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Abscisic acid (ABA) plays an important regulatory role in plants. It is very critical to obtain the dynamic changes of ABA in situ for botanical research. Herein, coupled with paper-based analysis devices, electrochemical immunoelectrodes based on disposable stainless steels sheet were developed for ABA detection in plants in situ. The stainless steel sheets were modified with carbon cement, ferrocene-graphene oxide-multi walled carbon nanotubes nanocomposites, and ABA antibodies. The system can detect the ABA in the range of 1 nM to 100 μM, with a limit of detection of 100 pM. The ABA content in tomato leaves under high salinity was detected in situ. The trend of ABA changes was similar to the expression of SlNCED1 and SlNCED2 . Overall, this study offers an approach for in situ detection of ABA in plants, which will help to study the regulation mechanism of ABA in plants and to promote the development of precision agriculture.

Background and Aims: Turions are vegetative, dormant overwintering organs formed in aquatic plants in response to unfavourable ecological conditions. Contents of cytokinin (CK), auxin metabolites and abscisic acid (ABA) as main growth and development regulators were compared in innately dormant autumnal turions of 22 aquatic plant species of different functional ecological or taxonomic groups with those in non-dormant winter apices in three aquatic species and with those in spring turions of four species after their overwintering., Methods: The hormones were analysed in miniature turion samples using ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry., Key Results: In innately dormant turions, the total contents of each of the four main CK types, biologically active forms and total CKs differed by two to three orders of magnitude across 22 species; the proportion of active CK forms was 0.18-67 %. Similarly, the content of four auxin forms was extremely variable and the IAA proportion as the active form was 0.014-99 %. The ABA content varied from almost zero to 54 µmol kg-1 dry weight and after overwintering it usually significantly decreased. Of all functional traits studied, hormone profiles depended most on the place of turion sprouting (surface vs bottom) and we suggest that this trait is crucial for turion ecophysiology., Conclusions: The key role of ABA in regulating turion dormancy was confirmed. However, the highly variable pattern of the ABA content in innately dormant and in overwintered turions indicates that the hormonal mechanism regulating the innate dormancy and its breaking in turions is not uniform within aquatic plants., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The presence of mycotoxins and other toxic metabolites in hops ( Humulus lupulus L.) was assessed for the first time. In total, 62 hop samples were sampled in craft breweries, and analyzed by a multi-toxin LS-MS/MS method. The study collected samples from craft breweries in all of the Croatian counties and statistically compared the results. Based on previous reports on Alternaria spp. and Fusarium spp. contamination of hops, the study confirmed the contamination of hops with these toxins. Alternaria toxins, particularly tenuazonic acid, were found in all tested samples, while Fusarium toxins, including deoxynivalenol, were present in 98% of samples. However, no Aspergillus or Penicillium metabolites were detected, indicating proper storage conditions. In addition to the Alternaria and Fusarium toxins, abscisic acid, a drought stress indicator in hops, was also detected, as well as several unspecific metabolites. The findings suggest the need for monitoring, risk assessment, and potential regulation of Alternaria and Fusarium toxins in hops to ensure the safety of hop usage in the brewing and pharmaceutical industries. Also, four local wild varieties were tested, with similar results to the commercial varieties for toxin contamination, but the statistically significant regional differences in toxin occurrence highlight the importance and need for targeted monitoring.

High-performance liquid chromatography with ultraviolet detection (HPLC-UV) is a common analysis technique due to its high versatility and simple operation. In the present study, HPLC-UV detection was integrated with immunoaffinity cleanup (IAC) of the sample extracts. The matrix effect was greatly reduced, and the limit of detection was as low as 1 ng/g of free abscisic acid (ABA) in fresh plant tissues. A monoclonal antibody 3F1 (mAb 3F1) was developed to specifically recognize free ABA but not ABA analogues. The mAb 3F1-immobilized immunoaffinity column exhibited a capacity of 850 ng/mL and an elution efficiency of 88.8-105% for standards. The extraction recoveries of the column for ABA ranged from 80.4 to 108.9%. ABA content was detected in various plant samples with IAC-HPLC-UV. The results were verified with ultraperformance liquid chromatography-electrospray tandem mass spectrometry. IAC-HPLC-UV can be a sensitive and cost-efficient method for plant hormone analysis.

The agricultural sector faces severe challenges owing to heavy metal (HM) contamination of farmlands, requiring urgent preventive measures. To address this, we investigated the impact of the synergistic application of Azospirillum brasilense, a growth-promoting rhizobacterium producing abscisic acid (ABA), and biochar to minimize HM accumulation in pak choi, using three distinct expression levels of the ABA transporter NRT1.2 in pak choi and three different types of contaminated soils as experimental materials. The results revealed that pak choi with low, medium, and high NRT1.2 expression intensity, when subjected to bacterial strain-biochar treatment, exhibited an increasing trend in ABA content compared to the control. Correspondingly, the aboveground HM content decreased by 1-49 %, 22-52 %, and 15-96 %, whereas the fresh weight increased by 12-38 %, 88-126 %, and 152-340 %, respectively, showing a significant correlation with NRT1.2 expression. Pearson correlation analysis demonstrated that NRT1.2 expression intensity was inversely associated with the combined treatment's reduction in HM accumulation and positively correlated with the promotional effect. Simultaneously, soil discrepancies significantly affected the combined treatment, which was likely associated with variations in the active forms of HM in each soil. Consequently, when employing ABA-producing bacteria for mitigating crop HM accumulation, selecting plants with higher relative NRT1.2 expression intensity, combined with biochar, is recommended., 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.)

Saffron ( Crocus sativus L.) is a valuable Chinese herb with high medicinal value. Saffron pistils are used as medicine, so increasing the number of flowers can increase the yield. Plant hormones have essential roles in the growth and development of saffron, as well as the response to biotic and abiotic stresses (especially in floral initiation), which may directly affect the number of flowers. Quantitative analysis of plant hormones provides a basis for more efficient research on their synthesis, transportation, metabolism, and action. However, starch (which interferes with extraction) is present in high levels, and hormone levels are extremely low, in saffron corms, thereby hampering accurate determination of plant-hormone levels in saffron. Herein, we screened an efficient and convenient pre-treatment method for plant materials containing abundant amounts of starch. Also, we proposed an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantification of abscisic acid (ABA) and auxin (IAA). Then, the method was applied for the detection of hormone-content differences between flowering and non-flowering top buds, as well as between lateral and top buds. Our method showed high sensitivity, reproducibility, and reliability. Specifically, good linearity in the range 2-100 ng ml -1 was achieved in the determination of ABA and IAA, and the correlation coefficient ( R 2 ) was >0.9982. The relative standard deviation was 2.956-14.51% (intraday) and 9.57-18.99% (interday), and the recovery range was 89.04-101.1% ( n = 9). The matrix effect was 80.38-90.50% ( n = 3). The method was thoroughly assessed employing various "green" chemistry evaluation tools: Blue Applicability Grade Index (BAGI), Complementary Green Analytical Procedure Index (Complex GAPI) and Red Green Blue 12 Algorithm (RGB12). These tools revealed the good greenness, analytical performance, applicability, and overall sustainability alignment of our method. Quantitative results showed that, compared with saffron with a flowering phenotype cultivated at 25 °C, the contents of IAA and ABA in the terminal buds of saffron cultivated at 16 °C decreased significantly. When cultivated at 25 °C, the IAA and ABA contents in the terminal buds of saffron were 1.54- and 4.84-times higher than those in the lateral buds, respectively. A simple, rapid, and accurate UPLC-MS/MS method was established to determine IAA and ABA contents. Using this method, a connection between the contents of IAA and ABA and the flowering phenotype was observed in the quantification results. Our data lay a foundation for studying the flowering mechanism of saffron.

Microbial-assisted phytoremediation has great potential to improve the efficiency of phytoremediation in heavy metal (HM)-contaminated soils. In this study, the synergistic effects of rhamnolipids and the abscisic acid (ABA)-metabolizing bacterium Rhodococcus qingshengii on the phytoremediation efficiency of Indian mustard (Brassica juncea) in HM-contaminated soils were investigated. The Cd, Zn, and Pb contents in plants treated with a combination of rhamnolipids and R. qingshengii were 48.4-77.1 %, 14.6-40.4 %, and 16.1-20.0 % higher, respectively, than in those treated with R. qingshengii alone, and 42.8-59.2 %, 13.1-48.2 %, and 7.3-67.5 % higher, respectively, than in those treated with rhamnolipids alone. In addition, the bioconcentration factors of each metal were improved, and the biomass further increased by 36.6-65.7 % compared to that of single treatments. Pearson's correlation analysis showed that rhamnolipids and R. qingshengii enhanced the accumulation of HMs in B. juncea by activating the available forms of HMs in the soil and regulating the ABA and indole-3-acetic acid in plants, respectively. The structural equation model indicated that R. qingshengii had a larger path coefficient than rhamnolipids in terms of HM content and plant biomass, suggesting that R. qingshengii may have a greater contribution to promoting the extraction of HMs from the soil under synergistic conditions. In conclusion, the combination of rhamnolipids and R. qingshengii has great potential to enhance the phytoremediation efficiency of hyperaccumulating plants in HM-contaminated soils., 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 © 2023 Elsevier B.V. All rights reserved.)

Arbuscular mycorrhizal fungi (AMF) are symbiotic fungi that colonize plant roots, and they are more common in Cd-polluted habitats. However, there is limited understanding of the response of root traits and cadmium (Cd) uptake to AMF in different crop varieties. Two maize varieties, Panyu 3 and Ludan 8, with high and low Cd uptake capacities, respectively, were cultivated as host plants in a pot experiment with Cd-polluted soil (17.1 mg/kg Cd). The effects of AMF on the growth, mineral nutrient concentration, root traits, phytohormone concentrations and Cd uptake of the two maize varieties and their comprehensive response to AMF fungal inoculation were investigated. AMF improved growth, mineral nutrient levels and root morphology and increased lignin and phytohormone concentrations in roots and Cd uptake in the two maize varieties. However, the two maize varieties, Panyu 3 and Ludan 8, had different responses to AMF, and their comprehensive response indices were 753.6% and 389.4%, respectively. The root biomass, branch number, abscisic acid concentrations, lignin concentrations and Cd uptake of maize Panyu 3 increased by 151.1%, 28.6%, 139.7%, 99.5% and 84.7%, respectively. The root biomass, average diameter, auxin concentration, lignin concentration and Cd uptake of maize Ludan 8 increased by 168.7%, 31.8%, 31.4%, 41.7% and 136.7%, respectively. Moreover, Cd uptake in roots presented very significant positive correlations with the average root diameter and abscisic acid concentration. A structural equation model indicated that the root abscisic acid concentration and root surface area had positive effects on Cd uptake by the Panyu 3 maize roots; the root abscisic acid concentration and root tip number had positive effects on Cd uptake by the Ludan 8 maize roots. Thus, AMF differentially regulated Cd uptake in the two maize varieties, and the regulatory effect was closely related to root traits and phytohormone concentrations., 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 © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

A novel, simple, rapid, 7-minute HPLC-DAD method for the determination of 10 phenolic compounds and abscisic acid commonly found in teas, wines, fruit and honey was successfully developed and validated according to the International Council of Harmonization (ICH) guidelines. This reverse-phase (RP) HPLC-DAD method boasts rapid separation and excellent resolution while introducing green chemistry techniques. The Agilent 1200 series diode array detector SL coupled with a reverse-phase Advanced Materials Technology Halo C18 column (100 × 3.0 mm i.d., 2.7 μm) contributed to the rapid analyses. This, together with a 0.1% formic acid in water (v/v) and methanol mobile phase, a flow rate of 0.8 mL/min and the utilization of a meticulous gradient elution resulted in a validated method for the determination of 10 phenolic compounds and abscisic acid commonly found in various foodstuffs. The resulting method proved to be rapid, accurate, precise and linear with sensitive detection limits from 0.025 μg/mL to 0.500 μg/mL and percentage recoveries of 98.07%-101.94%. Phenolic compounds have been acknowledged throughout literature for their therapeutic properties, interalia, antioxidant, anti-inflammatory and antiaging due to free radical scavenging potentials. However, resulting analysis, can be frequently complicated and long and very often discounts green chemistry techniques. The developed and validated method successfully and rapidly analyses, gallic acid, caffeic acid, trans-p-coumaric acid, rutin, myricetin, abscisic acid, trans-cinnamic acid, quercetin, luteolin, kaempferol and chrysin with excellent resolution and precision., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: The application of kaolin particle film is considered a short-term strategy against several environmental stresses in areas with a Mediterranean-like climate. However, it is known that temperature fluctuations and water availability over the season can jeopardize kaolin efficiency in many Mediterranean crops. Hence, this study aims to evaluate the effects of kaolin foliar application on berry phytohormones, antioxidant defence, and oenological parameters at veraison and harvest stages of Touriga-Franca (TF) and Touriga-Nacional (TN) grapevines in two growing seasons (2017 and 2018). The 2017 growing season was considered the driest (-147.1 dryness index) and the warmest (2705 °C growing degree days) of the study., Results: In 2017, TF kaolin-treated berries showed lower salicylic acid (-26.6% compared with unsprayed vines) and abscisic acid (ABA) (-10.5%) accumulation at veraison, whereas salicylic acid increased up to 28.8% at harvest. In a less hot season, TN and TF kaolin-treated grapevines showed a twofold in ABA content and a threefold increase in the indole-3-acetic acid content at veraison and lower ABA levels (83.8%) compared with unsprayed vines at harvest. Treated berries showed a decreased sugar content, without compromising malic and tartaric acid levels, and reactive oxygen species accumulation throughout berry ripening., Conclusion: The results suggest kaolin exerts a delaying effect in triggering ripening-related processes under severe summer stress conditions. Treated berries responded with improved antioxidant defence and phytohormone balance, showing significant interactions between kaolin treatment, variety, and developmental stage in both assessed years. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Plant hormones regulate numerous developmental and physiological processes. Abiotic stresses considerably affect production and distribution of phytohormones as the stress signal triggers. The homeostasis of plant hormones is controlled by their de novo synthesis and catabolism. The aim of this work was to analyse the contents of total and individual groups of endogenous cytokinins (CKs) as well as indole-3-acetic acid (IAA) in AtCKX overexpressing centaury plants grown in vitro on graded NaCl concentrations (0, 50, 100, 150, 200 mM). The levels of endogenous stress hormones including abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) were also detected. The elevated contents of total CKs were found in all analysed centaury shoots. Furthermore, increased amounts of all five CK groups, as well as enhanced total CKs were revealed on graded NaCl concentrations in non-transformed and AtCKX roots. All analysed AtCKX centaury lines exhibited decreased amounts of endogenous IAA in shoots and roots. Consequently, the IAA/bioactive CK forms ratios showed a significant variation in the shoots and roots of all AtCKX lines. In shoots and roots of both non-transformed and AtCKX transgenic centaury plants, salinity was associated with an increase of ABA and JA and a decrease of SA content., (© 2021. The Author(s).)

Background: Drought is a very important environmental stressor, which has negative effects on the growth of trees, decreasing their yield. The role of different-sized selenium nanoparticles (Se-NPs) in the mitigation of environmental stresses such as drought in crops has not yet been investigated., Results: Trees treated with Se-NPs displayed higher levels of photosynthetic pigments, a better nutrient status, better physical parameters (especially fruit cracking) and chemical parameters, a higher phenolic content, and higher concentrations of osmolytes, antioxidant enzymes, and abscisic acid than untreated trees under drought stress. Foliar spraying of 10 and 50 nm Se-NPs alleviated many of the deleterious effects of drought in pomegranate leaves and fruits and this was achieved by reducing stress-induced lipid peroxidation and H 2 O 2 content by enhancing the activity of antioxidant enzymes. Furthermore, the 10 nm Se-NPs treatment produced more noticeable effects than the treatment with 50 nm Se-NPs., Conclusion: Results confirm the positive effects of nanoparticle spraying, especially the role of 10 nm Se-NPs in the management of negative effects of drought not only for pomegranates but potentially also for other crops. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Plant response to osmotic stress is a complex issue and includes a wide range of physiological and biochemical processes. Extensive studies of known cultivars and their reaction to drought or salinity stress are very important for future breeding of new and tolerant cultivars. Our study focused on the antioxidant activity, accumulations of osmotica, and the content of abscisic acid in apple (cv. "Malinové holovouské", "Fragrance", "Rubinstep", "Idared", "Car Alexander") and cherry (cv. "Regina", "Napoleonova", "Kaštánka", "Sunburst", "P-HL-C") cultivated in vitro on media containing different levels of polyethylene glycol PEG-6000. Our results indicated that the studied genotypes responded differently to osmotic stress manifested as reduction in the leaf relative water content (RWC) and increment in the activities of antioxidant enzymes, proline, sugars, and abscisic acid content. Overall, cherry cultivars showed a smaller decrease in percentage RWC and enzymatic activities, but enhanced proline content compared to the apple plants cultivars. Cultivars "Rubinstep", "Napoleonova", and "Kaštánka" exhibited higher antioxidant capacity and accumulation of osmoprotectants like proline and sorbitol that can be associated with the drought-tolerance system.

Plant hormone abscisic acid (ABA) is essential for regulating plant growth and various stress responses. ABA-mediated signaling depends on local ABA levels rather than the overall cellular ABA concentration. While cellular concentration of ABA can be detected using Förster resonance energy transfer (FRET)-based ABA probes, direct imaging of subcellular ABA levels remains unsolved. Here, we modified the previously reported ABAleon2.1 and generated a new ABA sensor, named ABAleon2.1_Tao3. Via transient expression in tobacco (Nicotiana tabacum) protoplasts, we targeted ABAleon2.1_Tao3s to the endoplasmic reticulum (ER) membrane with the ABA sensing unit facing the cytosol and the ER, respectively, through a nanobody-epitope-mediated protein interaction. Combining FRET with fluorescence lifetime imaging microscopy, ABA-triggered-specific increases in the fluorescence lifetime of the donor mTurquoise in the ABAleon2.1_Tao3 were detected in both transient assays and stably transformed Arabidopsis plants. In tobacco protoplasts, ER membrane-targeted ABAleon2.1_Tao3s showed a generally higher basal level of ABA in the ER than that in the cytosol and ER-specific alterations in the level of ABA upon environmental cues. In ABAleon2.1_Tao3-transformed Arabidopsis roots, mannitol triggered increases in cytosolic ABA in the division zone and increases in ER ABA in the elongation and maturation zone within 1 h after treatment, both of which were abolished in the bg1-2 mutant, suggesting the requirement for BG1 in osmotic stress-triggered early ABA induction in Arabidopsis roots. These data demonstrate that ABAleon2.1_Tao3s can be used to monitor ABA levels in the cytosol and the ER, providing key information on stress-induced changes in the level of ABA in different subcellular compartments., (© American Society of Plant Biologists 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Both extreme usage of water in agriculture i.e., drought and flooding affect physiological and growth aspects of the plant as well as gene expression undertaken in water absorption. These affect depend on the stress duration i.e., shock or gradual stress exposer. The factorial experiment based on CRD with 10 replicates was conducted to investigate the physiological and water relation as well as aquaporin expression in (Capsicum annuum L.). Drought stress was applied gradually from - 2, - 3, - 4 to - 5 MPa during 8 days but in shock stress - 5 MPa applied at one time. The gradual flooding stress adjusted with changing the aeration duration from 15 to 0 min gradually every 2 days and for the shock- flooding, peppers keep in a nutrient solution without aeration in a sealed container. Results showed that both extreme water stress had a deleterious effect on the growth and physiological parameter of pepper for a longer duration. Antioxidant, proline, fluorescence chlorophyll stimulate in the gradual period except for ABA content, which is higher in shock stress. PIP1expression showed a reverse effect in leaf and root at flooding i.e., PIP1expression raised in root while it was reduced in leaf at shock-flooding. The highest PIP1expression was observed in gradual-drought of root and gradual duration of drought and flooding stress in leaf. In the physiological aspect of plant response to stress in pepper, results showed an enhanced in proline and phenol content to help osmotic adjustment and keep water status in moderate condition. Conclusively, shocked stress first, motivated these defense systems, and then in the next step, the other adaptive mechanism like gene expression activated to help pepper face stress. On the other hand, shock stress showed down-regulation, but when the stress lasted for a longer time results in up-regulation.

Plant hormones play important roles in plant growth and development and physiology, and in acclimation to environmental changes. The hormone signaling networks are highly complex and interconnected. It is thus important to not only know where the hormones are produced, how they are transported and how and where they are perceived, but also to monitor their distribution quantitatively, ideally in a non-invasive manner. Here we summarize the diverse set of tools available for quantifying and visualizing hormone distribution and dynamics. We provide an overview over the tools that are currently available, including transcriptional reporters, degradation sensors, and luciferase and fluorescent sensors, and compare the tools and their suitability for different purposes., (© 2020 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

In this study, we investigated the changes in the distribution and regulation of endogenous hormones in Phyllostachys edulis 'Pachyloen' during bamboo shooting. Enzyme-linked immunosorbent assay was used to measure the mass fractions of indole-3-acetic acid (IAA), gibberellic acid (GA), zeatin riboside (ZR), and abscisic acid (ABA) in rhizomes, shoots, and maternal bamboo organs during shoot sprouting, shoot growth, and new-bamboo formation. Measurements were compared among bamboo parts and developmental periods. The overall mass fractions of IAA and ABA were significantly higher than those of ZR and GA, driven by differences among bamboo parts and developmental periods. The abundance of each endogenous hormone varied among bamboo parts and developmental periods. During bamboo shooting, ABA had the highest mass fraction in all bamboo parts sampled, followed by IAA, GA, and ZR. Among bamboo parts, rhizomes had more IAA, ZR, and GA than the other parts, but significantly less ABA. Winter shoots had higher ZR: IAA and GA: IAA ratios than rhizomes and maternal bamboo organs. During shoot growth, ABA was the most abundant hormone in rhizomes and maternal bamboo organs, followed by IAA, ZR, and GA. In contrast, IAA was the most abundant hormone in spring shoots, followed by ABA, ZR, and GA. Maternal bamboo organs had a significantly higher ZR: GA ratio, and significantly lower IAA: ABA, ZR: ABA, and GA: ABA ratios than rhizomes. Spring shoots had significantly higher IAA: ABA, ZR: ABA, and GA: ABA ratios than rhizomes and maternal bamboo organs; significantly higher ZR mass fractions, and ZR: GA and ZR: IAA ratios and significantly lower ABA mass fractions than rhizomes; and significantly higher GA: IAA ratio than maternal bamboo organs. During new-bamboo formation, ABA was the most abundant hormone in rhizomes, winter shoots, and maternal bamboo organs, followed by IAA, ZR, and GA. Maternal bamboo organs had significantly lower IAA mass fractions and significantly higher ABA mass fractions than rhizomes and new bamboo tissue. IAA and ABA abundances exhibited an inverse relationship in rhizomes and maternal bamboo organs. GA: ABA and GA: IAA ratios decreased gradually and other hormone ratios exhibited parabolic trends over the bamboo-shooting period, with the highest ratios observed in new bamboo tissues. Overall, the coordination or antagonism among endogenous hormones plays a key regulatory role in bamboo shoot growth. The formation of thick walls in P. edulis 'Pachyloen', one of its major traits, may be partially attributed to the relatively high IAA and ZR and low GA mass fractions., Competing Interests: The authors have declared that no competing interests exist.

The hop ( Humulus lupulus L.) is an important specialty crop used in beer production. Untargeted UPLC-QTof-MS E metabolomics was used to determine metabolite changes in the leaves of hop plants under varying degrees of drought stress. Principal component analysis revealed that drought treatments produced qualitatively distinct changes in the overall chemical composition of three out of four genotypes tested (i.e., Cascade, Sultana, and a wild var. neomexicanus accession but not Aurora), although differences among treatments were smaller than differences among genotypes. A total of 14 compounds consistently increased or decreased in response to drought stress, and this effect was generally progressive as the severity of drought increased. A total of 10 of these marker compounds were tentatively identified as follows: five glycerolipids, glutaric acid, pheophorbide A, abscisic acid, roseoside, and dihydromyricetin. Some of the observed metabolite changes likely occur across all plants under drought conditions, while others may be specific to hops or to the type of drought treatments performed.

Fruit and leaf possess distinctly different metabolites. Here, metabolites and transcriptome were compared between mature leaves (ML) and juice sacs (JS) of Citrus grandis "Hirado Buntan" to investigate the possible reasons. Results indicated that the remarkable difference in starch, total flavonoids and carotenoids, l-ascorbate, and jasmonic acid between ML and JS was tightly related to the expression levels of their biosynthesis-related genes, while the significant difference in abscisic acid and citrate was mainly related to the gene expression level(s) of 9- cis -epoxycarotenoid dioxygenase and proton pump genes, respectively. In addition, ATP citrate lyase probably plays a key role in the levels of flavonoids between ML and JS via providing different levels of acetyl-CoA. Taken together, these results identified some key candidate genes responsible for the content of a given metabolite and will contribute to research in regulating such metabolite content in citrus fruits.

Agronomically important traits often develop during the later stages of crop growth as consequences of various plant-environment interactions. Therefore, the temporal physiological states that change and accumulate during the crop's life course can significantly affect the eventual phenotypic differences in agronomic traits among crop varieties. Thus, to improve productivity, it is important to elucidate the associations between temporal physiological responses during the growth of different crop varieties and their agronomic traits. However, data representing the dynamics and diversity of physiological states in plants grown under field conditions are sparse. In this study, we quantified the endogenous levels of five phytohormones - auxin, cytokinins (CKs), ABA, jasmonate and salicylic acid - in the leaves of eight diverse barley (Hordeum vulgare) accessions grown under field conditions sampled weekly over their life course to assess the ongoing fluctuations in hormone levels in the different accessions under field growth conditions. Notably, we observed enormous changes over time in the development-related plant hormones, such as auxin and CKs. Using 3' RNA-seq-based transcriptome data from the same samples, we investigated the expression of barley genes orthologous to known hormone-related genes of Arabidopsis throughout the life course. These data illustrated the dynamics and diversity of the physiological states of these field-grown barley accessions. Together, our findings provide new insights into plant-environment interactions, highlighting that there is cultivar diversity in physiological responses during growth under field conditions., (© The Author(s) 2020. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Apple trees grafted on different rootstock types, including vigorous rootstock (VR), dwarfing interstock (DIR), and dwarfing self-rootstock (DSR), are widely planted in production, but the molecular determinants of tree branch architecture growth regulation induced by rootstocks are still not well known. In this study, the branch growth phenotypes of three combinations of 'Fuji' apple trees grafted on different rootstocks (VR: Malus baccata; DIR: Malus baccata/T337; DSR: T337) were investigated. The VR trees presented the biggest branch architecture. The results showed that the sugar content, sugar metabolism-related enzyme activities, and hormone content all presented obvious differences in the tender leaves and buds of apple trees grafted on these rootstocks. Transcriptomic profiles of the tender leaves adjacent to the top buds allowed us to identify genes that were potentially involved in signaling pathways that mediate the regulatory mechanisms underlying growth differences. In total, 3610 differentially expressed genes (DEGs) were identified through pairwise comparisons. The screened data suggested that sugar metabolism-related genes and complex hormone regulatory networks involved the auxin (IAA), cytokinin (CK), abscisic acid (ABA) and gibberellic acid (GA) pathways, as well as several transcription factors, participated in the complicated growth induction process. Overall, this study provides a framework for analysis of the molecular mechanisms underlying differential tree branch growth of apple trees grafted on different rootstocks., Competing Interests: The authors have declared that no competing interests exist.

In the context of future climate change new habitats will be threatened and unique species will be forced to develop different strategies to survive. Saxifraga longifolia Lapeyr. is an endemic species from the Pyrenees with a very particular habitat. We explored the capacity and strategies of S. longifolia plants to face different severities of drought stress under both natural conditions and controlled water stress followed by a re-watering period of 20 days. Our results showed a role for abscisic acid (ABA), salicylic acid (SA) and cytokinins (CKs) in plant survival from drought stress, and as the stress increased, ABA lost significance and SA appeared to be more associated with the response mechanisms. Moreover, photo-oxidative stress markers revealed that both xanthophyll cycles played a photoprotection role with a stronger participation of the lutein epoxide cycle as the stress was more intense. Severe drought decreased the maximum efficiency of photosystem II (F v /F m ) below 0.45, being this the limit to survive upon rewatering. Overall, our results proved different strategies of S. longifolia plants to cope with drought stress and suggested a F v /F m threshold to predict plant survival in high-mountain environments., (© 2020 Scandinavian Plant Physiology Society.)

Background: Abscisic acid-, stress-, and ripening-induced (ASR) genes are a class of plant specific transcription factors (TFs), which play important roles in plant development, growth and abiotic stress responses. The wheat ASRs have not been described in genome-wide yet., Methods: We predicted the transmembrane regions and subcellular localization using the TMHMM server, and Plant-mPLoc server and CELLO v2.5, respectively. Then the phylogeny tree was built by MEGA7. The exon-intron structures, conserved motifs and TFs binding sites were analyzed by GSDS, MEME program and PlantRegMap, respectively., Results: In wheat, 33ASR genes were identified through a genome-wide survey and classified into six groups. Phylogenetic analyses revealed that the TaASR proteins in the same group tightly clustered together, compared with those from other species. Duplication analysis indicated that the TaASR gene family has expanded mainly through tandem and segmental duplication events. Similar gene structures and conserved protein motifs of TaASRs in wheat were identified in the same groups. ASR genes contained various TF binding cites associated with the stress responses in the promoter region. Gene expression was generally associated with the expected group-specific expression pattern in five tissues, including grain, leaf, root, spike and stem, indicating the broad conservation of ASR genes function during wheat evolution. The qRT-PCR analysis revealed that several ASRs were up-regulated in response to NaCl and PEG stress., Conclusion: We identified ASR genes in wheat and found that gene duplication events are the main driving force for ASR gene evolution in wheat. The expression of wheat ASR genes was modulated in responses to multiple abiotic stresses, including drought/osmotic and salt stress. The results provided important information for further identifications of the functions of wheat ASR genes and candidate genes for high abiotic stress tolerant wheat breeding.

Root plasticity is controlled by hormonal homeostasis and nutrient availability. In this work, we have determined the influence of different N regimens on growth parameters and on the expression of genes involved in auxin transport and N-assimilation in tomato seedlings. NH 4 + nutrition led to an inhibitory effect on root fresh weight (FW), lateral root (LR) number and root density, while an increase in the primary root (PR) length was observed. The expression of N assimilation genes GS2 and ASN1, is affected by NH 4 + nutrition. Moreover, in order to relieve the toxic effect of NH 4 + on root development, glucose or 2-oxoglutarate was supplied as a C source during NH 4 + treatment. The addition of 2-oxoglutarate improved root parameters compared to the NH 4 + regimen. N-assimilation gene analysis showed that NH 4 + -fed tomato plants try to alleviate the toxic effect by concurrently upregulating ASN1 and anaplerotic PEPC2 expression, whereas when 2-oxoglutarate is supplied, ASN1 induction was not observed. The addition of both C skeletons induced the expression of the ROS-scavenging genes GSH and SOD. In addition, since ABA plays a role in root development, the ABA-synthesis-defective mutant flacca was studied under NO 3 - and NH 4 + regimens. It displayed a decrease in LR number under NO 3 - conditions, whereas, the NH 4 + -fed seedlings showed a decrease solely in PR length that was reverted when ABA was exogenously supplied. Moreover, flacca seedlings displayed a reprogramming of the N/C assimilation genes. Altogether, these results reflect the importance of N and C sources and ABA homeostasis in root development of tomato seedlings., Competing Interests: Declaration of competing interest No conflict of interest exists., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Triazole fungicides have been used for seed treatment to control soilborne diseases of maize, but seedlings coming from triazole-coated seed show serious phytotoxicity under chilling stress. To understand this phytotoxic impact, maize seed was treated with four triazoles fungicides and the corresponding seedlings were analysed on growth and gene expression. We found that maize seed coated with difenoconazole and tebuconazole exhibited either no or increased effects on germination and growth of maize at 25 °C, regardless of chemical concentrations. When maize seedlings were subjected to chilling treatment, however, their growth was significantly inhibited, and the inhibition was positively correlated with the rate of triazole application. Mesocotyl length decreased by 32.19-44.73% by difenoconazole, and 23.53-32.08% by tebuconazolet at rates of 1:50 and 1:25, respectively. However, myclobutanil did not have any effects at any temperatures. The contents of the gibberellin GA12 and abscisic acid in maize seedlings developed from difenoconazole- or tebuconazole-coated seed were significantly increased under chilling stress. The expression of two key catabolic enzyme genes, GA2ox3 and GA2ox4, was significantly up-regulated immediately following chilling stress and 2 days after recovery at 25 °C in the seedlings treated with difenoconazole or tebuconazole. This imbalance in phytohormones may explain why difenoconazole- or tebuconazole-coated seed more likely results in the phytotoxicity of maize seedlings under a low temperature condition during seed emergence and seedling growth. Since myclobutanil did not have this negative effect, it can be applied for seed coating in areas where temperatures are low during early seedling growth., Competing Interests: Declaration of competing interest The authors have declared that no competing interest exist., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

A number of scientific reports published to date contain data on endogenous levels of various phytohormones in potato (Solanum tuberosum L.) but a complete cytokinin profile of potato tissues, that would include data on all particular molecular forms of cytokinin, has still been missing. In this work, endogenous levels of all analytically detectable isoprenoid cytokinins, as well as the auxin indole-3-acetic acid (IAA), and abscisic acid (ABA) have been determined in shoots and roots of 30 day old in vitro grown potato (cv. Désirée). The results presented here are generally similar to other data reported for in vitro grown potato plants, whereas greenhouse-grown plants typically contain lower levels of ABA, possibly indicating that in vitro grown potato is exposed to chronic stress. Cytokinin N-glucosides, particularly N7-glucosides, are the dominant cytokinin forms in both shoots and roots of potato, whereas nucleobases, as the bioactive forms of cytokinins, comprise a low proportion of cytokinin levels in tissues of potato. Differences in phytohormone composition between shoots and roots of potato suggest specific patterns of transport and/or differences in tissue-specific metabolism of plant hormones. These results represent a contribution to understanding the hormonomics of potato, a crop species of extraordinary economic importance.

Okra, Abelmoschus esculentus L. (Moench), also known as Lady's Fingers, gombo, or bamje, is an annual plant belonging to the Malvaceae family. Traditional olistic medicine since centuries directly associates this plant and its parts to a beneficial health hypoglycemic effect. Since the abscisic acid (ABA) has been associated to an interesting hypoglycemic effect, this triggered us to verify and quantify the presence of the abscisic acid in the okra phytocomplex. In particular, ABA, a plant derived hormone, has been proven by recent studies to be effective on mammals. To determine and quantify the ABA content, different parts of the Okra plant extracts have been evaluated, and HPLC-DAD analysis has been used allowing us to report for the first time the presence of this isoprenoid compound. Bioaccessibility has been also investigated using a simulated gastro intestinal (GI) digestion protocol with the aim of explore the possibility of okra extract as nutraceutical.

A multi-channel localized surface plasmon resonance system is described for absorptiometric determination of abscisic acid (ABA). The system is making use of gold nanoparticles and consists of a broadband light source, a multi-channel alignment device, and a fiber spectrometer. The method is based on the specific interaction between an ABA-binding aptamer and ABA. This induces the growth of gold nanoparticles (AuNPs) functionalized with a polyadenine-tailed aptamer that act as optical probes. Different concentrations of ABA give rise to varied morphologies of grown AuNPs. This causes a change of absorption spectra which is recorded by the system. ABA can be quantified by measurement of the peak wavelength shifts of grown AuNPs. Under optimized conditions, this method shows a linear relationship in the 1 nM to 10 μM ABA concentration range. The detection limit is 0.51 nM. The sensitivity of the ABA assay is strongly improved compared to the method based on salt-induced AuNP aggregation. This is attributed to the use of a poly-A-tailed aptamer and the catalytic ability of AuNPs. In the actual application, the ABA concentration of ABA in fresh leaves of rice is measured with the maximum relative error of 8.03% in comparison with the ELISA method. Graphical abstractSchematic representation of an absorptiometric approach for determination of abscisic acid based on the growth of polyA-tailed aptamer-AuNPs probes and a multi-channel localized surface plasmon resonance system.

Heavy metals such as cadmium and zinc constitute major pollutants in coastal areas and frequently accumulate in salt marshes. The wetland halophyte plant species Kosteletzkya pentacarpos is a promising species for phytostabilization of contaminated areas. In order to assess the role of the antisenescing phytohormone cytokinin in heavy metal resistance in this species, seedlings were exposed for two weeks to Cd (10 μM), Zn (100 μM) or Cd + Zn (10 μM + 100 μM) in the presence or absence of 50 mM NaCl and half of the plants were sprayed every two days with the cytokinin trans-zeatine riboside (10 μM). Zinc reduced the endogenous cytokinin concentration. Exogenous cytokinin increased plant growth, stomatal conductance, net photosynthesis and total ascorbate and reduced oxidative stress estimated by malondialdehyde in Zn-treated plants maintained in the absence of NaCl. Heavy metal induced an increase in the senescing hormone ethylene which was reduced by cytokinin treatment. Plants exposed to the mixed treatment (Cd + Zn) exhibited a specific hormonal status in relation to accumulation of abscisic acid and depletion of salicylic acid. Non-protein thiols (glutathione and phytochelatins) accumulated in response to Cd and Cd + Zn. It is concluded that toxic doses of Cd and Zn have different impacts on the plant behavior and that the simultaneous presence of the two elements induces a specific physiological constraint at the plant level. Salinity helps the plant to cope with heavy metal toxicities and the plant hormone cytokinin assumes key function in Zn resistance but its efficiency is lower in the presence of NaCl., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Honey is a natural product with a complex chemical composition consisting of sugars and other bioactive compounds. It is important in many traditional systems of medicine, exhibiting interesting bioactivities, in particular antimicrobial, anti-inflammatory and antioxidant effects. Authentication of botanical origin of honeys is particularly important in this context. Therefore, methods for quality control of honey and detection of its adulteration are very important. A HPTLC method for the quantitative determination of phenolic compounds in honey was developed for the first time. Seven phenolic compounds were detected and determined quantitatively in lime and acacia honey samples. The obtained results show that the HPTLC profiles of phenolic compounds and abscisic acid can be useful for determining the floral origin of honeys. Chromatographic tests were supplemented by statistical analysis (PCA and HCA) led to the successful separation of acacia and lime honey samples., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Mango byproducts, such as peels, contain high levels of antioxidants and fiber and represent important sources of nutraceuticals and pharmacological products. Fruit are collected at the mature green stage then stored and ripened, undergoing several structural and molecular changes over the course of this process. However, very little is known regarding the content and nature of antioxidant compounds in peels of elite and local cultivars during postharvest shelf life (PSL). We screened the phenolic compound content of six cultivars during PSL, including elite (Kent, Tommy, and Ataulfo) and local (Manila, Manililla, and Criollo) mangoes, using a targeted metabolomics approach. We determined that Ataulfo mangoes exhibited the highest content of phenolic compounds during PSL. Untargeted metabolomics and comparative proteomics in Ataulfo and Manililla showed these cultivars to be significant sources of phenolic and lipidic compounds, with the latter cultivar also representing an interesting candidate as a new source for nutraceutical products., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Main Conclusion: The possible molecular mechanisms regulating strawberry fruit ripening were revealed by plant hormone quantification, exogenous hormone application, and RNA-sequencing. Fruit ripening involves a complex interplay among plant hormones. Strawberry is a model for studies on non-climacteric fruit ripening. However, the knowledge on how plant hormones are involved in strawberry ripening is still limited. To understand hormonal actions in the ripening process, we performed genome-wide transcriptome and hormonal analysis for the five major hormones (abscisic acid and catabolites, auxins, cytokinins, gibberellins, and ethylene) in achenes and receptacles (flesh) at different ripening stages of the woodland strawberry Fragaria vesca. Our results demonstrate that the pre-turning stage (a stage with white flesh and red achenes defined in this study) is the transition stage from immature to ripe fruits. The combinatorial analyses of hormone content, transcriptome data, and exogenous hormone treatment indicate that auxin is synthesized predominantly in achenes, while abscisic acid (ABA), bioactive free base cytokinins, gibberellins, and ethylene are mainly produced in receptacles. Furthermore, gibberellin may delay ripening, while ethylene and cytokinin are likely involved at later stages of the ripening process. Our results also provide additional evidence that ABA promotes ripening, while auxin delays it. Although our hormone analysis demonstrates that the total auxin in receptacles remains relatively low and unchanged during ripening, our experimental evidence further indicates that ABA likely enhances expression of the endoplasmic reticulum-localized auxin efflux carrier PIN-LIKES, which may subsequently reduce the auxin level in nucleus. This study provides a global picture for hormonal regulation of non-climacteric strawberry fruit ripening and also evidence for a possible mechanism of ABA and auxin interaction in the ripening process.

Abscisic acid (ABA) has been advocated to play substantial role on ripening of non-climacteric fruit. Here we report that alginate oligosaccharide (AOS) postharvest treatments delayed the accumulation of ABA and ABA-conjugates and restrained the expression of ABA signaling genes, resulting enlarged storage life of strawberry. In addition, AOS postharvest treatments also increased the quality and reduced the degradation of cell wall components and repressed the expression of cell wall degradation genes. AOS treated fruits exhibited significant delays of hardness, decay percentage, titratable acidity, pH, total soluble solids and vitamin C content compared to untreated fruits. Moreover, AOS had a positive effect on retaining higher amount of anthocyanin, total phenol and flavonoids contents. The finding of this study suggests that AOS postharvest treatments are very useful for preserving fruit quality and enhancing shelf life by delayed ABA accretion, restrained the gene expression related to ABA signaling and cell wall degeneration., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The different wheat varieties have different tolerance to cadmium stress, while the mechanisms underlying the Cd tolerance are still poorly understood. A pot experiment was conducted to study the changes of antioxidant enzyme activities and endogenous hormones in wheat (Triticum aestivum) genotypes differing in cadmium (Cd) accumulation (low = Pingan 8 and high = Bainong 160) in different growth stages under Cd stress. The Cd treatment (3 mg/kg) increased the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) and concentrations of malondialdehyde (MDA) and abscisic acid (ABA); in contrast, it reduced the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), intercellular carbon dioxide concentration (Ci) and the concentrations of gibberellin (GA 3 ), auxin (IAA) and zeatin nucleoside (ZR) in wheat leaves compared to the CK (without Cd). The antioxidant enzyme activities were higher in Bainong 160 than Pingan 8 under Cd stress. In addition, the changes in endogenous hormone concentration were smaller in Bainong 160 than Pingan 8 leaves. The correlation coefficients of Bainong 160 and Pingan 8 were 0.87 and 0.66, respectively. Our results suggest that high Cd accumulation (greater Cd tolerance) in Bainong 160 is associated with higher photosynthetic parameters, higher activities of antioxidant enzyme and higher concentration of hormones than Pingan 8., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Introduction: Methanol utilization by bacteria is important for various industrial processes. Methylotrophic bacteria are taxonomically diverse and some species promote plant growth and induce stress tolerance. However, methylotrophic potential of bacterial endophytes is poorly understood., Objective: The current study aimed to evaluate the metabolomic and proteomic changes in endophytic Bacillus amyloliquefaciens RWL-1 caused by its methanol utilization and the resultant influence on its phytohormone production., Methods: B. amyloliquefaciens RWL-1 was grown in LB medium with different concentrations [0 (control), 0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4%) of methanol to examine its methylotrophic potential. SDS-PAGE analysis was carried out for bacterial protein confirmation. Moreover, the phytohormones (indole 3 acetic acid (IAA), gibberellins (GAs), abscisic acid (ABA)) produced by RWL-1 in methanol supplemented medium were quantified by GC-MS/SIM (6890N Network GC system, and 5973 Network Mass Selective Detector; Agilent Technologies, Santa Clara, CA, USA), while the antioxidants were estimated spectrophotometrically (T60 UV-VIS spectrophotometer, Leicester, UK). The amino acid quantification was carried out by amino acid analyzer (HITACHI L-8900, Japan). Furthermore, Nano-liquid chromatography (LC)-MS/MS analysis was performed with an Agilent system (Wilmington, DE, USA) for proteomic analysis while mascot algorithm (Matrix science, USA) was used to identify peptide sequences present in the protein sequence database., Results: RWL-1 showed significant growth in media supplemented with 2 and 3.5% methanol, when compared with other concentrations. Mass spectroscopy analysis revealed that RWL-1 utilizes methanol efficiently as a carbon source. In the presence of methanol, RWL-1 produced significantly higher levels of IAA but lower levels of ABA, when compared with the control. Further, enzymatic antioxidants and functional amino acids were significantly up-regulated, with predominant expression of glutamic acid and alanine. Nano-liquid chromatography, quadrupole time-of-flight analysis, and quantitative analysis of methanol-treated bacterial cells showed expression of eight different types of proteins, including detoxification proteins, unrecognized and unclassified enzymes with antioxidant properties, proteases, metabolism enzymes, ribosomal proteins, antioxidant proteins, chaperones, and heat shock proteins., Conclusion: Results demonstrate that RWL-1 can significantly enhance its growth by utilizing methanol, and could produce phytohormones when growing in methanol-supplemented media, with increased expression of specific proteins and different biochemicals. These results will be useful in devising strategies for utilizing methylotrophic bacterial endophytes as alternative promoters of plant growth. Understanding RWL-1 ability to utilize methanol. The survival and phytohormones production by Bacillus amyloliquefaciens RWL-1 in methanol supplemented media whistle inducing metabolic and proteomic changes.

Phytochemical defense responses of plants are often herbivore-specific and can be affected by a herbivore's feeding mode. However, comprehensive studies documenting the impact of multiple herbivores from different feeding guilds on induced phytochemical responses in distal leaves and its consequences for plant-mediated herbivore interactions are limited and findings are inconsistent. We investigated how herbivory by leaf-chewing caterpillars, cell-content feeding spider mites and phloem-feeding aphids and whiteflies affect secondary metabolomes and phytohormone levels in youngest, non-damaged cotton leaves (distal leaves). Furthermore, bioassays with caterpillars were conducted to assess their performance on distal leaves of plants infested with different herbivores. Caterpillars, and to a lesser degree spider mites, led to a systemic induction of terpenoids with negative consequences for caterpillar performance in the bioassays. Both herbivores reduced levels of various nutrients and potentially antioxidative compounds. Caterpillar damage increased levels of jasmonoyl-L-isoleucine and abscisic acid (ABA), whereas spider mite infestation had no effect on phytohormone levels. Aphid and whitefly infestation did not systemically affect secondary metabolites. Aphids decreased salicylic acid levels while whitefly-infested plants contained increased ABA levels. Neither aphid nor whitefly infestation affected caterpillar performance. In general, feeding mode of a herbivore can affect systemically induced changes in phytochemistry and plant-mediated indirect interactions even though the two phloem-feeding herbivores triggered different phytohormonal responses. The observed reduction of nutrients and potentially antioxidative compounds upon caterpillar and spider mite herbivory underlines the importance of further elucidating the role of resource sequestration as a potential systemic defensive response following herbivory by chewers and cell-content feeding herbivores.

Abscisic acid (ABA), indoleacetic acid (IAA) and jasmonic acid (JA) are plant hormones that were reported to play indispensable roles during seed germination. However, the interactions between these plant hormones during rice seed germination have still not been explored clearly. A sensitive method for determination of these plant hormones would be beneficial for the exploration of such interactions. Herein, we present a liquid chromatography coupled with mass spectrometry (LC-MS) method for the quantification of ABA, IAA and JA in a single tissue of rice seed to investigate the spatio-temporal distribution of these plant hormones during rice seed germination. To this end, an in silico strategy was developed in order to select a derivatization reagent with an ideal sensitivity of MS detection. This strategy was confirmed with experimental studies on three reagents N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC), N,N-dimethylethylenediamine (DMED), and N-(acridin-9-yl)-2-bromoacetamide (AYBA) and their formic acid derivatives. Our results from the in silico and LC-MS experiments show that AYBA is a good derivatization reagent for ABA, IAA and JA due to its reasonable ionization efficiency in electrospray ionization mass spectrometry (ESI-MS) and excellent hydrophobicity. Finally, a sensitive LC-MS method upon AYBA was established for the determination of ABA, IAA and JA in germinated seeds. Good linearities for ABA, IAA, and JA were obtained with correlation coefficients greater than 0.99. The limits of detection (LODs) were in the range of 0.14-0.16 pg mL -1 . The method exhibits good precisions with RSD 1.5%-13.8% (intra-day) and 1.2%-7.3% (inter-day) and acceptable recoveries (88.6%-102.9%, n = 6). Finally, the method was successfully employed in the spatio-temporal profiling of ABA, IAA and JA in a single tissue of rice seed during rice seed germination., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Plants can re-programme their transcriptome, proteome and metabolome to deal with environmental and biotic stress. It has been shown that the rhizosphere microbiome has influence on the plant metabolome and on herbivore behaviour. In the present study, Trichoderma gamsii was isolated from Arabidopsis thaliana rhizosphere soil. The inoculation of roots of Arabidopsis thaliana with T. gamsii significantly inhibited the feeding behaviour of Trichoplusia ni and affected the metabolome as well as the content of phytohormones in Arabidopsis leaves. T. gamsii-treated plant leaves had higher levels of amino acids and lower concentrations of sugars. In addition, T. gamsii-treated plant leaves had more abscisic acid (ABA) and lower levels of salicylic acid (SA) and indole-3-acetic acid (IAA) in comparison with the untreated plants. Furthermore, the inoculation with T. gamsii on different signalling mutants showed that the induction of defences were SA-dependent. These findings indicate that T. gamsii has potential as a new type of biocontrol agent to promote plant repellence to insect attacks., (© 2018 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.)

An efficient and selective pretreatment method of one-step hydrophilic interaction chromatography-based solid phase extraction (HILIC SPE) was developed using silica as the sorbent to quickly and sensitively detect endogenous ABA and its five catabolites in fresh Oryza sativa tissues. The extracted analytes were sensitively quantified with ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Under the optimized conditions, good linearity of the developed analytical method was obtained in the range of 0.2-1000 ng/mL with linear correlation coefficients ( r) greater than 0.9987. The limits of detection (LODs, signal/noise = 3) ranged from 0.01 to 0.74 ng/mL. The relative recoveries were between 83.3% and 112.0% with the relative standard deviations (RSDs) ranging from 0.5 to 15.0%. Using the proposed method, the concentration variations of ABA and its catabolites were monitored in the salt-stressed rice tissues.

A melamine-based porous organic polymer (mPMF) was synthesized as solid phase extraction (SPE) adsorbent for inline SPE-high performance liquid chromatography (HPLC)-ultraviolet detector (UV) analysis of five phytohormones. Melamine contributed to the rich π-electron and N-containing triazine structure for mPMF, which could form π-π interaction and intermolecular hydrogen bond with COOH containing phytohormones. The synthesized mPMF adsorbent shows extremely high extraction efficiency for five target analytes (>80%) including salicylic acid, indole-3-acetic acid, abscisic acid, 2,4-dichlorophenoxyacetic acid and 1-naphthalene acetic acid. The factors affecting extraction of the target phytohormones were investigated and the optimized experimental conditions were established. The linear range was 0.2-100 μg/L with the limits of detection in the range of 0.06-0.11 μg/L for five target phytohormones. The developed method of mPMF-inline-SPE-HPLC-UV was applied for the analysis of trace target phytohormones in tomato and grape juice samples, and the recovery in the range of 83.1-116% and 87.2-121% was obtained for the spiked tomato and grape juice, respectively. This method has the advantages of high sensitivity and good reproducibility, and it has good application potential for the analysis of phytohormones in fruit samples. Moreover, inline analysis avoided the problems of sample pollution and sample loss, and provided a sample throughput of 5/h., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Bisphenol A (BPA) is an endocrine disruptor that is ubiquitous in the environment. Previous studies have focused on the effects of BPA on plants to assess the ecological risk of BPA in the environment. To evaluate the effects of BPA on plant biological characters more systematically, we investigated the biological characters of above-ground and under-ground organs of soybean plants exposed to BPA. Meanwhile, the mechanisms for the observed changes were also analyzed from the view of hormone levels and photosynthesis. The results showed that after exposure to 0.8 mg L -1 BPA for three days, indole-3-acetic acid (IAA) and gibberellic acid levels in roots increased significantly, and the IAA level increased in leaves, so the character indices of roots and leaves both increased. The IAA and ethylene levels in stems increased, but the character indices of stems did not increased. With higher BPA concentrations, especially exposure to 17.2 mg L -1 BPA, the levels of IAA, gibberellic acid, and zeatin decreased (except for the increased zeatin in leaves), and abscisic acid and ethylene levels increased; thus, all of the character indices significantly decreased. By comparing the changes in various biological characters, we found that leaf area, root surface area, and root length changed most significantly. In addition, changes in photosynthetic parameters provided initial causes for plant growth changes, and impacted biological characters. The changes of character indices were stronger when the BPA exposure time was prolonged, and after the removal of BPA, the character indices showed some recovery. Therefore, BPA exposure can regulate the changes in plant characters by influencing hormone levels and photosynthesis, and root surface area, root length, and leaf area were the most sensitive to BPA., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Low temperature is one of the important limiting factors for growing season and geographical distribution of plants. Zoysiagrass (Zoysia Willd) is one of the widely used warm-season turfgrass that is distribute in many parts of the world. Zoysaigrass native to high latitude may have evolved higher cold tolerance than the ones native to low latitude. The objective of this study was to investigate the cold stress response in zoysiagrass native to diverse latitude at phenotypic, physiological and metabolic levels. Two zoysiagrass (Z. japonica) genotypes, Latitude-40 (higher latitude) and Latitude-22 (lower latitude) were subjected to four temperature treatments (optimum, 30/25°C, day/night; suboptimum, 18/12°C; chilling, 8/2°C; freezing, 2/-4°C) progressively in growth chambers. Low temperature (chilling and freezing) increased leaf electrolyte leakage (EL) and reduced plant growth, turf quality, chlorophyll (Chl) content, photochemical efficiency (Fv/Fm) and photosynthesis (Pn, net photosynthetic rate; gs, stomatal conductance; intercellular CO2; Tr, transpiration rate) in two genotypes, with more rapid changes in Latitude-22. Leaf carbohydrates content (glucose, fructose, sucrose, trehalose, fructan, starch) increased with the decreasing of temperature, to a great extend in Latitude-40. Leaf abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) content increased, while indole-3-acetic acid (IAA), gibberellic acid (GA3) and trans-zeatin ribside (t-ZR) content decreased with the reduction of temperature, with higher content in Latitude-40 than in Latitude-22. Chilling and freezing induced the up-regulation of C-repeat binding factor (ZjCBF), late embryogenesis abundant (ZjLEA3) and dehydration-responsive element binding (ZjDREB1) transcription factors in two genotypes, whereas those genes exhibited higher expression levels in Latitude-40, particularly under freezing temperature. These results suggested that zoysiagrass native to higher latitude exhibited higher freezing tolerance may attribute to the higher carbohydrates serving as energy reserves and stress protectants that stabilize cellular membranes. The phytohormones may serve signals in regulating plant growth, development and adaptation to low temperature as well as inducing the up-regulated ZjCBF, ZjLEA3 and ZjDREB1 expressions thus result in a higher cold tolerance., Competing Interests: The authors have declared that no competing interests exist.

Background and Aims: Predicting both plant water status and leaf gas exchange under various environmental conditions is essential for anticipating the effects of climate change on plant growth and productivity. This study developed a functional-structural grapevine model which combines a mechanistic understanding of stomatal function and photosynthesis at the leaf level (i.e. extended Farqhuhar-von Caemmerer-Berry model) and the dynamics of water transport from soil to individual leaves (i.e. Tardieu-Davies model)., Methods: The model included novel features that account for the effects of xylem embolism (fPLC) on leaf hydraulic conductance and residual stomatal conductance (g0), variable root and leaf hydraulic conductance, and the microclimate of individual organs. The model was calibrated with detailed datasets of leaf photosynthesis, leaf water potential, xylem sap abscisic acid (ABA) concentration and hourly whole-plant transpiration observed within a soil drying period, and validated with independent datasets of whole-plant transpiration under both well-watered and water-stressed conditions., Key Results: The model well captured the effects of radiation, temperature, CO2 and vapour pressure deficit on leaf photosynthesis, transpiration, stomatal conductance and leaf water potential, and correctly reproduced the diurnal pattern and decline of water flux within the soil drying period. In silico analyses revealed that decreases in g0 with increasing fPLC were essential to avoid unrealistic drops in leaf water potential under severe water stress. Additionally, by varying the hydraulic conductance along the pathway (e.g. root and leaves) and changing the sensitivity of stomatal conductance to ABA and leaf water potential, the model can produce different water use behaviours (i.e. iso- and anisohydric)., Conclusions: The robust performance of this model allows for modelling climate effects from individual plants to fields, and for modelling plants with complex, non-homogenous canopies. In addition, the model provides a basis for future modelling efforts aimed at describing the physiology and growth of individual organs in relation to water status.

Dormancy is an innate constraint on germination that occurs across all life forms. In this study, we investigated the seed dormancy release and germination characters of Taxus yunnanensis by exploring the seed morphology, permeability, germination inhibitors, endogenous hormones, and embryo germination in vitro during wet sand storage. Our results showed that seeds and embryos grew to a critical size to germination and permeability increased with the extension of storage. Seed coat and kernel methanol extracts reduced Brassica campestris seed vigor index. The in vitro embryo germination rate increased by 12.20% after storage for 30-360 d, whereas seed germination occurred after 450 d. Gibberellic acid and zeatin riboside contents were relatively stable, whereas abscisic acid (ABA) content decreased; indole acetic acid (IAA) content and the IAA/ABA ratio showed increasing trends. These results indicate that ABA is the key inhibitor of germination in Taxus. The chemical(s) in seed coat and kernel cause the inhibition of seed germination. Taken together, Taxus seeds have morphophysiological dormancy, in which the embryos can continue to grow and hormone imbalance inhibits further development and germination. Further, seed dormancy is active even during the middle of storage and shows "double peaks" during the entire dormancy process.

The methodology described here represents an improved strategy for analysis of a broad range of stress-related plant hormones including jasmonates, salicylic acid, abscisic acid, and auxin metabolites. The method conditions are optimized in order to reduce the background effect of complicated plant matrix, allow effective preconcentration and thus perform highly sensitive profiling of multiple plant hormones by ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS).

Main Conclusion: We cloned and characterized the ZmWRKY17 gene from maize. Overexpression of ZmWRKY17 in Arabidopsis led to increased sensitivity to salt stress and decreased ABA sensitivity through regulating the expression of some ABA- and stress-responsive genes. The WRKY transcription factors have been reported to function as positive or negative regulators in many different biological processes including plant development, defense regulation and stress response. This study isolated a maize WRKY gene, ZmWRKY17, and characterized its role in tolerance to salt stress by generating transgenic Arabidopsis plants. Expression of the ZmWRKY17 was up-regulated by drought, salt and abscisic acid (ABA) treatments. ZmWRKY17 was localized in the nucleus with no transcriptional activation in yeast. Yeast one-hybrid assay showed that ZmWRKY17 can specifically bind to W-box, and it can activate W-box-dependent transcription in planta. Heterologous overexpression of ZmWRKY17 in Arabidopsis remarkably reduced plant tolerance to salt stress, as determined through physiological analyses of the cotyledons greening rate, root growth, relative electrical leakage and malondialdehyde content. Additionally, ZmWRKY17 transgenic plants showed decreased sensitivity to ABA during seed germination and early seedling growth. Transgenic plants accumulated higher content of ABA than wild-type (WT) plants under NaCl condition. Transcriptome and quantitative real-time PCR analyses revealed that some stress-related genes in transgenic seedlings showed lower expression level than that in the WT when treated with NaCl. Taken together, these results suggest that ZmWRKY17 may act as a negative regulator involved in the salt stress responses through ABA signalling.

Environmental conditions profoundly affect plant disease development; however, the underlying molecular bases are not well understood. Here we show that elevated temperature significantly increases the susceptibility of Arabidopsis to Pseudomonas syringae pv. tomato (Pst) DC3000 independently of the phyB/PIF thermosensing pathway. Instead, elevated temperature promotes translocation of bacterial effector proteins into plant cells and causes a loss of ICS1-mediated salicylic acid (SA) biosynthesis. Global transcriptome analysis reveals a major temperature-sensitive node of SA signalling, impacting ~60% of benzothiadiazole (BTH)-regulated genes, including ICS1 and the canonical SA marker gene, PR1. Remarkably, BTH can effectively protect Arabidopsis against Pst DC3000 infection at elevated temperature despite the lack of ICS1 and PR1 expression. Our results highlight the broad impact of a major climate condition on the enigmatic molecular interplay between temperature, SA defence and function of a central bacterial virulence system in the context of a widely studied susceptible plant-pathogen interaction.

Abscisic acid (ABA) plays an important role in abiotic stress response and physiological signal transduction resisting to the adverse environment. Therefore, it is very essential for the quantitative detection of abscisic acid (ABA) due to its indispensable role in plant physiological activities. Herein, a new detection method based on localized surface plasmon resonance (LSPR) using aptamer-functionalized gold nanoparticles (AuNPs) is developed without using expensive instrument and antibody. In the presence of ABA, ABA specifically bind with their aptamers to form the ABA-aptamer complexes with G-quadruplex-like structure and lose the ability to stabilize AuNPs against NaCl-induced aggregation. Meanwhile, the changes of the LSPR spectra of AuNP solution occur and therefore the detection of ABA achieved. Under optimized conditions, this method showed a good linear range covering from 5×10-7 M to 5×10-5 M with a detection limit of 0.33 μM. In practice, the usage of this novel method has been demonstrated by its application to detect ABA from fresh leaves of rice with the relative error of 6.59%-7.93% compared with ELISA bioassay. The experimental results confirmed that this LSPR-based biosensor is simple, selective and sensitive for the detection of ABA. The proposed LSPR method could offer a new analytical platform for the detection of other plant hormones by changing the corresponding aptamer.

Background: The importance of ethylene in avocado ripening has been extensively studied. In contrast, little is known about the possible role of abscisic acid (ABA). The present work studied the effect of 1-methylcyclopropene (1-MCP) (0.3 μL L -1 ), e+® Ethylene Remover and the combination thereof on the quality of imported avocado cv. Hass fruit stored for 7 days at 12 °C. Ethylene production, respiration, firmness, colour, heptose (C7) sugars and ABA concentrations in mesocarp tissue were measured throughout storage., Results: Treatment with e+® Ethylene Remover reduced ethylene production, respiration rate and physiological ripening compared with controls. Fruit treated with 1-MCP + e+® Ethylene Remover and, to a lesser extent 1-MCP alone, had the lowest ethylene production and respiration rate and hence the best quality. Major sugars measured in mesocarp tissue were mannoheptulose and perseitol, and their content was not correlated with ripening parameters. Mesocarp ABA concentration, as determined by mass spectrometry, increased as fruit ripened and was negatively correlated with fruit firmness., Conclusions: Results suggest a relationship between ABA and ethylene metabolism since blocking ethylene, and to a larger extent blocking and removing ethylene, resulted in lower ABA concentrations. Whether ABA influences avocado fruit ripening needs to be determined in future research. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

We have developed a new liquid chromatography-electrospray ionization tandem mass spectrometry methodology based on 2-picolylamine derivatization and positive ion mode detection for abscisic acid (ABA) identification. The selected reaction leads to the formation of an amide derivative which contains a highly active pyridyl group. The enhanced ionization allows for a 700-fold increase over commonly monitored unmodified ABA, which in turn leads to excellent limits of detection and quantification values of 0.03 and 0.15ngmL -1 , respectively. This method has been validated in the highly complex matrix of a red blood cell extract. In spite of the high sensitivity achieved, ABA could not be detected in Plasmodium falciparum-infected red blood cells, suggesting that, if present, it will be found either in ultratrace amounts or as brief bursts at defined time points within the intraerythrocytic cycle and/or in the form of a biosynthetic analogue., (Copyright © 2017 Elsevier B.V. All rights reserved.)