Be prepared: how does discontinuous hydration in Tabebuia heterophylla seeds induce stress tolerance in seedlings?

Discontinuous hydration and dehydration (HD) cycles refer to controlled imbibition followed by dehydration before seed germination. Here, we investigated whether the level of imbibition before HD cycles affects the physiology of Tabebuia heterophylla seeds and seedlings. Seeds were imbibed for 10 h (T1; phase I of imbibition) or 35 h (T2; phase II), dehydrated, and progressively rehydrated one to four times (HD cycles). Germination and biochemical parameters (membrane integrity; total soluble, reducing, and nonreducing (NRS) sugars; proteins, amino acids, proline, H2O2, catalase, ascorbate peroxidase, and glutathione reductase activity) were quantified at the last rehydration step of each cycle. Biometric and biochemical parameters (including pigments) were analysed in seedlings 60 days after germination. HD cycles at T1 led to reduced seed germination and greater plasma membrane damage, higher enzyme activity (catalase and glutathione reductase) and accumulation of NRS, total amino acids, and proline compared to the controls and T2 treatment. Cellular damage became more severe with more HD cycles. HD cycles at T2 synchronized germination regardless of the number of cycles and also had a priming effect. T2 seeds had less NRS, total amino acids, and proline content than T1. HD cycles at T1 produced seedlings with higher carotenoid and total chlorophyll content than controls and T2, while seedlings from HD cycles at T2 had higher amounts of osmoprotectants. HD cycles at T2 benefited seeds and seedlings more than at T1. This suggests that the physiological and biochemical effects of HD cycles in seeds modulate seedling plasticity, depending on water availability, potentially promoting increased tolerance to recurrent droughts that will be intensified with ongoing climate changes.

Desiccation Tolerance of Epiphytic Macrolichens in an Evergreen Temperate Rain Forest (Alerce Costero National Park, Chile).

The Valdivian region has a temperate rainy climate with differences in rainfall throughout the year. This heterogeneity results in periods of summer drought that expose the poikilohydric epiphytes to desiccation. With this research, we aim to answer different research questions related to phorophyte preference, response to desiccation, and response to radiation. How does the diversity of macrolichens vary at a local and microclimate scale in three tree species within an evergreen forest? What is the tolerance limit of macrolichens against prolonged desiccation, according to evaluation of the maximum efficiency of PSII (Fv/Fm) and pigment concentration? What is the tolerance limit against a potential increase in radiation? We found that macrolichen communities are determined by tree species, which regulate the suitability of the substrate by modifying the temperature and humidity conditions. In addition, our results show a rapid photosynthetic alteration in temporal exposure to desiccation, measured through Fv/Fm and pigment concentration. Our results showed that the most sensitive lichens to radiation and desiccation are not coincident. We confirm the low tolerance of macrolichen species to high radiation, reflected in the saturation profile obtained for the set studied. The lichen community in the evergreen forest showed high complexity and vulnerability, pointing to the importance of more research.

Hydrogel capsules as new delivery system for Trichoderma koningiopsis Th003 to control Rhizoctonia solani in rice (Oryza sativa).

The incorporation of biological control agents (BCAs) such as Trichoderma spp. in agricultural systems favors the transition towards sustainable practices of plant nutrition and diseases control. Novel bioproducts for crop management are called to guarantee sustainable antagonism activity of BCAs and increase the acceptance of the farmers. The encapsulation in polymeric matrices play a prominent role for providing an effective carrier/protector and long-lasting bioproduct. This research aimed to study the influence of biopolymer in hydrogel capsules on survival and shelf-life of T. koningiopsis. Thus, two hydrogel capsules prototypes based on alginate (P1) and amidated pectin (P2), containing conidia of T. koningiopsis Th003 were formulated. Capsules were prepared by the ionic gelation method and calcium gluconate as crosslinker. Conidia releasing under different pH values of the medium, survival of conidia in drying capsules, storage stability, and biocontrol activity against rice sheath blight (Rhizoctonia solani) were studied. P2 prototype provided up to 98% survival to Th003 in fluid bed drying, faster conidia releasing at pH 5.8, storage stability greater than 6 months at 18 °C, and up to 67% of disease reduction. However, both biopolymers facilitate the antagonistic activity against R. solani, and therefore can be incorporated in novel hydrogel capsules-based biopreparations. This work incites to develop novel biopesticides-based formulations with potential to improve the delivery process in the target site and the protection of the active ingredient from the environmental factors.

Hydrogel capsules as new approach for increasing drying survival of plant biostimulant gram-negative consortium.

Several plant growth-promoting bacteria (PGPB) are gram-negative, and their cell viability is affected during the bio-inoculant production. Hence, formulation-drying processes provide challenges that limit the adoption of these beneficial microorganisms in sustainable agricultural production. Among delivery system strategies for gram-negative PGPB, the encapsulating cells in biopolymeric materials are emerging as a promising alternative. This research aims to evaluate the effect of additives and crosslinking agents on the survival of the consortium of Herbaspirillum frisingense AP21, Azospirillum brasilense D7, and Rhizobium leguminosarum T88 in hydrogel capsules. Three crosslinkers and diverse potential drying protectors were tested. Calcium gluconate provides notable consortium survival advantages regarding colony-forming units (CFUs) (losses of up to 4 log CFU) compared to calcium lactate and calcium chloride (up to 6 log CFU). Additives such as skimmed milk, whey protein, and Gelita® EC improve the recovery of viable cells after the drying process, demonstrating an increase in cell survival of the three bacteria by up to 4 log CFU. The combination of these substances into a capsule prototype extends the storage stability of bacterial consortium up to 3 months at 18 ± 2 °C. This study expands the knowledge for formulating gram-negative PGPB consortium, regarding the crosslinker and drying protector relationship on encapsulation processes with drying survival and further storage stability performance. KEY POINTS: • Hydrogel immobilization formulation approach for PGPB consortium • Enhancing drying survival of gram-negative PGPB consortium • Increasing storage stability of PGPB consortium at 18 °C.

Impact of osmotic stress on production, morphology, and fitness of Beauveria bassiana blastospores.

Beauveria bassiana is a cosmopolitan entomopathogenic fungus that can infect over 1000 insect species. During growth inside the host, B. bassiana transitions from hyphal to yeast-like unicellular growth as blastospores. Blastospores are well suited as an active ingredient in biopesticides due to their ease of production by liquid fermentation. Herein, we investigated the impact of hyperosmotic growth environments mediated by ionic and non-ionic osmolytes on two strains of B. bassiana (ESALQ1432 and GHA) relevant to growth morphology, blastospore production, desiccation tolerance, and insecticidal activity. Polyethylene glycol (PEG200) increased osmotic pressure in submerged cultures leading to decreased blastospore size but higher blastospore yields for one strain. Morphologically, decreased blastospore size was linked to increased osmotic pressure. However, smaller blastospores from PEG200 supplemented cultures after air-drying exhibited delayed germination. Ionic osmolytes (NaCl and KCl) generated the same osmotic pressure (2.5-2.7 MPa) as 20% glucose and boosted blastospore yields (> 2.0 × 109 blastospores mL-1). Fermentation performed in a bench-scale bioreactor consistently promoted high blastospore yields when using NaCl (2.5 MPa) amended media within 3 days. Mealworm larvae (Tenebrio molitor) were similarly susceptible to NaCl-grown blastospores and aerial conidia in a dose-time-dependent manner. Collectively, these results demonstrate the use of hyperosmotic liquid culture media in triggering enhanced yeast-like growth by B. bassiana. Understanding the role of osmotic pressure on blastospore formation and fitness will hasten the development of viable commercial fungal biopesticides. KEY POINTS: • Osmotic pressure plays a critical role in submerged fermentation of B. bassiana. • Ionic/non-ionic osmolytes greatly impact blastospore morphology, fitness, and yield. • Desiccation tolerance and bioefficacy of blastospores are affected by the osmolyte.

Sex Differences in Desiccation Tolerance Varies by Colony in the Mesic Liverwort Plagiochila porelloides.

Water scarcity, a common stress factor, negatively impacts plant performance. Strategies to cope with it, such as desiccation tolerance, are becoming increasingly important to investigate. However, phenomena, such as intraspecific variation in stress responses have not received much attention. Knowledge of this variability and the environmental drivers can be leveraged to further investigate the mechanisms of desiccation tolerance. Here we tested for variation in desiccation tolerance in Plagiochila porelloides among colonies and sexes within the same riparian zone. Field-collected dehardened plants were subjected to a desiccation event, under controlled conditions and then rehydrated. Plant water status, photosynthetic rates, net carbon gain, and efficiency of photosystem II (PSII) were assayed to evaluate tissue desiccation, basic metabolic processes and plant recovery. To establish a linkage between plant response and environmental factors, field light conditions were measured. We detected intraspecific variation, where a more exposed colony (high percentage of open sky, large temporal range of light quantity, and high red/far-red ratio) showed sex differences in desiccation tolerance and recovery. Overall, PSII recovery occurred by 72 h after rehydration, with a positive carbon gain occurring by day 30. This within species variation suggests plastic or genetic effects, and likely association with light conditions.

Dehydration tolerance and storage sensitivity of Campomanesia adamantium seeds

Cultivating native fruit species depends on the existence of technical and scientific information. In this context, studies on propagation are the basis for other investigations. Campomanesia adamantium is propagated by seeds, and quality seeds guarantee the formation of high-performance plants in the field. This study aimed to analyze the dehydration tolerance and storage sensitivity of Campomanesia adamantium seeds. Experiment 1 evaluated the physiological quality of seeds with moisture contents of 43, 30, 26, 23, 18, 15, and 5%. Experiment 2 determined the seed moisture content, germination, and vigor of seeds depulped and immediately stored at 16 and 25°C, for 25 and 50 days. Seed viability was not affected by dehydration up to 15% of moisture content but decreased at 5%. Drying was accompanied by reduced seed vigor and seedling mass. The germination of unstored seeds was superior to stored ones. Storage at 16°C for 25 days was promising to maintain seed viability. The seeds showed intermediate recalcitrant behavior. Dehydration below 30% reduced vigor, while 5% affected viability. Campomanesia adamantium seeds can be stored for 25 days at 16°C in permeable packaging that maintains approximately 10% of moisture, with germination higher than 60%.

A comparative genomics examination of desiccation tolerance and sensitivity in two sister grass species.

Desiccation tolerance is an ancient and complex trait that spans all major lineages of life on earth. Although important in the evolution of land plants, the mechanisms that underlay this complex trait are poorly understood, especially for vegetative desiccation tolerance (VDT). The lack of suitable closely related plant models that offer a direct contrast between desiccation tolerance and sensitivity has hampered progress. We have assembled high-quality genomes for two closely related grasses, the desiccation-tolerant Sporobolus stapfianus and the desiccation-sensitive Sporobolus pyramidalis Both species are complex polyploids; S. stapfianus is primarily tetraploid, and S. pyramidalis is primarily hexaploid. S. pyramidalis undergoes a major transcriptome remodeling event during initial exposure to dehydration, while S. stapfianus has a muted early response, with peak remodeling during the transition between 1.5 and 1.0 grams of water (gH2O) g-1 dry weight (dw). Functionally, the dehydration transcriptome of S. stapfianus is unrelated to that for S. pyramidalis A comparative analysis of the transcriptomes of the hydrated controls for each species indicated that S. stapfianus is transcriptionally primed for desiccation. Cross-species comparative analyses indicated that VDT likely evolved from reprogramming of desiccation tolerance mechanisms that evolved in seeds and that the tolerance mechanism of S. stapfianus represents a recent evolution for VDT within the Chloridoideae. Orthogroup analyses of the significantly differentially abundant transcripts reconfirmed our present understanding of the response to dehydration, including the lack of an induction of senescence in resurrection angiosperms. The data also suggest that failure to maintain protein structure during dehydration is likely critical in rendering a plant desiccation sensitive.

Seed coating as a delivery system for the endophyte Trichoderma koningiopsis Th003 in rice (Oryza sativa).

Seed coating is a technique to cover seeds with external agents to upgrade their performance, handling, and plant establishment. Plant beneficial microbes (PBMs), such as plant growth-promoting bacteria, mycorrhizal fungi, and other fungi (e.g., Trichoderma spp.), decrease agrochemical inputs, enhance tolerance to biotic-abiotic stresses, and increase essential plant nutrition. The demand for pre-treated seeds as delivery systems for biological agents is advancing. Here, a seed coating formulation containing Trichoderma koningiopsis is presented. The physicochemical and biological characterization of the seed coating prototypes included drying protector screening, the effect of the inoculum concentration on survival, the assessment of microbial release profiles in soil extract, and plant tissue colonization capability under semi-controlled conditions. Gelatine and pectin, two of the tested drying protectors, maintained fungus germination after 60 days at 18 °C with significantly higher values of up to 38% compared with the control. The initial concentration of 106 colony-forming units (CFU) per seed undergoes a positive effect on survival over time. Regarding plant tissue colonization, the fungus establishes endophytically in rice. In conclusion, seed coating is a promising alternative for the formulation of beneficial microbial agents such as Trichoderma sp., maintaining cell survival and further promoting the establishment in rice systems.Key points• Enhancing drying survival of T. koningiopsis formulates• Seed coating formulation approach for T. koningiopsis in rice• Colonization capacity of formulated T. koningiopsis in rice tissue.

An Animal Able To Tolerate D2 O.

It is possible to gain a deeper insight into the role of water in biology by using physicochemical variant molecules, such as deuterium oxide (D2 O); however, D2 O is toxic to multicellular organisms in high concentrations. By using a unique desiccation-rehydration process, we demonstrate that the anhydrobiotic nematode Panagrolaimus superbus is able to tolerate and proliferate in 99 % D2 O. Moreover, we analysed P. superbus' water-channel protein (aquaporin; AQP), which is associated with dehydration/rehydration, by comparing its primary structure and modelling its tertiary structure in silico. Our data evidence that P. superbus' AQP is an aquaglyceroporin, a class of water channel known to display a wider pore; this helps to explain the rapid and successful organismal influx of D2 O into this species. This is the first demonstration of an animal able to withstand high D2 O levels, thus paving a way for the investigation of the effects D2 O on higher levels of biological organization.

Comparative transcriptome analysis suggests convergent evolution of desiccation tolerance in Selaginella species.

BACKGROUND: Desiccation tolerant Selaginella species evolved to survive extreme environmental conditions. Studies to determine the mechanisms involved in the acquisition of desiccation tolerance (DT) have focused on only a few Selaginella species. Due to the large diversity in morphology and the wide range of responses to desiccation within the genus, the understanding of the molecular basis of DT in Selaginella species is still limited. RESULTS: Here we present a reference transcriptome for the desiccation tolerant species S. sellowii and the desiccation sensitive species S. denticulata. The analysis also included transcriptome data for the well-studied S. lepidophylla (desiccation tolerant), in order to identify DT mechanisms that are independent of morphological adaptations. We used a comparative approach to discriminate between DT responses and the common water loss response in Selaginella species. Predicted proteomes show strong homology, but most of the desiccation responsive genes differ between species. Despite such differences, functional analysis revealed that tolerant species with different morphologies employ similar mechanisms to survive desiccation. Significant functions involved in DT and shared by both tolerant species included induction of antioxidant systems, amino acid and secondary metabolism, whereas species-specific responses included cell wall modification and carbohydrate metabolism. CONCLUSIONS: Reference transcriptomes generated in this work represent a valuable resource to study Selaginella biology and plant evolution in relation to DT. Our results provide evidence of convergent evolution of S. sellowii and S. lepidophylla due to the different gene sets that underwent selection to acquire DT.

Decoding Gene Networks Modules That Explain the Recovery of Hymenoglossum cruentum Cav. After Extreme Desiccation.

Hymenoglossum cruentum (Hymenophyllaceae) is a poikilohydric, homoiochlorophyllous desiccation-tolerant (DT) epiphyte fern. It can undergo fast and frequent dehydration-rehydration cycles. This fern is highly abundant at high-humidity/low-light microenvironments within the canopy, although rapid changes in humidity and light intensity are frequent. The objective of this research is to identify genes associated to desiccation-rehydration cycle in the transcriptome of H. cruentum to better understand the genetic dynamics behind its desiccation tolerance mechanism. H. cruentum plants were subjected to a 7 days long desiccation-rehydration process and then used to identify key expressed genes associated to its capacity to dehydrate and rehydrate. The relative water content (RWC) and maximum quantum efficiency (F v/F m) of H. cruentum fronds decayed to 6% and 0.04, respectively, at the end of the desiccation stage. After re-watering, the fern showed a rapid recovery of RWC and F v/F m (ca. 73% and 0.8, respectively). Based on clustering and network analysis, our results reveal key genes, such as UBA/TS-N, DYNLL, and LHC, orchestrating intracellular motility and photosynthetic metabolism; strong balance between avoiding cell death and defense (CAT3, AP2/ERF) when dehydrated, and detoxifying pathways and stabilization of photosystems (GST, CAB2, and ELIP9) during rehydration. Here we provide novel insights into the genetic dynamics behind the desiccation tolerance mechanism of H. cruentum.

Water stress tolerance tracks environmental exposure and exhibits a fluctuating sexual dimorphism in a tropical liverwort.

Water shortage events negatively impact plant productivity, threaten ecosystem functioning, and are predicted to increase dramatically in the coming years. Consequently, building a detailed understanding of how plants respond to water stress is critical for improving predictions of ecological processes and species range shifts under climate change. Here, we characterized patterns of intraspecific variation in dehydration tolerance (DhT, also dehydration tolerant) across a variable landscape in the tropical plant, Marchantia inflexa. DhT enables tissues to survive substantial drying (below an absolute water content of - 10 MPa) and despite the ecological significance of DhT, many questions remain. We tested if DhT was correlated with an environmental exposure gradient, if male and female plants had contrasting DhT phenotypes, and if variation in DhT had a genetic component. To do so, we collected plants from five populations, spanning an environmental exposure gradient in the forests of northern Trinidad, Republic of Trinidad and Tobago. We measured DhT immediately after collection, and after growing plants for ~ 1 year in a common garden. We found that DhT varied significantly among populations and tracked the characterized exposure gradient. Additionally, we showed that phenotypic differences among populations in DhT were maintained in the common garden, suggesting that underlying genetic differences contribute to DhT variability. Finally, we detected a fluctuating sexual dimorphism where males had lower DhT than females in less exposed sites, but not in more exposed sites. Interestingly, this fluctuating sexual dimorphism in DhT was driven primarily by male variation (females exhibited similar DhT across sites).

Desiccation tolerance in the Antarctic moss Sanionia uncinata.

BACKGROUND: One of the most extreme environments on our planet is the Maritime Antarctic territory, due to its low-water availability, which restricts the development of plants. Sanionia uncinata Hedw. (Amblystegiaceae), the main colonizer of the Maritime Antarctic, has effective mechanisms to tolerate this environment. It has been described that the tolerance to desiccation is mediated by the hormone abscisic acid (ABA), antioxidants systems, accumulation of compatible solutes and proteins of the late embryogenesis abundant (LEA). However, to date, these mechanisms have not been described in S. uncinata. Therefore, in this work, we postulate that the tolerance to desiccation in the Antarctic moss S. uncinata is mediated by the accumulation of ABA, the osmolytes proline and glycine betaine, and dehydrins (an LEA class 11 proteins). To demonstrate our hypothesis, S. uncinata was subjected to desiccation for 24 h (loss in 95% of water content), and the effects on its physiological, photosynthetic, antioxidant and biochemical parameters were determined. RESULTS: Our results showed an accumulation of ABA in response to water loss, and the activation of protective responses that involves an increment in levels of proline and glycine betaine, an increment in the activity of antioxidant enzymes such as SOD, CAT, APX and POD, and the accumulation of dehydrins proteins. CONCLUSION: The results showed, suggest that S. uncinata is a  desiccation-tolerant moss, property mediated by high cellular plasticity regulated by ABA.

Genomic Features for Desiccation Tolerance and Sugar Biosynthesis in the Extremophile Gloeocapsopsis sp. UTEX B3054.

For tolerating extreme desiccation, cyanobacteria are known to produce both compatible solutes at intracellular level and a copious amount of exopolysaccharides as a protective coat. However, these molecules make cyanobacterial cells refractory to a broad spectrum of cell disruption methods, hindering genome sequencing, and molecular studies. In fact, few genomes are already available from cyanobacteria from extremely desiccated environments such as deserts. In this work, we report the 5.4 Mbp draft genome (with 100% of completeness in 105 contigs) of Gloeocapsopsis sp. UTEX B3054 (subsection I; Order Chroococcales), a cultivable sugar-rich and hardly breakable hypolithic cyanobacterium from the Atacama Desert. Our in silico analyses focused on genomic features related to sugar-biosynthesis and adaptation to dryness. Among other findings, screening of Gloeocapsopsis genome revealed a unique genetic potential related to the biosynthesis and regulation of compatible solutes and polysaccharides. For instance, our findings showed for the first time a novel genomic arrangement exclusive of Chroococcaceae cyanobacteria associated with the recycling of trehalose, a compatible solute involved in desiccation tolerance. Additionally, we performed a comparative genome survey and analyses to entirely predict the highly diverse pool of glycosyltransferases enzymes, key players in polysaccharide biosynthesis and the formation of a protective coat to dryness. We expect that this work will set the fundamental genomic framework for further research on microbial tolerance to desiccation and to a wide range of other extreme environmental conditions. The study of microorganisms like Gloeocapsopsis sp. UTEX B3054 will contribute to expand our limited understanding regarding water optimization and molecular mechanisms allowing extremophiles to thrive in xeric environments such as the Atacama Desert.

Effect of the desiccation tolerance and cryopreservation methods on the viability of Citrus limon L. Burm cv. Eureka seeds.

Cryopreservation of the germplasm for long-term periods is of great importance to maintain the genetic resource. Argentina is one of the world's highest lemon producing country. The performance of different cooling/warming rates in the cryopreservation method of Citrus limon L. Burm cv. Eureka seeds and their influence on the interval of optimal moisture content in the desiccation stage were analyzed. Water sorption isotherm was determined and modeled using D'Arcy & Watt equation; it provided important information concerning the amounts of water associated to strong, weak and multimolecular binding sites along the sorption isotherm. Seeds tolerated a wide range of desiccation conditions (0.180%), however desiccation to 0.0526 g H2O g-1 d.b. (aw = 0.0901) produced a significant loss of viability. Differential Scanning Calorimetry was used to identify the thermal transitions of lipids and water in the seed; enthalpies were used to calculate the unfrozen water fraction (0.19 g H2O g-1 d.b. corresponding to aw = 0.64). Two cooling/warming rates were tested on desiccated seeds (0.110.64 corresponding to the unfrozen water fraction. The use of higher cooling/warming rates enables a wider range of desiccation conditions (0.33

Desiccation tolerance in the Antarctic moss Sanionia uncinata

BACKGROUND: One of the most extreme environments on our planet is the Maritime Antarctic territory, due to its low-water availability, which restricts the development of plants. Sanionia uncinata Hedw. (Amblystegiaceae), the main colonizer of the Maritime Antarctic, has effective mechanisms to tolerate this environment. It has been described that the tolerance to desiccation is mediated by the hormone abscisic acid (ABA), antioxidants systems, accumulation of compatible solutes and proteins of the late embryogenesis abundant (LEA). However, to date, these mechanisms have not been described in S. uncinata. Therefore, in this work, we postulate that the tolerance to desiccation in the Antarctic moss S. uncinata is mediated by the accumulation of ABA, the osmolytes proline and glycine betaine, and dehydrins (an LEA class 11 proteins). To demonstrate our hypothesis, S. uncinata was subjected to desiccation for 24 h (loss in 95% of water content), and the effects on its physiological, photosynthetic, antioxidant and biochemical parameters were determined. RESULTS: Our results showed an accumulation of ABA in response to water loss, and the activation of protective responses that involves an increment in levels of proline and glycine betaine, an increment in the activity of antioxidant enzymes such as SOD, CAT, APX and POD, and the accumulation of dehydrins proteins. CONCLUSION: The results showed, suggest that S. uncinata is a desiccation-tolerant moss, property mediated by high cellular plasticity regulated by ABA.

Slow and Fast Desiccation of Single-cell Thick Fronds of Filmy Ferns.

Filmy ferns can desiccate and recover after rehydration to resume photosynthesis. Slow and fast desiccation rates were compared in filmy fern fronds to determine whether structural or physiological differences may occur between desiccation rates. Slow desiccation is considered to be more similar to natural conditions experienced by plants that grow under the forest canopy. A fast desiccation rate will help to understand whether slow desiccation is important for recovery and viability.

Comparative transcriptome profile of the leaf elongation zone of wild barley (Hordeum spontaneum) eibi1 mutant and its isogenic wild type

Abstract The naturally occurring wild barley mutant eibi1/hvabcg31 suffers from severe water loss due to the permeable leaf cuticle. Eibi1/HvABCG31 encodes a full ATP-binding cassette (ABC) transporter, HvABCG31, playing a role in cutin deposition in the elongation zone of growing barley leaves. The eibi1 allele has pleiotropic effects on the appearance of leaves, plant stature, fertility, spike and grain size, and rate of germination. Comparative transcriptome profile of the leaf elongation zone of the eibi1 mutant as well as its isogenic wild type showed that various pathogenesis-related genes were up-regulated in the eibi1 mutant. The known cuticle-related genes that we analyzed did not show significant expression difference between the mutant and wild type. These results suggest that the pleiotropic effects may be a compensatory consequence of the activation of defense genes in the eibi1 mutation. Furthermore, we were able to find the mutation of the eibi1/hvabcg31 allele by comparing transcript sequences, which indicated that the RNA-Seq is useful not only for researches on general molecular mechanism but also for the identification of possible mutant genes.

Physiological, cellular and molecular aspects of the desiccation tolerance in Anadenanthera colubrina seeds during germination / Aspectos fisiológicos, celulares e moleculares da tolerância à dessecação em sementes de Anadenanthera colubrina durante a germinação

During germination, orthodox seeds become gradually intolerant to desiccation, and for this reason, they are a good model for recalcitrance studies. In the present work, physiological, biochemical, and ultrastructural aspects of the desiccation tolerance were characterized during the germination process of Anadenanthera colubrina seeds. The seeds were imbibed during zero (control), 2, 8, 12 (no germinated seeds), and 18 hours (germinated seeds with 1 mm protruded radicle); then they were dried for 72 hours, rehydrated and evaluated for survivorship. Along the imbibition, cytometric and ultrastructural analysis were performed, besides the extraction of the heat-stable proteins. Posteriorly to imbibition and drying, the evaluation of ultrastructural damages was performed. Desiccation tolerance was fully lost after root protrusion. There was no increase in 4C DNA content after the loss of desiccation tolerance. Ultrastructural characteristics of cells from 1mm roots resembled those found in the recalcitrant seeds, in both hydrated and dehydrated states. The loss of desiccation tolerance coincided with the reduction of heat-stable proteins.(AU)

Durante a germinação, sementes ortodoxas tornam-se gradualmente intolerantes à dessecação, e por isso podem ser utilizadas como modelo para o estudo da recalcitrância. No presente trabalho realizou-se uma caracterização dos aspectos fisiológicos, bioquímicos e ultraestruturais da perda da tolerância à dessecação de sementes de Anadenanthera colubrina em processo germinativo. Para isso as sementes foram embebidas durante 0 (controle), 2,8,12 e aproximadamente 18 horas (sementes germinadas com 1 mm de radícula), secas por 72 horas, reidratadas e a sobrevivência avaliada. Ao longo da embebição foram realizadas análises citométricas, ultraestruturais e extração de proteínas resistentes ao calor e após embebição e secagem foram avaliados danos ultraestruturais. A tolerância à dessecação foi totalmente perdida após a protrusão radicular. Não houve aumento do conteúdo de DNA 4C quando a tolerância à dessecação foi perdida. Características ultraestruturais de células de radículas de 1 mm assemelharam-se às encontradas em sementes recalcitrantes tanto no estado hidratado quanto desidratado. A perda da tolerância à dessecação coincidiu com a redução do conteúdo de proteínas resistentes ao calor.(AU)