IDENTIFICATION OF PROCAMBIUM IN THE PRIMARY ROOT OF TRIFOLIUM PRATENSE (FABACEAE)

Histochemical and morphometric analyses were used to identify and define an early stage of procambial differentiation in 1.5–2.0-cm-long (48 hr after germination) primary roots of Trifolium pratense L. Esterase activity was used as a histochemical marker for early differentiation of procambium. Morphometric analysis of cell length and width as a function of distance from the root cap junction was performed on the same tissue using brightfield and Nomarski DIC optics. This combination of techniques allowed the identification of esterase activity in both the cell wall and cytoplasm and permitted the determination of the exact location, size, and shape of the histochemically stained cells within the apex. Esterase activity identified the proendodermis and procambial cylinder (six to seven cells in diameter) two to three cells proximal to the root cap junction. In this system, esterase activity proved to be an earlier marker for procambial differentiation than morphometric or cytological changes. It is suggested that these techniques will be useful in characterizing procambial pattern development in more complex shoot systems.     

Studies on the Fine Structure of the Scutellum of the Oat Seed During Germination

Structural changes in the seutellar parenchyma and epithelial cells of oats during the first 3 days of germination were followed by electron microscopy. The seutellar parenchyma cells contain more protein bodies than the epithelial cells, otherwise the general fine structures of the two types of cells arc quite similar: When the seed starts to germinate the protein bodies change into vacuoles and the proteins inside the protein bodies gradually disappear. Spherosomes are in abundance ill the seutcllar cells of the dry seed. Few disappeared during germination. Other cellular organelles, such as the mitochondria, endoplasmie reticulum, plastids, Golgi apparatus and glyoxysomes are scarcely seen in the seutellar cells of the dry seed. They become more obvious and easily recognizable after germination. In the dry seed, the walls of the epithelial cell that abut the endospernl show a two layered structure, consisted of an inner and outer layer. The outer layer becomes hydrolysed during seed germination, but the inner layer remains intact. The scutetlar epithelial cells are known for their ability to secret enzymes ute and absorb nutrients from the endosperm. But in the fine structural studies we have not been able to locate any specific strurcture that could be related to their known functions of enzyme secretion and nutrient absorption.     

Accumulation of Storage Materials,Precocious Germination and Development of Desiccation Tolerance During Seed Maturation in Mustard (Sinapis alba L.)

Under defined environmental conditions (20°C, continuous light of 15 klx) development of mustard seeds from artificial pollination to maturity takes about 60 d. After surpassing the period of embryo cell division and histodifferentiation (12–14d after pollination = dap), the seed enters into a maturation period. The time courses of various physiological, biochemical, and structural changes of embryo and testa during seed maturation were analyzed in detail (dry and fresh mass changes, osmotic and water potential changes, respiration, DNA amplification by endomitosis, total ribosome and polysome formation, storage protein synthesis and accumulation, storage lipid accumulation). In addition to the final storage products protein and lipid, embryo and testa accumulate transiently large amounts of starch within the chloroplasts during early maturation. Concomitantly with the subsequent total breakdown of the starch, the plastids lose most of their internal structure and chlorophyll and shrink into proplastids, typical for the mature seed. At about 30 dap the seeds shift from a desiccation-sensitive to a desiccation-tolerant state and are able then to germinate rapidly upon drying and reimbibition. If isolated from the immature fruit and sown directly on water, the seeds demonstrate precocious germination from about 13 dap onwards. Young seeds (isolated ≦ 38 dap) germinate only after surpassing a lag-phase of several days (after-ripening) during which the embryo continues to accumulate storage protein and lipid at the expense of the surrounding seed tissues. We conclude from these results that the maturing seed represents a rather closed developmental system which is able to continue its development up to successful germination without any specific regulatory influence from the mother plant. Immature seeds are able to germinate without a preceding dehydration treatment, which means that partial or full desiccation does not serve as an environmental signal for reprogramming seed development from maturation to germination. Instead, it is argued that the water relations of the seed are a critical element in the control of maturation and germination: during maturation on the mother plant the embryo is subject to a considerable turgor pressure (of the order of 12 bar) accompanied by a low water potential (of the order of ?12 bar). This turgor permits maturation growth but is subcritical for germination growth. However, upon imbibition in water, the low water potential provides a driving force for a burst of water uptake overcoming the critical turgor threshold and thereby inducing germination.     

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.     

Histochemical localization of alcohol dehydrogenase during the aging of seeds]

Alcohol dehydrogenase activity has been demonstrated in section of fresh and aged pea seeds, both air-dried and during germination. The activity in the various tissues has been observed using nitro-blue tetrazolium. From the comparison a general decrease appear in aged pea seeds and a tendency to a shift in the peaks of alcohol deydrogenase activity in the considered tissues (epidermis upper and lower, hypodermis, storage parenchyma and vascular system). The procambial zone forms the tracheary elements during germination, but in aged seeds this occur only rarely. The experiments here reported confirm the hypothesis that seed ageing includes changes in enzyme activity and in morphogenetic pathways.     

Bimodal changes in germinability of pea seeds under the influence of low doses of gamma-radiation

Pea seeds (cv. 'Nemchinovskii-85', harvest of 2002, 80%-germination percentage) were exposed to gamma-radiation with doses ranging from 19 cGy to 100 Gy. One week after the irradiation with doses of 19 cGy and 3 Gy. the germination percentage decreased to 58 and 45%, respectively; at doses of 7 and 10 Gy it was 73 and 70% respectively. At greater doses (25, 50, and 100 Gy), germination percentage decreased in proportion. Anomalous changes in seed germination percentage (as a function of irradiation dose) were caused by the redistribution of irradiated seeds between fractions I and II. The measurements of room temperature phosphorescence in air-dry seeds and the phosphorescence of endogenous porphyrines of imbibing seeds showen that the germination decrease after the irradiation with low doses (19 cGy and 3 Gy) was caused by the increase in the number of weak seeds of fraction II, which had high rates of water uptake and suffered from hypoxia under seed coat. Some of these seeds suffocated from hypoxia, and other seeds produced seedlings with morphological defects (such seeds were considered incapable of germination). During storage of seeds irradiated at doses 19 cGy-10 Gy, the recovery of germination percentage (after its initial decrease) was caused by the decrease in seed number in fraction II. The subsequent germination decrease was caused by seed death. The higher was the irradiation dose, the faster were changes in germination percentage during storage of irradiated seeds. Bimodal changes in pea seed germination with the increase of y-irradiation dose has apparently the same origin as the changes in seed germination during accelerated ageing.     

白花蛇舌草种子发芽及化感部位的研究

目的:研究白花蛇舌草种子发芽的最佳条件及化感部位。方法:采用正交实验寻找种子发芽的最佳条件,用生物工程技术研究不同部位对种子萌发的影响。结果:白花蛇舌草种子在25℃,24 h、20%的光照强度,采用机械破碎贮藏12个月发芽率最高,并且光照强度的变化对种子发芽的差异具有显著性;白花蛇舌草提取液中醇提组、水提组、乙酸乙酯组均存在化感作用。结论:寻找到白花蛇舌草种子发芽的最佳条件,白花蛇舌草化感作用可能是多组分共同作用的结果。     

The initiation of growth in isolated lima bean axes. Physiological and fine structural effects of actinomycin D, cycloheximide and chloramphenicol.

Excised lima bean axes were germinated for 24 hr in water, variousconcentrations of CHI, CAP or actinomycin D. None of the inhibitorsaffected water uptake before onset of growth. Growth was completelyinhibited by CHI and CAP, but only partly by actinomycin D.Rate of ³H-leucine incorporation rose during the lag phase inwater uptake and increased further during growth. The incorporationwas inhibited by CAP and CHI at all stages, but by actinomycinD only during growth. Actinomycin D prevented developmentalfine structural changes. In CHI treated axes number of mitochondriaincreased, O₂2 uptake was reduced, and higher cytochrome oxidaseactivity induced. CAP caused changes in the appearance of proteinvacuoles, but apparently prevented loss of storage material.The relation between the physiological and structural effectsof the inhibitors is discussed. (Received June 13, 1970; )     

Rapporti Tra Assunzione D'acqua,Metabolismo e Sintesi di Enzimi Nell'endosperma Di Semi Germinanti di Ricinus Communis

Abstract

Water uptake, activation of metabolism and enzyme synthesis in germinating castor bean seeds. — During the first days of germination water uptake by the castor bean seed endosperm is accompanied by a rapid rise of respiratory activity and of the « in vitro » detectable activity of a number of enzymes. The finding that the increase of enzyme activity is strongly inibited by protein synthesis inhibitors suggests an « ex novo » synthesis of enzymes in the endosperm of the germinating seed. The present investigation on the relationship between water uptake, metabolic activity and enzyme activity level lead to the following conclusions:

I - The increase of enzyme activity is strictly dependent on the availability of water, and on the rate of water uptake. When water uptake is depressed by incubation of the seed in high osmolarity media, enzyme activation is also severely depressed.

This is also observed when the seeds are germinating in contact with an amount of water consistently lower then the one they would taken up, in a given time (24 h), under conditions of unlimeted water availability.

II - The temperature coefficient of water uptake is close to 1.5 during the first 24 h, higher than 2 in the following 3 days. Low temperature almost completely inhibits the increase of enzyme activities in the endosperm.

III - Anaerobiosis inhibits the rate of water uptake by about 50%, in the first 24 h, and almost completely, in the following 3 days. Also the rise of enzyme activities is severely inhibited by lack of oxygen. The effect of protein synthesis inhibitors on water uptake is somewhat smaller, and the one on enzyme activity is somewhat larger than that of anaerobiosis.

These results are interpreted as indicating that during the early period of germination water uptake becomes more and more dependent on the metabolic activities of the endosperm cells, in as much the latter lead to the appearance of osmotically active substances and, possibly, to changes of the cell wall properties.

On the other hand, the level of hydration of the cytoplasm represents a limiting factor for the development of the mechanism involved in enzyme synthesis and metabolic activation.     

Effects of osmotic priming and ageing on the germination and emergence of carrot and leek seed

Carrot and leek seed was osmotically primed in polyethylene glycol solution (273 g/kg water and 342 g/kg water respectively) for 10, 14 or 17 days before accelerated ageing for 0, 24, 48, 72 or 96 h. Priming reduced the germination time compared with non-primed seed. Accelerated ageing increased germination and emergence times and decreased percentage germination and emergence to a greater extent for the primed seeds than for non-primed seeds in both species. Primed and dried but non-aged seed from both species was stored at 10°C for 12 months. There was no loss of viability and improvements in germination time due to priming were maintained throughout the storage period for all the priming treatments in leek, and for the 10 and 14 day priming treatments in carrot. Carrot seed primed for 17 days lost some viability after 12 months storage compared with non-stored seed.     

Water uptake and distribution in germinating lupine seeds studied by magnetic resonance imaging and NMR spectroscopy

Magnetic resonance imaging (MRI) was used to study temporal and spatial water uptake and distribution in germinating lupine ( Lupinus luteus L.) seeds. During 24 h of imbibition, water was unevenly distributed within the seed and some anatomical parts were more hydrated than others. Water entered the seed through the hilum and micropyle. The embryonic axis was the first to show hydration followed by seed coat and later cotyledons. The changes in water status were characterized by NMR spectroscopy. Analyses of T₂ relaxation times revealed a three-component water proton system (structural, intracellular and extracellular water) in germinating lupine seeds. The data on the components of transverse relaxation time studies indicated the complex exchange processes taking place between water components inside lupine seed over first 2.5 h of hydration, with a distinguished increase in structural water and decrease in other components. This speaks in favor of the high water-absorbing capacity of lupine seeds as related to high protein content. Germination was accompanied by swelling of protein bodies and changes in the organization of stored reserves with gradual disappearance of protein from the cells.     

Differences in the activity and distribution of peroxidases from three different portions of germinating Brassica oleracea seeds

Peroxidase (POD, EC 1.11.1.7) activity, cellular localization and isozyme patterns were investigated in the seed integument, cotyledon and embryo axis of Brassica oleracea cv. Cappuccio during pregermination and seedling growth. Seeds started to germinate after 24 h of imbibition. POD activity was localized in the pigmented layer of the integument and in procambial strands of the cotyledon and embryo axis in the first 24 h of imbibition. It was localized in the integumental cells of palisade, pigmented and aleurone layers and in epidermal, meristematic, procambial cells and xylem elements of the root and hypocotyl after 48 h of imbibition. POD activity increased during germination and early seedling growth: in the integument, it reached a maximum value after 72 h of imbibition, in the embryo axis and cotyledons, it increased up to 144 h of imbibition. The increase in peroxidase activity was accompanied by the appearance of new isozymes correlated with the development of seedling tissues. The isozyme profile was characterized by nine peroxidases: isoperoxidase of 50 kDa peculiar to integuments, that of 150 kDa to cotyledons and that of 82 kDa to the embryo axis. During pregerminative phase isozymes of 84 kDa were detected in the integument and cotyledons, of 48.5 kDa in the embryo axis. After germination, peroxidase activity and the complexity of the isozyme pattern increased, suggesting that they play a relevant role after rupture of the integument.     

CHANGES IN SENSITIVITY OF MAIZE CHROMOSOMES TO X RAYS DURING SEED GERMINATION

Latterell , Richard L., and Dale M. Steffensen . (Brookhaven Natl. Lab., Upton, L. I.. N. Y.) Changes in sensitivity of maize chromosomes to X rays during seed germination. Amer. Jour. Bot. 49(5): 472–478. Illus. 1962.—Changes in the radiosensitivity of maize seeds during early stages of germination were studied by means of somatic-mutation techniques. Seeds heterozygous for the yg₂ (yellow-green) locus were irradiated with 800 r of X rays after soaking in running tap water up to 42 hr. Yellow-green sectors, representing mutations affecting the dominant yg₂ locuss in leaves 4 and 5 of seedling plants were used as a criterion of radiosensitivity. The frequency of somatic sectors was virtually nil for dry seed and for seeds soaked up to 16 hr. Sector frequencies underwent a marked (9- to 15-fold) rise from 16 to 28 hr, reached a plateau of sensitivity and subsequently declined. Manometric studies were conducted on seeds soaked under the same conditions as those irradiated. The rate of oxygen consumption rose rapidly from 0 to 7 hr, remained essentially constant from 7 to 16 hr, then underwent an approximately 2-fold increase from 16 to 24 hr, after which the rate of progressive increase was retarded. The fact that the marked rise in frequency of X-ray-induced somatic sectors coincided with the major increase in oxygen consumption suggests that radiosensitivity of soaked seeds is conditioned by metabolic changes during seed development. Changes in radiosensitivity that followed attainment of peak sector frequencies were apparently governed by factors that influenced the rate of seed development.     

莲种子萌发和幼苗生长时期营养物质的代谢变化

莲子叶细胞中储存了丰富的营养物质,主要为蛋白质、淀粉和淀粉质体DNA.这些贮藏物质为种子萌发和幼苗的生长提供必需的能量和养料.通过组织化学和显微镜观察,研究莲从种子萌发到植株生长至具有4个节时,子叶中贮藏物质消耗的全过程.在此过程中,子叶中的贮藏物质不断降解,营养物质发生转运.蛋白体首先发生降解,其大量降解主要发生在幼苗三叶期.淀粉质体降解时会聚集成团,之后体积逐渐减小,最后完全降解.种子萌发后65天是子叶贮藏物质消耗末期,淀粉质体DNA的含量比萌发后20天的三叶期明显减少.细胞壁的形态结构发生多种形式的变化,细胞壁发生的这些变化与子叶细胞间物质的运输有关.含多糖的球形颗粒通过维管束在子叶中运输.     

莲种子萌发和幼苗生长时期营养物质的代谢变化

莲子叶细胞中储存了丰富的营养物质, 主要为蛋白质、淀粉和淀粉质体DNA。这些贮藏物质为种子萌发和幼苗的生长提供必需的能量和养料。通过组织化学和显微镜观察, 研究莲从种子萌发到植株生长至具有4个节时, 子叶中贮藏物质消耗的全过程。在此过程中, 子叶中的贮藏物质不断降解,营养物质发生转运。蛋白体首先发生降解, 其大量降解主要发生在幼苗三叶期。淀粉质体降解时会聚 集成团, 之后体积逐渐减小, 最后完全降解。种子萌发后65天是子叶贮藏物质消耗末期, 淀粉质体DNA的含量比萌发后20天的三叶期明显减少。细胞壁的形态结构发生多种形式的变化, 细胞壁发生的这些变化与子叶细胞间物质的运输有关。含多糖的球形颗粒通过维管束在子叶中运输。     

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.     

Characterization of water distribution and activities of enzymes during germination in magnetically-exposed maize (Zea mays L) seeds

Magnetic seed treatment is one of the physical pre-sowing seed treatments to enhance the performance of crop plants. In our earlier experiment, we found significant increase in germination and vigour characteristics of maize (Zea mays L.) seeds subjected to magnetic fields. Among various combinations of magnetic field (MF) strength and duration, best results were obtained with MF of 100 mT for 2 h and 200 mT for 1 h exposure. The quicker germination in magnetically-exposed seeds might be due to greater activities of germination related enzymes, early hydration of membranes as well as greater molecular mobility of bulk and hydration water fractions. Thus, in the present study, changes in water uptake during imbibition and its distribution and activities of germinating enzymes during germination were investigated in maize seeds exposed to static magnetic fields of 100 and 200 mT for 2 and 1 h respectively by nuclear magnetic resonance (NMR) spectroscopy. The magnetically-exposed seed showed higher water uptake in phase II and III than unexposed seed. The longitudinal relaxation time T1 of seed water showed significantly higher values and hence greater molecular mobility of cellular water in magnetically-exposed seeds as compared to unexposed. Component analysis of T2 relaxation times revealed the early appearance of hydration water with least mobility and higher values of relaxation times of cytoplasmic bulk water and hydration water in magnetically-exposed over unexposed seeds. Activities of alpha-amylase, dehydorgenase and protease during germination were higher in magnetically-exposed seeds as compared to unexposed. The quicker germination in magnetically-exposed seeds might be due to greater activities of germination related enzymes, early hydration of membranes as well as greater molecular mobility of bulk and hydration water fractions.     

甘蔗健康种子发芽率影响因素分析

该研究主要通过温室沙培试验,分析了甘蔗健康种子单芽材料各因素(单芽长度、不同芽位、泡水时间)及甘蔗基因型、种衣剂及保存天数三种因素对甘蔗健康种子发芽率的影响。结果表明:不同浸泡时间、不同单芽长度和芽位对健康种子单芽发芽率的影响极显著。随着单芽长度的增加,发芽率有明显的提高;随着泡水时间的增加,发芽率不断降低,但各处理之间差异不显著;顶部芽位的发芽率较高,但其他芽位之间发芽率的差异不明显。甘蔗健康种子发芽率具有明显的基因型差异,其中GT28和ROC22的发芽率较高,发芽率在70%左右。种衣剂包衣处可显著提高甘蔗健康种子的发芽率,与其他种衣剂相比扑力猛包衣处理的甘蔗健康种子发芽率最高,且差异显著。随着保存天数的增加,甘蔗健康种子的发芽率不断下降,且差异显著,当保存时间超过8 d时,发芽率低于60%。     

The presence of a p53-like protein during pea seed maturation and germination

ABSTRACT

The mechanisms that allow monitoring of DNA damage and the activation of repair systems in plants are poorly known. In mammalian cells the tumor suppressor protein p53 plays an important role in the checkpoint pathway induced by DNA damage. In this work, we investigated the presence and distribution of the p53-like protein in pea root tip nuclei and its role during early germination in relation to DNA damage. In pea seed, PFGE and TdT assays show that DNA fragmentation occurs during maturation and dry seed storage, and that this DNA fragmentation is repaired at the beginning of germination before the onset of proliferation. In the same seeds, the p53-like protein was found during maturation and germination. Immunoblotting characterization of this protein led to the identification of a single specific protein of about 94 kDa, more abundant at the beginning of the hydration process than in actively cycling cells. Furthermore, the p53-like protein revealed different nuclear distribution patterns, probably in relation to the formation of DNA fragments in dry seeds, and to the reactivation of repair mechanisms during early germination. These data suggest that the presence of a p53-like protein in quiescent or proliferating pea embryo cells is related to DNA damage, and serves for the maintenance of genetic information and the development of normal seedlings.     

Water uptake in germinating pecan (Carya illinoinensis) seed

• Water uptake is the fundamental and essential requirement for seed germination. Pecan seed has a hard woody endocarp that plays an important role during water uptake.
• To explore water uptake during germination, the spatiotemporal pattern of water and effect of the endocarp were analysed using high-field MRI, dye-tracing, wax blocking and SEM of water uptake.
• Isolated seeds completed water uptake in 8 h while whole seeds required 6 days, hence, cracking the endocarp plays an important role. The hilum is the channel through which water enters the seed, while the remainder of the seed coat consist of cells covered with a waxy layer that act as a barrier to water absorption. The region with the highest water content in pecan seed is the edge of the U-shaped region, and water can progressively diffuse from this U-shaped region into the whole kernel.
• We report a new water absorption stage between phase II and phase III of the triphasic model of water uptake of pecan seeds. Cracking the endocarp changed water distribution in pecan seeds, which may trigger further water absorption and radicle elongation.