Pepper seed germination assessed by combined X-radiography and computer-aided imaging analysis

A lot of pepper seeds having 87 % germination were subjected to X-ray inspection using a non lethal dose of radiation. Seeds with less than 2.7 % (on the basis of total seed area) of free space area, i.e. the spaces between embryo and endosperm, were classified as highly viable seeds (97–100 % germination) with the lowest level of abnormal seedlings. Seeds X-ray classified as good were subjected to a computerised image analysis to study seed imbibition and radicle elongation. The patterns of seed area increase, chosen as the most accurate indicator of seed swelling, resembled the triphasic curve of water uptake. The first phase was completed at 9 h followed by a second phase that varied widely in time until completion of germination between 52 and 96 h. The proportion of seeds with radicle protrusion between 52–56 h and 64–72 h assessed with the image analysis was significantly higher than that recorded using a conventional germination test. In addition, the rate of increase of seed area during the third phase of imbibition, mostly due to protrusion of the radicle tip and its growth, was highly correlated with the corresponding radicle elongation rate.     

The Effect of Low Temperature during Imbibition on Germination Characteristics of Tetracentron sinense (Tetracentraceae) Seeds

In this paper, the effects of low temperature, soil moisture and freezing time on the germination of Tetracentron sinense Oliv. seeds were studied by simulating wild environment conditions, and the factors influencing the natural regeneration of Tsinense population were discussed. The results are as follows: (1) The imbibition process of Tsinense seed can be divided into four stages, the water absorbing capacity of Tsinense seed after imbibition would increase with the increase of soil moisture, and the seeds would be fully imbibed under the condition of 30% soil moisture; (2) The germination rate, germination force and water content of Tsinense seed increased with the increase of storage time, and the vigor index gradually decreased after increasing before; (3) At different low temperature, the germination rate of Tsinense seed reached the highest when the soil moisture is 20% or 30%; (4) When treated at 20% soil moisture for 2-4 days or 21-28 days, 0℃ is best for the germination rate of Tsinense seed; when treated at 30% soil moisture for 14-28 days, -7℃ is best fort he germination rate of Tsinense seed; (5) When treated at the same condition of freezing time, the germination rate of seeds in fallen leaves had no significant difference with the decrease of low temperature; under the same condition of low temperature (0℃ low temperature except), the germination rate increased after frozen 28 days. The results showed that there is no significant influence of low temperature during imbibition on germination characteristics of Tetracentron sinense seeds and indicating that low temperature during imbibition is not the main factor influencing the natural regeneration of Tsinense population.     

Effect of Salinity on Germination and Seedling Growth of twoAtriplexspecies (Chenopodiaceae)

Salinity is one of the environmental factors that has a criticalinfluence on the germination of halophyte seeds and plant establishment.Salinity affects imbibition, germination and root elongation.However, the way in which NaCl exerts its influence on thesevital processes, whether it is through an osmotic effect ora specific ion toxicity, is still not resolved. Dimorphic seedsof the halophytesAtriplex prostrataandA. patulawere treatedwith various iso-osmotic solutions of NaCl and polyethyleneglycol (PEG). For each treatment, imbibition, germination rate,percent germination, germination recovery and nuclear area ofroot tip cells were compared. Higher concentrations of NaCl(-1.0 MPa) were more inhibitory to imbibition, germination andseedling root elongation than iso-osmotic PEG solutions. Allseeds recovered from a pre-treatment with -2.0 MPa NaCl andPEG solutions, except large seeds ofA. prostratawhich failedto germinate following transfer from -2.0 MPa NaCl. NaCl causeda greater increase in nuclear volume than iso-osmotic PEG solutions.These data suggest that the influence of NaCl is a combinationof an osmotic effect and a specific ion effect.Copyright 1998Annals of Botany Company Atriplex patula,Atriplex prostrata,cytophotometry, osmotic potential, salinity, seed germination.     

Proteomic analysis of rice (Oryza sativa) seeds during germination

Although seed germination is a major subject in plant physiological research, there is still a long way to go to elucidate the mechanism of seed germination. Recently, functional genomic strategies have been applied to study the germination of plant seeds. Here, we conducted a proteomic analysis of seed germination in rice (Oryza sativa indica cv. 9311) - a model monocot. Comparison of 2-DE maps showed that there were 148 proteins displayed differently in the germination process of rice seeds. Among the changed proteins, 63 were down-regulated, 69 were up-regulated (including 20 induced proteins). The down-regulated proteins were mainly storage proteins, such as globulin and glutelin, and proteins associated with seed maturation, such as "early embryogenesis protein" and "late embryogenesis abundant protein", and proteins related to desiccation, such as "abscisic acid-induced protein" and "cold-regulated protein". The degradation of storage proteins mainly happened at the late stage of germination phase II (48 h imbibition), while that of seed maturation and desiccation associated proteins occurred at the early stage of phase II (24 h imbibition). In addition to alpha-amylase, the up-regulated proteins were mainly those involved in glycolysis such as UDP-glucose dehydrogenase, fructokinase, phosphoglucomutase, and pyruvate decarboxylase. The results reflected the possible biochemical and physiological processes of germination of rice seeds.     

激素浸种对薄壳山核桃种子萌发的影响及其种子吸胀过程中SOD和POD活性变化

薄壳山核桃﹝Carya illinoinensis ( Wangenh.) K. Koch﹞为胡桃科( Juglandaceae)山核桃属( Carya Nutt.)植物,又名美国山核桃、长山核桃,是世界上著名的经济树种之一[1-2]。目前,主要采用本砧嫁接的方法培育薄壳山核桃良种嫁接苗,而培育优质实生苗作为砧木是该技术的关键环节之一,因此,了解其种子萌发特性极为重要。目前,对薄壳山核桃种子萌发特性的研究主要集中在植物激素催芽方面[3-5],而有关激素催芽过程中的适宜浸泡天数、种子吸胀过程中相关的酶活性变化等问题尚不明确。     

Development of combined imbibition and hydrothermal threshold models to simulate maize (Zea mays) and chickpea (Cicer arietinum) seed germination in variable environments

* The ability of hydrothermal time (HTT) and virtual osmotic potential (VOP) models to describe the kinetics of maize (Zea mays) and chickpea (Cicer arietinum) seed germination under variable conditions of water potential was investigated with a view to gaining an improved understanding of the impact of on-farm seed priming on seedling establishment through simulation. * Germination and/or imbibition time courses were recorded over a wide range of constant temperatures and water potentials and simple stepwise changes in water potential. * Both models adequately described germination under constant environmental conditions, but not conditions of water potential that varied. To test the hypothesis that this inaccuracy resulted from the use of ambient water potential, a parsimonious model of seed imbibition was developed to calibrate the HTT and VOP models (IHTT and IVOP) and drive them with estimates of seed water potential. * The IHTT and IVOP models described germination during stepwise changes in water potential more accurately than the conventional models, and should contribute to improved predictions of germination time in the field.     

Germination and Dormancy of Reed Canary-Grass Seeds (Phalaris arundinacea)

Effects of various chemical and physical factors on the germination of several seed lots of reed canary-grass (Phalaris arundinacea L.) have been studied. Germination at the optimum constant temperatures of 24 to 27°C was significantly stimulated by the following treatments: moist chilling in light, red light given during the first 3 days of imbibition, three 2-h periods at 12°C given during the second day of imbibition, ethylene, increased oxygen tension and soaking in aerated water for 4 days. Dry storage at 20–30°C had no effect on the germination ability of the seeds. No significant quantities of germination inhibitors were found either in water or methanol extracts of seed dispersal units. By comparing three cultivars with various degrees of seed dormancy, respiration measurements showed that there was a significant positive correlation between oxygen uptake prior to visible germination and germination capacity. Similarly, germination-stimulating treatment significantly enhanced oxygen uptake prior to visible germination.     

NaCl胁迫对2种表型盐地碱蓬种子萌发的渗透效应和离子效应研究

采用高低2个浓度的NaCl、LiCl及等渗甘露醇溶液处理紫红色表型(紫色型)和绿色表型(绿色型)盐地碱蓬种子,通过测定它们的种子萌发率、吸胀速率和胚内离子含量,研究NaCl胁迫对2种表型种子萌发的离子效应和渗透效应.结果表明:(1)2种表型盐地碱蓬种子萌发率在高浓度(300 mmol/L)和低浓度(100 mmol/L)NaCl处理下均显著降低,紫色型种子萌发率在低浓度下显著低于绿色型,而在高浓度下却显著高于绿色型;绿色型种子萌发率在高浓度(30 mmol/L)和低浓度(10 mmol/L)LiCl处理下均未受到显著影响,但紫色型种子萌发率却均极显著降低;2种表型盐地碱蓬种子萌发率在低浓度等渗甘露醇处理下均极显著低于低浓度NaCl处理,而高浓度等渗甘露醇处理却均与高浓度NaCl处理无显著差异.(2)2表型种盐地碱蓬种子的吸胀速率在低浓度NaCl处理下没有受到显著影响,但高浓度NaCl处理及与之等渗的高浓度甘露醇处理下都显著降低,而且紫色型种子的吸胀速率在等渗甘露醇处理时显著高于绿色型.(3)2种表型盐地碱蓬种子胚中的Na 含量和Na /K 在对照和低浓度NaCl处理下无显著差异,但紫色型种子胚中的Na 、K 含量在高浓度NaCl处理时都显著高于对照,且K 含量增加的幅度远大于Na 含量,导致紫色型种子胚中的Na /K 显著低于绿色型.研究发现,盐地碱蓬种子萌发在低浓度NaCl胁迫下主要受离子效应抑制,而高浓度NaCl胁迫下则主要受渗透效应抑制,紫色型种子萌发率在高浓度NaCl胁迫下高于绿色型的原因之一是前者能维持更低的Na /K 比.     

The nuclear protein Poly(ADP‐ribose) polymerase 3 (AtPARP3) is required for seed storability in Arabidopsis thaliana

The deterioration of seeds during prolonged storage results in a reduction of viability and germination rate. DNA damage is one of the major cellular defects associated with seed deterioration. It is provoked by the formation of reactive oxygen species (ROS) even in the quiescent state of the desiccated seed. In contrast to other stages of seed life, DNA repair during storage is hindered through the low seed water content; thereby DNA lesions can accumulate. To allow subsequent seedling development, DNA repair has thus to be initiated immediately upon imbibition. Poly(ADP‐ribose) polymerases (PARPs) are important components in the DNA damage response in humans. Arabidopsis thaliana contains three homologues to the human HsPARP1 protein. Of these three, only AtPARP3 was very highly expressed in seeds. Histochemical GUS staining of embryos and endosperm layers revealed strong promoter activity of AtPARP3 during all steps of germination. This coincided with high ROS activity and indicated a role of the nuclear‐localised AtPARP3 in DNA repair during germination. Accordingly, stored parp3‐1 mutant seeds lacking AtPARP3 expression displayed a delay in germination as compared to Col‐0 wild‐type seeds. A controlled deterioration test showed that the mutant seeds were hypersensitive to unfavourable storage conditions. The results demonstrate that AtPARP3 is an important component of seed storability and viability.     

曼陀罗种子休眠机理与破眠方法研究

通过对曼陀罗种子生活力测定、发芽试验、吸水率测定及种子萌发抑制物研究,揭示曼陀罗种子休眠机理,并利用物理、化学法处理曼陀罗种子,以探寻打破曼陀罗种子休眠的最佳方法.结果表明:(1)新采收的曼陀罗种子为综合休眠,休眠原因包括:种皮障碍、缺少萌发所需激素以及种皮和种仁中存在萌发抑制物,其中种皮障碍是限制种子萌发的首要因素.(2)室温存储6个月可解除曼陀罗种子种仁的休眠,但种皮障碍始终是其种子萌发的限制因素.(3)机械摩擦、浓H2SO4处理和NaOH处理均可打破除曼陀罗种皮的休眠障碍,促进种子萌发,其中用10% NaOH处理90 min为破除曼陀罗种皮休眠障碍的最佳方法,且发芽率比对照提高了83%.     

The use of an ELISA to quantitate the extent of 11S globulin mobilization in untreated and primed sugar beet seed lots

Seed priming (controlled imbibition) is a widely used technique for improving crop establishment, because it allows a reduction of the time to radicle emergence following seed imbibition and synchronization of individual seeds within seed lots with respect to germination timing. The major problem encountered in seed priming is the control of seed imbibition to a level permitting pre-germinative processes to proceed but that blocks radicle emergence. If not, the consequence of drying back the seeds to initial moisture content for storage purposes could be a total loss of the treated batch. This is because, as long as radicle growth has not begun, seeds may be re-dried without any permanent deleterious effects upon subsequent germination or growth. Recently, we reported the discovery of a molecular marker of sugar beet seed priming, corresponding to the basic B-subunit of the seed storage protein 11S globulin. An ELISA based upon this molecular marker has been used to analyse how different sugar beet seed lots respond to a priming treatment. The results demonstrate that this ELISA allows us to readily distinguish between the primed seeds and the corresponding untreated seeds.     

Cauliflower (Brassica oleracea L.) Seed Vigour: Imbibition Effects

By means of a machine vision facility, the process of water-imbibitionin a small seeded brassica species was recorded as the visibleincrease in seed volume. Dry cauliflower seed (Brassica oleracea)showed an immediate rapid phase of imbibition upon the additionof water. This initial phase was associated with the reductionin seedling root growth resulting from the imbibition of coldwater, and the rate of uptake of water at 20?C was negativelycorrelated with subsequent seedling growth. Damage to the testaof dry seed resulted in an increased rate of imbibition anda corresponding decrease in seedling root growth measured onslant boards. This showed that the intact testa of cauliflowerseed is capable of acting as a barrier to water influx and thata high rate of water uptake is damaging to the embryo. Testadamage reduced percentage soil emergence of seeds, as did raisingthe soil moisture content during the imbibition period by wateringimmediately after sowing. Both these treatments were believedto increase imbibition rate. Conditions which encouraged a lowrate of water uptake also improved the rate and uniformity ofemergence. Correlation of mean imbibition rates (measured ina laboratory test) with soil-emergence indicated that the sensitivityto imbibition damage varied between seed lots and interactedwith the absolute rate of water influx to determine the finalpercentage emergence. This factor prevented reliable predictionof seedling performance from the ranked-order of measured imbibitionrates. The significance of these findings to the seed productionand modular transplant raising industries is discussed. Key words: Machine vision, imbibition rate, testa, soil moisture, vigour     

Viability of onion (Allium cega) seed as influenced by temperature during seed growth

The yield and performance of seeds from crops of winter-hardy, bolting-resistant onion grown at temperatures of 15–16, 18–19 and 22–23°C in 1979, 1980 and 1982 were compared. Yields of seed from crops grown at 22–23°C were lower than those from crops grown at lower temperatures but the seeds ripened between 11 and 32 days earlier. Seeds from crops grown at mean temperatures of above 18°C gave higher percentage germination when imbibed at 30°C than 20°C and they also gave higher percentage seedling emergence than those from crops grown at lower temperatures. Seedlings from seeds produced at mean temperatures above 18°C were heavier than those from seeds of a similar weight but produced at lower temperatures. None of these differences were associated with differences in seed weight, embryo weight or seed dormancy but were positively correlated with differences in seed N-concentration. The differences were also associated with the rate of imbibition of water as high germination, high N-content seeds had a slower rate of imbibition than low germination, low N-content seeds of the same weight.     

Effect of high temperature on moisture loss,imbibition and germination of seeds ofTrianthema Govindia Buch.-Ham. ex DC

Summary Seeds of erect and prostrate plants ofTrianthema govindia Buch. ham. ex DC., growing in shade and open respectively, differed significantly in seed weight and percentage germination. Effect of high temperature exposure to these seeds has been studied in view of water depletion, imbibition and seed germination. The seeds of both the types were subjected to temperatures of 40, 50, 60, and 70° C for 24, 48, 96, and 144 hours. The three factors viz., loss of water, water imbibition and germination of seeds were positively correlated to the duration of treatment at different temperatures. A highly significant positive correlation was also observed between moisture depletion and imbibition, and between imbibition and germination. The percentage germination was favoured at 40° C in both the types of seeds and was increased with the increase of treatment duration. However, at higher temperatures (50 and 60° C) the percentage declined while at 70° C the seeds lost their vitality.     

ATP-levels of germinating seeds in relation to vigor

Determination of seed vigor was attempted by comparing ATP-levels of deteriorating seed to germination percentage and production of dry matter. Immediately after imbibition of any seed lot investigated, a production of ATP took place. This ATP-accumulation invariably reached a plateau after 6 h of imbibition. Two well germinating seed lots of rape, one of cauliflower and one of sugar beet, were artificially aged by means of elevated storage temperature and humidity. Every second week through 16 weeks of deterioration the levels of ATP, ADP and AMP after 7 h of imbibition were compared with the germination percentage. While ADP- and AMP-contents of germinating seed displayed no change (when imbibed 7 h) during the period of artificial aging, seed deterioration was reflected in the ATP-levels long before loss of viability could be detected by the conventional germination test.
When ATP-levels per seed were related to germination percentage throughout the aging, all four seed lots displayed similar patterns although the absolute figures differed. In contrast to the conventional "per seed' basis, however, ATP per gram seed not only displayed similar deterioration patterns, but the absolute values were also of the same magnitude.     

一氧化氮供体硝普钠浸种缓解盐胁迫对小麦种子萌发的抑制作用

以小麦品种‘德抗961＇为材料,用NO供体硝普钠（SNP）浸种研究外源NO对盐胁迫下小麦种子萌发的影响.结果表明：0.06 mmol/L的SNP浸种24 h后对盐胁迫下小麦种子发芽率、发芽指数、活力指数和吸胀速率的下调都有显著缓解作用;SNP浸种对盐胁迫下α-淀粉酶的活性无明显影响,但能显著提高盐胁迫下β-淀粉酶的活性;进一步研究表明,SNP浸种预处理对盐胁迫下的α-淀粉酶同工酶变浅的条带有所恢复（尤其是条带3）,同时使盐胁迫下变浅的β-淀粉酶同工酶的条带有明显的恢复（尤其是d、e、f、g）.并且SNP能显著降低盐胁迫下小麦地上部分和根中的Na^＋含量,提高其K＋含量,从而使K^＋/Na^＋显著提高.以上结果表明：SNP浸种预处理提高盐胁迫下小麦种子的萌发,主要是通过提高β-淀粉酶的活性来实现的.     

大豆种子萌发过程中的差异蛋白质组研究

运用蛋白质组学技术对大豆(Glycinemax)N2899种子萌发0h、8h、36h、60h4个时期蛋白质的差异表达情况进行了研究.结果发现,在考马斯亮蓝染色的双向电泳pH3～10胶上,PDQuest图像分析软件可识别的点约350个,其中表达量变化2.5倍以上的蛋白质点有24个,而绝大部分大豆种子贮藏蛋白在萌发期尚未降解.在萌发的第一阶段,24个差异表达蛋白中有10个蛋白质的丰度发生变化.第二阶段,差异表达蛋白的种类和量增加,其中15个蛋白质是动态变化的,14个蛋白质在胚根突破种皮时表达量达到峰值,表明吸胀后种子内的生命活动越来越强.对这24个蛋白质点进行胶内酶解,用基质辅助激光解析电离飞行时间质谱测定均获得肽质量指纹图谱.搜索大豆的UniGene库初步鉴定出6个蛋白质,分别是核苷二磷酸激酶、热激蛋白、硫氧还蛋白、35ku种子成熟蛋白及种子成熟蛋白PM36.对这些蛋白质在种子萌发过程中可能的作用进行了讨论.     

beta]-Tubulin Accumulation and DNA Replication in Imbibing Tomato Seeds

The activation of the cell cycle in embryo root tips of imbibing tomato (Lycopersicon esculentum Mill. cv Lerica) seeds was studied by flow cytometric analyses of the nuclear DNA content and by immunodelection of [beta]-tubulin. With dry seeds, flow cytometric profiles indicated that the majority of the cells were arrested at the G1 phase of the cell cycle. In addition, [beta]-tubulin was not detectable on western blots. Upon imbibition of water, the number of cells in G2 started to increase after 24 h, and a 55-kD [beta]-tubulin signal was detected between 24 and 48 h. Two-dimensional immunoblots revealed at least three different [beta]-tubulin isotypes. Thus, [beta]-tubulin accumulation and DNA replication were induced during osmotic priming. These processes, as well as seed germination rate, were enhanced upon subsequent imbibition of water, compared with control seeds that imbibed but were not primed. By contrast, when aged seeds imbibed, DNA replication, [beta]-tubulin accumulation, and germination were delayed. In all cases studied, both DNA replication and [beta]-tubulin accumulation preceded visible germination. We suggest that activation of these cell-cycle-related processes is a prerequisite for tomato seed germination. Furthermore, [beta]-tubulin expression can be used as a parameter for following the initial processes that are activated during seed imbibition.     

Dynamic analysis of Arabidopsis seed shape reveals differences in cellulose mutants

In a previous work, the shape of Arabidopsis seed was described as a cardioid modified by a factor of Phi. In addition, J index was defined as the similarity of the seed (in an orthogonal, bi-dimensional image) to a cardioid, thus allowing the quantitative comparison of seed shape in seeds of varieties and mutants at different stages of development. Here, J index is used for modeling changes in seed morphology during the dynamic process of seed imbibition before germination. The analysis was carried out by means of a general linear model with two fixed factors (genotype and time) applied to two Arabidopsis varieties: Columbia and Wassilewskija and two mutants in cellulose synthesis: prc1-1 (CESA6 in Columbia) and kor1-1 (in Wassilewskija). Equations representing the changes in seed form during imbibition are given. The analysis of changes in seed shape by this procedure provides (1) a quantitative method to record changes in seed shape and to compare between genotypes or treatments showing the time points with maximum differences, and (2) the observation of remarkable differences between wild-type seeds and mutants in cellulose biosynthesis, indicating new phenotypic characteristics previously unknown in the latter. While wild-type seeds increase their J index values during imbibition, in the cellulose mutants J index values decrease. In addition, shape comparisons were done with other mutants. Seeds of ga1-1 mutants behave like cellulose mutants, whereas different ethylene mutants present varied responses. Quantitative analysis of seed morphology is a new basis for the record of differences between wild-type and mutants as well as for phenotypic characterization.     

Proteolytic and Trypsin Inhibitor Activity in Germinating Jojoba Seeds (Simmondsia chinensis)

Changes in proteolytic activity (aminopeptidase, carboxypeptidase, endopeptidase) were followed during germination (imbibition through seedling development) in extracts from cotyledons of jojoba seeds (Simmondsia chinensis). After imbibition, the cotyledons contained high levels of sulfhydryl aminopeptidase activity (APA) but low levels of serine carboxypeptidase activity (CPA). CPA increased with germination through the apparent loss of a CPA inhibitor substance in the seed. Curves showing changes in endopeptidase activity (EPA) assayed at pH 4, 5, 6, 7, and 8 during germination were distinctly different. EPA at pH 4, 5, 6, and 7 showed characteristics of sulfhydryl enzymes while activity at pH 8 was probably due to a serine type enzyme. EPA at pH 6 was inhibited early in germination by one or more substances in the seed. Activities at pH 5 and later at pH 6 were the highest of all EPA throughout germination and increases in these activities were associated with a rapid loss of protein from the cotyledons of the developing seedling.