Morphological and biochemical changes of Aesculus chinensis seeds in the process of maturation

Morphological and biochemical changes of Chinese horse chestnut (Aesculus chinensis Bge.) seeds were studied during the process of maturation. Fruit and seed diameter increased linearly from DPA (Days Post Anthesis) 60 to DPA 150 and were stable thereafter. The weight of fruit and seed increased sigmoidally between DPA 100 and DPA 160. The water contents of whole seeds, axes and cotyledons had a sharp reduction from DPA 110 to DPA 130 but changed little after DPA 130. The water contents of whole seeds, axes and cotyledons were 1.42, 1.93 and 1.30?g?g^?1DW, respectively at the end of maturation. The starch content of cotyledons from mature seeds was high on a dry weight basis (31.4%) while the total soluble sugar content varied between 12 and 20%. In both axes and cotyledons, the total soluble sugar content decreased during the process of maturation, as did all five soluble sugar components measured (glucose, sucrose, trehalose, raffinose and stachyose). Sucrose and stachyose contents remained relatively high at the time of seed maturation, followed by glucose and a trace level of trehalose. The oligosaccharide/disaccharide (O/D) ratio in the axes was 3.1 at DPA 140. The high level of sucrose and stachyose might contribute to the greater desiccation tolerance of Aesculus chinensis seeds relative to typical recalcitrant tropical seeds. High levels of ABA content in seed coats and a low GA₃/ABA ratio in both seed coats and axes might also influence the desiccation tolerance and shallow dormancy of mature Aesculus chinensis seeds.     

色木槭和拧筋槭种子脱水耐性分析——基于含水量、发芽率和ABA含量变化的结果

通过采集不同发育阶段种子并对其进行自然干燥处理,以含水量、发芽率和ABA含量变化为指标,研究了色木槭和拧筋槭种子在自然发育和干燥过程中的脱水耐性变化.结果表明,色木槭和拧筋槭种子虽经历了成熟脱水,但种子仍不耐脱水;其发芽率和活力指数随发育进程而逐渐增加,但随自然脱水进程而逐渐降低;色木槭和拧筋槭种子最大脱水耐性分别出现在花后100 d和120 d;在整个发育过程中,两树种种子的ABA(脱落酸)含量均较低;拧筋槭种子中ABA含量在脱水过程中呈下降趋势,而色木槭种子中ABA含量在脱水过程中虽有升高但变化不显著.推测ABA含量低可能为色木槭和拧筋槭种子不耐脱水的主要原因之一.     

Effects of desiccation on the physiology and biochemistry of Quercus alba acorns

Seeds that lose viability when dried to a water content of less than 12% are said to be recalcitrant. We subjected acorns of Quercus alba L., a species with recalcitrant seeds, to desiccation to determine the effects of drying on lipids, proteins and carbohydrates of the embryonic axis and cotyledon tissues. Samples of fresh seed and seed dried for selected intervals were analyzed for water content and germination, and for lipids, proteins and carbohydrates by Fourier transform-infrared (FT-IR) spectroscopy. Carbohydrates were further analyzed by gas chromatography (GC). The FT-IR analysis revealed that membrane lipid structure initially exhibited reversible shifts between gel and liquid crystalline phases in response to drying and rehydration; however, reversibility declined as viability was lost. Changes in carbohydrate concentration were observed based on peak height comparisons; sucrose concentration in the embryonic axis increased dramatically after 5 days of drying. The most sensitive indicator of desiccation damage was the irreversible change in protein secondary structure in embryonic axes and cotyledon tissue. These changes were illustrated by shifts in amide absorbance near 1650 cm(-1). Gas chromatography indicated an abundance of sucrose in both the embryonic axes and the cotyledon tissue. Although sucrose concentrations in these tissues were initially similar, sucrose concentration in the embryonic axes became significantly greater than in the cotyledons as the acorns dried. We hypothesize that, in drying acorns, increased concentration of sucrose does not prevent loss of viability, but acts as a glycoprotectant against cell collapse and cell wall membrane damage as water stress increases.     

Effect of desiccation on viability and biochemical changes in Knema attenuata seeds

We studied effect of desiccation on Knema attenuata seeds in the laboratory conditions(28 ± 2℃) for 8 days.We also monitored biochemical changes during germination by following standard procedures.Seeds lost their viability in 8 days by decreasing to 18% of moisture content from its original value of 31%.The total sugar,protein and lipid contents decreased during seed ageing.An increase was noticed in the protease activity during desiccation.The phenolic content in seed exudates was increased during ageing.The result confirms the degradation of stored biomolecules in seeds during ageing.Desiccation sensitivity and biochemical changes during dehydration confirm its recalcitrant nature.     

自然脱水过程中七叶树种子生理特性的变化

于 2003 年研究了自然脱水过程中七叶树种子生理特性的变化。结果表明:七叶树种子对脱水高度敏感,为顽拗性种子。种子成熟采收时含水量高达 60.3%,种子在室温下自然干燥 30 天后,含水量下降到 30.2%,此时种子生活力完全丧失。在脱水初期,种子发芽率有所上升,但随后种子发芽率迅速降低。随着含水量的下降,种子浸出液相对电导率上升,但当含水量在 53.7% 到 50.9%之间时,种子浸出液相对电导率出现异常升高。同时种子超氧化物歧化酶(SOD)的活性随脱水时间的延长而下降,只有种子含水量在 53.7% 到 50.9%之间时出现例外。丙二醛(MDA)含量在脱水初期缓慢上升,在含水量下降到低于 50.9%时,丙二醛(MDA)含量快速上升。可溶性糖含量随脱水时间的延长缓慢上升。当种子含水量为 47%–60%时,种子发芽率较高,说明这一含水量区间有利于种子生活力的保持。图 6 参 13。     

Seed storage behavior of <Emphasis Type="Italic">Knema attenuata</Emphasis>, an endemic species of Western Ghats,India

We performed desiccation and storage trials to better under- stand storage behavior of Knema attenuata seeds. Mature seeds with moisture content (MC) of 31% exhibited 73% germination. During the period of desiccation (open lab condition) seeds with MC 23% showed 40% germination. After further drying to MC 21% germination was reduced to 16%. Complete loss in viability resulted when seed moisture was reduced to 18%. The seeds stored at -10°C, 0°C, 10°C and 28±2°C (open lab condition) lost their viability within 10 days. Seeds stored in sealed polythene bags and moist sand retained viability for more days than did seeds stored under all other storage conditions. Sensitivity of seeds to lower temperature and desiccation suggest that the storage be- havior of K. attenuata seeds is recalcitrant. Seeds stored in moist condi- tions can, at best, be stored for a period of two months.     

Germination and seedling growth of Quercus vulcanica: effects of stratification, desiccation, radicle pruning, and season of sowing

Effects of stratification, desiccation, radicle pruning, and season of sowing on Quercus vulcanica germination and growth were studied to identify optimum nursery procedures for artificial regeneration of this species. Following stratification (0, 2, 4 and 8 weeks), acorns were germinated, and acorn moisture content and germination performance were also determined at various times during desiccation. In early December unsprouted acorns were planted in containers, and in early April sprouted acorns, which had been stored in polyethylene bags at 4 °C, with radicles left intact or with radicles pruned back to 1.0 cm were planted in containers.Stratification for 2 and 4 weeks did not increase germination percentage but significantly increased germination rate. Germination percentage of the seeds dropped when the moisture content of the seeds was reduced by desiccation, and the critical minimum moisture content of the recalcitrant Q. vulcanica acorns was found to be 11–16%. Spring sowing of sprouted acorns altered the morphology the containerized seedlings and caused the formation of significantly more main roots, but resulted in significantly less shoot height and shoot dry weight. Spring-sown sprouted acorns had also a significant advantage over the fall-sown unsprouted acorns in seedling survival, and thus nursery personnel should not be concerned if acorns sprout before sowing.     

Abscisic acid and the regulation of embryo dormancy in the genus Acer

Germination of Acer pseudoplatanus L. and Acer platanoides L. seeds was increased by moist storage of the intact fruits (seed + pericarp) at 5 degrees C, but not at 17 degrees C. In both species, germinative capacity of isolated embryos (seeds with testa removed) was increased by moist storage of fruits at either 5 or 17 degrees C. Thus dormancy loss by intact seeds and excised embryos was not necessarily correlated. Loss of dormancy in isolated embryos was associated with a reduction in free abscisic acid (ABA) content. Embryos from freshly harvested fruits of A. pseudoplatanus had a higher germinative capacity and a lower free ABA content than embryos from freshly harvested fruits of A. platanoides. Morever, germinative capacity increased, and free ABA content decreased, more rapidly with length of fruit storage in A. platanoides embryos than in A. pseudoplatanus embryos.     

Abscisic acid in leaves and roots of willow: significance for stomatal conductance

Excised leaves and roots of willow (Salix dasyclados Wimm.) accumulated abscisic acid (ABA) in response to desiccation. The accumulation of ABA was greater in young leaves and roots than in old leaves and roots. In mature leaves, ABA accumulation was related to the severity and duration of the desiccation treatment. Water loss equal to 12% of initial fresh weight caused the ABA content of mature leaves to increase measurably within 30 min and to double in 2.5 h. The drying treatment caused significant (P = 0.05) reductions in leaf water potential and stomatal conductance. Recovery of leaf water potential to the control value occurred within 10 min of rewatering the dehydrated leaves, but recovery of stomatal conductance took an hour or longer, depending on the interval between dehydration and rewatering. The addition of ABA to the transpiration stream of well-watered excised leaves was sufficient to cause partial stomatal closure within 1 h and, depending on ABA concentration, more or less complete stomatal closure within 3 h. When the ABA solution was replaced with water, stomatal conductance increased at a rate inversely related to the concentration of the ABA solution with which the leaves had been supplied.     

Effects of radiational heating at low air temperature on water balance, cold tolerance, and visible injury of red spruce foliage

Recent studies have shown that winter needle mortality in red spruce (Picea rubens Sarg.) is increased by exposure to direct solar radiation, possibly as a result of photo-oxidative damage, accelerated winter desiccation, or reduced cold tolerance due to heating of sun-exposed needles. In an experiment at controlled subfreezing air temperatures of -10 to -20 degrees C, visible radiation was less effective than infrared radiation in producing needle desiccation and visible injury during freeze-thaw cycles. However, visible radiation produced a red-brown color in injured needles, similar to natural winter injury, whereas injured needles exposed to infrared radiation were yellow and injured needles kept in darkness were dark brown. Thus, visible radiation was necessary to produce the red-brown color of damaged needles, but not the injury itself. Needle desiccation was not strongly correlated with visible injury, but the pattern of variation in visible injury among trees and the positive correlation between electrolyte leakage and visible injury suggested that freezing damage following freeze-thaw cycles might cause the visible injury. This was confirmed by a second experiment that showed loss of cold hardiness in needles thawed by radiational heating for six consecutive days. Even with a constant nighttime temperature of -10 degrees C, six days of radiational heating of needles to above freezing caused a small (2.8 degrees C) mean decrease in needle cold tolerance, as measured by electrolyte leakage. Continuous darkness at -10 degrees C for six days resulted in an estimated 5.6 degrees C mean increase in needle cold tolerance. Freezing injury stimulated desiccation: cooling at 4 degrees C h(-1) to -43 or -48 degrees C increased the dehydration rate of isolated shoots by a factor of two to three during the first day after thawing. Within three days at 15 to 22 degrees C and 50% relative humidity, the mean water content of these shoots fell to 60% or lower, compared to 90% or greater for unfrozen controls or shoots subject to less severe freezing stress. In some but not all severely freeze-stressed shoots, accelerated needle desiccation and abscission were accompanied by a red-brown color typical of red spruce winter needle injury. We conclude that severe winter desiccation in red spruce may often be due to prior freezing injury, increased as a result of exposure to direct solar radiation. Furthermore, freezing injury in red spruce may sometimes cause desiccation and abscission of green needles.     

Roles of gibberellins and abscisic acid in dormancy and germination of red bayberry (Myrica rubra) seeds

Intact seeds from freshly harvested fruits of Myrica rubra (Sieb et Zucc.) were dormant and required 8 weeks of warm stratification followed by 12 weeks of cold stratification for germination. Exogenous application of gibberellic acid (GA(3)) to intact fresh seeds was effective in breaking dormancy, with > 70% of seeds germinating when treated with 5.2 mM GA(3) and incubated at a day/night temperature of 30/20 degrees C for 20 weeks. Removing the hard endocarp or endocarp plus seed coat of fresh seeds promoted germination, and addition of GA(3) to the embryo accelerated germination. The gibberellins GA(1) and GA(4) were more effective than GA(3) in promoting germination of seeds with the endocarp removed. Endogenous contents of GA(1), GA(3), GA(4), GA(7) and GA(20) were quantified by gas chromatography-mass spectrometry-selected ion monitoring in the endocarps, seed coats and embryos of fresh seeds treated with 5.2 mM GA(3). The content of GA(3) decreased in the endocarp during incubation, whereas GA(1) contents increased in the endocarp and seed coat. A high GA(1) content was detected in the endocarps and embryos of newly germinated seeds. We speculate that GA(3) was converted to GA(1) during incubation and that GA(1) is involved in seed germination. Endogenous abscisic acid (ABA) contents were measured in fresh seeds and in warm and cold stratified seeds. The ABA content in fresh seeds was distributed in the order endocarp > seed coat > embryo, with the content in the endocarp being about 132-fold higher than in the seed coat and embryo. Total ABA content of seeds subjected to warm or cold stratification, or both, was 8.7- to 14.0-fold lower than that of fresh seeds. Low contents of endogenous GA(1), GA(3), GA(7) and GA(20), but elevated contents of GA(4), were found in the seed coats and endocarps of warm plus cold stratified seeds and in the seed coats and embryos of newly germinated seeds. These observations, coupled with the finding that GA stimulated germination of dormant Myrica seeds, provide evidence that endogenous ABA inhibited release of dormancy and that endogenous gibberellins, especially GA(4) or GA(1), or both, are involved in germination.     

In situ experimental freezing produces symptoms of winter injury in red spruce foliage

Two mechanisms have been proposed to explain winter injury to needles of red spruce (Picea rubens Sarg.): (1) desiccation, which is characterized by net loss of foliar water from the needle to the environment, with cell injury resulting from dehydration; and (2) freezing, which is characterized by direct injury to cells resulting from intracellular or extracellular ice formation during exposure to low temperature. To compare the separate and combined effects of freezing and desiccation, branches of a mature red spruce at 1160 m were (a) experimentally frozen in situ to -50 degrees C; (b) cut and tied in their original orientation and allowed to desiccate passively; or (c) both frozen in situ and cut and tied in their original orientation. Needle water content, electrolyte leakage (an index of cell injury), and needle color were monitored for 60 days after treatment. Freezing resulted in immediate increases in electrolyte leakage, rapid water loss, and reddening necrosis of needles similar to that of naturally injured needles. Cutting resulted in more gradual water loss, no significant changes in electrolyte loss until severe desiccation had occurred, and a change in the color of the needles to a dull green. Because freezing produced reddening necrosis, a key symptom of winter injury, whereas desiccation did not, we conclude that freezing is probably the primary cause of winter injury in red spruce, and that desiccation is a secondary effect.     

Effects of presowing treatments, desiccation and storage conditions on germination of Strychnos nux-vomica seeds, a valuable medicinal plant

Seeds of Strychnos nux-vomica have slow and erratic germination; thus different presowing treatments were applied to enhance the germination of its seeds collected from Tamaraikulam, Tamil Nadu, India. In addition, the effects of desiccation and different storage conditions on the germination of S. nux-vomica seeds were investigated. The results show that soaking in 500 ppm gibberellic acid (GA₃) for 24 h, incubation of seeds at 40 °C for 3 days and alternate water soaking (16 h) and drying (8 h) for 14 days significantly increased the percentage germination compared to the control. Desiccation of seeds down to 10% moisture content resulted in better germination. Germination of S. nux-vomica seeds differed significantly between different storage periods, moisture contents of the seed and for first and second order interactions (p<0.001). The highest germination (92%) was achieved when seeds with 10% moisture content were stored at ambient temperature for 30 weeks. Evidence from the present study indicates that S. nux-vomica seeds possess physiological dormancy that can be broken effectively by after-ripening. As seeds of S. nux-vomica are found to be desiccation tolerant, dry seed (10% moisture content) can be hermitically stored at ambient temperature for 30 weeks without losing their viability.     

A probabilistic model for tropical tree seed desiccation tolerance and storage classification

Knowledge of seed desiccation tolerance is fundamental for conservation and use of forest species. The protocol used for classification of seed desiccation tolerance and storage is time consuming and many times limited by the lack of information about optimum conditions for seed germination and treatments to overcome seed dormancy. This study evaluated 66 Brazilian tree species aiming to correlate seed characteristics with desiccation tolerance. For this purpose, a model was established to explain the relationship of tegument/seed mass ratio (SCR), seed mass, and water content of embryo + endosperm with desiccation tolerance. The principal component analysis showed the establishment of two groups, indicating the interaction between desiccation tolerance and seed characteristics. Recalcitrant seeds are more often associated with the water content of embryo + endosperm and water content of tegument + endocarp, while orthodox seeds are more associated with SCR and number of seeds per kilogram. The classification found using the model proposed was significantly correlated with desiccation tolerance and storage, with 92% confidence for the analyzed species. Seeds morphological characteristics can be used for prediction of desiccation tolerance and storage behavior; however, the use of a model that combines more variables increases the chance of accurate classification.     

Effect of Seed Maturity and Pretreatment on Dormancy and Germination of Sorbus mougeotii Seeds

The effect of harvest time on seed germination, desiccation tolerance and level of dormancy is not known in Sorbus mougeotii (Soyer-Will. & Godron). Dry weight (DW), moisture content (MC) and leakage conductivity (LC) were tested on seeds from six harvest times. Chilling requirement and germinability were tested on seeds without drying and after drying and rehydration. The effect of postharvest ripening of seeds with or without drying was also tested. The DW was stable during the last 6 weeks of maturation, whereas MC initially decreased but later increased simultaneously with an increase in LC. Seeds were desiccation tolerant, deeply dormant and after a chilling treatment germinated to a high percentage at all harvest times. The chilling requirement decreased significantly with later harvest, suggesting a decrease in level of dormancy. Drying and postharvest ripening reduced and synchronized the chilling requirement.     

Seed Development, Maturation and Storage Behaviour of Mimusops elengi L.

Mass maturity (end of the seed-filling phase) occurred at about 72 days after flowering (DAF) in developing seeds of Mimusops elengi, at which time seed moisture content had declined to about 55%. The onset of ability to germinate was detected at 56 DAF and seeds showed 98% germination by 84 DAF. Tolerance of desiccation to 10% moisture content was first detected at 70 DAF and was maximal by 84 DAF. Delaying collection by a further 14 days to 98 DAF, when fruits began to be shed, reduced seed viability, particularly for seeds first dried to 10% moisture content. Hence the best time for seed collection appears to be about 14 days before fruits shed. In a separate investigation with six different seed lots, desiccation below about 8–12% moisture content reduced viability (considerably in some lots). The viability of dry seeds (below about 10% moisture content) stored hermetically was reduced at cool temperatures (5 °C and below), and none survived storage at sub-zero temperatures. The results suggest that Mimusops elengi shows intermediate seed storage behaviour and that the optimal hermetic seed storage environment is about 10% moisture content at 10 °C, while short-term, moist, aerated storage at high (40%) moisture content is also feasible.     

Effect of ABA on freezing resistance of Betula papyrifera and Alnus incana woody plant cell suspensions

Treatment of birch (Betula papyrifera Marsh) and alder (Alnus incana (L.) Moench) cell suspension cultures with ABA increased the freezing resistance of the cells. After 7 days of treatment with 10(-5) M ABA, birch cells grown at 23 and 4 degrees C attained an LT(50) of -16.9 and -14.1 degrees C, respectively, whereas control cells had an LT(50) of -9.1 degrees C. In alder cell suspensions, treatment with 10(-5) M ABA at 23 degrees C induced a small increase in freezing resistance from -7.3 to -10.8 degrees C. Exposure to 4 degrees C alone did not induce a significant increase in hardiness in birch cell suspensions. Addition of 10(-5) M ABA to the medium inhibited fresh weight increase over 10 days of 3-g inocula of birch and alder by 70 and 52%, respectively. With the same concentration of ABA in the medium we found different intracellular ABA concentrations in 3- and 6-g inocula. We conclude that the concentration of ABA in the medium does not reflect the intracellular concentration of tissue cultures, and that cultural conditions may influence ABA accumulation by cell cultures.     

Germination of Eucalyptus sieberi L. Johnson seeds. II. Internal water relations

At 15-20 degrees C, seeds of Eucalyptus sieberi L. Johnson need to maintain a water content equal to 30% or more of dry weight for approximately 60 h in order to germinate. Several shorter periods of imbibition at 30% water content separated by between 6 and 120 h desiccation were as effective as an unintenupted imbibition in bringing seeds to the point of germination. Even after 8 months' desiccation an earlier imbibition shortened the time required for the completion of germination, although it did not hasten the onset of germination. The water potential of seeds sufficiently hydrated to germinate was about -0.6 MPa, whereas that of the embryo was only about -4.5 MPa. The difference was caused by the inhibition of water uptake by the inner integument of the seed coat. During germination, reserve protein in a small group of cells above the collet was hydrolyzed. Subsequent expansion of these cells caused rupture of the inner integument thereby allowing further water uptake by the embryo. The balance of water absorption and evaporative loss by the outer integument was such that most seeds on a moist substrate but exposed to dry air (6.5-8 x 10(-4) MPa) did not maintain the 30% water content necessary for germination. Thus although able to germinate on seedbeds subject to intermittent drying, E. sieberi seeds have the capacity to avoid germination except under humid conditions.     

Effects of logging, liana tangles and pasture on seed fate of dry forest tree species in Central Brazil

We assessed seed fate of six tree species in a seasonally deciduous forest of Central Brazil. Seed germination, predation, removal and death caused by pathogens or dessication were followed in an undisturbed forest site, a logged forest site, and an active pasture. In both forest types we sowed seeds under patches of liana tangles (Low Forest habitat) and under patches of mature forest (High Forest habitat). Seeds were monitored weekly for up to 8 months. Overall, small-seeded species (Astronium fraxinifolium and Tabebuia impetiginosa) had high germination, although germination was lower in the pasture due to insect predation. The hard-coated seeds of Erythrina sp. had higher germination rates in the pasture. The large-seeded Cavanillesia arborea and Swartzia multijuga had high seed removal (ca. 90%) in the pasture and under High Forest, but only 23–34% under Low Forest in the logged site. Lower removal of large seeds in Low Forest suggests that this habitat conceals the seeds and constrains the movement of large mammals. Both species were favored in Low Forest, suggesting that caution should be used when managing lianas. When protected from predation, seeds with a soft coat and high water content, such as the seeds of Eugenia dysenterica and S. multijuga, died from desiccation in the pasture. Restoration efforts must take into consideration high mortality caused by desiccation or insect predation in abandoned pastures of dry forests.     

Changes of Plasma Membrane H~+-ATPase Activities of Glycine max Seeds by PEG Treatment

The soybean (Glycine max) Heihe No. 23 is sensitive to imbibitional chilling injury. Polyethylene glycol (PEG) treatment can improve chilling tolerance of soybean seeds to a certain extent. The changes of hydrolytic ATPase in plasma membranes and H+-pumping responses in soybean seeds were investigated during PEG treatments. Effects of exogenous calcium and exogenous ABA on the hydrolytic ATPase were also examined in order to understand the mechanism of chilling resistance. Highly purified plasma membranes were isolated by 6.0% aqueous two-phase partitioning from soybean seeds, as judged by the sensitivity of hydrolytic ATPase to sodium vanadate. PEG treatment resulted in a slight increase of the hydrolytic ATPase activity in 12 h. Then the activity decreased gradually, but still higher than the control. The H+-pumping activity increased steadily during PEG treatment. Exogenous calcium had both activating and inhibiting effects on the hydrolytic ATPase, but the activity was inhibited in soybean seeds treated with exogenous ABA. Results suggested that PEG treatment, not the exogenous calcium and ABA, up-regulated H+-ATPase activities in soybean seeds.