ABA Levels Research Articles

Combined effect of partial root drying and elevated atmospheric CO2 on the physiology and fruit quality of two genotypes of tomato plants with contrasting endogenous ABA levels

Elevated CO2 concentration (e[CO2]) could induce the decline of nutrient concentration in tomato fruit, although it enhances the yield. Alternate partial root drying (PRD) irrigation is a water-saving irrigation strategy, which may enhance tomato fruit quality including both organic and inorganic attributes. Yet, it remains unknown whether PRD could alleviate the negative impact of e[CO2] on the quality attributes of tomato fruit. To investigate that, two genotypes of tomato plants differing in their endogenous abscisic acid level, i.e. Ailsa Craig (wild type, WT) and its correspondent ABA-deficient mutant (flacca), were grown in ambient CO2 (a[CO2], 400 ppm) and e[CO2] (800 ppm) environment, respectively. And three irrigation regimes including full irrigation (FI), deficit irrigation (DI) and PRD were applied during anthesis to fruit ripping stages. The results showed that e[CO2] increased net photosynthetic rate, decreased stomatal conductance and transpiration rate. Tomato yield was more sensitive to irrigation regimes than [CO2], and the reduction of fruit yield caused by PRD was attenuated under e[CO2], especially in flacca. Both e[CO2] and PRD enhanced water use efficiency. The concentration of total soluble solid, total sugar and total acid, as well as fruit firmness were improved in PRD. A negative effect of e[CO2] on fruit mineral nutrition was seen only in flacca, and as expected, such impact was attenuated by the reduced irrigation regimes (DI and PRD). Collectively, it is suggested that the responses of tomato fruit quality to e[CO2] and reduced irrigation regimes are genotypic-dependent, where the endogenous ABA level of the plants plays an important role, and this must be considered in climate management in greenhouse tomato cultivation.

Source-Sink Manipulation Affects Accumulation of Zinc and Other Nutrient Elements in Wheat Grains.

To better understand the source–sink flow and its relationships with zinc (Zn) and other nutrients in wheat (Triticum aestivum L.) plants for biofortification and improving grain nutritional quality, the effects of reducing the photoassimilate source (through the flag leaf removal and spike shading) or sink (through the removal of all spikelets from one side of the spike, i.e., 50% spikelets removal) in the field of the accumulation of Zn and other nutrients in grains of two wheat cultivars (Jimai 22 and Jimai 44) were investigated at two soil Zn application levels. The kernel number per spike (KNPS), single panicle weight (SPW), thousand kernel weight (TKW), total grain weight (TGW) sampled, concentrations and yields of various nutrient elements including Zn, iron (Fe), manganese (Mn), copper (Cu), nitrogen (N), phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg), phytate phosphorus (phytate-P), phytic acid (PA) and phytohormones (ABA: abscisic acid, and the ethylene precursor ACC: 1-aminocylopropane-1-carboxylic acid), and carbon/N ratios were determined. Soil Zn application significantly increased the concentrations of grain Zn, N and K. Cultivars showing higher grain yields had lower grain protein and micronutrient nutritional quality. SPW, KNPS, TKW (with the exception of TKW in the removal of half of the spikelets), TGW, and nutrient yields in wheat grains were most severely reduced by half spikelet removal, secondly by spike shading, and slightly by flag leaf removal. Grain concentrations of Zn, N and Mg consistently showed negative correlations with SPW, KNPS and TGW, but positive correlations with TKW. There were general positive correlations among grain concentrations of Zn, Fe, Mn, Cu, N and Mg, and the bioavailability of Zn and Fe (estimated by molar ratios of PA/Zn, PA/Fe, PA × Ca/Zn, or PA × Ca/Fe). Although Zn and Fe concentrations were increased and Ca was decreased in treatments of half spikelet removal and spike shading, the treatments simultaneously increased PA and limited the increase in bioavailability of Zn and Fe. In general, different nutrient elements interact with each other and are affected to different degrees by source–sink manipulation. Elevated endogenous ABA levels and ABA/ACC ratios were associated with increased TKW and grain-filling of Zn, Mn, Ca and Mg, and inhibited K in wheat grains. However, the effects of ACC were diametrically opposite. These results provide a basis for wheat grain biofortification to alleviate human malnutrition.

Gold nanoparticles-mediated ratiometric fluorescence aptasensor for ultra-sensitive detection of Abscisic Acid

Herein, a novel ratiometric aptasensor based on carbon quantum dots@2-Methylimidazole zinc salt (CQDs@ZIF-8) and aptamer-functionalized gold nanoparticles (Apt-AuNPs) was developed for highly sensitive detection of ABA by fluorescence spectrometry. The CQDs@ZIF-8 nanomaterials displayed dual-emission properties at 490 nm and 657 nm with excitation at 420 nm were synthesized for the first time. ZIF-8 not only served as an anchor point for CQDs but also acted as a modulator to regulate fluorescence signals of CQDs. Interestingly, introduction of ZIF-8 changed the quenching properties of the AuNPs on CQDs. The AuNPs quenched the fluorescence of CQDs@ZIF-8 at 490 nm but not at the second peak of 657 nm. Few studies have been reported on the ineffectiveness of AuNPs in fluorescence quenching as far as we know. In this study, we found that incorporation of ABA triggered the aggregation of AuNPs due to the specific ABA-aptamer recognition and this changed the fluorescence intensity of the ratiometric probe (CQDs@ZIF-8@Apt-AuNPs). The proposed probe increased the sensitivity and selectivity of determining ABA levels in rice seeds in the range of 0.100–150 ng/mL with an LOD of 30.0 ng/L. Importantly, the method proposed here offers a new unique strategy for the construction of ratiometric probes and ultra-sensitive measurement of biomolecules.

Influence of Endogenous Plant Hormones on Physiological and Growth Attributes of Kinnow Mandarin Grafted on Nine Rootstocks

Citrus holds the key position in horticulture sector of Pakistan in terms of area and production. Kinnow is considered as the trademark of Pakistan’s citriculture industry. Cultivation experiment was conducted to evaluate the endogenous plant hormones, leaf gas exchange, photosynthetic pigments, vegetative performance, and yield of Kinnow mandarin grafted on nine rootstocks (Rough lemon, cox mandarin, Fraser hybrid, Troyer citrange, Cleopatra mandarin, Poncirus trifoliata, Benton, C-35, and Carrizo citrange). Maximum indole acetic acid (IAA), gibberellic acid (GA3), and zeatin (ZT) levels were recorded in scion and rootstock of Poncirus trifoliata followed by Fraser hybrid. Troyer citrange rootstock depicted maximum amount of abscisic acid in scion and rootstock. Maximum leaf gas exchange in Kinnow mandarin was recorded on Fraser hybrid rootstock. Kinnow grafted on Troyer citrange rootstock had minimum stomatal conductance and photosynthetic activity because of high ABA level. Photosynthetic pigments of Kinnow mandarin were maximum on Poncirus trifoliata and minimum on Troyer citrange. A positive correlation was observed among the levels of IAA, GA, ZT, and vegetative growth of Kinnow mandarin. It is concluded that endogenous hormones modulated the growth of Kinnow mandarin. Poncirus trifoliata and Fraser hybrid rootstocks could be an alternate rootstocks of traditional Rough lemon for Kinnow mandarin.

Morphogenesis, pigment content, phytohormones and productivity of sweet pepper under the action of gibberellin and tebuconazole

One of the main tasks of modern plant physiology is regulation of growth and development of cultivated plants in order to optimize the productive process. The attention of the scientific community is focused on the use of natural activators and growth inhibitors. We investigated the effect of foliar treatment with 0.005% solution of gibberellic acid (GA3) and 0.025% solution of the antigibberellic preparation tebuconazole (EW-250) on morphogenesis, leaf mesostructure, the content of photosynthetic pigments, the balance of endogenous phytohormones and productivity of Capsicum annuum L., Antey variety. The vegetation experiment was carried out in the conditions of soil-sand culture in vessels with a volume of 10 L. Treatment of plants was carried out in the budding phase. Morphometric parameters were determined every 10 days. The mesostructure of the middle tier leaves was studied in the fruit formation phase, and the chlorophyll content was determined in the raw material by spectrophotometric method. Analytical determination of endogenous phytohormones – indolyl-3-acetic (IAA), gibberellic (GA3) and abscisic (ABA) acids and cytokinins – zeatin (Z), zeatin-O-glucoside (ZG), zeatinribozide (ZR), isopentenyladenine (iP) and isopentenyladenosine (iPA) were performed by high performance liquid chromatography – mass spectrometry (HPLC-MS). With GA3 treatment, plant height increased considerably, while with EW-250, it decreased. Both regulators led to an increase in the number of leaves on the plant, the leaf raw biomass, stems and roots and the dry matter of the whole plant, the area of a single leaf blade and the total area of leaves on the plant. Under the action of EW-250, the chlorophyll content in the leaves surged, while under the action of GA3 it tended to decrease or did not change at all. Both regulators thickened the chlorenchyma and boosted the volume of the columnar parenchyma cells. GA3 treatment induced a rise in the thickness of the upper and lower epidermis, and EW-250 led, on the contrary, to a decrease. It is shown that after treatment with exogenous GA3, the content of endogenous IAA and ABA decreased and GA3 in plant stems increased. Instead, EW-250 caused a decrease in the levels of GA3, IAA and ABA in the stems. Exogenous GA3 enhanced the accumulation of endogenous GA3 and IAA and inhibited ABA in the leaves. Under the action of the retardant, the level of ABA in the leaves did not change, while GA3 and IAA decreased. Treatment of plants with the studied growth regulators caused a decrease in the pool of cytokinins (CK) in stems. EW-250 showed a significant rise in the hormone content in the leaves. After spraying with GA3 solution, the level of ZG, Z and ZR grew. Under the action of the retardant, the increase in the CK pool occurred exclusively due to the iP. Growth regulators optimized the productivity of sweet pepper plants: under the action of GA3 there was an increase in the number of fruits per plant, and after the use of EW-250 there was a rise in the average weight of one fruit. The obtained results showed that anatomical-morphological and structural-functional rearrangements of sweet pepper plants under the action of exogenous gibberellic acid and EW-250 took place against the background of changes in the balance and distribution of endogenous hormones. Increased photosynthetic activity, stimulation of growth processes of some plant organs and inhibition of others enlarged biological productivity of the culture.

Halotolerant bacteria mitigate the effects of salinity stress on soybean growth by regulating secondary metabolites and molecular responses

BackgroundSalinity is a major threat to the agriculture industry due to the negative impact of salinity stress on crop productivity. In the present study, we isolated rhizobacteria and evaluated their capacities to promote crop growth under salt stress conditions.ResultsWe isolated rhizospheric bacteria from sand dune flora of Pohang beach, Korea, and screened them for plant growth-promoting (PGP) traits. Among 55 bacterial isolates, 14 produced indole-3-acetic acid (IAA), 10 produced siderophores, and 12 produced extracellular polymeric and phosphate solubilization. Based on these PGP traits, we selected 11 isolates to assess for salinity tolerance. Among them, ALT29 and ALT43 showed the highest tolerance to salinity stress. Next, we tested the culture filtrate of isolates ALT29 and ALT43 for IAA and organic acids to confirm the presence of these PGP products. To investigate the effects of ALT29 and ALT43 on salt tolerance in soybean, we grew seedlings in 0 mM, 80 mM, 160 mM, and 240 mM NaCl treatments, inoculating half with the bacterial isolates. Inoculation with ALT29 and ALT43 significantly increased shoot length (13%), root length (21%), shoot fresh and dry weight (44 and 35%), root fresh and dry weight (9%), chlorophyll content (16–24%), Chl a (8–43%), Chl b (13–46%), and carotenoid (14–39%) content of soybean grown under salt stress. Inoculation with ALT29 and ALT43 also significantly decreased endogenous ABA levels (0.77-fold) and increased endogenous SA contents (6–16%), increased total protein (10–20%) and glutathione contents, and reduced lipid peroxidation (0.8–5-fold), superoxide anion (21–68%), peroxidase (12.14–17.97%), and polyphenol oxidase (11.76–27.06%) contents in soybean under salinity stress. In addition, soybean treated with ALT29 and ALT43 exhibited higher K+ uptake (9.34–67.03%) and reduced Na+ content (2–4.5-fold). Genes involved in salt tolerance, GmFLD19 and GmNARK, were upregulated under NaCl stress; however, significant decreases in GmFLD19 (3–12-fold) and GmNARK (1.8–3.7-fold) expression were observed in bacterial inoculated plants.ConclusionIn conclusion, bacterial isolates ALT29 and ALT43 can mitigate salinity stress and increase plant growth, providing an eco-friendly approach for addressing saline conditions in agricultural production systems.

Methane-rich water induces bulblet formation of scale cuttings in Lilium davidii var. unicolor by regulating the signal transduction of phytohormones and their levels.

Previous studies have shown that methane (CH4 ) has promoting roles in the adventitious root (AR) and lateral root (LR) formation in plants. However, whether CH4 could trigger the bulblet formation in scale cutting of Lilium davidii var. unicolor has not been elucidated. To gain insight into the effect of CH4 on the bulblet formation, different concentrations (1, 10, 50, and 100%) of methane-rich water (MRW) and distilled water were applied to treat the scale cuttings of Lilium. We observed that treatment with 100% MRW obviously induced the bulblet formation in scale cuttings. To explore the mechanism of CH4 -induced bulblet formation, the transcriptome of scales was analyzed. A total of 2078 differentially expressed genes (DEGs) were identified. The DEGs were classified into different metabolic pathways, especially phenylpropanoid biosynthesis, starch and sucrose metabolism, and plant signal transduction. Of these, approximately 38 candidate DEGs involved in the plant signal transduction were further studied. In addition, the expression of AP2-ERF/ERF, WRKY, GRAS, ARF, and NAC transcription factors (TFs) was changed by MRW treatment, suggesting their potential involvement in bulblet formation. As for hormones, exogenous IAA, GA, and ABA could induce the bulblet formation. Additional experiments suggested that MRW could increase the endogenous IAA, GA, and JA levels, but decrease the levels of ABA during bulblet formation, which showed that higher IAA, GA, JA levels and lower ABA content might facilitate bulblet formation. In addition, the levels of endogenous hormones were consistent with the expression level of genes involved in phytohormone signal transduction. Overall, this study has revealed that CH4 might improve the bulblet formation of cutting scales in Lilium by regulating the expression of genes related to phytohormone signal transduction and TFs, as well as by changing the endogenous hormone levels.

Effect of 4-chloro-2-methylphenoxy acetic acid on tomato gene expression and rhizosphere bacterial communities under inoculation with phosphate-solubilizing bacteria

The herbicide 4-chloro-2-methylphenoxy acetic acid (MCPA) is widely used to control the spread of broad-leaved weeds in agricultural soils, though it remains unclear how tomato plants cope with the phytotoxic effects of MCPA at the molecular level. In this study, RNA-seq and Illumina MiSeq were used to sequence bacterial communities in tomato rhizosphere soils treated with MCPA and the phosphate-solubilizing bacterial strain N3. The results showed that MCPA induced abnormal growth of lateral roots in tomato seedlings and reduced uptake of the nutrients N, P, and K as well as the hormone (ABA and GA3) levels. Inoculation with strain N3 increased nutrient uptake by roots and increased levels of the hormones ABA, ZEA, and JA in tomato seedlings and also increased the abundance of the phyla Proteobacteria and Gemmatimonadetes in soil under MCPA treatment. GO functional groups in which differentially expressed genes (DEGs) are involved included DNA binding transcription factor activity, transcriptional regulator activity, enzyme inhibitor activity, and cell wall biogenesis. The highest numbers of DEGs are annotated to ribosome, photosynthesis, and carbon metabolism categories. Our findings provide valuable information for the application of strain N3, which is beneficial for reducing the toxic effect of MCPA on vegetable plants.

Phytohormone and integrated mRNA and miRNA transcriptome analyses and differentiation of male between hermaphroditic floral buds of andromonoecious Diospyros kaki Thunb

BackgroundPersimmon (Diospyros kaki Thunb.) has various labile sex types, and studying its sex differentiation can improve breeding efficiency. However, studies on sexual regulation patterns in persimmon have focused mainly on monoecy and dioecy, whereas little research has been published on andromonoecy. In order to reveal the sex differentiation regulation mechanism of andromonoecious persimmon, we performed histological and cytological observations, evaluated OGI and MeGI expression and conducted phytohormones assays and mRNA and small RNA transcriptome analyses of the male and hermaphroditic floral buds of the andromonoecious persimmon ‘Longyanyeshi 1’.ResultsStages 2 and 4 were identified as the critical morphological periods for sex differentiation of ‘Longyanyeshi 1’ by histological and cytological observation. At both stages, OGI was differentially expressed in male and hermaphroditic buds, but MeGI was not. This was different from their expressions in dioecious and monoecious persimmons. Meantime, the results of phytohormones assays showed that high IAA, ABA, GA3, and JA levels at stage 2 may have promoted male floral bud differentiation. However, high JA levels at stage 4 and high ZT levels at stages 2 and 4 may have promoted hermaphroditic floral bud differentiation. In these phytohormone biosynthesis and signaling pathways, 52 and 54 differential expression genes (including Aux/IAA, ARFs, DELLA, AHP, A-ARR, B-ARR, CYP735A, CRE1, PP2C, JAZ, MYC2, COI1, CTR1, SIMKK, ACO, and MPK6) were identified, respectively. During the development of male floral buds, five metacaspases genes may have been involved in pistil abortion. In addition, MYB, FAR1, bHLH, WRKY, and MADS transcription factors might play important roles in persimmon floral bud sex differentiation. Noteworthy, miR169v_1, miR169e_3, miR319_1, and miR319 were predicted to contribute to phytohormone biosynthesis and signaling pathways and floral organogenesis and may also regulate floral bud sex differentiation.ConclusionThe present study revealed the differences in morphology and phytohormones content between male and hermaphroditic floral buds of ‘Longyanyeshi 1’ during the process of sex differentiation, and identified a subset of candidate genes and miRNAs putatively associated with its sex differentiation. These findings can provide a foundation for molecular regulatory mechanism researching on andromonoecious persimmon.

The Effects of Different Planting Densities and Pruning Methods on Changes of Endogenous Hormone Levels in Shoot Tips and Flowering in ‘Gemlik’ Olive Cultivar

Olive cultivation tends to adopt new planting density and pruning systems in domestic olive types in Turkey. The effect of different planting densities and pruning types on the changes of some endogenous plant hormones in shoot tips and flowering in ‘Gemlik’ olive cultivar was studied. The two-year-old seedlings were planted at 5 × 5 m, 4 × 3 m and 4 × 1.5 m densities, and were pruned as either open vase or vertical axe styles in addition to unpruned control. The shoots were taken monthly during the two successive years for hormone analysis. The levels of ABA, IAA, GA3 and Z were determined in the shoot tips. GA3 was determined to be the highest on each month whereas Z was the lowest in shoot tip samples at most of the sampling dates. Pruning types affected ABA, GA3 and IAA, but not Z levels. Planting densities had significant effect on endogenous ABA and GA3 content. There were important differences among monthly levels of ABA, GA3, IAA, and Z. In the first year, IAA and Z content in shoot tips were higher than that of in the second year, whereas ABA level was lower in the first year. GA3 was not changed in both years. Results show that planting density, pruning style, and monthly and yearly changes in endogenous level of hormones in shoot tips play significant effect on phenological development and flowering in ‘Gemlik olive’ cultivar. High planting density reduced juvenility period and induced flower formation in olive.

Removal of superior wheat kernels promotes filling of inferior kernels by changing carbohydrate metabolism and sink strength

Poor filling and low weight of inferior kernels limit the further improvement of wheat yield. Two cultivars, Shuangda 1 and Xinong 538, with different grain weights, were selected to investigate the physiological changes of inferior kernels by removal of superior kernels (RS) at the flowering stage. iTRAQ combined with physiological indexes was used to identify factors limiting the filling of inferior kernels. Removal of superior kernels significantly increased the mean filling rate of inferior kernels and thus increased their weight. A set of 6012 proteins in inferior wheat kernels were differentially expressed between the RS and control. These differentially expressed proteins were involved mainly in carbon metabolism and energy metabolism. The main reason for the promoting effect of RS on the filling of inferior kernels may be that RS downregulated proteins involved in glycolysis and pyruvate metabolism while upregulating proteins involved in carbon fixation and photosynthesis. Consequently, RS greatly increased the ATP content in inferior kernels, supplying energy for them to absorb photosynthetic assimilates. Removal of superior kernels increased the activities of sucrose synthase, soluble starch synthase, adenosine diphosphate glucose pyrophosphorylase, and starch branching enzyme in inferior kernels and promoted starch accumulation in them. Thus, RS promoted the filling of inferior kernels and increased their weight. The promoting effect of RS on starch synthesis in inferior kernels was associated with their endogenous IAA and ABA levels.

Biochemical differences in the skin of two blueberries (Vaccinium corymbosum) varieties with contrasting firmness: Implication of ions, metabolites and cell wall related proteins in two developmental stages

The pursuit of firmer and better-quality blueberries is a continuous task that aims at a more profitable production. To this end it is essential to understand the biological processes linked to fruit firmness, which may diverge among tissues. By contrasting varieties with opposing firmness, we were able to elucidate events that, taking place at immature stage, lay the foundation to produce a firmer ripe fruit. A deep analysis of blueberry skin was carried out, involving diverse comparative approaches including proteomics and metabolomics coupled to immunolocalization assays. In‘O'Neal’ (low firmness) enhanced levels of aquaporins, expansins and pectin esterases at the green stage were found to be critical in distinguishing it from ‘Emerald’ (high firmness). The latter featured higher levels of ABA, low methyl esterified pectins in tricellular junctions and high levels of catechin at this stage. Meanwhile, in ‘Emerald’ ‘s ripe fruit epicarp, several mechanisms of cell wall reinforcement such as calcium and probably boron bridges, appear to be more prominent than in ‘O'Neal’. This study highlights the importance of cell wall reorganization and structure, abundance of specific metabolites, water status, and hormonal signalling in connection to fruit firmness. These findings result particularly valuable in order to improve the fertilization procedures or in the search of molecular markers related with firmness.

Characterization of salicylic acid- and abscisic acid-mediated photosynthesis, Ca2+ and H2O2 accumulation in two distinct phases of drought stress intensity in Brassica napus

Drought modifies the endogenous hormonal status, which is involved in the regulation of stress responses and tolerance. This study aimed to characterize the drought-responsive hormone-mediated regulation of stress responses based on the time-course of drought intensity. Alterations in H2O2, Ca2+ and their signaling genes with decreasing leaf water potential (Ψw) were interpreted as being linked to the endogenous hormonal level. Drought responses in relation to SA and ABA balance represented two distinct phases. The early phase, during the first 6 days when the Ψw was greater than -0.78 MPa, was characterized by a rapid decrease in photosynthetic activity with a progressive accumulation of H2O2 and Ca2+ in the strong correlation with the increases in both SA and ABA level. The late phase, days 6–14, was distinguished by ABA-mediated stress symptomatic responses (e.g., the maintenance of the depressed photosynthesis with severe leaf wilting) with highly enhanced expression of ABA-related genes (NCED3 and MYC2) and concomitant depression in the SA level and SA-related genes (ICS1 and NPR1). The regression analysis revealed that both SA and ABA levels as being positively correlated with Ca2+ and H2O2 were closely related to the depression of photosynthesis for the early phase (days 0–6), whereas the correlations were significant only with ABA for the whole period of drought (days 0–14). These results indicate that SA-stimulated H2O2 accumulation and SA responses during the early drought phase are part of upstream H2O2-stimulated ABA accumulation, which causes ABA signaling and responses, leading to severe drought symptoms during the late phase.

OsJAZ9 is involved in water-deficit stress tolerance by regulating leaf width and stomatal density in rice

Drought stress poses a severe threat to grain yield in rice. Our previous report demonstrated the role of OsJAZ9 in potassium homeostasis by modulating Jasmonic Acid (JA) signalling. While both potassium (K) and JA are known to have an important role in drought stress response, JA's repressor, i.e., JAZs' role in drought stress, remains elusive. Here we report that OsJAZ9 plays a critical role in rice water-deficit stress tolerance via influencing JA and ABA signalling. Overexpression of OsJAZ9 led to the enhanced ABA and JA levels. Our data further revealed that exogenous JA application antagonises the ABA-mediated inhibition of seed germination. Further, OsJAZ9 overexpression reduces leaf width and stomata density, leading to lower leaf transpiration rates than WT. This reduced transpiration and higher K content as osmoticum improved the water-deficit stress tolerance in OsJAZ9 overexpression lines. On the contrary, OsJAZ9 RNAi lines displayed enhanced sensitivity towards water-deficit stress. Our data provide new insights on the role of JA signalling repressors in rice response to water-deficit stress.

Profiles of endogenous ABA, bioactive GAs, IAA and their metabolites in Medicago truncatula Gaertn. non-embryogenic and embryogenic tissues during induction phase in relation to somatic embryo formation

Main conclusionDuring the 3-week-long induction phase, when M. truncatula cells leaf explants from non-embryogenic genotype (M9) and embryogenic variant (M9-10a) were forming the callus, biosynthesis and degradation of ABA, Gas and IAA proceeded at different levels. Induction of embryo formation is related to a lower ABA content, compared to the content of IAA and that of total bioactive GAs.Endogenous phytohormones are involved in the regulation of zygotic embryogenesis, but their role, especially of ABA, a plant growth inhibitor, in inducing somatic embryogenesis (SE) in angiosperms is still incompletely known. To arrive a better understanding of the ABA role in the process, we analyzed simultaneously and in detail changes in the contents of both ABA and five bioactive GAs (GA4, GA7, GA1, GA3, GA6) and IAA in M. truncatula non-embryogenic M9 (NE) and embryogenic M9-10a (E) genotypes. The initial leaf explants of both genotypes, and particularly NE, contained many times more ABA compared to the total bioactive GAs or IAA. In tissues during the entire 21-day induction all the hormones mentioned and their metabolites or conjugates were present; however, their contents were found to differ between the lines tested. The ABA level in primary explants of NE genotype was more than two times higher than that in E genotype. An even larger difference in the ABA content was found on the last day (day 21) of the induction phase (IP); the ABA content in E callus was over six times lower than in NE callus. In contrast, the IAA and GAs contents in primary explants of both genotypes in relation to ABA were low, but the contents of IAA and GAs exceeded that of ABA in the M9-10a tissues on the last day of IP. It is shown for the first time that endogenous ABA together with endogenous bioactive GAs and IAA is involved in acquisition of embryogenic competence in Medicago truncatula leaf somatic cells. These findings have a strong functional implication as they allow to improve the SE induction protocol.

Plant-Specific AtS40.4 Acts as a Negative Regulator in Abscisic Acid Signaling During Seed Germination and Seedling Growth in Arabidopsis.

Abscisic acid (ABA) is an important phytohormone regulating plant growth, development and stress responses. A multitude of key factors implicated in ABA signaling have been identified; however, the regulation network of these factors needs for further information. AtS40.4, a plant-specific DUF584 domain-containing protein, was identified previously as a senescence regulator in Arabidopsis. In this study, our finding showed that AtS40.4 was negatively involved in ABA signaling during seed germination and early seedling growth. AtS40.4 was highly expressed in seeds and seedlings, and the expression level was promoted by ABA. AtS40.4 was localized both in the nucleus and the cytoplasm. Moreover, the subcellular localization pattern of AtS40.4 was affected by ABA. The knockdown mutants of AtS40.4 exhibited an increased sensitivity to ABA, whereas the overexpression of AtS40.4 decreased the ABA response during seed germination and seedling growth of Arabidopsis. Furthermore, AtS40.4 was involved in ABRE-dependent ABA signaling and influenced the expression levels of ABA INSENTIVE (ABI)1-5 and SnRK2.6. Further genetic evidence demonstrated that AtS40.4 functioned upstream of ABI4. These findings support the notion that AtS40.4 is a novel negative regulator of the ABA response network during seed germination and early seedling growth.

Comparative analysis of the gene expression profile under two cultivation methods reveals the critical role of ABA in grape quality promotion

Root restriction is a critical cultivation technique that accelerates ripening and improves fruit quality, by a still unknown mechanism. ABA is an important phytohormone in both ripening and stress responses. To elucidate the relationship between ABA content and root restriction-induced fruit quality promotion, the endogenous ABA level was investigated in five-year-old ‘Muscat Hamburg’ grapevines from the pre-anthesis stage to maturity. The results showed that ABA production was significantly enhanced by root restriction. A transcriptomics-based approach was taken to identify transcriptionally regulated genes associated with ABA biosynthesis, catabolism and signal transduction in the four development periods of grape. A total of 2873 (884 upregulated and 1989 downregulated) DEGs were identified in all stages. Those related to metabolic processes, cellular processes, responses to stimuli, biological regulation and signal transduction were abundant in the biological process GO category. KEGG analysis revealed that plant hormone signal transduction and carotenoid biosynthesis were enriched out of the 127 total pathways. 12 DEGs involved in ABA biosynthesis and catabolism and 69 DEGs involved in plant hormone signal transduction were significantly regulated by root restriction. Twelve DEGs related to the biosynthesis and catabolism of ABA or plant hormone signal transduction were selected to validate the data obtained by RNA-Seq, and the results showed that genes such as PP2C, NCED, AAO3, ABA3, and CYP707A1, involved in ABA biosynthesis and signal transduction, may play important roles in the promotion of fruit quality by root restriction. This study suggests a potential ABA-mediated mechanism by which root-restriction cultivation promotes fruit ripening.

Transcriptome analysis of Chongyi wild mandarin, a wild species more cold-tolerant than Poncirus trifoliata, reveals key pathways in response to cold

Much effort has been made on the dissection of cold response in citrus, but relevant study in wild germplasm has been rare. Based on the result of the membership function value analysis with 12 physiological indexes, we reported a wild citrus germplasm, Chongyi wild mandarin (Citrus reticulata), which was more cold-tolerant than a cold-hardy rootstock, Poncirus trifoliata. To gain an overview of transcriptome changes under cold, leaves of Chongyi wild mandarin were sampled at 0, 1, 5 d after 0 °C treatment, and sequenced using an Illumina sequencing platform. Totally, 3921 and 5995 differentially expressed genes (DEGs) were identified in 1 d and 5 d, respectively. And then, RNA-seq results were further validated via real-time quantitative PCR with 17 DEGs. Many DEGs related to various cold signaling pathways were identified, especially those related to brassinosteroid cascade. Around 10 % of the total DEGs were transcription factors from 67 families, suggesting that transcriptional regulation, both C-repeat/dehydration responsive element-binding factor-dependent/independent pathways, was crucial for the cold tolerance of Chongyi wild mandarin. Our data also revealed that the transcriptional levels of many DEGs related to anti-oxidant enzymes and osmoprotectants metabolism were altered, which were consistent with the results of physiological experiments. Moreover, a number of DEGs involved in ABA, BR, GA metabolisms were isolated. Accordingly, the levels of ABA and BR were significantly increased under cold, but GAs were decreased. Taken together, this work provided a comprehensive view of physiological and transcriptional alterations in cold response of Chongyi wild mandarin.

Dynamics of Water and Abscisic Acid During Embryogeny and Embryo Drying in the Recalcitrant Seeds of Vateria indica L.

Vateria indica L. is a critically endangered tree species in South-Western Ghats of India, commercially exploited for its valuable resins. Seed recalcitrance is a major problem hindering the natural regeneration of this species and it poses a great challenge in seed storage and conservation. There was a continuous import of water from the maternal tissues to seed tissues till maturity and the seeds were released in a fully hydrated state. Differential accumulation of water has been noticed in the cotyledons and embryonal axis. There was a positive correlation between seed moisture content and rate of germination which is a character of recalcitrant seeds. The critical moisture content was found to be 40% in the axis and 23.5% in the cotyledons, below which the embryo will not germinate. Loss of germination ability as a result of desiccation was attributed to the cell membrane damage, expressed as the electrolyte leakage exceeding 0.79 μS/cm. ABA peaked in the mid embryogenesis, then dropped drastically and maintained a lower level till seed maturity. On desiccation, ABA started to increase but gradually dropped down. Both cotyledons and embryonal axis had differential ABA content but exhibited a general pattern of ABA level during embryogeny. Due to the thin seed coat/embryo ratio and low investment in the seed coat, this recalcitrant seed could not hold water as efficient as orthodox seeds. Thus, it germinated as soon as it was shed from the mother plant. On desiccation, ABA shot up and moisture content decreased along with electrolyte leakage and cell membrane damage. All these hindered germination of the seed. Thus, we can see a clear interplay between moisture content and ABA levels during embryogeny and desiccation. Since the seed biology of this species has not been well documented, the present work is mainly intended to study the dynamics of water and ABA during embryogeny and embryo drying. This study can surely contribute to the long-term storage and conservation of recalcitrant seeds which is a less explored area.

Foliar applied 24-epibrassinolide alleviates salt stress in rice (Oryza sativa L.) by suppression of ABA levels and upregulation of secondary metabolites

ABSTRACT Salt stress hinders plant growth and disrupts many physio-biochemical and molecular processes. The present study evaluated the potential role of foliar applied 24-epibrassinolide (EBR) for mitigating the adverse effects of salinity in rice plants. Our findings showed that salt stress reduced several growth and physiological parameters, and substantially increased oxidative stress in rice seedlings. The foliar application of EBR replenished the negative effects of salt stress by improving plant growth and physiological attributes in rice plants. Moreover, application of EBR substantially reduced oxidative injuries by enhancing antioxidant enzymes and phenolic compounds in rice plants during salt stress. Interestingly, the endogenous ABA levels were declined in stressed plants by EBR treatment, also validated by expression analysis of ABA-related genes. In addition, application of EBR upregulated carotenoids and flavonoids pathway genes, thus showing its function to activate secondary metabolism in rice plants during salt stress. The principal component analysis (PCA) indicated that, under salt stress, the response of EBR-treated rice plants was strongly correlated with the accumulation of phenolic compounds and flavonoids. Collectively, this study presents useful insight into the distinct role of EBR by activating underlying enzymatic and non-enzymatic defense mechanisms to mitigate salinity in rice plants.