Involvement Of Abscisic Acid Research Articles

Changes in Hormonal Profiles and Corresponding Gene Expressions During the Initiation and Development of Juice Sac Primordia in Citrus Ovaries and Fruitlets

The edible part of citrus fruit is composed of juice vesicles/sacs which develop from the endocarp, the two to three inner cell layers of the white spongy peel termed albedo. Juice sac primordia usually appear 1 week after anthesis. Hormones, especially auxin and gibberellin, play a role in pericarp development during the ovary-to-fruit transition, but their effect on juice vesicle induction has not been studied. Here, hormone profiling in the pericarp and changes in the expression of their corresponding genes in the endocarp and pericarp were compared between two citrus cultivars: Calabria citron, in which juice sacs develop normally, and Yemenite citron, in which juice sac formation does not initiate. Most of the identified hormones, abscisic acid, gibberellin A4, indole-3-acetic acid, isopentenyladenine, jasmonic acid and zeatin riboside, were at higher levels in Yemenite than in Calabria. Overall, changes in abscisic acid levels in the pericarp were very well correlated with changes in the expression of abscisic acid-related genes in the endocarp. However, the application of various hormones, including abscisic acid, to Calabria flowers failed to arrest juice sac initiation. The possible involvement of abscisic acid and other hormones in the process of juice vesicle initiation and pericarp growth is discussed.

Involvement of abscisic acid on antioxidant enzymes activity and gene expression in Lavandula angustifolia cv. Munstead under drought stress

Involvement of abscisic acid on antioxidant enzymes activity and gene expression in Lavandula angustifolia cv. Munstead under drought stress

Involvement of Abscisic Acid in Transition of Pea (Pisum sativum L.) Seeds from Germination to Post-Germination Stages.

The transition from seed to seedling represents a critical developmental step in the life cycle of higher plants, dramatically affecting plant ontogenesis and stress tolerance. The release from dormancy to acquiring germination ability is defined by a balance of phytohormones, with the substantial contribution of abscisic acid (ABA), which inhibits germination. We studied the embryonic axis of Pisum sativum L. before and after radicle protrusion. Our previous work compared RNA sequencing-based transcriptomics in the embryonic axis isolated before and after radicle protrusion. The current study aims to analyze ABA-dependent gene regulation during the transition of the embryonic axis from the germination to post-germination stages. First, we determined the levels of abscisates (ABA, phaseic acid, dihydrophaseic acid, and neo-phaseic acid) using ultra-high-performance liquid chromatography-tandem mass spectrometry. Second, we made a detailed annotation of ABA-associated genes using RNA sequencing-based transcriptome profiling. Finally, we analyzed the DNA methylation patterns in the promoters of the PsABI3, PsABI4, and PsABI5 genes. We showed that changes in the abscisate profile are characterized by the accumulation of ABA catabolites, and the ABA-related gene profile is accompanied by the upregulation of genes controlling seedling development and the downregulation of genes controlling water deprivation. The expression of ABI3, ABI4, and ABI5, which encode crucial transcription factors during late maturation, was downregulated by more than 20-fold, and their promoters exhibited high levels of methylation already at the late germination stage. Thus, although ABA remains important, other regulators seems to be involved in the transition from seed to seedling.

Light Quality and Sucrose-Regulated Detached Ripening of Strawberry with Possible Involvement of Abscisic Acid and Auxin Signaling.

The regulation of detached ripening is significant for prolonging fruit shelf life. Although light quality and sucrose affecting strawberry fruit ripening have been widely reported, little information is available about how they co-regulate the strawberry detached ripening process. In this study, different light qualities (red light-RL, blue light-BL, and white light-WL) and 100 mM sucrose were applied to regulate the ripening of initial red fruits detached from the plant. The results showed RL-treated samples (RL + H2O, RL + 100 mM sucrose) had brighter and purer skin color with a higher L*, b*, and C* value, and promoted the ascorbic acid. Almost all light treatments significantly decreased TSS/TA (total soluble solid/titratable acid) and soluble sugar/TA ratio, which is exacerbated by the addition of sucrose. Blue or red light in combination with sucrose notably increased total phenolic content and decreased malondialdehyde (MDA) accumulation. In addition, blue or red light combined with sucrose increased abscisic acid (ABA) content and promoted ABA signaling by inducing ABA-INSENSITIVE 4 (ABI4) expression and inhibiting SUCROSE NONFERMENTING1-RELATED PROTEIN KINASE 2.6 (SnRK2.6) expression. The strawberries exposed to blue and red light significantly improved auxin (IAA) content compared to the control (0 d), whereas the addition of sucrose inhibited IAA accumulation. Moreover, sucrose treatment suppressed the AUXIN/INDOLE-3-ACETIC ACID 11 (AUX/IAA11) and AUXIN RESPONSE FACTOR 6 (ARF6) expression under different light qualities. Overall, these results indicated that RL/BL + 100 mM sucrose might promote the detached ripening of strawberries by regulating abscisic acid and auxin signaling.

Hydrogen Peroxide and GA3 Levels Regulate the High Night Temperature Response in Pistils of Wheat (Triticum aestivum L.).

High night temperature (HNT) impairs crop productivity through the reproductive failure of gametes (pollen and pistil). Though female gametophyte (pistil) is an equal partner in the seed-set, the knowledge of the antioxidant system(s) and hormonal control of HNT tolerance or susceptibility of pistils is limited and lacking. The objectives of this study were to determine the antioxidant mechanism for homeostatic control of free radicals, and the involvement of abscisic acid (ABA) and gibberellic acid (GA3) in HNT stress protection in the wheat pistils of contrasting wheat genotypes. We hypothesized that HNT tolerance is attributed to the homeostatic control of reactive oxygen species (ROS) and hormonal readjustment in pistils of the tolerant genotype. The ears of two contrasting wheat genotypes-HD 2329 (susceptible) and Raj 3765 (tolerant) were subjected to two HNTs (+5 °C and +8 °C) over ambient, in the absence and presence of dimethylthiourea (DMTU), a chemical trap of hydrogen peroxide (H2O2). Results showed that HNTs significantly increased ROS in pistils of susceptible genotype HD 2329 to a relatively greater extent compared to tolerant genotype Raj 3765. The response was similar in the presence or absence of DMTU, but the H2O2 values were lower in the presence of DMTU. The ROS levels were balanced by increased activity of peroxidase under HNT to a greater extent in the tolerant genotype. Cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC) activity was inversely related to H2O2 production within a critical range in Raj 3765, indicating its modulation by H2O2 levels as no change was observed at the transcriptional level. The hormonal status showed increased ABA and decreased GA3 contents with increasing temperature. Our study elucidates the role of H2O2 and GA3 in stress tolerance of pistils of tolerant genotype where GAPC acts as a ROS sensor due to H2O2-mediated decrease in its activity.

The suppressed expression of a stress responsive gene 'OsDSR2' enhances rice tolerance in drought and salt stress

Rice is a crucial staple food crop in many countries, yet, abiotic factors like salt and drought impact its growth. The Domain of Unknown Function 966 (DUF966) gene family may be crucial in how rice plants respond to abiotic stress. Our earlier research showed that overexpression of OsDSR2 (DUF966-stress repressive gene 2 in Oryza sativa) decreased resistance to salt and drought stress. To further understand how OsDSR2 negatively affects rice tolerance to salt and drought stress, transgenic rice plants with decreased OsDSR2 expression levels were created employing the RNAi technique. We investigated alterations in rice phenotype, physiology, and differentially expressed genes (DEGs) using a combination of physio-biochemical measurement and RNA-seq analysis. The results of the study demonstrated that rice seedling lines with OsDSR2 knockdown exhibited improved salt and drought stress tolerance. Statistical analysis revealed that the transgenic plants' survival rate (56–68%) was higher than the control plants (30%), in addition to a roughly 3 fold, 3.5 fold, 20% and 10.5% reduction in cell membrane permeability, malondialdehyde (MDA), superoxide anion radical (O2−) and hydrogen peroxide (H2O2) contents, respectively. However, the proline content and antioxidant enzymes (superoxide dismutase (SOD) and peroxidase (POD)) activities were considerably increased by about 5.5 fold, 3.5 fold, and 4.5 fold, respectively, at physiological levels. There were 115 up-regulated and 173 down-regulated DEGs in the leaves of the transgenic lines on the transcriptional regulation under the combined salt-drought stress. Among these, both up-regulation DEGs (e.g., OsHAK5, OsIAA25) and the down-regulation DEGs (e.g., OsbZIP23, OsERF48, OsAP2-39, etc.) may be related to the enhanced tolerance of the transgenic lines under combined salt-drought stress. This possibly depended on the involvement of abscisic acid (ABA) and indoleacetic acid (IAA) signaling pathways. These findings further confirmed that OsDSR2 negatively affected rice's ability to withstand salt and drought, suggesting that it could be a helpful gene for CRISPR-Cas9 technology-based genetic modification of rice's ability to withstand abiotic stress.

Involvement of abscisic acid in silicon-mediated enhancement of copper stress tolerance in Artemisia annua

Heavy metal (HM) toxicity is a well-known hazard which causes deleterious impact on the growth and development of plants. The impact of abscisic acid (ABA) in presence of silicon (Si) on plant development and quality traits has largely gone unexplored. The effects of ABA and Si on the growth, yield, and quality characteristics of Artemisia annua L. plants growing under copper (Cu) stress (20 and 40 mg kg−1) were investigated in a pot experiment. During this investigation, Cu stress caused severe damage to the plants but exogenous administration of Si and ABA ameliorated the harmful effects of Cu toxicity, and the plants displayed higher biomass and improved physio-biochemical attributes. Copper accumulated in the roots and shoots and its toxicity caused oxidative stress as demonstrated by the increased 2-thiobarbituric acid reactive substance (TBARS) content. It also resulted in the increased activity of antioxidant enzymes, however, the exogenous Si and ABA supplementation decreased the buildup of reactive oxygen species (ROS) and lipid peroxidation, alleviating the oxidative damage produced by HM stress. Copper toxicity had a considerable negative impact on glandular trichome density, ultrastructure as well as artemisinin production. However, combined Si and ABA enhanced the size and density of glandular trichomes, resulting in higher artemisinin production. Taken together, our results demonstrated that exogenous ABA and Si supplementation protect A. annua plants against Cu toxicity by improving photosynthetic characteristics, enhancing antioxidant enzyme activity, protecting leaf structure and integrity, avoiding excess Cu deposition in shoot and root tissues, and helping in enhanced artemisinin biosynthesis. Our results indicate that the combined application of Si and ABA improved the overall growth of plants and may thus be used as an effective approach for the improvement of growth and yield of A. annua in Cu-contaminated soils.

The potential of seed germination inhibition test for early pregnancy detection and improved reproductive efficiency of cattle in Zambia

Early pregnancy diagnosis is an important management practice for reducing calving interval, increasing cattle reproductive efficiency, and the overall herd productivity. This study was undertaken to assess the viability of seed germination inhibition technique (Punyakoti test) for early pregnancy detection in cattle under the tropical rearing conditions. Twenty-four randomly selected cows were used for the experiment. Urine samples were collected and subjected to Punyakoti test, using maize seeds, within 6 hours of collection. Descriptive statistics employing means and standard error were used to analyse data, also, inferential statistics including analysis of variance and t-tests were employed to ascertain differences between the variables under study. Urine from pregnant cows had the highest (80.03±3.99) inhibitory effect while the distilled water group had the least (7.50±3.81) mean seed germination inhibition. The means of germinated maize seeds in pregnant and non-pregnant cow urine treatment groups were significantly different. The means of germinated maize seeds in unstripped and stripped pregnant cow urine were significantly different, while the mean shoot length values for the same treatments were not significantly different. Urine from 42 days’ pregnant cows had the highest germination inhibition (80.21±3.59) while the least (25.00±4.35) was observed on day 10. The Punyakoti test reliably detected pregnancy starting from 26 days after insemination. In conclusion, this technique can be used for detecting pregnancy as early 26 days after insemination. The potential role of steroids and involvement of abscisic acid (ABA) in seed germination inhibition requires further investigation.

Is ABA the exogenous vector of interplant drought cuing?

ABSTRACT We have recently demonstrated that root cuing from drought-stressed plants increased the survival time of neighboring plants under drought, which came at performance costs under benign conditions. The involvement of abscisic acid (ABA) was implicated from additional experiments in which interplant drought cuing was greatly diminished in ABA-deficient plants. Here, we tested the hypothesis that ABA is the exogenous vector of interplant drought cuing. Pisum sativum plants were grown in rows of three split-root plants. One of the roots of the first plant was subjected to either drought of benign conditions in one rooting vial, while its other root shared its rooting vial with one of the roots of an unstressed neighbor, which in turn shared its other rooting vial with an additional unstressed neighbor. One hour after subjecting one of the roots of the first plant to drought, ABA concentrations were 106% and 145% higher around its other root and the roots of its unstressed neighbor, compared to their respective unstressed controls; however, the absolute concentrations of ABA found in the rooting media were substantially low. The results may indicate that despite its involvement in interplant drought and the commonly observed exchange of ABA between drought-stressed plants and their rhizospheres, ABA is not directly involved in exogenous interplant drought cuing. However, previous studies have shown that even minute concentrations of ABA in the rhizosphere can prevent ABA leakage from roots and thus to significantly increase endogenous ABA levels. In addition, under drought conditions, plants tend to accumulate ABA, which could markedly increase internal ABA concentrations over time and ABA concentrations in close proximity to the root surface might be significantly greater than estimated from entire rooting volumes. Finally, phaseic acid, an ABA degradation product, is known to activate various ABA receptors, which could enhance plant drought tolerance. It is thus feasible that while the role of ABA is limited, its more stable degradation products could play a significant role in interplant drought cuing. Our preliminary findings call for an extensive investigation into the identity and modes of operation of the exogenous vectors of interplant drought cuing.

The Effects of Rhizosphere Inoculation with Pseudomonas mandelii on Formation of Apoplast Barriers, HvPIP2 Aquaporins and Hydraulic Conductance of Barley.

Pseudomonas mandelii strain IB-Ki14 has recently been shown to strengthen the apoplastic barriers of salt-stressed plants, which prevents the entry of toxic sodium. It was of interest to find out whether the same effect manifests itself in the absence of salinity and how this affects the hydraulic conductivity of barley plants. Berberine staining confirmed that the bacterial treatment enhanced the deposition of lignin and suberin and formation of Casparian bands in the roots of barley plants. The calculation of hydraulic conductance by relating transpiration to leaf water potential showed that it did not decrease in bacteria-treated plants. We hypothesized that reduced apoplastic conductivity could be compensated by the higher conductivity of the water pathway across the membranes. This assumption was confirmed by the results of the immunolocalization of HvPIP2;5 aquaporins with specific antibodies, showing their increased abundance around the areas of the endodermis and exodermis of bacteria-treated plants. The immunolocalization with antibodies against auxins and abscisic acid revealed elevated levels of these hormones in the roots of plants treated with bacteria. This root accumulation of hormones is likely to be associated with the ability of Pseudomonas mandelii IB-Ki14 to synthesize these hormones. The involvement of abscisic acid in the control of aquaporin abundance and auxins—in the regulation of and formation of apoplast barriers—is discussed.

The involvement of abscisic acid in glucose promoted adventitious root development in cucumber

Several lines of evidence suggest that glucose (Glc) and abscisic acid (ABA) are important signal molecules involved in plant growth and development. In the study, cucumber (Cucumis sativus ‘Xinchun NO. 4′) was used to understand the roles and relationships of Glc and ABA in adventitious root development. The results show that the effect of Glc and ABA on adventitious rooting in cucumber was dose-dependent, with a maximal biological response at 0.10 mM Glc and 0.05 μM ABA, respectively. However, the promotive roles of Glc in adventitious rooting were suppressed by ABA inhibitor fluridone (FLU), suggesting that endogenous ABA might be involved in Glc-induced adventitious rooting. Glc increased endogenous ABA content and the activity of ABA biosynthesis-related enzyme zeaxanthin epoxidase (ZEP), 9-cis-epoxy-carotenoid dioxigenase (NCED) and aldehyde oxidase (AO) during adventitious rooting. Glc decreased the activity of ABA degradation-related enzyme ABA 8′-hydroxylase (ABA8’-H). Glc significantly up-regulated the expression of ABA biosynthesis-related genes CsNCED1 and CsNCED2 and ABA signal transduction-related genes CsPYR1, CsPYR2, CsPYL8 and CsSRK2E, and significantly down-regulated the expression of ABA degradation-related genes CsCYP707A1 and CsCYP707A2. However, Glc-enhanced ABA content, ABA-related enzyme activity and ABA-related gene expression were reversed by FLU. Therefore, Glc might promote adventitious rooting by enhancing ABA accumulation, in which the activity of ABA biosynthesis enzyme (ZEP, NCED and AO), the expression of ABA biosynthesis-related genes (CsNCED1 and CsNCED2) and ABA signal transduction-related genes (CsPYR1, CsPYR2, CsPYL8 and CsSRK2E) were increased by Glc, and the activity of ABA degradation enzyme ABA8’-H and the expression of ABA degradation-related genes (CsCYP707A1 and CsCYP707A2) were inhibited by Glc.

The involvement of abscisic acid in hydrogen gas-enhanced drought resistance in tomato seedlings

Hydrogen gas (H2) regulates plant responses to abiotic and biotic stresses. Abscisic acid (ABA) might alleviate the adverse effects of drought stress. However, the mechanisms by which ABA and H2 ameliorate plant drought stress are unclear. Here, tomato “Micro-Tom” seedlings were used to investigate the interaction between H2 and ABA under drought conditions. The results showed that both hydrogen-rich water (HRW) and ABA increased plant height, stem diameter and root activity of tomato seedlings under drought stress, with optimal concentrations of 75% and 150 μM, respectively. HRW or ABA treatment was able to enhance drought tolerance by increasing photosynthesis, antioxidant enzyme activity and gene expression of antioxidant enzymes under drought stress. Fluridone (FLU), a synthetic inhibitor of ABA, significantly diminished the positive effects of HRW on plant height, stem diameter and root activity under drought stress, suggesting that ABA might play a crucial role in H2-enhanced drought resistance in tomato seedlings. The results revealed that the ABA content in the PEG + HRW treatment was 18% higher than that in the polyethylene glycol (PEG) treatment alone. Furthermore, we found that H2 enhanced endogenous ABA content by increasing zeaxanthin epoxidase (ZEP), 9-cis-epoxycarotenoid dioxygenase (NCED) and abscisic aldehyde oxidase (AAO) activities and the expression of SlZEP, SlNCED, and SlAAO. PEG + HRW treatment increased ZEP and NCED activities, which were 13% and 40% higher than those of PEG treatment, respectively. Meanwhile, the transcription levels of SlSnRK2 and SlAREB were upregulated by HRW or ABA under drought stress, whereas this upregulation was reversed by FLU. Thus, our results demonstrate that H2 was able to enhance drought resistance by regulating ABA biosynthesis and the expression of ABA signal transduction genes in tomato seedlings.

Identification of ABA-Mediated Genetic and Metabolic Responses to Soil Flooding in Tomato (Solanum lycopersicum L. Mill).

Soil flooding is a compound abiotic stress that alters soil properties and limits atmospheric gas diffusion (O2 and CO2) to the roots. The involvement of abscisic acid (ABA) in the regulation of soil flooding-specific genetic and metabolic responses has been scarcely studied despite its key importance as regulator in other abiotic stress conditions. To attain this objective, wild type and ABA-deficient tomatoes were subjected to short-term (24 h) soil waterlogging. After this period, gas exchange parameters were reduced in the wild type but not in ABA-deficient plants that always had higher E and gs. Transcript and metabolite alterations were more intense in waterlogged tissues, with genotype-specific variations. Waterlogging reduced the ABA levels in the roots while inducing PYR/PYL/RCAR ABA receptors and ABA-dependent transcription factor transcripts, of which induction was less pronounced in the ABA-deficient genotype. Ethylene/O2-dependent genetic responses (ERFVIIs, plant anoxia survival responses, and genes involved in the N-degron pathway) were induced in hypoxic tissues independently of the genotype. Interestingly, genes encoding a nitrate reductase and a phytoglobin involved in NO biosynthesis and scavenging and ERFVII stability were induced in waterlogged tissues, but to a lower extent in ABA-deficient tomato. At the metabolic level, flooding-induced accumulation of Ala was enhanced in ABA-deficient lines following a differential accumulation of Glu and Asp in both hypoxic and aerated tissues, supporting their involvement as sources of oxalacetate to feed the tricarboxylic acid cycle in waterlogged tissues and constituting a potential advantage upon long periods of soil waterlogging. The promoter analysis of upregulated genes indicated that the production of oxalacetate from Asp via Asp oxidase, energy processes such as acetyl-CoA, ATP, and starch biosynthesis, and the lignification process were likely subjected to ABA regulation. Taken together, these data indicate that ABA depletion in waterlogged tissues acts as a positive signal, inducing several specific genetic and metabolic responses to soil flooding.

Involvement of abscisic acid and other stress indicators in taro (Colocasia esculenta (L.) Schott) response to drought conditions

Taro (Colocasia esculenta (L.) Schott) is a staple food and represents an important food security role in most tropical regions. It is, unfortunately, susceptible to prolonged drought conditions. Abscisic acid (ABA) is a well-documented stress-induced phytohormone that tolerant crops usually accumulate in leaves to induce stomatal closure, preventing water loss through inhibition of transpiration. Hitherto, exists very scarce information regarding the ABA role in taro response to drought. Here, we determined the ABA content in the shoots and corms of taro subjected to seven months of water scarcity and linked ABA to other drought resilience traits, including carbon isotopic discrimination (Δ13C), oxalic acid (OA), chlorophyll content index (CCI), water use efficiency (WUE), and biomass (B). The Δ13C-shoot content showed partially open stomata in all accessions, and significant correlation with Δ13C-corm, CCI, and WUE. The osmotically active OA-shoot decrease seemed not to interfere with the stomatal aperture. The tolerant accessions subjected to drought stress had higher B-corm, ABA-shoot, Δ13C-shoot, CCI, OA, and WUE. However, the observed under drought conditions increase of ABA in the shoots, and its decrease in the corms were not significantly correlated, nor with other traits, suggesting that ABA was not the main regulator of taro physiological processes under stress. The information gained should be considered in breeding programs to predict taro’s response to climate change.

Investigation of physiological and molecular mechanisms conferring diurnal variation in auxinic herbicide efficacy.

The efficacy of auxinic herbicides, a valuable weed control tool for growers worldwide, has been shown to vary with the time of day in which applications are made. However, little is known about the mechanisms causing this phenomenon. Investigating the differential in planta behavior of these herbicides across different times of application may grant an ability to advise which properties of auxinic herbicides are desirable when applications must be made around the clock. Radiolabeled herbicide experiments demonstrated a likely increase in ATP-binding cassette subfamily B (ABCB)-mediated 2,4-D and dicamba transport in Palmer amaranth (Amaranthus palmeri S. Watson) at simulated dawn compared to mid-day, as dose response models indicated that many orders of magnitude higher concentrations of N-1-naphthylphthalamic acid (NPA) and verapamil, respectively, are required to inhibit translocation by 50% at simulated sunrise compared to mid-day. Gas chromatographic analysis displayed that ethylene evolution in A. palmeri was higher when dicamba was applied during mid-day compared to sunrise. Furthermore, it was found that inhibition of translocation via 2,3,5-triiodobenzoic acid (TIBA) resulted in an increased amount of 2,4-D-induced ethylene evolution at sunrise, and the inhibition of dicamba translocation via NPA reversed the difference in ethylene evolution across time of application. Dawn applications of these herbicides were associated with increased expression of a putative 9-cis-epoxycarotenoid dioxygenase biosynthesis gene NCED1, while there was a notable lack of trends observed across times of day and across herbicides with ACS1, encoding 1-aminocyclopropane-1-carboxylic acid synthase. Overall, this research indicates that translocation is differentially regulated via specific protein-level mechanisms across times of application, and that ethylene release, a chief phytotoxic process involved in the response to auxinic herbicides, is related to translocation. Furthermore, transcriptional regulation of abscisic acid involvement in phytotoxicity and/or translocation are suggested.

Stomatal closure response to soil drying at different vapor pressure deficit conditions in maize

A plant transpiration rate under progressive soil drying remains constant until a threshold fraction of transpirable soil water (FTSW) is reached, and subsequently decreases linearly. The sensitivity of this function and the involvement of abscisic acid (ABA) and aquaporins in such responses have not been compared at various levels of atmospheric evaporative demand conditions. This study was conducted in controlled environment chambers with a drought-tolerant maize hybrid imposing progressive drought stress under three levels of vapor pressure deficit (VPD- 1.2, 2.3, and 3.5 kPa). A shift in threshold-FTSW from 1.2 kPa (FTSW-0.42) VPD to 3.5 kPa(FTSW-0.51) VPD was observed, showing an effect of VPD on stomatal closure response under soil drought conditions. Foliar ABA showed a substantial rise approximately at the same time as of stomatal closure initiated (FTSW-threshold), indicating ABA involvement. As the drought progressed, an increase in plasma membrane intrinsic protein and a decrease in tonoplast intrinsic protein expression levels were observed. Overall, this study suggests the influence of evaporative demand on the initiation of stomatal closure of drought-tolerant maize subjected to soil drying. The sensitivity of stomatal closure was associated with foliar ABA under drought stress but not under high evaporative demand conditions, indicating alternative water conservative mechanisms.

Gene Expression Analysis and Metabolite Profiling of Silymarin Biosynthesis during Milk Thistle (Silybum marianum (L.) Gaertn.) Fruit Ripening.

Mature fruits (i.e., achenes) of milk thistle (Silybum marianum (L.) Gaertn., Asteraceae) accumulate high amounts of silymarin (SILM), a complex mixture of bioactive flavonolignans deriving from taxifolin. Their biological activities in relation with human health promotion and disease prevention are well described. However, the conditions of their biosynthesis in planta are still obscure. To fill this gap, fruit development stages were first precisely defined to study the accumulation kinetics of SILM constituents during fruit ripening. The accumulation profiles of the SILM components during fruit maturation were determined using the LC-MS analysis of these defined developmental phases. The kinetics of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and peroxidase (POX) activities suggest in situ biosynthesis of SILM from l-Phenylalanine during fruit maturation rather than a transport of precursors to the achene. In particular, in contrast to laccase activity, POX activity was associated with the accumulation of silymarin, thus indicating a possible preferential involvement of peroxidase(s) in the oxidative coupling step leading to flavonolignans. Reference genes have been identified, selected and validated to allow accurate gene expression profiling of candidate biosynthetic genes (PAL, CAD, CHS, F3H, F3’H and POX) related to SILM accumulation. Gene expression profiles were correlated with SILM accumulation kinetic and preferential location in pericarp during S. marianum fruit maturation, reaching maximum biosynthesis when desiccation occurs, thus reinforcing the hypothesis of an in situ biosynthesis. This observation led us to consider the involvement of abscisic acid (ABA), a key phytohormone in the control of fruit ripening process. ABA accumulation timing and location during milk thistle fruit ripening appeared in line with a potential regulation of the SLIM accumulation. A possible transcriptional regulation of SILM biosynthesis by ABA was supported by the presence of ABA-responsive cis-acting elements in the promoter regions of the SILM biosynthetic genes studied. These results pave the way for a better understanding of the biosynthetic regulation of SILM during the maturation of S. marianum fruit and offer important insights to better control the production of these medicinally important compounds.

Involvement of abscisic acid, ABI5, and PPC2 in plant acclimation to low CO2.

Phosphoenolpyruvate carboxylase (PEPC) plays a pivotal role in the photosynthetic CO2 fixation of C4 plants. However, the functions of PEPCs in C3 plants are less well characterized, particularly in relation to low atmospheric CO2 levels. Of the four genes encoding PEPC in Arabidopsis, PPC2 is considered as the major leaf PEPC gene. Here we show that the ppc2 mutants suffered a growth arrest when transferred to low atmospheric CO2 conditions, together with decreases in the maximum efficiency of PSII (Fv/Fm) and lower levels of leaf abscisic acid (ABA) and carbohydrates. The application of sucrose, malate, or ABA greatly rescued the growth of ppc2 lines under low CO2 conditions. Metabolite profiling analysis revealed that the levels of glycine and serine were increased in ppc2 leaves, while the abundance of photosynthetic metabolites was decreased under these conditions. The transcript levels of encoding enzymes involved in glycine or serine metabolism was decreased in ppc2 in an ABI5-dependent manner. Like the ppc2 mutants, abi5-1 mutants had lower photosynthetic rates and Fv/Fm compared with the wild type under photorespiratory conditions (i.e. low CO2 availability). However, the growth of these mutants was similar to that of the wild type under non-photorespiratory (low O2) conditions. The constitutive expression of ABI5 prevented the growth arrest of ppc2 lines under low CO2 conditions. These findings demonstrate that PPC2 plays an important role in the acclimation of Arabidopsis plants to low CO2 availability by linking photorespiratory metabolism to primary metabolism, and that this is mediated, at least in part, through ABA- and ABI5-dependent processes.

Abscisic acid induced cellular responses of sub1A QTL to aluminium toxicity in rice (Oryza sativa L.)

Involvement of abscisic acid (ABA) was studied for aluminium (Al) sensitivity through functioning of sub1A quantitative trait loci in rice cultivars. sub1A quantitative trait loci bearing cv. Swarna Sub1 was found almost compatible with non sub1A quantitative trait loci bearing cv. Swarna for abscisic acid accumulation all through the aluminium concentrations. However, abscisic acid was self inductive by over expression of its biosynthetic gene in nine-cis-epoxycarotenoid dioxygenase 3 (NCED3) more in cv. Swarna than other. The effect of abscisic acid pretreatment was variable for specific leaf weight, leaf mass ratio and others for the cultivars. Bio-accumulation of aluminium had revealed the sensitivity of toxicity more in cv. Swarna than other. In connection to oxidative stress generation of reactive oxygen species was detected by both histochemical and in vitro assays through hematoxylin, Evans blue and schiff's reactions. Abscisic acid pretreatment had significantly reversed the effects of aluminium toxicity for reactive oxygen species generation. Regardless of varieties sensitivity of aluminium was more prone in shoot than root as detected by nitro blue tetrazolium and 3,3′-diaminobenzidine mediated signalling. Activity in metal chelation in extra cellular spaces monitored through esterase activity and that also established independence of abscisic acid pretreatment for cv. Swarna Sub1. The specific polymorphism of esterase at protein level strengthened the bio-monitoring of aluminium through the rice varieties as well its modulation with abscisic acid. Abscisic acid has been discussed an important effectors to modulate the tolerance pathway of rice cultivars through intrusion of sub1A quantitative trait loci.

Involvement of abscisic acid in the resistance of citrus fruit to Penicillium digitatum infection

The involvement of hormone abscisic acid (ABA) in citrus fruit resistance to Penicillium digitatum infection was investigated. To this end, the effect of both exogenous ABA and inhibitors of ABA biosynthesis in plants on the resistance of sweet oranges to be infected by this major postharvest pathogen, and also on in vitro fungal growth, was examined. The ability of both P. digitatum to infect wild-type Navelate orange fruit (Citrus sinesis (L.) Osbeck) and its spontaneous ABA-deficient mutant Pinalate was compared. The results showed that the Pinalate orange was more susceptible than its parental to infection, and exogenous ABA reduced the percentage of infected fruits and also the maceration zone of this mutant without affecting in vitro fungal growth. The global results indicated that the ABA of the fruit plays a defensive role in citrus fruit against P. digitatum infection. This work demonstrates, for the first time, the ability of P. digitatum to produce ABA. The hormone decreased in fruit in early stages as a consequence of infection. Thereafter when tissue maceration became evident, ABA increased, and this rise seemed related to the P. digitatum-induced rise in ethylene production. This was suggested by the fact that ABA increased in Navelate orange, but not in its ABA-deficient mutant Pinalate in this infection stage, while exogenous ethylene barely increased ABA in the mutant. Finally, the sharp marked rise in free ABA in later stages was likely related to free ABA being released from conjugated ABA, and also to fungal ABA. However, fungal ABA was not a primary virulence factor in citrus fruit infection.