Abscisic acid (ABA) is a plant hormone well known to regulate abiotic stress responses. ABA is also recognised for its role in biotic defence, but there is currently a lack of consensus on whether it plays a positive or negative role. Here, we used supervised machine learning to analyse experimental observations on the defensive role of ABA to identify the most influential factors determining disease phenotypes. ABA concentration, plant age and pathogen lifestyle were identified as important modulators of defence behaviour in our computational predictions. We explored these predictions with new experiments in tomato, demonstrating that phenotypes after ABA treatment were indeed highly dependent on plant age and pathogen lifestyle. Integration of these new results into the statistical analysis refined the quantitative model of ABA influence, suggesting a framework for proposing and exploiting further research to make more progress on this complex question. Our approach provides a unifying road map to guide future studies involving the role of ABA in defence.

“Autism” refers to a wide variety of disabilities, with numerous possible clinical signs and heterogeneous origins. Nevertheless, beyond those diverse clinical signs, a consensus exists concerning the necessity of interventions, particularly educational interventions, for people with autism . In a society such as ours, turned towards digital technologies and tactile devices, (e.g. ), it is important to question the use of these technologies with people with developmental disabilities. Especially as some authors pointed the utility of these devices for teaching skills . Various theoretical corpuses and elements of the scientific literature were taken into account and integrated to develop the LearnEnjoy applications, created initially for children with autism. These facts and knowledge, linked to the peculiarities of people with autism, to the functional approach of language, to the fundamental principles of learning, to an ABA approach, were integrated from the first stages of the development of the applications, to create tools having solid scientific foundations. This way, the LearnEnjoy apps give the users (i.e. the “teachers”) the possibilities of teaching in a progressive and coherent way, different skills such as language (receptive, expressive), imitation, play and motricity, cognitive, academic skills or even independence skills. They also allow the progress in each area to be shared with the parents and the whole team, a necessary feature for the implementation of global and coordinated interventions. Finally, and maybe more importantly, these applications were created so as to specifically foster the contact between the person with autism and the “teacher”. This way the apps, at the same time, reduce the risk of pervasiveness of the tactile tablet, while favouring, just as much for the person with autism than for the accompanying person, the development of a positive, structured and structuring social relationship.

The understanding of the genetic basis of grain dormancy in wheat has rapidly improved in the last few years, and a number of genes have been identified related to that trait. We recently identified the wheat genes TaPM19-A1 and -A2 and we have now taken the first step towards understanding the role of this class of genes in seeds. By investigating the Arabidopsis homologous PM19-Like 1 (PM19L1) we have found that it has a seed-specific expression pattern and, while its expression is higher in dormant than in non-dormant seeds, knock-out mutations produced seeds with increased dormancy. Not only primary dormancy, but also secondary dormancy in response to high temperature was increased by the loss-of-function. We have also examined the function of PM19L1 by localizing the PM19 protein primarily to the cotyledon cells in seeds, possibly in membranes. By investigating the co-expression network of this gene we have found that it is connected to a small group of abscisic acid (ABA)-induced seed maturation and storage-related genes. The function of PM19L1 represents a good opportunity to explore the interactions of key factors that can influence seed dormancy such as ABA, temperature and membrane properties.

In Arabidopsis, seed germination is a biphasic process involving rupture of the seed coat followed by emergence of the radicle through the micropylar endosperm. Embryo expansion results in seed coat rupture and removal of seed coat imposed dormancy with DELLA proteins blocking embryo expansion in the absence of gibberellins. Exogenous abscisic acid (ABA) treatment does not block seed coat rupture but does block radicle emergence. We used this limited effect of exogenous ABA to further investigate the mechanism by which it blocks the onset of germination marked by seed coat rupture. We show that physical nicking of the seed coat results in exogenous ABA treatment blocking both seed coat and endosperm rupture and this block requires the transcription factors ABI3 and ABI5, but not ABI4. Furthermore, we show that the repression of expression of several EXPANSIN genes (EXPA1, EXPA2, EXPA3, EXPA9 and EXPA20) by exogenous ABA requires ABI5. We conclude that ABI5 plays an important role in the ABA-mediated repression of germination through prevention of seed coat rupture and propose that this involves EXPANSIN related control of cell wall loosening.

The AixMICADAS facility is in part dedicated to research on radiocarbon (14C) calibration by means of various archives. For this purpose, we are improving upon the capacity to accurately date subfossil wood. In the current study, nine chemical pretreatment protocols are tested on six wood samples of known ages. The optimization based on 14C ages, 13C/12C ratios, carbon % and overall mass yield % leads us to favor the acid-base-acid-bleaching pretreatment (ABA-B). This efficient method is shown to provide a residue of holocellulose with optimal blanks equivalent to an age of 51,300 14C BP with a standard deviation of 1500 yr based on 25 analyses. The seven wood samples from the Sixth International Radiocarbon Intercomparison (SIRI) are then analyzed as a further verification of the accuracy of our method. As a first scientific contribution, we studied two tree-ring sequences from subfossil pines (Barb12 and Barb17) collected in the southern French Alps. New 14C analyses were performed at high resolution (every third year) and are shown to agree well with results obtained previously by high precision β-counting on CO2 from large samples at lower resolution for Barb17 and accelerator mass spectrometry (AMS) data for Barb12. The new 14C series are then matched to the Kauri and YDB chronologies: the new sequence of Barb12-17 tentatively corresponds to the interval between 12,836 and 12,594 cal BP within the Younger Dryas cold period. The 14C comparison between the Barb12-17 sequence from France and the Kauri sequence from New Zealand allows calculating the 14C Inter-Hemispheric Gradient (IHG), with an average value of ca. 57 yr. The IHG stayed relatively high throughout the studied period. Interestingly, the IHG exhibits a transient maximum value (ca. 100 yr) during the period of rapid Δ14C rise (12,750–12,720 cal BP), a behavior that could be due to a delayed response of the Southern Hemisphere.

Induction of nine-cis-epoxycarotenoid dioxygenase 6 (NCED6), an abscisic acid (ABA) biosynthesis gene, alone is sufficient to suspend germination in testa-ruptured seeds, which are at the final step of germination. Molecular consequences of NCED6 induction in imbibed seeds were investigated by RNA sequencing. The analysis identified many unknown and uncharacterized genes that were up-regulated by NCED6 induction, in addition to the major regulators of ABA signalling. Interestingly, other NCEDs were up-regulated by NCED6 induction, suggesting that the major rate-limiting enzymes in the ABA biosynthesis pathway are subject to positive-feedback regulation. ZEAXANTHIN EPOXIDASE and ABSCISIC ALDEHYDE OXIDASE3, which function upstream and downstream of NCED, were also up-regulated in seeds by NCED6 induction, which suggests that the distinct layers of positive feedback loops are coordinately operating in the NCED6-induced seeds. SOMNUS (SOM), which was also up-regulated by NCED6 induction, was the major mediator of enhanced ABA signalling in NCED6-induced seeds. SOM exerted negative effects on GA biosynthesis, which also contributes to a high ABA–GA ratio and reinforces the suppressive state of germination. Besides these coding genes, long intergenic non-coding RNAs (lincRNAs) were also up-regulated upon NCED6 induction (termed N6LINCRs). Conditional expression of N6LINCR1 altered gene expression profiles in seeds. Twenty-six genes were up-regulated and 66 genes were down-regulated by the induction of N6LINCR1. These results suggest that some of N6LINCRs have a regulatory role in gene expression in seeds, which potentially contributes to the regulation of germination by ABA.

Among domesticated traits, pre-harvest sprouting (PHS) caused by the early breakage of dormancy leads to severe economic losses. Therefore, regulating PHS is important for cereal crop improvement against changes in climate. In this study, we surveyed naturally occurring variations in seed germination in diverse rice germplasm for the available resources of this trait, and investigated the changes of abscisic acid (ABA) levels during grain development by the distinguished PHS-resistant groups. We discovered wide variations in germination among the 205 rice accessions examined and found that 90 accessions are resistant (germination <20%) to PHS. Tropical and subtropical accessions, which are subjected to long wet periods, are more resistant to PHS than the other accessions. We detected an increase in germination of detached seeds from the panicle compared with intact seeds in panicle at harvesting time. This might be attributed to a weakening of the mechanical barrier that prevents water imbibition and radical emergence. ABA levels were maximal at 10 d after flowering and decreased thereafter. Interestingly, PHS-susceptible accessions maintained higher or similar ABA levels compared with PHS-resistant accessions, suggesting that the key factors for seed dormancy and its breakage are ABA perception and signal transduction rather than total ABA content. The diversity of germination ability detected in this study could be sustainably used for crop improvement and to help unveil the genetic and physiological basis of this quantitative trait.

Charcoal is widely used for radiocarbon dating in archaeological and paleoenvironmental studies. Reliable 14C dating requires appropriate chemical treatment to remove postdeposition contamination from the charcoal samples. This study assesses two pretreatments: acid-base-acid (ABA) and acid-base-oxidation with stepped combustion (ABOx-SC). In addition to 14C, the effects of the treatments on the chemical structure and composition of charcoal were studied using Fourier transform infrared spectroscopy (FTIR) and C/H/O elemental analysis. Samples of pine wood charred in the laboratory at 270, 300, 400, 500, and 600°C, and environmental samples of charred pine wood from pyroclastic flow deposits in southern Kyushu, Japan, were tested. The laboratory-charred samples showed that NaOH treatment removed highly hydrophilic organic components derived from endogenous and exogenous organic materials in the samples and that oxidation treatment caused the oxidative degradation of molecules in samples starting from its edges. The ABA-treated environmental charcoal yielded younger 14C dates than the ABOx-treated samples, probably owing to the effects of remaining organic contaminants bound to the edges of the aromatic molecular structures produced by the original pyrolysis. Meanwhile, it was found that ABA-SC treatment can reduce contaminants as effectively as ABOx-SC treatment. This implies that the stepped combustion (SC), not the chemical oxidation, is the key to reduce contaminant residue left after ABA and ABOx treatments. The results in this study indicate that the investigation of the structural and compositional changes of charcoal during its pretreatment is useful for assessment of the reliability of the 14C ages.

Physiological dormancy has been described as a physiological inhibiting mechanism that prevents radicle emergence. It can be caused by the embryo (embryo dormancy) as well as by the structures that cover the embryo. One of its functions is to time plant growth and reproduction to the most optimal season and therefore, in nature, dormancy is an important adaptive trait that is under selective pressure. Dormancy is a complex trait that is affected by many loci, as well as by an intricate web of plant hormone interactions. Moreover, it is strongly affected by a multitude of environmental factors. Its induction, maintenance, cycling and loss come down to the central paradigm, which is the balance between two key hormonal regulators, i.e. the plant hormone abscisic acid (ABA), which is required for dormancy induction, and gibberellins (GA), which are required for germination. In this review we will summarize recent developments in dormancy research (mainly) in the model plant Arabidopsis thaliana, focusing on two key players for dormancy induction, i.e. the plant hormone ABA and the DELAY OF GERMINATION 1 (DOG1) gene. We will address the role of ABA and DOG1 in relation to various aspects of seed dormancy, i.e. induction during seed maturation, loss during dry seed afterripening, the rehydrated state (including dormancy cycling) and the switch to germination.

The endogenous ABA contents of dormant and nondormant barley grains were determined following application of different compounds to break dormancy. The chemicals used for breaking of dormancy in intact dormant grains were weak and strong acids, alcohols, hydrogen peroxide, cyanide, nitrate, salicylic acid, gibberellic acid and fusicoccin. The dormancy-breaking compounds could be classified into two major groups: compounds that caused a decrease in endogenous ABA (class I) and compounds which did not affect endogenous ABA (class II). Class I compounds included gibberellic acid, ethanol, hydrogen peroxide, nitrate, salicylic acid; class II compounds were fusicoccin, acid (H2SO4), sodium azide, n-caproic acid. In addition, these dormancy-breaking compounds were able to stimulate the germination rate when applied to embryos isolated from dormant grains. The concentrations necessary for stimulation of germination of isolated embryos were much lower than the concentrations for breaking the dormancy of intact grains. After embryos were isolated from dormant grains and incubated in water, ABA was determined in both embryos and in the incubation media. The class I compounds stated above also reduced ABA content in the incubation medium of isolated embryos, while class II compounds had no effect on ABA content of the medium. External application of ABA could overcome the effect of dormancy-breaking compounds of class I but not of class II. The results suggest that in the presence of the agents belonging to class II, ABA responsiveness of isolated embryos from dormant grains is decreased, compared to nontreated embryos.

The fruits of Quercus robur are shed containing seeds at high moisture contents which remain desiccation sensitive and exhibit recalcitrant storage behaviour. Little is known of the control of seed development in these and other recalcitrant seeds. In the present work the changing concentrations of four plant growth regulators during seed reserve accumulation was studied over five years on seeds from the same tree. The pattern of reserve accumulation and changing ABA content in seeds differed between years. Although ABA content in the cotyledons increased in line with increasing dry weight to different contents at shedding, similar concentrations existed in cotyledons in each year. Thus ABA did not appear to be influencing dry weight accumulation. However, unlike orthodox seeds the decline in ABA concentration prior to shedding was limited and consistent with a continuing role for ABA in preventing precocious germination. An earlier peak in ABA concentration was associated with greater desiccation tolerance at shedding across years. The concentrations of zeatin and zeatin riboside in cotyledons were similar in each year and declined during reserve accumulation in a similar fashion to that reported for orthodox seeds. By contrast, IAA concentration increased in both the cotyledons and axes in the latter stages of seed development, opposite to that reported for orthodox seeds. It is possible that the increasing IAA concentration in cotyledons and axes and the stable concentration of zeatin and zeatin riboside throughout the latter stages of development in the axes of Q. robur are linked to the maintenance of active metabolism for the rapid initiation of germination upon shedding observed in seeds of this species.

The contents of the cytokinins zeatin (Z), zeatin riboside (ZR), and isopentenyladenine (IPA), the combined contents of gibberellins1 + 3 (GA), and the contents of indoleacetic acid (IAA) and abscisic acid (ABA) were measured during the development of the desiccation-sensitive seeds of Avicennia marina (Forssk.) Vierh. During the stage of histodifferentiation the amounts of these plant growth regulators (PGRs) were measured on whole fruits. During the phase of seed growth and reserve accumulation measurements were made on the embryonic axis, cotyledons and pericarp separately. Patterns in the amounts of PGRs present during histodifferentiation were similar to those reported for desiccation-tolerant seeds and it suggested that this process is under similar ‘hormonal’ control in A. marina as in orthodox seeds. Very high contents of cytokinins, particularly ZR, were present in both axes and cotyledons during reserve accumulation. This is thought to be related to the nature of the reserves accumulated (soluble sugars), rather than to the phenomenon of desiccation sensitivity. With the exception of ABA, embryonic contents of PGRs were relatively high at seed shedding, consistent with the rapid germination of this highly recalcitrant seed. ABA contents in the embryo were low during reserve accumulation, but concentrations in the pericarp increased throughout this development stage. ABA in the perticap could act to prevent precocious germination. The low concentrations of ABA in the embryo could be related to the desiccation-sensitivity of the seeds of A. marina.

Plants undergo a series of physiological, biochemical and molecular changes in response to adverse environmental conditions or stresses such as drought, low temperature or high salt. Several genes and their corresponding proteins have been described that may play a role in withstanding water-deficit-related stresses or full desiccation. In particular, sugars and late-embryogenesis-abundant (LEA) proteins have received the most attention. Plant responses to water-deficit and desiccation have been well-characterized at the molecular level; however, pinpointing the precise roles of the gene products in protecting cells under conditions of water deficit remains a challenging task. While few plants are capable of withstanding full desiccation, most seeds undergo this event as a pre-programmed and final stage in their development. These are the so-called ‘orthodox’ seeds. In contrast to seeds of orthodox species, those of recalcitrant species do not acquire desiccation tolerance during their development and are shed from the parent plant at relatively high water contents. The essential components of desiccation tolerance of seeds are likely to involve the ability to effect repair upon subsequent rehydration as well as the ability to accumulate protective substances that limit the amount of damage which otherwise would be caused by water loss. Studies have begun to examine whether the desiccation sensitivity of recalcitrant seeds is at least partially the result of an insufficient accumulation of LEA-type proteins, or whether other factors (including a lack of protective sugars) are more important. This review assesses some of these studies as well as recent research to understand gene and protein function using transgenic host plant systems.

The development of techniques for somatic embryogenesis in conifers has led to rapid advances in the ability to culture conifer tissue in vitro. Somatic embryogenesis now provides a means to clonally propagate commercially valuable conifers of several species and offers an in vitro experimental system for studying embryogenesis. As a result of these developments, the conifer zygotic and somatic embryo system has recently attracted the attention not only of the cell biologist but also of biochemists and molecular biologists. In this review I have attempted to consolidate the recent information on ultrastructure, biochemical and molecular aspects of conifer zygotic and somatic embryogenesis. In the first section, salient features of zygotic embryogenesis are highlighted followed by biochemical and ultrastructural characterization of development with reference to storage reserve deposition. In the second section, detailed characterization of seed storage proteins, changes in gene expression with emphasis on storage protein genes and Lea genes during zygotic and somatic embryogenesis are described. Effect of culture treatments such as abscisic acid, osmoticum and desiccation on storage protein gene expression in somatic embryos is discussed. Based on these studies, comparisons are presented between angiosperm and gymnosperm embryogeny regarding the unique morphological and developmental patterns of conifer embryogenesis. Finally, a brief discussion of recent progress in biochemical and molecular characterization of conifer seed germination is presented.

Dormancy in rice (Oryza sativa L.) seed is imposed by certain physical and chemical factors associated with its covering structures, i.e.hull and pericarp. The nature of these germination blocks, their mode ofaction, and processes regulating the release of dormancy are not fully understood. Of nine rice cultivars studied, Ching-shi 15, Stejaree 45, PTB10, and Mahsuri are weakly dormant, and Bansphul, Benaful, Kataktara, Dular, and N22 are dormant. Release of seed dormancy in rice by various treatments, oxidative processes and enzymic changes associated with dormancy, and parallelism between natural and artificially imposed dormancy patterns were examined. The influence of the hull in imposing dormancy was stronger and more prolonged than that of the pericarp. Application of GA3 was effective in inducing germination only in weakly dormant cultivars. Dormancy was completely released in all cultivars by subjecting the seeds to moist heat treatment, by removing the hull and pericarp, and by applying GA3 after dehulling. Dormant cultivars had higher O2 uptake rate and peroxidase activity and lower amylase and dehydrogenase activities than the weakly dormant ones. Hull removal substantially decreased peroxidase activity but enhanced amylase and dehydrogenase activities. Nonanoic acid (C90), a short-chain saturated fatty acid (SCSFA), when exogenously applied to non-dormant seeds imposed dormancy. Dry heat treatment or presoaking in 0.01 m KNO3 or 0.1 m H2O2 was very effective in releasing SCSFA-imposed dormancy. Amylase activity was greatly reduced by treatments with nonanoic acid (C90) or ABA. Considering earlier reports and results of the present study, it is proposed that seed dormancy in rice is regulated both by the presence of SCSFAs and ABA in the hull and the pericarp. The relative significance of these substances in cultivars of tropical and temperate origins and its implications in terms of ecogeographic adaptability are discussed.

Seeds of Oryza sativa L. (desiccation tolerant, orthodox) completed each of five distinct stages of development approximately 7 days earlier than seeds of Porteresia coarctata (Roxb.) Tateoka (desiccation intolerant, recalcitrant), despite the fact that O. sativa plants matured under cooler conditions. Isolated embryos of O. sativa were more sensitive to rapid desiccation at 6 days post anthesis (DPA) compared with naked caryopses. More than 90% of the latter were desiccation tolerant at 8 DPA and at all stages tested the germination capacity and/or rate of germination was greater following drying. In contrast, drying resulted in a complete loss of viability in seeds of P. coarctata at all stages tested. In both species, abscisic acid (ABA) began to accumulate in embryos mid-way through development. At the respective peaks, the concentration of ABA in P. coarctata was twice that recorded in O. sativa confirming that ABA levels per se do not determine desiccation tolerance in seeds. The possibility that desiccation tolerance is linked to sensitivity to ABA is discussed.

Wheat which was grown in acid-washed sand and irrigated with a molybdenum-free nutrient solution was treated with various concentrations of molybdenum (Mo) as a foliar spray at the flag leaf stage. At maturity, dormancy levels and abscisic acid (ABA) content of the seed were determined. Seed dormancy and ABA content increased with increasing rates of Mo application. In a field experiment, wheat (cv. SST 66) was treated with 100 ppm Mo by foliar application at the flag leaf stage. Embryos were tested for sensitivity to exogenously applied ABA. Embryos from Mo-treated plants showed enhanced sensitivity to ABA-induced inhibition of germination. The Mo application also resulted in significantly higher levels of endogenous ABA and Mo in the seeds. It is postulated that Mo deficiency leads to a lack of dormancy in wheat via reduced synthesis of ABA.

The role of abscisic acid (ABA) in seed germination of two cultivars of lettuce (Lactuca sativa L.; light-sensitive Ritsa and light-insensitive Strada) was investigated. The inhibition of Ritsa seed germination by exogenous ABA was higher than that of Strada seeds, the extent of inhibition of both cultivars being reduced by a short light break. At 25°C the sensitivity of both cultivars to exogenous ABA was higher than at 15°C. The endogenous level of ABA was similar in dry seeds of both cultivars, increasing temporarily in Ritsa seeds during the first 4 h of imbibition in darkness but not in Strada seeds, nor in Ritsa seeds exposed to a short light break. The transitory increase of ABA content in Ritsa seeds imbibed in darkness was accompanied by increased expression of the gene responsive to ABA (Rab) under these conditions. Zorial (Norflurazone), an ABA-biosynthesis inhibitor, decreased ABA content and allowed dark germination of the light-requiring Ritsa seeds. A short light break induced germination of Ritsa seeds when applied at 24 and 48 h after imbibition onset, i.e. after the transitory increase of ABA. GA3, on the other hand was effective when applied at the beginning of imbibition. It seems that light induces germination of the photoblastic Ritsa seeds by both inhibiting ABA synthesis and decreasing seed sensitivity to ABA and inhibitory processes induced by it.

The influence of calcium and ABA on ethylene biosynthesis was investigated at different stages and patterns of tomato (Lycopersicon esculentum cv. Lichun) fruit to analyse the relationship between calcium and ABA in relation to the ethylene biosynthesis system. The ethylene production of discs excised from mature green tomato fruits increased rapidly with 100 μmol/l ABA or 100 mmol/l calcium treatment. The increase with the two treatments combined was more than that after application of a single chemical, which suggests that these two chemicals could play synergistic roles in the ethylene synthesis of mature green tomato fruits. It was also found that application of the calcium chelator EGTA inhibited ethylene synthesis, and the production of ethylene after application of EGTA plus ABA was higher than that after treatment with EGTA alone. However, the effect changed when immature fruits were treated with these chemicals. ABA inhibited ethylene synthesis obviously and calcium still promoted it. When we treated immature fruit discs with both calcium and ABA simultaneously, ethylene production was more than that of ABA and less than that of calcium, which indicates that ABA and calcium play agonistic roles in ethylene biosynthesis of the immature tomato fruit discs. Furthermore, ethylene release from transgenic antisense ACS immature tomato fruit discs with application of both calcium and ABA was inhibited markedly, but was stimulated when the mature fruits were treated with ABA and calcium. All of these data suggest that there are different relationships between calcium and ABA in the ethylene synthesis system in different patterns and stages of tomato fruit ripening.

Deschampsia antarctica Desv. is one of two vascular plants from the Maritime Antarctic. It is usually exposed to cold, salt, and desiccating winds. We hypothesize that D. antarctica has genes that encode dehydrin proteins and their expression is regulated by low temperature, salt or osmotic stress. To test this hypothesis a fragment of a dehydrin gene from D. antarctica was identified and used as a probe to study dehydrin expression under low temperature, salt, and osmotic stress, and exogenous ABA (abscisic acid) treatments. An anti-dehydrin antibody was also used to study dehydrin protein accumulation under the same treatments. Southern analysis of genomic DNA treated with different endonucleases showed more than four bands recognized by the probe, suggesting that D. antarctica has several dehydrin genes. Northern analysis showed two putative dehydrin transcripts of 1.0 kb accumulated only under exogenous ABA and 1.6 kb under osmotic and salt treatments, suggesting that D. antarctica would have ABA-dependent and - independent pathways for regulation of dehydrin expression. Western analysis showed seven dehydrin proteins (58, 57, 55, 53, 48, 30 and 27 kDa) under the different stress treatments. Cold-accumulated dehydrin proteins were immunolocalized, showing that they are associated with vascular and epidermal tissue, which are preferential ice nucleation zones.