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.     

AN ULTRASTRUCTURAL AND MICROSPECTROPHOTOMETRIC STUDY OF THE SHOOT APEX DURING THE INITIATION OF THE FIRST LEAF IN GERMINATING PINUS BANKSIANA

The morphology and anatomy of the shoot apex in germinating Pinus banksiana seeds is described by using scanning and transmission electron microscopy and microspectrophotometry, with special attention given to events preceding the appearance of the first leaf primordia at about 72 hr post-imbibition. The 2C nuclei begin DNA synthesis at about 43 hr. RNA increases until 52 hr and is followed by a reduction related to cytokinesis. Protein drops after 36 hr, apparently related to digestion of storage protein bodies, which by 48 hr are about 50% digested. The resulting protein body vacuoles do not enlarge. Starch is digested just prior to appearance of the leaves and may be mediated by α-amylase production from stacks of endoplasmic reticulum. Heterochromatin increases in the nuclei during germination and coincides with an increase in repeated nucleotide sequences. Golgi bodies increase in number after the first mitoses.     

THE UPTAKE OF MERCURY BY SEEDS TREATED WITH MERCURIC CHLORIDE SOLUTION, AND ITS SUBSEQUENT RELEASE

SUMMARY: When sweet pea seeds were soaked in mercuric chloride solution a considerable quantity of mercury was taken up and was not removable by a washing procedure even more thorough than that usually employed in the technique for the surface sterilization of plant tissues. To remove the bulk of it, soaking the seed for 48 hr was needed.     

The level of proteins and leakage of solutes in germinating fresh and stored seeds ofCicer arietinum L.

One-year-old seeds of chickpea (Cicer arietinum L. cv. C-235) lost about 23 % germinability and leaked larger quantities of N, P, K, saccharides and proteins into the soaking medium in the first 48 h, as compared with fresh seeds. The protein content in stored seeds decreased more than in fresh seeds, as the soaking progressed.     

Imbibition and Germination: Influence of Hard Seed Coats on RNA Metabolism

RNA metabolism of Indian rice grass seeds was studied in relation to imbibition and germination. These seeds germinate only after scarification of the seed coats. The hard seed coats restrict germination but not water intake or changes in the quantity and quality of RNA formed during early hours of soaking. These changes differ markedly from those in scarified (H₂SO₄ treated) seeds which are able to germinate. Gibberellic acid hastens germination of scarified seeds and causes changes in the population of RNA transcribed.     

赤霉素和温水浸种对紫斑牡丹种子下胚轴萌发及其种胚解剖结构的影响

以紫斑牡丹种子为试验材料,采用不同浓度(0、100、300、500 mg/L)赤霉素(GA3)浸种和不同温度温水浸种(25、30、40和50℃)处理,考察各处理种子生根率的变化,并通过透射电镜观察不同生根发育时期种子种胚超微结构,探究赤霉素和温水浸种处理解除紫斑牡丹种子休眠进程中种胚超微结构变化以及这种变化与休眠解除的相关性。结果表明:(1)GA3和温水浸种均明显提前了紫斑牡丹种子生根时间,并以300 mg/L GA3和40℃温水处理的生根效果最好,分别较对照提前14.7 d和16.0 d,生根率分别达62.33%和67.00%。(2)光学显微镜观察发现,GA3、温水浸种处理较对照组紫斑牡丹种胚子叶的上、下表皮细胞形状及排列方式无明显影响,但其厚度均变薄,维管束结构明显,以300 mg/L GA3和40℃温水浸种处理生根效果最佳。(3)透射电镜观察发现,在各GA3浸种和温水浸种处理下,紫斑牡丹种子萌发前期,种胚子叶脂类物质出现降解现象,脂体呈大小不一的圆形或块状分布在细胞壁周边,数量明显减少,细胞核明显,核仁清晰,出现少量线粒体,细胞质内开始有蛋白质积累,在液泡的周围形成蛋白质沉积物;在种子萌发后期,种胚子叶细胞内物质稠密,脂体已降解融合成较大块状,细胞中储存的营养物质基本降解完全,内质网、高尔基体、线粒体等细胞器出现,结构更加完整,表明此时细胞内物质代谢活动加强,种子休眠解除,且300 mg/L GA3、40℃温水浸种处理种子种胚子叶结构变化最为明显。研究发现,300 mg/L GA3浸种和40℃温水浸种均可显著提高种子的萌发率,且40℃温水浸种的效果更好,在实际生产中建议使用温水浸种法。     

Glutamic acid decarboxylase in different areas of the developing chick central nervous system

The temporal course of the development of GAD activity in GABAergic neurons was studied in the chick retina, optic lobe and cerebellum. The developmental pattern of GAD activity was similar in the three areas studied, showing typical sigmoideal curves, which reached a maximal value at the 3^rd post-hatching day. Kinetic studies during development revealed that K_m remained unchanged while V_max increased 3-fold in the retina (48.99±0.84 nmol/hr/mg protein), almost 4-fold in the optic lobe (162.77±4.32 nmol/hr/mg protein) and 3.5 fold in the cerebellum (69.30±1.26 nmol/hr/mg protein). The developmental pattern of GAD activity in homogenates of the three areas studied from dark-reared and light-reared chicks with respect to normal light-dark cycle animals showed no significant differences. These results indicate that the increase in GAD activity during development are not due to a change in the affinity for its substrate but rather to changes in the concentration of the enzyme. The developmental pattern of GAD activity in the chick visual system was not affected by environmental conditions suggesting that the developmental profile is lightindependent.     

Eco-physiological studies on desert plants

Summary The O₂ uptake and RQ of germinating negatively photoblastic Zygophyllum coccineum seeds were studied during the first fourty hours after soaking in water under dark or light conditions. Five phases could be distinguished in the course of O₂ uptake in water in dark. The respiration course in light did not differ from that in dark till the 12th hour after soaking, then it deviated showing a low level, The RQ values in dark and light are nearly identical; both had sharp decline between the 8th and the 16th hour after soaking. Light, by blocking germination, affected the O₂ uptake.Moisture stress simulated by mannitol solutions caused a decrease in the O₂ uptake of seeds and seedlings. Na₂SO₄ caused a reduction and a lag in the time of maximum O₂ uptake. NaCl showed a particular effect on respiration of germinating seeds causing earlier rise and decrease in O₂ uptake than in water. NaCl was effective in rising the O₂ uptake of seedlings specially those supplied with glucose. Its effect was not pronounced on starved seedlings. O₂ uptake of seedlings germinated in NaCl was considerably high when compared with that of seeds germinating in the same medium 40 hours after soaking.The decreasing effect of moisture stress caused by mannitol on O₂ uptake was reversed by diluting the medium or replacing it with water. Values of O₂ uptake on number of seedlings basis are different from those calculated on fresh weight basis due to the difference in water content of seedlings germinated under various conditions.     

Effect of temperature of imbibition on phospholipid metabolism in pea embryonic axes

Pea seeds were imbibed in radioactive choline and then germinated. Treatments were either at 5° or 25° and the seeds were imbibed for 5 hr at one temperature and then transferred to the other. [$\text{Me}$-¹⁴C]Choline incorporation into phosphatidyl choline in the ER and the plasma membrane obtained from the embryonic axes after germination was measured. Seeds kept constantly at 25° had a very rapid initial incorporation of choline followed by a loss of label. Seeds kept at 5° had a very much lower rate of incorporation. However, seeds transferred from 5 to 25° behaved for at least 48 hr as if continuously kept at 5°, while in seeds transferred from 25 to 5° incorporation stopped after 15 hr. The seeds apparently respond to transient exposure to temperature by a changed metabolism of phospholipid. Data are also given for the choline content of the seeds under the different treatments and for the changes in total phospholipid.     

Development of protein bodies and accumulation of carbohydrates in a soybean (Leguminosae) shriveled seed mutant

The soybean seed mutant T311, when grown under specific environmental conditions, produces shriveled seed. This research investigated changes in development of protein bodies and accumulation of carbohydrates during seed development by comparing the mutant with P2180 seeds. The shriveled seeds contained larger protein bodies but fewer protein bodies per cell than round seeds. Protein bodies in T311 seeds included more dispersed crystals and less globoid regions than P2180 seeds. The elemental compositions of the crystals and of whole seeds in T311 were different from that in P2180 seeds. Starch breakdown was reduced with concomitant lower soluble sugar content in T311 seeds after the D11 stage (10.0-11.9 mm long seeds). The reduced starch breakdown and lowered soluble sugar content were consistent with lower a-amylase activity and earlier and greater water loss in T311 seeds. Changes in development of protein bodies and accumulation of carbohydrates were associated with the development of the shriveled seeds.     

The reserve proteins in the cells of mature cotyledons ofLupinus albus var.Lucky. I. Quantitative ultrastructural study of the protein bodies

Summary A quantitative ultrastructural study of the protein bodies of the lupin cotyledonary cells was performed to determine the protein content per cell. Two kinds of protein body were observed by transmission and scanning electron microscopy whatever the cellular type within the cotyledon. Some of these were conventional spherical structures, entirely filled with a dense protein matrix, others exhibited one large or several small light inclusions within the dense matrix. Even when a few light areas contained globoids, the majority remained of unknown nature, but could not be considered proteinaceous since they never reacted with specific protein dyes. The reserve protein content per cell was determined by image analysis on two seeds (L₁ and L₂) selected because they had a markedly different total protein content. The volume occupied by the dense matrix of the intracellular protein bodies was considered representative of the reserve protein quantity. The protein content per cell increased from the periphery to the centre of the cotyledon in the two seeds studied. The protein content per cell of the L₂ seed was generally found to be greater than the L₁ seed, in particular in the abaxial zone where it was markedly higher. The difference in total protein content of the two seeds was demonstrated to be primarily due to a differential alveolation of the protein bodies. These results will be used to study the relationship between the protein content of the cotyledonary cells and their nuclear DNA content.     

油菜素内酯浸种对盐胁迫番茄种子萌发的影响及其生理机制

为揭示油菜素甾醇类化合物提高作物耐盐的效应和机理,研究了10^-11、10^-10、10^-9、10^-8、10^-7、10^-6、10^-5 mol/L 2,4-表油菜素内酯（EBL）浸种处理对0、50、100、150、175 mmol/L NaCl胁迫7 d的番茄种子萌发、生长、溶质积累、抗氧化代谢的影响。结果显示：NaCl浓度越高的盐胁迫下,10^-9 mol/L EBL浸种可体现出越显著的促进番茄种子萌发的效应;在所有处理下,EBL浸种浓度过高,即10^-6、10^-5 mol/L EBL,均表现出对种子萌发的抑制效应。盐胁迫下种子萌发后,一定浓度的EBL浸种可表现出明显的增加种子胚根和下胚轴长,提高萌发种子鲜重和种子活力指数,其中10^-9 mol/L EBL浸种处理促进效果最适;EBL浸种浓度过高,则表现出抑制效应。150 mmol/L NaCl胁迫或非盐胁迫下,10^-9 mol/L EBL浸种均可降低萌发种子体内的O₂^·-、H₂O₂、丙二醛（MDA）和脯氨酸（Pro）含量;盐胁迫下,10^-9 mol/L EBL浸种可显著提高萌发种子可溶性糖（SS）和可溶性蛋白（SP）的含量。150 mmol/L NaCl胁迫或非盐胁迫下,10^-9 mol/L EBL处理可不同程度促进番茄种苗超氧化物歧化酶（SOD）和过氧化物酶（POD）活性的上升。综上所述,盐胁迫下,一定浓度范围内的EBL浸种可明显促进番茄种子萌发或成苗,其中以10^-9 mol/L EBL浸种的效果最好,主要是因为EBL施用可积极促进番茄种子萌发中物质转化,SS和SP等溶质积累增多,增强其渗透调节能力;同时SOD和POD酶活增强,缓解盐胁迫导致番茄种子萌发中的次生氧化胁迫。     

Interrelations between Carbon Dioxide and Ethylene on the Stimulation of Cocklebur Seed Germination

Interrelations between CO₂ and C₂H₄ on promotion of seed germination were examined in more detail at 23°C with presoaked upper seeds of Xanthium pennsylvanicum Wallr. The germination-promoting effect of C₂H₄ decreased gradually as its application time was delayed during a soaking period, whereas CO₂ was most promotive in application at 5 days of soaking, then its effect declined. CO₂ and C₂H₄ were additive in earlier soaking periods and synergistic in later periods. Such changes in germination behavior in response to CO₂ and/or C₂H₄ during a soaking period were closely associated with growth responsiveness of the axial tissues, but not of the cotyledonary ones. Growth responsiveness of axial tissues to CO₂ or C₂H₄ disappeared finally during a soaking period, but their extinct responsiveness to any one of these gases was almost fully restored in the simultaneous presence of the other. The extinct responsiveness to CO₂ was partially recovered by a preexposure to C₂H₄. This suggests that in the later period of soaking, unlike the case in a very early period of soaking, the C₂H₄-sensitive phase for seed germination precedes the CO₂-sensitive phase in which CO₂ potentiated axial growth. The restoration of CO₂ responsiveness in axial growth occurred not only after C₂H₄ treatment but also after exposure to 8 or 33°C or after KCN treatment. Thus, secondarily dormant Xanthium seeds could germinate in response to CO₂ alone, when they were previously exposed for shortterms not only to C₂H₄ but also 8°C, 33°C, or KCN.     

Immunocytochemical localization of concanavalin A in developing jack-bean cotyledons

The lectin, concanavalin A (Con A), was localized in the cotyledon of developing jack beans (Canavalia ensiformis (L.) DC) by electron-microscope immunocytochemistry. In mature seeds, Con A was present in protein-storage vacuoles (protein bodies) of storage-parenchyma cells. Although protein bodies could be seen in other cell types, only protein bodies in storage-parenchyma cells contained Con A. During seed development, Con A was also localized on the endoplasmic reticulum and Golgi apparatus, presumably en route toward deposition within the protein bodies. The intensity of labeling of the endoplasmic reticulum was much greater during the developmental stage of protein-body filling (66% final seed weight) than in mature seeds.Abbreviations Con A concanavalin A - ER endoplasmic reticulum - IgG immunoglobulin G     

Timing of the first cleavage post‐insemination affects cryosurvival of in vitro–produced bovine blastocysts

The time of the first cleavage of bovine zygotes during in vitro culture can affect the rate of development and cell number of the blastocysts. The aim of this work was to study the effect of the timing of first cleavage on the cryosurvival of the resulting blastocysts. Following standard IVM and IVF, zygotes were cultured in modified synthetic oviduct fluid (SOF), with 10% fetal calf serum (FCS) added 48 hr post insemination, in a humidified atmosphere of 5% CO_2, 5% O₂ and 90% N₂. Embryos which cleaved by 24, 27 30, 33, or 36 hr after insemination (IVF) were harvested and further cultured to the blastocyst stage (day 7 or day 8 post IVF). All developing blastocysts on days 7 and 8 were classified into three groups and were cryopreserved by vitrification. Group A consisted of blastocysts (<150 μm, small blastocysts); group B consisted of expanded or hatching blastocysts (>150 μm, large blastocysts); and group C consisted of morphologically poor quality blastocysts. The vitrification solution consisted of 6.5 M glycerol and 6% bovine serum albumin in PBS (VS3a). Thawed embryos were cultured further and survival was defined as the re‐expansion and maintenance of the blastocoel over 24, 48, and 72 hr, respectively. Overall survival and hatching at 72 hr post‐thawing was higher in blastocysts formed by day 7 than those formed by day 8 (60% vs. 40% survival; 63% vs. 45% hatching). Large blastocysts from day‐7 and day‐8 groups survived significantly better than small or poor quality blastocysts (76% vs. 63% and 31%; 72% vs. 30% and 26%, respectively; P < 0.05). Day‐7 blastocysts from the 27‐ and 30‐hr cleavage groups survived significantly better than those from the 36‐hr group (63% and 66% vs. 25%, P < 0.05). Day‐8 blastocysts from later cleaved (30 hr) zygotes had a higher survival than the 27‐hr cleavage groups (52% vs. 26%, P < 0.05). These results indicate that the day of blastocyst appearance, developmental stage, and timing of the first cleavage post‐insemination can influence the cryosurvival of bovine blastocysts following vitrification. Mol. Reprod. Dev. 53:318–324, 1999. © 1999 Wiley‐Liss, Inc.     

Immunocytochemical localisation of lectins in cells of Phaseolus vulgaris L. seeds

Antibodies against pure E₄- and L₄-lectins from the seeds of Phaseolus vulgaris L. raised in rabbits were made monospecific by immunoaffinity chromatography on E₄- or L₄-lectin Sepharose 4B columns. Localisation of lectins in bean seeds was investigated by indirect immunofluorescence and by electron microscopy on sections stained with colloidal gold particles coated with monospecific anti-E₄- and anti-L₄-IgG. In parenchyma cells from the cotyledons both E- and L-type lectins were found inside the protein bodies. Apparently the matrix of all protein bodies contained both types of lectins. On the other hand in vascular and in axis cells the two types of lectins were localised in the cytoplasm, outside the protein bodies. Thus these findings suggest different roles for the lectins: in cotyledons this may be a specific form of N storage, while in vascular and axis cells lectins may have a more direct metabolic part to play.     

Developmental changes in the subcellular structure of the seeds of Atropa belladonna

Subcellular changes in the embryo and endosperm of Atropa belladonna were studied at four developmental stages. The endosperm cells turn to storage cells much earlier than those of the embryo, which matures later. Cells of the cotyledon and radicle are very similar in structure. The young cells contain large osmiophilic spherosomes. The cytoplasm is filled with ribosomes but dictyosomes are very rare. Some proplastids, containing starch, and mitochondria are present in the early developmental stages but do not occur in the dormant cells. During ripening, the vacuoles of the endosperm cells and embryo develop into protein bodies. They become filled with protein material without any recognisable transport mechanism. Protein bodies have several electron-translucent globoid cavities and the protein mass contains a roundish or crystalline body. This body does not stain with potassium iodide but with periodic acid Schiff-reagent and protein stains, indicating that it contains glycoproteins. The embryo and endosperm cells of ripe Atropa seeds are very similar and filled with protein bodies and small spherosomes.     

Synthèse et translocation de RNA dans les cellules radiculaires de Zea mays au début de la germination

Summary RNA synthesis in root of germinating corn during the period extending from the 4th to the 8th hr after soaking is localized exclusively in the chromatin. This unusual situation allowed us to follow easily by section autoradiography the fate of the chromatin-synthesized RNA. The embryo was pulse-labeled with ³H-uridine from the 6th to the 8th hr after soaking and chased with cold uridine (2 hr) and then left in water (40 hr). It was found that during the chase a large amount of the chromatin-synthesized RNA moves to the nucleolus, where it is accumulated, and that a small amount finally reaches the cytoplasm.     

Plasma catecholamines and plasma corticosterone following restraint stress in juvenile alligators

Ten juvenile alligators, mean body mass 793 g, hatched from artificially incubated eggs and raised under controlled conditions, were held out of water with their jaws held closed for 48 hr. An initial blood sample was taken and further samples collected at 1, 2, 4, 8, 24, and 48 hr. Epinephrine, norepinephrine, and dopamine were measured in plasma aliquots of 1.5 ml using high pressure liquid chromatography with electrochemical detection. Corticosterone was measured by radioimmunoassay. Plasma glucose was measured using the Trinder method and plasma calcium, cholesterol, and triglycerides were measured in an autoanalyzer. Epinephrine was about 4 ng/ml at the initial bleed, but declined steadily to < 0.4 ng/ml by 24 hr. Norepinephrine was also about 4 ng/ml at the initial bleed, but rose to over 8 ng/ml at 1 hr, and then declined to < 0.2 ng/ml at 24 hr. A second, but smaller increase in plasma norepinephrine was seen at 48 hr. Plasma dopamine was low at the initial bleed (< 0.7 ng/ml), rose to over 8 ng/ml at 1 hr, then declined to < 0.2 ng/ml. Plasma corticosterone rose progressively for the first 4 hr, declined at 8 hr and 24 hr, then rose again at 48 hr. Plasma glucose rose significantly by 24 hr and remained elevated for 48 hr. Plasma calcium increased at 1, 2, and 4 hr then returned to levels not significantly different from the initial sample at 24 and 48 hr. The white blood cells showed changes indicating immune system suppression. By the end of the treatment the hetorophil/lymphocyte ratio increased to 4.7. These results suggest that handling alligators, taking multiple blood samples, and keeping them restrained for more than 8 hr is a severe stress to the animals.