This work aimed to characterize the re-induction of desiccation tolerance (DT) in germinated seeds, using polyethylene glycol (PEG 8000). Cell changes were investigated through cytological assays (cell viability and transmission electronic microscopy) as well as DNA integrity during loss and re-establishment of DT. The loss of DT was characterized by drying germinated seeds with different radicle lengths (1, 2, 3, 4 and 5 mm) in silica gel, decreasing the moisture content to ten percentage points intervals, followed by prehumidification (100% RH / 24 h) and rehydration. To re-induce DT, germinated seeds were treated for 72 h with PEG (-2.04 MPa) and PEG (-2.04 MPa) + ABA (100 μM) before dehydration. Germinated seeds did not tolerate desiccation to 10% moisture content, irrespectively of the radicle length. However, when incubated in PEG, those with 1 and 2 mm long radicle attained 71% and 29% survival, respectively. The PEG+ABA treatment was efficient to re-establish DT in seeds with 1 mm long radicles (100% survival). The ultrastructural assays of the cells of germinated seeds with 2 and 5 mm length confirmed the obtained physiological results. Germinated seeds of C. fissilis constitute a useful tool for desiccation tolerance investigations.

Genipa americana (Rubiaceae) is important for restoration of riparian forest in the Brazilian Cerrado. The objective was to characterize the mechanism and control of germination of G. americana to support uniform seedling production. Morphology and morphometrics of seeds, embryo and endosperm were assessed by light and scanning electron microscopy during germination. Imbibition and germination curves were generated and over the same time interval endosperm digestion and resistance were measured by puncture force analysis and activity assay of endo-β-mannanase (EBM) in water and in abscisic acid (ABA). The gene encoding for EBM was partially cloned and its expression monitored by quantitative real-time-polymerase chain reaction. Embryos displayed growth prior to radicle protrusion. A two-phase increase in EBM activity coincided with the two stages of weakening of the micropylar endosperm. The second stage also coincided with growth of the embryo prior to radicle protrusion. Enzyme activity was initiated in the micropylar endosperm but spread to the lateral endosperm. ABA completely inhibited germination by inhibiting embryo growth, the second stage of weakening and expression of the EBM gene, but EBM activity was not significantly inhibited. This suggests that a specific isoform of the enzyme is involved in endosperm weakening. EBM may cause a general 'softening' of micropylar endosperm cell walls, allowing the embryo to puncture the endosperm as the driving force of the decrease in puncture force.

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Erythrina velutina (“mulungu”) is a legume tree from Caatinga that associates with rhizobia but the diversity and symbiotic ability of “mulungu” rhizobia are poorly understood. The aim of this study was to characterize “mulungu” rhizobia from Caatinga. Bacteria were obteined from Serra Talhada and Caruaru in Caatinga under natural regeneration. The bacteria were evaluated to the amplification of nifH and nodC and to metabolic characteristics. Ten selected bacteria identified by 16S rRNA sequences. They were tested in vitro to NaCl and temperature tolerance, auxin production and calcium phosphate solubilization. The symbiotic ability were assessed in an greenhouse experiment. A total of 32 bacteria were obtained and 17 amplified both symbiotic genes. The bacteria showed a high variable metabolic profile. Bradyrhizobium (6), Rhizobium (3) and Paraburkholderia (1) were identified, differing from their geographic origin. The isolates grew up to 45 °C to 0.51 mol L−1 of NaCl. Bacteria which produced more auxin in the medium with l-tryptophan and two Rhizobium and one Bradyrhizobium were phosphate solubilizers. All bacteria nodulated and ESA 90 (Rhizobium sp.) plus ESA 96 (Paraburkholderia sp.) were more efficient symbiotically. Diverse and efficient rhizobia inhabit the soils of Caatinga dry forests, with the bacterial differentiation by the sampling sites.

ABSTRACT. This research aimed to investigate the cellular alterations during the loss and reestablishment of desiccation tolerance (DT) in germinated Sesbania virgata seeds. The loss of DT was characterized in germinated seeds with increasing radicle lengths (1, 2, 3, 4 and 5 mm) when subjected to dehydration in silica gel, followed by rehydration. To re-establish DT, the germinated seeds were incubated for 72h in polyethylene glycol (PEG, -2.04 MPa) with or without ABA (100 μM) before dehydration in silica gel. Cell viability was assessed by seedling survival, and DNA integrity was evaluated by gel electrophoresis. Seeds with 1 mm radicle length survived dehydration to the original moisture content (MC) of the dry seed (approximately 10%). PEG treatment was able to re-establish DT, at least partially, with 2, 3 and 4 mm but not in 5 mm radicle lengths. Germinated seeds treated with PEG+ABA performed better than those treated only with PEG, and DT was re-established even in germinated seeds with a 5 mm radicle length. Among the PEG-treated germinated seeds dehydrated to 10% MC, DNA integrity was maintained only in those with a 1 mm radicle length.Keywords: abscisic acid, cytological alteration, osmotic stress.Perda e restabelecimento da tolerância à dessecação em sementes germinadas de Sesbania virgata (Cav.) (Pers.) RESUMO. Esta pesquisa objetivou investigar as alterações celulares durante a perda e o restabelecimento da tolerância à dessecação (TD) em sementes germinadas de Sesbania virgata. A perda da TD foi caracterizada em sementes germinadas com 1, 2, 3, 4 e 5 mm de comprimento de radícula, submetidas à desidratação em sílica gel seguida de reidratação. Para restabelecer a TD, as sementes germinadas foram incubadas por 72h em PEG (-2,04 MPa) com e sem ABA (100 μM) antes da secagem em sílica gel. A viabilidade celular foi avaliada pela sobrevivência de plântulas e a integridade do DNA foi avaliada por meio de eletroforese em gel. Sementes com 1 mm de radícula sobreviveram à secagem até o teor de água original (aproximadamente 10%). O tratamento com PEG foi eficiente para restabelecer a TD, parcialmente, em sementes com 2, 3 e 4 mm, exceto com 5 mm de radícula. Sementes germinadas tratadas com PEG+ABA apresentaram melhor desempenho em relação às sementes sem ABA, sendo que a TD foi restabelecida em sementes com 5 mm de radícula. Dentre as sementes tratadas com PEG e secas até 10% de teor de água, a integridade do DNA foi mantida em sementes com 1 mm de radícula.Palavras-chave: ácido abscísico, alteração citológica, estresse osmótico.

Legume-rhizobia symbiosis is a cross-kingdom association that results in large amounts of nitrogen incorporated in food webs. For the Brazilian semi-arid region, data on genetic variability and symbiotic efficiency of Papilionoidae rhizobial communities are very scarce. The aim of this study was to evaluate the genetic variability and the symbiotic efficiency of eight rhizobial isolates obtained from "mulungu" (Erythrina velutina Willd.) nodules. For 16S rRNA gene sequencing, the genomic DNA was extracted using a commercial kit, amplified with universal primers, and subjected to sequencing reactions. For the isolate ESA 71, PCR amplifications for nodC and nodA genes were attempted. Rhizobial efficiency was assessed by two greenhouse experiments. The first assay was carried out under gnotobiotic conditions, with sterile sand as a substrate; the second experiment was conducted in a non-sterile soil. For both experiments, the inoculation treatments consisted of a single inoculation of each isolate, in addition to a treatment with Bradyrhizobium elkanii BR 5609 as a reference strain. Furthermore, two non-inoculated control treatments, supplied and not supplied with mineral N, were also evaluated. Bacterial identification indicated that both α and β-rhizobia could be found in "mulungu" root nodules. Three isolates where classified within the Rhizobium genus, four bacteria belonged to Bradyrhizobium and one isolate clustered with Burkholderia. Positive amplification of an intragenic fragment of the nodA gene using a primer set to β-rhizobia could be found for ESA 71 (Burkholderia). All bacterial isolates were effective in colonizing "mulungu" roots. In the first experiment, all inoculated treatments and N fertilization increased the N concentration in "mulungu" shoot tissues. For total N in the shoots, the isolates ESA 70, ESA 72, and ESA 75 stood out. In the non-sterile substrate experiment, the isolates ESA 70, ESA 71, ESA 72, and ESA 75, together with the reference strains, induced increases in the shoot N concentration and total accumulation compared to the absolute control. The results indicate that "mulungu" is able to establish associations with efficient α and β-rhizobia in Brazilian semi-arid soils.

OSMOPRIMING ON Sesbania virgata (CAV.) PERS (FABACEAE) SEEDS potentials-0.2; -0.4; -0.6 and -0.8

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