Embryonic Dormancy Research Articles

Embryonic dormancy in Aedes aegypti and Aedes albopictus (Diptera: Culicidae): a survival and dispersal mechanism

Aedes aegypti and Aedes albopictus are the main vectors of arboviruses such as dengue, Zika virus, and chikungunya. Ae. aegypti is a widely spread mosquito in tropical and subtropical regions, whereas Ae. albopictus is a culicid of Asian origin that shows exophilic behavior and can be found in subtropical and temperate areas. Climatic factors could influence the distribution of both species, making them use genetic and environmental resources to adapt to the environment, activating survival mechanisms (embryonic dormancy) that increase the developmental period and keep their offspring in the environment. From this perspective, this review aimed to compare the different physiological mechanisms of embryonic dormancy between Ae. aegypti and Ae. albopictus and their impact on the development and environmental adaptability of these two species. A total of 62 articles were identified in the PubMed, Scopus, and Web of Science databases corresponding to the period from 1981 to 2021. In diapause, the results mentioned above are indirectly linked to temperature and directly linked to photoperiod variations. With regard to quiescence, temperature and humidity are directly related to the activation of this mechanism. In conclusion, it is essential to highlight the expansion of arboviruses such as dengue, chikungunya, yellow fever, and Zika virus and their relationship with embryonic dormancy, diapause and quiescence, extremely important strategies for Ae. aegypti and Ae. albopictus to keep their offspring in the environment under adverse conditions.

Seed germination, morphology and fruit phenology insight of Cylicomorpha solmsii (Urb.) Urb: a step towards sustainable restoration planning

Cylicomorpha solmsii (Urb.) Urb (Caricaceae) is a wild relative of domesticated Carica papaya native to the humid tropical forest of Cameroon. C. solmsii is becoming extinct due to rapid urbanization of its habitat. There is currently no restoration planning, no available data on seed germination, details on morphological description and fruit phenology. We investigated the effects of light and soil on seed germination, updated its morphological description and provided cues of its fruit phenology. In two series of experiments, a germination test was first conducted under light and dark conditions with three seed pre-treatments (scarification, drying and cold). Secondly, pre-treated seeds were sown in native soils of C. solmsii habitat collected at Eloumden I and II, two ex-situ and mixtures soil with sand. Qualitative and quantitative data were collected on different part of the plant and analyzed using R package version 4.3.2. Our findings showed that C. solmsii seeds can germinate only under light. The seeds manifested a physiological embryonic dormancy. The native soils showed the highest germination percentage and seedling establishment. The dioicy of C. solmsii was clearly described with incomplete staminate and pistillate unisexual flower whorls. C. solmsii was observed to produce fruits throughout the year at varying intensity. This information is a vital cue to species restoration and policy makers towards C. solmsii conservation.

TET activity safeguards pluripotency throughout embryonic dormancy.

Dormancy is an essential biological process for the propagation of many life forms through generations and stressful conditions. Early embryos of many mammals are preservable for weeks to months within the uterus in a dormant state called diapause, which can be induced in vitro through mTOR inhibition. Cellular strategies that safeguard original cell identity within the silent genomic landscape of dormancy are not known. Here we show that the protection of cis-regulatory elements from silencing is key to maintaining pluripotency in the dormant state. We reveal a TET-transcription factor axis, in which TET-mediated DNA demethylation and recruitment of methylation-sensitive transcription factor TFE3 drive transcriptionally inert chromatin adaptations during dormancy transition. Perturbation of TET activity compromises pluripotency and survival of mouse embryos under dormancy, whereas its enhancement improves survival rates. Our results reveal an essential mechanism for propagating the cellular identity of dormant cells, with implications for regeneration and disease.

The Response of Hormones, Reactive Oxygen Species and Nitric Oxide in the Polyethylene-Glycol-Promoted, Salt-Alkali-Stress-Induced Embryo Germination of Sorbus pohuashanensis.

Polyethylene glycol can abrogate plant seed dormancy and alleviate salt-alkali stress damage to plants, but its role in embryonic dormancy abrogation and germination in Sorbus pohuashanensis is not yet clear. The mechanism by which polyethylene glycol promotes the release of embryonic dormancy may be related to the synthesis and metabolism of endogenous hormones, reactive oxygen species and reactive nitrogen. In this article, germination in indoor culture dishes was used, and the most suitable conditions for treating S. pohuashanensis embryos, with polyethylene glycol (PEG) and sodium carbonate (Na2CO3), were selected. Germination was observed and recorded, and related physiological indicators such as endogenous hormones, reactive oxygen species and reactive nitrogen were measured and analyzed to elucidate the mechanism of polyethylene glycol in alleviating salt-alkali stress in S. pohuashanensis embryos. The results showed that soaking seeds in 5% PEG for 5 days is the best condition to promote germination, which can increase the germination rate of embryos under salt-alkali stress by 1-2 times and improve indicators such as germination speed and the germination index. Polyethylene glycol led to an increase in gibberellin (GA), indole-3-acetic acid (IAA), ethylene (ETH), cytokinin (CTK), nitric oxide (NO), soluble protein and soluble sugar in the embryos under salt-alkali stress; increased activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), nitrate reductase (NR) and nitric oxide synthase (NOS) in the embryos; a reduction in the accumulation of abscisic acid (ABA), hydrogen peroxide (H2O2) and malondialdehyde (MDA). Therefore, it is suggested that the inhibitory effect of polyethylene glycol on the salt-alkali-stress-induced germination of S. pohuashanensis embryos is closely related to the response of endogenous hormones, reactive oxygen species and nitric oxide signalling.

FOXO1-mediated lipid metabolism maintains mammalian embryos in dormancy

Mammalian developmental timing is adjustable in vivo by preserving pre-implantation embryos in a dormant state called diapause. Inhibition of the growth regulator mTOR (mTORi) pauses mouse development in vitro, yet how embryonic dormancy is maintained is not known. Here we show that mouse embryos in diapause are sustained by using lipids as primary energy source. In vitro, supplementation of embryos with the metabolite l-carnitine balances lipid consumption, puts the embryos in deeper dormancy and boosts embryo longevity. We identify FOXO1 as an essential regulator of the energy balance in dormant embryos and propose, through meta-analyses of dormant cell signatures, that it may be a common regulator of dormancy across adult tissues. Our results lift a constraint on in vitro embryo survival and suggest that lipid metabolism may be a critical metabolic transition relevant for longevity and stem cell function across tissues.

Analysis of the transcriptional pathways associated with the induction of quiescent embryonic arrest in the calanoid copepod Acartia tonsa

The copepod species Acartia tonsa (Dana)(Crustacea) have the unique ability to induce quiescent embryonic dormancy if adverse environmental conditions occur; a characteristic shared by 41 other species belonging to the superfamily Centropagoida in the Calanoida order. However, the transcriptional changes characterizing this process are not known. Here, we compare the transcriptome of embryos in arrested quiescence with the normal development to identify pathways and differentially regulated transcripts involved in quiescent embryogenesis. Quiescence was induced by incubating eggs at 4 °C with anoxia for 26 h(hr), while eggs undergoing normal immediate development were incubated at 16.9 °C in normoxia for 7 h (where gastrulation occurs) or 14 h (where organogenesis occurs) before collecting for RNA extraction and analysis by RNA-sequencing. Results indicate that the expression profile of the quiescent embryo is not as different from the normal embryonic gastrulation as initially expected: None of the mapped transcripts is uniquely expressed in quiescence. Moreover, in quiescence a large proportion of the annotated transcripts display expression values halfway in-between the normal, immediate developmental stages of gastrulation and organogenesis. In depth comparison between the organogenesis stage and quiescent samples, reveal a high degree of divergence, confirming that a developmental arrest has been induced through quiescence. Specifically: Stress response transcripts are prominent in the quiescent phase with a transcript like the mammalian autophagy gene Sequestosome-1/p62 (SQSTM) being upregulated. The present analysis provides a better understanding of the molecular mechanisms characterizing the quiescent embryonic state of A. tonsa.

Differential regulations of abscisic acid-induced desiccation tolerance and vegetative dormancy by group B3 Raf kinases in liverworts.

Phytohormone abscisic acid (ABA) plays a key role in stomata closure, osmostress acclimation, and vegetative and embryonic dormancy. Group B3 Raf protein kinases (B3-Rafs) serve as positive regulators of ABA and osmostress signaling in the moss Physcomitrium patens and the angiosperm Arabidopsis thaliana. While P. patens has a single B3-Raf called ARK, specific members of B3-Rafs among six paralogs regulate ABA and osmostress signaling in A. thaliana, indicating functional diversification of B3-Rafs in angiosperms. However, we found that the liverwort Marchantia polymorpha, belonging to another class of bryophytes, has three paralogs of B3-Rafs, MpARK1, MpARK2, and MpARK3, with structural variations in the regulatory domains of the polypeptides. By reporter assays of the P. patens ark line and analysis of genome-editing lines of M. polymorpha, we found that these B3-Rafs are functionally redundant in ABA response, with respect to inhibition of growth, tolerance to desiccation and expression of stress-associated transcripts, the majority of which are under the control of the PYR/PYL/RCAR-like receptor MpPYL1. Interestingly, gemmae in gemma cups were germinating only in mutant lines associated with MpARK1, indicating that dormancy in the gametophyte is controlled by a specific B3-Raf paralog. These results indicated not only conservation of the role of B3-Rafs in ABA and osmostress response in liverworts but also functional diversification of B3-Rafs, which is likely to have occurred in the early stages of land plant evolution.

A comparative analysis of Aedes albopictus and Aedes aegypti subjected to diapause-inducing conditions reveals conserved and divergent aspects associated with diapause, as well as novel genes associated with its onset.

Aedes albopictus and Aedes aegypti are mosquito species that are distributed worldwide and transmit diverse arboviruses of medical importance, such as those causing yellow fever, dengue, chikungunya and Zika. A. albopictus embryos may remain viable for long periods in the environment due to their ability to become dormant through quiescence or diapause, a feature that contributes to their dispersion and hinders control actions. Diapause incidence can vary among natural populations of A. albopictus, but metabolic and genetic parameters associated with its induction still need to be better defined. The present study aimed to investigate the effect of exposure to diapause-inducing conditions on several biological parameters in different populations of A. albopictus (from tropical and temperate areas) and the diapause-refractory A. aegypti (tropical and subtropical populations). As expected, only the A. albopictus populations exhibited diapause, but with a lower incidence for the population from a tropical area. Exposure to diapause-inducing conditions, however, led to a sharp reduction in fecundity for both A. albopictus and A. aegypti tropical populations, with no effect on fertility (>90%). It also led to a prolonged period as pupae for the progeny of all induced groups, with a further delay for those from temperate climates. In all those induced groups, the lipid contents in eggs and adult females were higher than in the non-induced controls, with the highest values observed for both A. albopictus groups. Three genes were selected to have their expression profile investigated: cathepsin, idgf4, and pepck. Upon exposure to diapause-inducing conditions, all three genes were upregulated in the A. albopictus embryos from the tropical region, but only idgf4 was upregulated in the temperate climate embryos. This represents a new gene associated with diapause that can be used as a target to evaluate and prevent embryonic dormancy, a possible new vector control strategy for mosquito species from temperate areas, such as A. albopictus.

Copepod Embryonic Dormancy: "An Egg Is Not Just an Egg".

Long-lasting embryonic dormancy in invertebrates defies our understanding of what constitutes life because, for example, eggs of some copepods can delay hatching for decades or even centuries. Copepods, often millimeter-sized crustaceans, are some of the most numerous multicellular organisms on earth and are key organisms in most aquatic food webs. Some important free-living marine and estuarine species overwinter or oversummer by arrested embryogenesis in dormancy. The present contribution discusses the complex mechanisms behind embryonic dormancy by compiling knowledge from the 42 calanoid copepods from the superfamily Centropagoidea with well-described embryonic dormancy, which has been of scientific interest for decades. However, the determination of categories of copepod resting eggs-that is, diapause and quiescence, transitions between categories, the mechanisms controlling arrested development by the embryos, and how they interact with their surroundings-is not fully understood. Moreover, a clear link between the presence of the free-swimming population and their resting eggs in sediments is still not convincingly demonstrated. Here I evaluate the relative significance of potential cues driving the production of and the phase shift between egg categories. Understanding the initiation and termination of embryonic dormancy is of great importance for fundamental science-that is, population and food web ecology as well as climate science, aquaculture live feed, and ballast water research. Molecular techniques are developing rapidly, especially within health sciences, thus providing relevant tools applicable for plankton research. Here I suggest that applying molecular methods in addition to traditional physiological approaches in future research will lead to greater understanding of copepod embryonic dormancy, one of nature's wonders.

Ecology and Physiology of Dormancy in a Changing World: Introduction to a Virtual Symposium in The Biological Bulletin.

Ecology and Physiology of Dormancy in a Changing World: Introduction to a Virtual Symposium in The Biological Bulletin.

Adaptation to temperate climates: Evidence of photoperiod-induced embryonic dormancy in Aedes aegypti in South America

Dormancy is a developmental arrest in arthropods, in response to unfavorable conditions in temporally varying environments. In Aedes aegypti, the supposed inability of eggs to inhibit hatching has been used to explain the restriction of this species to tropical and subtropical regions. However, the geographic range of Ae. aegypti is constantly expanding towards temperate regions. Thus, the aim of the present study was to assess the ability of Ae. aegypti individuals from a temperate region (Buenos Aires City, Argentina) to enter photoperiod induced dormancy. To this end, we exposed both the parental generation and the eggs to short-day (SD: 10L:14D) and long-day (LD: 14L:10D) photoperiods, and studied the temporal variation in egg hatching. The experiment consisted of 28 treatment combinations of three factors: parental photoperiod (SD or LD), egg storage photoperiod (SD or LD), and age of eggs (14, 28, 42, 56, 70, 91, and 112 days). The results showed a lower hatching response with the SD parental photoperiod, and a trend to higher hatching with longer egg storage time in all photoperiod treatment combinations. The egg storage photoperiod showed no effect on egg hatching. In both parental photoperiod treatments, egg replicates of most ages from different females showed a large variability, with some replicates with lowest hatching response and others with highest hatching response. Our results show the ability of Ae. aegypti to inhibit egg hatching in response to a short-day photoperiod, which could allow the further expansion of this species to regions with colder winters.

Resting eggs in free living marine and estuarine copepods

Marine free living copepods can survive harsh periods and cope with seasonal fluctuations in environmental conditions using resting eggs (embryonic dormancy). Laboratory experiments show that temperature is the common driver for resting egg production. Hence, we hypothesize (i) that seasonal temperature variation, rather than variation in food abundance is the main driver for the occurrence of the resting eggs strategy in marine and estuarine copepod species; and (ii) that the thermal boundaries of the distribution determine where resting eggs are produced and whether they are produced to cope with warm or cold periods. We compile literature information on the occurrence of resting egg production and relate this to spatio-temporal patterns in sea surface temperature and chlorophyll a concentration obtained from satellite observations. We find that the production of resting eggs has been reported for 42 species of marine free living copepods. Resting eggs are reported in areas with high seasonal variation in sea surface temperature (median range 11°C). Temporal variation in chlorophyll a concentrations, however, seems of less importance. Resting eggs are commonly produced to cope with both warm and cold periods and, depending on the species, they are produced at the upper or lower thermal boundaries of a species’distribution.

Population dynamics and seasonal variation in the embryonic dormancy of Pilophorus gallicus (Hemiptera: Miridae): ‘don't put all your eggs in one basket'

AbstractPilophorus gallicus Remane is a generalist predator in southern European pear orchards. Nymphs and adults are present in orchards from March to November; their winter absence suggests either migration to other hosts or embryonic dormancy on pear trees to overcome the adverse period. In addition, it has been hypothesized that aestivation takes place to cope with extreme summer conditions. The present study aimed to investigate the reproductive strategy of P. gallicus to overcome unfavourable periods. Accordingly, (i) its population dynamics were followed during several years and (ii) females were sampled in three different seasons to study the condition (diapausing or nondiapausing) of the laying. The results obtained show that nymphs were always the first mobile instar to show up in pear orchards and the presence of adults was delayed, indicating egg overwintering. Nondiapausing eggs prevailed in spring and summer, whereas the autumn eggs were mainly diapausing. Aestivation was rejected. Worthy of note are: (i) the presence of diapausing eggs under favourable conditions and (ii) the existence of females laying both diapausing and nondiapausing eggs; thus, ‘not laying all the eggs in one basket’. The reproductive strategy of P. gallicus is considered as bet‐hedging for the short‐ and long‐term survival of the species.

Embryonic dormancy in seeds of Bactris gasipaes Kunth (peach-palm)

Abstract: Bactris gasipaes is a domesticated palm whose fruits are of great importance for the Amazonian people and whose heart of palm is also receiving economic interest in other brazilian and Latin America regions. The aim of this study was verify embryonic dormancy and its correlation with first cataphyll emergence in B. gasipaes seeds collected from four plants at Manaus city and four others at Coari city, both in the Amazonas state, Brazil. After extraction and cleaning, some of the seeds (4 replications of 25 per plant) were sown in a seedbed with a sawdust and sand mixture as substrate, and embryos (4 replications of 10 per plant), after extraction, were inoculated into half strength Murashige and Skoog cultures. Were used 100 seeds and 40 embryo per treatment. Whole seed and embryo germination varied between the different source plants and locations, with the greatest difference observed for the emergence of first cataphyll from seeds in the seedbed. For the most part of variables, results of seed and embryo were positively associated, namely, as one went up the other also, and vice versa. These results suggesting that, at least in part, seed dormancy in Bactris gasipaes is associated with embryonic dormancy.

Nitric oxide-polyamines cross-talk during dormancy release and germination of apple embryos

Nitric oxide (NO) and polyamines (PAs) belong to plant growth and development regulators. These compounds play a key role in numerous physiological processes e.g. seed germination. Based on the suggestion of overlapping of NO and PAs biosynthetic pathways, we demonstrated a cross-talk of NO and PAs in regulation of embryonic dormancy release. The aim of the work was to investigate an impact of PAs (Put, Spd and Spm) or NO short-term fumigation on nitrite, urea, Arg and ornithine (Orn) content, NO synthase-like (NOS-like) and arginase activity in axes of apple (Malus domestica Borkh.) embryos during dormancy alleviation and at the stage of termination of germination sensu stricto. NO, Put/Spd induced dormancy breakage and germination of apple embryos corresponded to stimulation of urea cycle and high free Arg pool in seedlings roots. After two days of the culture Put and Spd stimulated Arg dependent NO formation, inhibition of which was observed after Spm application. Put or Spd application as well as NO short-term pretreatment of apple embryos influenced level of ubiquitin-conjugated proteins. Higher abundance of such modified proteins correlated well to the declined content of nitrated proteins, suggesting their important role in regulation of embryo germination. NO led to stimulation of embryos germination by increasing level of free PAs (mostly Put). While transcriptomic approach showed down regulation of Spm synthesis and up-regulation of Spm degradation by NO, confirming negative role of Spm over-accumulation in embryo dormancy removal. Our data clearly indicate positive relationship of NO-Put/Spd acting as dormancy removing factors.

Extreme tolerance and developmental buffering of UV-C induced DNA damage in embryos of the annual killifish Austrofundulus limnaeus.

Free-living aquatic embryos are often at risk of exposure to ultraviolet radiation (UV-R). Successful completion of embryonic development depends on efficient removal of DNA lesions, and thus many aquatic embryos have mechanisms to reverse DNA lesions induced by UV-R. However, little is known of how embryos that are able to enter embryonic dormancy may respond to UV-R exposure and subsequent DNA damage. Embryos of the annual killifish Austrofundulus limnaeus are unique among vertebrates because their normal embryonic development includes (1) a complete dispersion of embryonic blastomeres prior to formation of the definitive embryonic axis, and (2) entry into a state of metabolic depression and developmental arrest termed diapause. Here, we show that developing and diapausing embryos of A. limnaeus have exceptional tolerance of UV-C radiation and can successfully complete embryonic development after receiving substantial doses of UV-C, especially if allowed to recover in full-spectrum light. Recovery in full-spectrum light permits efficient removal of the most common type of DNA lesion induced by UV-R: cyclobutane pyrimidine dimers. Interestingly, whole-mount embryo TUNEL assays suggest that apoptosis may not be a major contributor to cell death in embryos UV-C irradiated during dispersion/reaggregation or diapause. We also observed embryo mortality to be significantly delayed by several weeks in diapausing embryos irradiated and allowed to recover in the dark. These atypical responses to UV-R induced DNA damage may be due to the unique annual killifish life history and provide insight into DNA damage repair and recognition mechanisms during embryonic dormancy.

Growth of apple-tree seedlings (Malus domesticd) Borkh cv. Antonówka obtained from non-stratified embryos

The growth of seedlings (<i>Malus domestica</i> Borkh cv. Antonówka) obtained from embryos isolated from non-stratified and stratified (90 days at 4-5°C) seeds was compared. The lack of differences in the germination power of embryos isolated from non stratified and stratified seeds, and the observed normal growth of plants obtained from non-stratified seeds, point to the fact, that the seeds of the autumn apple 'Antonówka' do not pass the stage of embryonal dormancy.

Short communication. Effects of the duration of after-ripening period on seed germinations and seedling size in three fescue species

After-ripening is a complex enzymatic and biochemical process resulting in the breakage of non-deep physiological dormancy. The experiment was set up to identify the minimum duration of storage after harvest for completion of afterripening in meadow (Festuca pratensis Huds.), tall (Festuca arundinacea Schreb.) and red fescue (Festuca rubra L.). The seeds were stored under ware-house conditions for 0, 30, 60, 90, 120, 150, 180, 210 and 240 days after harvest (DAH). After the storage periods, the seeds were tested for germination and the primary root length, shoot length and seedlings biomass were measured. The final counts in all three species amounted to 60-65% germination immediately after harvest, suggesting a medium level of embryonic dormancy. The after-ripening during storage improved the final germination in all three species with decreasing intensity. In meadow fescue, the final germination was significantly improved up to 150 DAH, while the seedling size did not change significantly. In tall and red fescue, after ripening significantly improved the final germination up to 120 DAH, while the seedling size variables reached maximum values after 150 DAH. The obtained data can serve for the determination of proper storage duration management between harvest and sowing (autumn/spring) of the tested fescue species under agro-ecological conditions of south-eastern Europe.

Metabolic control of embryonic dormancy in apple seed: seven decades of research

Embryonic dormancy is defined as a set of blocks imposed upon a process(es) cardinal for growth. In apple seeds, all these blocks are removed as a result of cold treatment (stratification), but some of them are also affected by light and/or hormonal treatments. This review summarizes published data related to the modes of action of above factors on the changes in the levels of endogenous hormones and some other plant growth regulators (e.g., hydrogen cyanide), and on certain enzymes involved in mobilization of seed reserves and catabolism of their hydrolysis products. Phytochrome and activities of acid lipase and a protease have been indicated as receptors of light and low temperature, respectively. Several chains of events initiated by these two environmental factors and leading to dormancy removal are proposed, and the sites of their control by hormones and HCN are indicated. These chains are postulated to contribute to the elimination of particular blocks that hinder germination and therefore to be involved in the mechanisms of dormancy breakage.

ROS production and protein oxidation as a novel mechanism for seed dormancy alleviation

At harvest, sunflower (Helianthus annuus L.) seeds are dormant and unable to germinate at temperatures below 15 degrees C. Seed storage in the dry state, known as after-ripening, is associated with an alleviation of embryonic dormancy allowing subsequent germination at suboptimal temperatures. To identify the process by which dormancy is broken during after-ripening, we focused on the role of reactive oxygen species (ROS) in this phenomenon. After-ripening entailed a progressive accumulation of ROS, namely superoxide anions and hydrogen peroxide, in cells of embryonic axes. This accumulation, which was investigated at the cellular level by electron microscopy, occurred concomitantly with lipid peroxidation and oxidation (carbonylation) of specific embryo proteins. Incubation of dormant seeds for 3 h in the presence of hydrogen cyanide (a compound that breaks dormancy) or methylviologen (a ROS-generating compound) also released dormancy and caused the oxidation of a specific set of embryo proteins. From these observations, we propose a novel mechanism for seed dormancy alleviation. This mechanism involves ROS production and targeted changes in protein carbonylation patterns.