Cell cycle activity during seed priming is not essential for germination advancement in tomato

Seed priming is a technique of controlled hydration and drying that results in more rapid gemination when the seeds are reimbibed. Advancement of radicle meristem cells into the S and G2 phases of the cell cycle, increasing the percentage of nuclei having a 4C DNA content, has been reported to occur during priming. It has been suggested that the efficiency of priming is related to the accumulation of 4C nuclei in the radicle meristem, but the extent of cell cycle activity varied among different treatments and seed lots. A wide range of priming treatments across temperatures, water potentials and durations can be compared on a common basis using the hydrothermal priming time model. Flow cytometry was used to monitor cell cycle activity in a number of tomato (Lycopersicon esculentum Mill.) seed lots during priming in relation to the accumulation of hydrothermal priming time and the subsequent germination rate response. In some seed lots, the percentage of 4C nuclei in the radicle meristems prior to emergence increased in proportion to accumulated hydrothermal priming time, while in other lots, no increase in nuclear DNA content was detected. All lots, however, demonstrated rapid radicle emergence following priming. Thus, replicative DNA synthesis in radicle meristem nuclei often occurred during seed priming, but an increase in the percentage of 4C nuclei was not essential for germination advancement.     

Hydrothermal time analysis of tomato seed germination responses to priming treatments

Controlled hydration of seeds followed by drying (seed priming) is used to break dormancy, speed germination, and improve uniformity of radicle emergence. To date, empirical trials are used to predict optimal priming conditions for a given seed lot. Since priming is based upon seed water relations, it was hypothesized that the sensitivity of germination to reduced water potential before priming might be mechanistically related to, and therefore predictive of, priming responsiveness. Analyses of germination of 13 tomato (Lycopersicon esculentum Mill.) seed lots at two temperatures (15C and 20C) and three water potentials (0, -0.28 and -0.43 MPa) showed that seed lot germination responses could be quantitatively characterized by parameters derived from thermal time, hydrotime, and hydrothermal time models (R²0.73-0.99). Six of the seed lots were primed at two temperatures (15°C and 20°C) and three water potentials (-1.0, -1.5 and -2.0 MPa) for various durations, dried, and their subsequent germination rates analysed according to hydropriming time and hydrothermal priming time models. The responses of germination rates to priming were characterized by hydropriming time (HP) and hydrothermal priming time (HTP) constants and the minimum water potential (min) and temperature (Tmin) for achieving a priming effect. The values of min and Tmin varied relatively little among tomato seed lots, and the generalized values of min=2.39 MPa and Tmin=9.10°C accounted for 74% (15°C), 57% (20°C), and 62% (across both temperatures) of the increase in germination rates following priming. Nonetheless, while the hydrothermal time models described germination patterns both before and after priming, there was relatively little predictive relationship between them.     

Sugar-beet seed steep treatments to improve germination under cold,wet conditions

Modifying the present commercial sugar-beet steep treatment (12h in 0.2% thiram suspension, 25°C) to include an initial 2h steep in 0.3M hydrochloric acid, decreased fruit weight and cortex density and improved the performance of the inherently slower part of the population under cold, wet conditions. Adding gibberellins (GA_4/7) or an N-substituted phthalimide (AC 94377) to the thiram steep was also beneficial whereas kinetin of N⁶-benzyladenine gave no improvement. Germination was even more rapid and better synchronised following a 4-day seed advancement sequence, particularly when this started with the acid steep. Overall, it was possible to increase the proportion of seeds which gave a root or produced hypocotyls at least 2cm tall by 9% and 14% respectively; the thermal time needed for the 90th seed to germinate was reduced from 73 to 30 day degrees and synchrony could be improved at least two-fold.     

Interactions between seed priming treatments and nine seed lots of carrot, celery and onion. I. Laboratory germination

Samples of three seed lots of each of three cultivars of carrot, celery and onion were primed in polyethylene glycol solution for 2 weeks at 15 °C. Priming reduced the mean germination times (recorded at 15 °C) of all seed lots (compared to the untreated control) by 3–4 days in carrot, 6–10 days in celery and 3–5 days in onion. The largest reductions in mean germination time occurred in the slowest-germinating seed lots. There were highly significant interactions between priming and cultivars, and between priming and seed lots within cultivars for each species. Drying back the primed seeds at 15 °C increased the mean germination times (compared to primed seed which had not been dried) by 0·6 day in carrot and 1·4 days in celery, and there was no interaction with cultivars or seed lots. The corresponding increase for onion was either 1·0 or 1·8 days, according to the cultivar, but this variation was largely attributable to differences in time taken for the dried seeds to re-imbibe. Seeds dried back at 30 °C germinated 0·2·0·7 day (depending on the species) later than those dried at 15 °C. Percentage germination was not affected by either priming or drying back. Priming reduced the spread of germination times in all cultivars. For primed and dried-back seed, the spread of germination times was larger than that of primed seed in certain cultivars, but was always smaller than that of untreated seeds.     

Integrating low water potential seed hydration with other treatments to improve cold tolerance

Matriconditioning improved the performance of pepper, tomato, sweet corn, snap bean, table beet, sugar beet and watermelon seeds in early field plantings at suboptimal temperatures (averaged over 10 d after planting) ranging from 12 to 18 °C. Reduction in the time to 50% (T(50)) emergence in conditioned seeds ranged from 0·6 d in watermelon to 3·3 d in pepper and improvement in emergence from 10% in sugar beet to 30% in table beet. Further improvement in emergence occurred by inclusion of pesticides and/or gibberellin during conditioning. A 4 d conditioning of pepper at 25 °C was superior to 7 d conditioning at 15 °C in seeds germinated at 15 °C on filter paper, but 15 °C conditioning was superior in improving percentage emergence in early field plantings. Tomato seeds conditioned at 15 or 25 °C performed equally well in the field. A 2 d conditioning was superior to 1 d conditioning in improving the performance of supersweet sweet corn cultivars grown in a growth chamber at 10/20 °C. The water uptake rate in the presence of Micro-Cel E during matriconditioning of sweet corn seeds was slower than when the seeds were exposed to the same amount of water in absence of the carrier. Electrolyte leakage was greater in supersweet 'Challenger' sweet corn seeds carrying the sh(2) gene compared to the sugary type sweet corn 'More', and in both cases matriconditioning reduced the leakage. Lettuce seeds matriconditioned for 24 h had higher 1-aminocyclopropane-l-carboxylic acid (ACC) content, developed greater ACC oxidase activity and performed better at 10 °C (germinated earlier and had higher percentage germination) than the untreated seeds. Matriconditioning appears to bring about beneficial physical, physiological and biochemical changes that seemingly improve embryo growth potential and tolerance to low temperatures.     

Cell cycle: sustained ERK signalling represses the inhibitors

The duration of ERK pathway signalling determines the proliferative response of cells. A recent study has shown that cell cycle re-entry requires the downregulation of an anti-proliferative gene cluster following sustained activation of the ERK pathway.     

Effects of combining priming and plant growth regulator treatments on the synchronisation of carrot seed germination

Osmotic priming of carrot seeds for 2 wk in polyethylene glycol (PEG, — 10 MPa) at 15 °C led to more rapid and synchronous germination at 20 °C compared to untreated seeds. These responses were enhanced by a 24 h pre-priming soak in water or a change of solution after the first 24 h of priming to remove leachate. The inclusion of 200 mg litre^-1N-substituted phthalimide in the pre-priming soak and/or in the PEG further enhanced the results of priming. Leachate removal combined with phthalimide inclusion gave 79% and 86% germination from seeds of two carrot cultivars during the first day in 20°C water following priming. In contrast, cumulative germination of untreated seeds of the same cultivars was 18% and 61% respectively after 3 days in 20°C water. Seeds primed in PEG containing 200 mg phthalimide litre^-1with the solution replaced after the first 24 h germinated earlier and more synchronously than untreated seeds over a range of germination temperatures (5, 10, 15, or 20°C), but the effects of priming were most marked at 5°C.     

Interactions between seed priming treatments and nine seed lots of carrot, celery and onion. II. Seedling emergence and plant growth

Samples of three seed lots of each of three cultivars of carrot, celery and onion were primed in polyethylene glycol solution for two weeks at 15 °C. Seedling emergence was recorded in the field for carrot and onion and in the glasshouse for celery. Compared to the untreated control, priming increased the percentage seedling emergence in certain poorly-emerging seed lots of carrot and celery, but had no effect on onion. Mean emergence times were reduced by priming in all seed lots, by 3–5, 5–8 and 3–9 days in carrot, celery and onion respectively. The largest effects occurred in the slowest-emerging seed lots. There were significant interactions between priming and seed lots within cultivars in carrot and celery and between priming and cultivars in celery and onion. Priming generally reduced the spread of emergence times, but the effects were not statistically significant in carrot. Drying back the primed seeds had no effect on percentage emergence in onion, but reduced it (compared to primed seed which had not been dried-back) in certain carrot and celery seed lots. Primed and dried-back seeds emerged later than primed seeds, by up to 1·5, 2·6 and 2·6 days in carrot, celery and onion respectively. The spread of emergence times was generally larger for primed and dried-back seeds than for primed seeds, but the differences were not always statistically significant. Plant fresh weights were recorded 9, 7 and 12 wk after sowing for carrot, celery and onion, respectively. In each species, mean plant weight was inversely related to seedling emergence time; thus plants grown from primed seed were always heavier than the controls, by up to 33%, 182% and 47% in carrot, celery and onion respectively.     

Genetic analysis of seed-soluble oligosaccharides in relation to seed storability of Arabidopsis

Seed oligosaccharides (OSs) and especially raffinose series OSs (RSOs) are hypothesized to play an important role in the acquisition of desiccation tolerance and consequently in seed storability. In the present work we analyzed the seed-soluble OS (sucrose, raffinose, and stachyose) content of several Arabidopsis accessions and thus identified the genotype Cape Verde Islands having a very low RSO content. By performing quantitative trait loci (QTL) mapping in a recombinant inbred line population, we found one major QTL responsible for the practically monogenic segregation of seed stachyose content. This locus also affected the content of the two other OSs, sucrose, and raffinose. Two candidate genes encoding respectively for galactinol synthase and raffinose synthase were located within the genomic region around this major QTL. In addition, three smaller-effect QTL were identified, each one specifically affecting the content of an individual OS. Seed storability was analyzed in the same recombinant inbred line population by measuring viability (germination) under two different seed aging assays: after natural aging during 4 years of dry storage at room temperature and after artificial aging induced by a controlled deterioration test. Thus, four QTL responsible for the variation of this trait were mapped. Comparison of the QTL genetic positions showed that the genomic region containing the major OS locus did not significantly affect the seed storability. We concluded that in the studied material neither RSOs nor sucrose content had a specific effect on seed storability.     

Effect of priming treatments on seed germination and seedling growth in bamboo [Dendrocalamus strictus (Roxb.) Nees]

Instances of flowering of bamboo species Dendrocalamus strictus are few and far between which are taken as opportunity by nurserymen to collect seeds for propagation. Germination of seeds is reported to be poor. Therefore, different seed priming treatments were applied to D. strictus seeds collected from Ranchi in order to obtain uniform and high germination. Under laboratory conditions, dehusking of seeds before sowing ensured cent percent germination. Seed priming with KNO₃ 1% solution resulted in 80.4% increase in germination followed by hydropriming by 16 h (73.1% increase). In field conditions, dehusked seeds gave 23.0% germination without any priming treatment. Priming treatment with KNO₃ 1% gave the highest rise in germination (39.1%) followed by hydropriming for 16 h (26.1%). Seeds with their seed coats intact could give germination of 9.5% when germinated without any treatment. A rise of 115.8% in germination was obtained by priming with KNO₃ 1% (final germination count 20.5%). The next best treatment was hydropriming for 16 h (final germination 18.5%, a rise of 94.7%). KNO₃ 1% also induced the earliest and the most rapid germination. When seedlings germinated in laboratory were transferred to soil, all seedlings from all treatments established successfully without any mortality whatsoever. Therefore, it is recommended that seeds should be primed for 8 h with 1% KNO₃ and germinated in laboratory or in farm house under normal atmospheric condition before transplanting the seedlings to soil.     

A comparison of two large-scale seed priming techniques

Leek seed lots of high (91%) and low (82%) viability were primed in aerated polyethylene glycol (PEG) solutions in Bubble-columns and by a non-osmotic priming technique in both 1987 and 1988 and the seeds were then sown in the field. Both methods of priming established a similar seed moisture content during treatment and, in the laboratory, produced seeds with more rapid and uniform germination than untreated seeds, with a greater advantage for Drum compared with Bubble-column priming in PEG. In the field, both priming techniques gave seedling emergence responses similar to those from priming in PEG by the laboratory-scale technique on filter paper. Both large-scale priming methods gave earlier and more uniform emergence than untreated seeds and gave similar or slightly higher levels of seedling emergence, except on one sowing occasion when seeds were stored before sowing followed by sowing into a drying seedbed.     

Proteomic analysis of arabidopsis seed germination and priming

To better understand seed germination, a complex developmental process, we developed a proteome analysis of the model plant Arabidopsis for which complete genome sequence is now available. Among about 1,300 total seed proteins resolved in two-dimensional gels, changes in the abundance (up- and down-regulation) of 74 proteins were observed during germination sensu stricto (i.e. prior to radicle emergence) and the radicle protrusion step. This approach was also used to analyze protein changes occurring during industrial seed pretreatments such as priming that accelerate seed germination and improve seedling uniformity. Several proteins were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry. Some of them had previously been shown to play a role during germination and/or priming in several plant species, a finding that underlines the usefulness of using Arabidopsis as a model system for molecular analysis of seed quality. Furthermore, the present study, carried out at the protein level, validates previous results obtained at the level of gene expression (e.g. from quantitation of differentially expressed mRNAs or analyses of promoter/reporter constructs). Finally, this approach revealed new proteins associated with the different phases of seed germination and priming. Some of them are involved either in the imbibition process of the seeds (such as an actin isoform or a WD-40 repeat protein) or in the seed dehydration process (e.g. cytosolic glyceraldehyde-3-phosphate dehydrogenase). These facts highlight the power of proteomics to unravel specific features of complex developmental processes such as germination and to detect protein markers that can be used to characterize seed vigor of commercial seed lots and to develop and monitor priming treatments.     

Intermittent hypoxia treatments cause cellular priming in human microglia

Obstructive sleep apnoea syndrome (OSAS) is a sleep-disordered breathing characterized by nocturnal collapses of the upper airway resulting in cycles of blood oxygen partial pressure oscillations, which lead to tissue and cell damage due to intermittent hypoxia (IH) episodes. Since OSAS-derived IH may lead to cognitive impairment through not fully cleared mechanisms, herein we developed a new in vitro model mimicking IH conditions to shed light on its molecular effects on microglial cells, with particular attention to the inflammatory response. The in vitro model was set-up and validated by measuring the hypoxic state, HIF-1α levels, oxidative stress by ROS production and mitochondrial activity by MTS assay. Then, the mRNA and protein levels of certain inflammatory markers (NF-κB and interleukin 6 (IL-6)) after different IH treatment protocols were investigated. The IH treatments followed by a normoxic period were not able to produce a high inflammatory state in human microglial cells. Nevertheless, microglia appeared to be in a state characterized by increased expression of NF-κB and markers related to a primed phenotype. The microglia exposed to IH cycles and stimulated with exogenous IL-1β resulted in an exaggerated inflammatory response with increased NF-κB and IL-6 expression, suggesting a role for primed microglia in OSAS-driven neuroinflammation.     

Mechanism of seed priming in circumventing thermodormancy in lettuce

Lettuce (Lactuca sativa L. cv Minetto) seeds were primed in aerated solutions of 1% K₃PO₄ or water at 15°C in the dark for various periods of time to determine the manner by which seed priming bypasses thermodormancy. Seeds which were not primed did not germinate at 35°C, whereas those which were primed for 20 h in 1% K₃PO₄ or distilled H₂O had up to 86% germination. The rate of water uptake and respiration during priming were similar regardless of soak solution. Cell elongation occurred in both water and 1% K₃PO₄, 4 to 6 h prior to cell division. Both processes commenced sooner in water than K₃PO₄. Radicle protrusion (germination) occurred in the priming solution at 21 h in water and 27 h in 1% K₃PO₄.

Respiration, radicle protrusion and cell division consistently occurred sooner in primed (redried) seeds compared to nonprimed seeds when they were imbibed at 25°C. Cell division and elongation commenced after 10 h imbibition in primed (redried) seeds imbibed at 35°C. Neither process occurred in nonprimed seeds. Respiratory rates were higher in both primed and nonprimed seeds imbibed at 35°C compared to those imbibed at 25°C, although radicle protrusion did not occur in nonprimed seeds which were imbibed at 35°C. It is apparent that cell elongation and division are inhibited during high temperature imbibition in nonprimed lettuce seeds. Seed priming appears to lead to the irreversible initiation of cell elongation, thus overcoming thermodormancy.

     

Drought resistance in rice seedlings conferred by seed priming

Seed priming is a method by which seeds are subjected to different stress conditions to impart stress adaptation in seedlings germinating and growing under stressful situations. Drought stress is a major reason behind failure of crops. We studied the effects of hydropriming, dehydration priming (induced by PEG), and osmopriming (induced by NaCl and KH₂PO₄) on subsequent germination, growth and anti-oxidant defense mechanisms of 2-week-old rice seedlings under continuing dehydration stress. Unprimed seeds grown in PEG showed significantly lower germination and growth along with significantly higher reactive oxygen species (ROS) and lipid peroxidation levels. Among the priming methods, 5 % PEG priming was found to be the best in terms of germination and growth rate along with the lowest amount of ROS and lipid peroxidation (malondialdehyde [MDA]) values. MDA levels were reduced significantly by all of the priming methods. Hence, reduction of lipid peroxidation may be a key factor underlying the drought tolerance produced by the priming treatments. Glutathione peroxidase (GPX) activity seemed to bear an excellent correlation with oxidative stress resistance through seed priming. The PEG priming produced minimum peroxidative damage and superior germination and growth rate along with efficient GPX activity, overexpressed MnSOD and maintenance of HSP70 expression in normal as well as in drought condition. Therefore, in PEG-primed seeds the existence of robust protective mechanisms is definitely indicated.     

Improving the performance of transplanted rice by seed priming

Transplanting is the major method of rice cultivation in the world, in which seedlings are raised in nursery and then transplanted into well puddle and prepared fields. The traditional nursery sowing method is tedious and produces week seedlings that reduce the final yield due to high mortality. The potential of seed priming to improve the nursery seedlings and thus the transplanted rice was evaluated in the present study. The experiment was conducted in the rice growing area (31.45° N, 73.26° E, and 193 m) of Pakistan, during 2004–2005. Seed priming tools employed during the investigation included traditional soaking, hydropriming for 48 h, osmohardening with KCl or CaCl₂ (Ψ_s −1.25 MPa) for 24 h (one cycle), 10 ppm ascorbate for 48 h or seed hardening for 24 h. Priming improved the initial seedling vigor and resulted in improved growth, yield and quality of transplanted fine rice while traditional soaking behaved similar to that of untreated control. Osmohardening with CaCl₂ resulted in the best performance, followed by hardening, ascorbate priming and osmohardening with KCl. Osmohardening with CaCl₂ produced 3.75 t ha⁻¹ (control: 2.87 t ha⁻¹) kernel yield, 11.40 t ha⁻¹ (control: 10.03 t ha⁻¹) straw yield and 24.57% (control: 22.27%) harvest index. The improved yield was attributed due to increase in the number of fertile tillers. Significant positive correlation was found between mean emergence time of nursery seedlings and kernel yield, nursery seedling dry weight and kernel yield, fertile tillers and kernel yield, and leaf area duration and kernel yield.