosensors and reporters allows following hormone signaling activity distribution at high spatiotemporal resolution without chemical fixation, an approach that that can also be applied to follow the dynamics of expression in vivo of any fluorescent marker.

Plant models may be useful as test organisms for initial screening of potential toxicity of personal care products. The objective of the present study was to assess the efficacy of the Allium cepa (common onion) test system as a bioanalytical tool for screening potential cytotoxicity and genotoxicity of herbal-based hair dye formulations. Exposure of black hair dye formulations for 48 hours resulted in root growth retardation and mitosis suppression in the root meristems of A. cepa bulbs indicating concentration dependent cytotoxicity. At the 72 hour post exposure, cytotoxic effects on the roots were reduced but not recovered completely signifying prolong toxic action of the hair dyes. The condensed nuclei was the most frequent nuclear abnormality found in the dye exposed root meristematic cells indicating the cell death process. Induction of micronuclei and chromosomal aberrations in the root meristematic cells even at the post exposure stage indicates persistent genotoxicity of the hair dyes which may be attributed to the interactive effects of chemical mixtures present in the commercial hair dye formulations. The results revealed that A. cepa test system is an effective bioanalytical tool for screening cytogenotoxicity of commercial hair dye formulations.

The benefits of in vitro plant cultivation are mainly due to very high multiplication rate. Cultivation of plant material in vitro can be carried out during the whole year regardless of the time of the year or weather conditions. We create artificial conditions in the lab (heat, light, humidity), and we can regulate these conditions at any time. For the preservation of cultivar identity, we recommend establishing in vitro cultures from shoot tips usually larger than 0.2 mm. In practice, in vitro cultivation of plants uses these growth regulators to achieve organogenesis, for example, root formation, prolonged growth, or multiplication. During each subculture, these cultures are then transferred on a solid agar medium in the form of actively growing multiple shoots with a well-differentiated shoot tip containing meristematic area. Cytokinins are important for cell division and causes branching of plants. Auxins, both endogenous and exogenous, act at as a trigger for the differentiation and formation of root primordia. Morphological characteristics (formation of leaves or callus) and shoot development should be observed during in vitro multiplication and after transfer to ex vitro conditions.

The root meristem-one of the plant's centers of continuous growth-is a conveyer belt in which cells of different identities are pushed through gradients along the root's longitudinal axis. An auxin gradient has long been implicated in controlling the progression of cell states in the root meristem. Recent work has shown that a PLETHORA (PLT) protein transcription factor gradient, which is under a delayed auxin response, has a dose-dependent effect on the differentiation state of cells. The direct effect of auxin concentration on differential transcriptional outputs remains unclear. Genomic and other analyses of regulatory sequences show that auxin responses are likely controlled by combinatorial inputs from transcription factors outside the core auxin signaling pathway. The passage through the meristem exposes cells to varying positional signals that could help them interpret auxin inputs independent of gradient effects. One open question is whether cells process information from the changes in the gradient over time as they move through the auxin gradient., (© 2020 Elsevier Inc. All rights reserved.)

Excessive copper (Cu) exposure ultimately results in toxicosis in all organisms. The protective potential of resveratrol compound against the CuCl 2 toxicity was evaluated in Allium cepa L. root tip cells. For this aim, A. cepa bulbs were divided into six groups and the groups were treated with tap water, 400 mg/L resveratrol, 800 mg/L resveratrol, 20 μM CuCl 2 , 400 mg/L resveratrol + 20 μM CuCl 2 , and 800 mg/L resveratrol + 20 μM CuCl 2 solutions, respectively for 72 h. The first group irrigated with tap water was accepted as control. All groups were screened for the germination percentage, root elongation, total bulb weight gain, micronucleus (MN) frequency, mitotic index (MI), chromosomal aberrations, and anatomical changes. Furthermore, superoxide dismutase (SOD) and catalase (CAT) activities as well as malondialdehyde (MDA) level as an indicator of lipid peroxidation were investigated. It was found that CuCl 2 exposure alone triggered a definite negative effects on all parameters examined. On the other hand, the groups treated with resveratrol did not have statistically different values compared to the control group. Resveratrol doses induced a remarkable recovery in growth parameters including germination percentage, root elongation, and total bulb weight gain when applied with CuCl 2 . In these groups, MN frequency, chromosomal aberrations, and anatomical abnormalities were alleviated, whereas the MI levels increased significantly. Moreover, lipid peroxidation level and antioxidant enzyme activities showed a marked amelioration when resveratrol and CuCl 2 were applied together. Copper compounds have become common pollutants due to their direct uses as pesticides in agricultural areas as well as their spreading to natural areas from industrial fields. The study clearly demonstrated the therapeutic potential of resveratrol against the harmful effects of CuCl 2 exposure in Allium roots. So, resveratrol could be considered as a plant-derived restorative agent for the reduction of the risks from the other copper compounds.