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%.

Intrinsically disordered proteins (IDPs) and the impact on cell stress resistance / 生物工程学报

Intrinsically disordered proteins (IDPs) are proteins or protein regions that fail to get folded into definite three-dimensional structures but participate in various biological processes and perform specific functions. Defying the traditional protein "sequence-structure-function" paradigm, they enrich the protein "structure-function" diversity. Ubiquitous in organisms, they show extreme hydrophilicity, charged amino acids, and highly repetitive amino acid sequences, with simple arrangement. As a result, they feature highly variable binding affinities and high coordination, which facilitate their functions. IDPs play an important role in cell stress response, which can improve the tolerance to a variety of stresses, such as freezing, high salt, heat shock, and desiccation. In this study, we briefed the characteristics, classifications, and identification of IDPs, summarized the molecular mechanism in improving cell stress resistance, and described the potential applications.

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.

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

Abstract 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.

Resumo 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.

Seedling survival of Handroanthus impetiginosus (Mart ex DC) Mattos in a semi-arid environment through modified germination speed and post-germination desiccation tolerance / Sobrevivência de plântulas de Handroanthus impetiginosus (Mart ex DC) Mattos em ambiente semi-árido ocorre por modificação da velocidade de germinação e da tolerância à dessecação pós germinação

Abstract Uniform rapid seed germination generally forms a great risk for the plant population if subsequent intermittent precipitation causes desiccation and seedling death. Handroanthus impetiginosus can be found commonly in a wide range of biomes within Brazil including those that are semi-arid. Germination and early growth was studied to understand how germinated seeds survive under these stringent conditions. Accessions were sampled from four seasonally dry biomes in Brazil. Precipitation at the start of the rainy season in the Caatinga, a semi-arid biome, is less predictable and the number of successive dry days per dry interval in the first four months of the rainy season was higher than in the other studied biomes. Plants from the Caatinga produced thicker seeds and this trait concurred with slow germination and stronger osmotic inhibition of germination across the accessions, forming a stress avoidance mechanism in the Caatinga. Post-germination desiccation tolerance was high in the Caatinga accession, could be re-induced in accessions from biomes with more regular precipitation (Cerrado and transition zone), but remained poor in the Cerradão accession; thus forming a stress tolerance mechanism. Production of adventitious roots ascertained survival of all tested individuals from all four locations, even if protruded radicles did not survive desiccation, forming an additional stress tolerance mechanism. A sequence of stress avoidance and stress tolerance mechanisms in seeds and germinated seeds was associated with precipitation patterns in different biomes. These mechanisms purportedly allow rapid seedling establishment when conditions are suitable and enable survival of the young seedling when conditions are adverse.

Resumo A germinação rápida e uniforme geralmente pode apresentar riscos para a população de plantas caso a precipitação ocorra de maneira intermitente, provocando seca e morte das plântulas. Handroanthus impetiginosus pode ser encontrado em uma grande variedade de biomas no Brasil. A germinação e o crescimento inicial das plântulas de Handroanthus impetiginosus foram estudadas para compreender como as sementes germinadas sobrevivem nestas condições. Acessos de quatro biomas que apresentam secas sazonais foram amostrados. A precipitação, no início da estação chuvosa na Caatinga, um bioma semi-árido, é menos previsível e o número de dias secos consecutivos por intervalo de seca, nos primeiros quatro meses da estação chuvosa, foi maior do que nos outros biomas estudados. Plantas do bioma Caatinga produzem sementes mais espessas e essa característica está associada com uma germinação lenta e com a inibição osmótica maior da germinação através dos acessos, formando um mecanismo para evitar o estresse. Tolerância à dessecação em sementes germinadas foi superior no bioma Caatinga e que podem ser reinduzida em acesso com uma precipitação mais regular (Cerrado e na zona de transição), mas manteve-se baixa no acesso Cerradão, formando assim um mecanismo para evitar o estresse. Produção de raízes adventícias foi observada nos indivíduos estudados de todos os quatro biomas, mesmo quando as raízes primárias não sobreviveram à dessecação, formando um mecanismo adicional de tolerância ao estresse. Assim, foi observado estratégias para evitar o estresse e mecanismos de tolerância ao estresse em sementes e sementes germinadas associada com padrões de precipitação nos diferentes biomas estudados. Estes mecanismos podem favorecer o estabelecimento das plântulas de forma rápida quando as condições são adequadas e permitirem a sobrevivência de plântulas quando as condições são adversas.

Water potential and viability of seeds of Eugenia (Myrtaceae), a tropical tree species, based upon different levels of drying

The aim of the present work was to study the water potential and viability of Eugenia based upon different levels of drying. The seeds showed high water content at shedding and were sensitive to desiccation; however, the level of this sensitivity (water content) varied among the species. Five target drying levels were defined according to the critical levels of hydration. Results showed that water potential seemed to be more reliable than water content. It was hypothesize that the differences in desiccation tolerance, as well as recalcitrance, among the seeds of different species could be related to the maturity level of these seeds during shedding from the mother plant. Consequently, this could be a result of the evolution on how far the maturation process could represent an advantage for the species propagation. This could also mean that Eugenia seeds were dispersed at a maturity stage similar to that observed in immature orthodox seeds.

Anhydrobiosis in bacteria: From physiology to applications.

Anhydrobiosis is a phenomenon related to the partial or total desiccation of living organisms, keeping their vital functions after rehydration. The desiccated state in prokaryotes has been widely studied, mainly due to the broad spectrum of the anhydrobiosis applications. In this review, we present the basic theoretical concepts related to anhydrobiosis, focusing on bacterial species. An update about desiccation tolerance in bacteria is given; and the general mechanisms of desiccation tolerance and desiccation damage are described. In addition, we show how the study of anhydrobiosis in prokaryotes has established the theoretical and practical basis for the development of the drying technologies. With regard to the desiccation tolerance in bacteria, although many mechanisms remain undiscovered at the molecular level, important research about the physiology of the anhydrobiotic state and its applications has been performed, and here we provide the most recent information about this subject. On the other hand, the most widely used drying technologies and their particular applications in several fields are described (e.g. medicine, agriculture and food industry). Finally, topics on the stability of desiccated bacterial cells are treated, concluding with the necessity of focusing the research on the mathematical modelling of the desiccated state in bacteria.

Protein expression upon desiccation and imbibition of Magnolia ovata A. St.-Hil seeds

The effect of seed drying and imbibition was studied by differential protein expression using two dimensional gel electrophoresis. After drying to a range of water contents, seeds were germinated to assess the viability. Seeds of Magnolia ovata did not withstand the desiccation down to 0.10 g H2O · g-1 dw. The critical water content below which the desiccation sensitivity became apparent was around 0.18 g H2O · g-1 dw (-26.5 MPa). Total protein was extracted and separated by 2D electrophoresis from fresh seeds (0.28 g H2O · g-1 dw), mild dried seeds (0.25 g H2O · g-1 dw) and seeds at low water content (0.10 g H2O · g-1 dw) before and after imbibition for 10 days. The proteome profile revealed the presence of 588 spots on each silver stained gel, from which 21 showed differential expression, correlated with desiccation and germination, by increased or decreased expression. After MS/MS sequencing, three protein spots produced spectra that matched to a Magnolia salicifolia legumin precursor. Results suggested an involvement of this protein in the events taking place during the drying and subsequent imbibitions of the dried seeds.

Reserve carbohydrates and lipids from the seeds of four tropical tree species with different sensitivity to desiccation

Considering the importance of water content for the conservation and storage of seeds, and the involvement of soluble carbohydrates and lipids for embryo development, a comparative study was carried out among the seeds of Inga vera (ingá), Eugenia uniflora (pitanga), both classified as recalcitrant, and Caesalpinia echinata (brazilwood) and Erythrina speciosa (mulungu), considered as orthodox seeds. Low concentrations of cyclitols (0.3-0.5 percent), raffinose family oligosaccharides (ca. 0.05 percent) and unsaturated fatty acids (0-19 percent) were found in the seeds of ingá and pitanga, while larger amounts of cyclitols (2-3 percent) and raffinose (4.6-13 percent) were found in brazilwood and mulungu, respectively. These results, in addition to higher proportions of unsaturated fatty acids (53-71 percent) in orthodox seeds, suggested that sugars and lipids played important role in water movement, protecting the embryo cell membranes against injuries during dehydration.

Os compostos de reserva das sementes, além de suprirem energia para o desenvolvimento embrionário, desempenham importantes funções relacionadas à proteção celular contra secagem e congelamento. Considerando a importância do teor de água para a conservação e para o armazenamento de sementes de espécies arbóreas e o envolvimento dos carboidratos solúveis e de lipídios nesses processos, foi realizada uma análise comparativa desses compostos em sementes de Inga vera (ingá) e Eugenia uniflora (pitanga), consideradas recalcitrantes, e em Caesalpinia echinata (pau-brasil) e Erythrina speciosa (mulungu), com comportamento ortodoxo. Baixas concentrações de ciclitóis (0,3-0,5 por cento), de oligossacarídeos da série da rafinose (0,05 por cento) e de ácidos graxos insaturados (0-19 por cento) foram encontradas em sementes de ingá e pitanga, enquanto maiores quantidades de ciclitóis (2-3 por cento) e de rafinose (4,6-13 por cento) foram encontradas nas sementes de pau-brasil e mulungu, respectivamente. Estes resultados, juntamente com as altas proporções de ácidos graxos insaturados (53-71 por cento) presentes em sementes de pau-brasil e mulungu, sugerem que os açúcares e os lipídios poderiam desempenhar papel importante no movimento de água, protegendo as membranas celulares embrionárias contra injúrias provocadas durante a dessecação.

Changes in soluble carbohydrates during storage of Caesalpinia echinata LAM. (Brazilwood) seeds, an endangered leguminous tree from the Brazilian Atlantic Forest

Caesalpinia echinata seeds stored in laboratory environmental conditions lose their viability in one month whilst under low temperatures germination is maintained for 18 months of storage. These seeds are tolerant to desiccation, keeping their viability up to 0.08 gH2O.gDW-1. Since soluble carbohydrates are believed to be involved with desiccation tolerance and seed storability, the aim of this work is to analyze the content and composition of soluble carbohydrates in C. echinata seeds during storage in paper bags (PB) and glass flasks (GF) at laboratory room (RT) and cool (CT) temperatures. In freshly harvested seeds, total soluble carbohydrates comprised approximately 10% of the dry weight, decreasing to ca. 8% over 18 months of storage at RT. In seeds stored at CT, sugars varied differently decreasing initially and being restored at the end of the analysis period. The main neutral sugars in seeds from all treatments were sucrose, fructose and glucose. Raffinose and stachyose were present as traces. Free myo-inositol and other cyclitols were also detected. The main tendency observed was the variation in levels of both glucose and fructose in relation to sucrose, the highest levels of monosaccharides which were found in seeds stored at CT. The values of glucose and fructose were practically constant in seeds stored in paper bags for 18 months at CT, decreasing consistently in the other treatments, mainly at RT. Sucrose contents remained relatively stable. Changes in soluble sugars during storage suggest that the loss of germinability of seeds of C. echinata could be associated with low levels of glucose and fructose in relation to sucrose.

Sementes de Caesalpinia echinata (pau-brasil) perdem a viabilidade em um mês quando armazenadas no ambiente de laboratório, enquanto a capacidade germinativa é mantida quando armazenadas sob temperturas baixas. O presente trabalho teve como objetivos analisar o conteúdo e a composição dos carboidratos de sementes de C. echinata armazenadas em câmara fria (CT) e em temperatura ambiente do laboratório (RT), em duas embalagens distintas (permeável e impermeável), visando a avaliar o envolvimento desses compostos com a capacidade germinativa das sementes. Os resultados mostraram que os carboidratos solúveis são constituídos principalmente de sacarose, glicose, frutose, myo-inositol e traços de rafinose e estaquiose, totalizando cerca de 10% da massa seca das sementes. As variações nos carboidratos solúveis foram semelhantes nos dois tipos de embalagem, mas diferentes quanto à temperatura de armazenamento. Em CT, as proporções dos monossacarídeos encontradas nas sementes recém-colhidas foram mantidas por cerca de 18 meses de armazenamento, coincidindo com alta porcentagem de germinação (80%). Nas armazenadas em RT houve redução expressiva nas proporções de glicose e frutose e perda completa da germinabilidade. O conteúdo de sacarose se manteve relativamente estável durante todo o período de análise. Os resultados indicam que a perda da germinabilidade de sementes de C. echinata está associada à diminuição dos níveis de glicose e frutose em relação aos níveis de sacarose.