RNA synthesis and the germination of light-sensitive lettuce seeds

Summary The germination of lettuce seeds is inhibited by the nucleotide base analogue 6-methylpurine. RNA synthesis has been measured during imbibition and germination as ³²P-phosphate incorporation into RNA species as fractionated by polyacrylamide gel electrophoresis. Seeds were surface sterilized and imbibed in the presence of various antibiotics. RNA preparations from lettuce seeds were coelectrophoresed with ³H-RNA prepared from bacteria to check for bacterial contamination of the seeds. There is a much higher rate of RNA synthesis in illuminated, germinating seeds as compared to dark, non-germinating seeds. This difference does not develop until after 12 hours of imbibition at 27°, which is the time of onset of germination and radicle growth.This investigation was supported by a contract from the United States Department of Agriculture (No. 616-15-3). Journal paper of the Purdue Agriculture Experiment Station.     

Analysis of DNA methylation during the germination of wheat seeds

DNA methylation is known to play a crucial role in regulating plant development and organ or tissue differentiation. Here, we focused on the DNA methylation dynamics during the germination of wheat seeds using the adapted AFLP technique so called methylation-sensitive amplified polymorphism (MSAP). The MSAP profiles of genomic DNA in embryo and endosperm tissues of germinating seeds, as well as dry seeds were characterized and notable changes of cytosine methylation were detected. Comparisons of MSAP profiles in different tissues tested showed that the methylation level in dry seeds is the highest. The alteration analysis of cytosine methylation displayed that the number of demethylation events were three times higher than that of de novo methylation, which indicated that the demethylation was predominant in germinating wheat seeds, though the methylation events occurred as well. Sixteen differentially displayed DNA fragments in MSAP profiles were cloned and the sequencing analysis confirmed that nine of them contained CCGG sites. The further BLAST search showed that four of the cloned sequences were located in coding regions. Interestingly, three of the sixteen candidates were homologous to retrotransposons, which indicated that switches between DNA methylation and demethylation occurred in retrotransposon elements along with the germination of wheat seeds.     

The relationships between ethylene production and germination ofCicer arietinum seeds

The germination percentage of chick-pea (Cicer arietinum) Seeds was greatly reduced by temperatures of 30°C and 35°C. This thermoinhibition was overcome by ethylene (ethrel). Both ABA and PEG diminished ethylene production and germination percentage in a parallel way. FC, MGBG and CHA stimulated both ethylene production and germination. AVG reduced ethylene production to some extent but did not inhibit germination. CoCl₂ and PG completely prevented both ethylene production and germination; this effect was reversed by ethylene but not by its immediate precursor ACC. NBE prevented both germination and ethylene production. Our results suggest that high ethylene production rates are not essential for germination of chick-pea seeds but that certain quantities of ethylene may be required.     

Mitochondrial DNA synthesis during fungal spore germination

Summary Germinating spores of the fungus Botryodiplodia theobromae incorporated guanine-8-C¹⁴ into both the nuclear DNA and mitochondrial DNA fractions. Ethidium bromide inhibited the synthesis of mitochondrial DNA without having a significant effect on nuclear DNA synthesis or on the rate and extent of spore germination. Rates of leucine and uracil incorporation and of oxygen uptake were not significantly affected by ethidium bromide until germination was nearly completed. Mitochondrial DNA synthesis is apparently not required for germination of the spores of B. theobromae but is probably essential to continued vegetative growth.Abbreviations DNA deoxyribonucleic acid - mit-DNA mitochondrial DNA - nuc-DNA nuclear DNA - RNA ribonucleic acid - EB ethidium bromide - Tris tris (hydroxymethyl)aminomethane Published with the approval of the Director as Paper No. 3331, Journal Series, Nebraska Agricultural Experiment Station. Research reported was conducted under Project No. 21-17. Paper No. 7877, Scientific Journal Series, Minnesota Agricultural Experiment Station.     

Early DNA synthesis during the germination of wheat embryos

Both [³H]thymidine and [³H]deoxyadenosine were found to be incorporated into the nuclear DNA of wheat embryos immediately after dry embryos were allowed to imbibe aqueous solutions of the radioactive precursors. The early labelled DNA sedimented in a manner suggesting that replicative intermediates were already formed within the first 90 min of germination. However, aphidicolin remained without any effect on this early DNA synthesis. Likewise, a cell-free system derived from early embryos incorporated [³H]dCTP into DNA independently of the presence of aphidicolin. On the contrary, dideoxyTTP inhibited the DNA synthesis considerably. It is concluded that a proportion of the resting wheat embryo cells is able to initiate a replicative DNA synthesis immediately upon imbibition. The synthesis seems, however, to proceed with the participation of a γ-like, rather than an α-like, DNA polymerase.     

DNA and histone synthesis are uncoupled during germination of maize embryos

The rate of synthesis of DNA and histones was studied in germinating maize embryos as a function of the length of the germination period. To that end excised embryos from seeds germinated for different periods of time were pulse labelled either with [¹⁴C]protein hydrolysate or with [³H]TdR. Specific activities were determined for the total cellular proteins and the total histone fraction obtained by acid-extraction of the cellular homogenate and BioRex70 ion exchange chromatography. The results show that the early germination period is characterized by a lack of coupling between the histone synthesis and that of the nuclear DNA. The early histone synthesis peak might be necessitated by the reprogramming of the embryo genome that takes place during germination.     

Interactions between hydrostatic pressure and oxygen concentration on the germination of lettuce seeds

Lettuce seeds were germinated at different hydrostatic pressures and O₂ concentrations. Some combinations of pressure and O₂ concentration stimulated while others retarded germination. The highest germination rates which occurred with pressure application and added O₂ were equalled at atmospheric pressure if the O₂ concentration was increased sufficiently. Increasing pressure generally retarded germination, but the pressure effect was partly offset by added O₂. Sustained pressures affected subsequent germination after pressure was released, but only when O₂ was present. Seeds to which pressure was applied in the absence of O₂ germinated as well after pressure exposure as untreated seeds.     

Synchronization of germination of celery seeds

The effects of osmotic pre-treatments on the germination response of celery seeds were studied in a series of experiments designed to optimize that treatment. The experimental variables were different osmotica (polyethylene glycol (PEG) or a mixture of potassium nitrate and tri-potassium orthophosphate), levels of osmotic potential, temperature and duration of treatment. The rate and synchronization of germination were very significantly improved by many of these pre-treatments but the maximum effects on cv. Lathom Blanching were obtained by treating the seeds with a solution of the salt mixture of c.–10 bars potential at 15 °C for 21 days. This treatment resulted in 50 % germination of the viable seeds within 1–4 days at a temperature of 20 °C compared with 13-7 days from the untreated seeds. There were indications that the optimum pre-treatment may vary somewhat between varieties and seed stocks. The results showed that the effects of the two osmotica on germination response were similar, thus enabling a technique to be developed for treating, with the salt solution, larger quantities of seed and germinating them prior to fluid drilling in the field.     

Ecology of germination of weed seeds

Summary Merremia gangetia Linn. (Cuf) is one of the most important weeds invading cultivated lands of India. Temperature, both high (45° C) and low (15° C) exhibit a definite role in development of the weed, particularly during germination. Higher tempeeratures (45° C) during dry storage appeared to be the main factor in laboratory experiments as well as in nature. Low temperature induced secondary dormancy and its removal by incubation at 35° C helps to explain the ecological mechanism through which the weed endures winter conditions. Germination of buried seeds confirm that the atmospheric conditions were found to be more effective in upper layers of soil. Ecological adaptations of this weed via germination requirements are assessed in the paper.     

Treatment of cotton seeds for germination

Summary Effects of H₂SO₄, H₂O₂, C₂H₅OH and of acetone on germination of cotton seeds and on the growth of seedling root were studied under laboratory conditions. H₂SO₄ or H₂O₂ hastened germination but decreased the ultimate number of seeds germinated. Higher temperature (27°C) hastened germination and increased the number of seeds germinated. Root length remained unaffected by treatments at room temperature while H₂SO₄ or H₂SO₄/H₂O₂ substantially decreased it at 27°C.     

Isoenzyme changes accompanying germination of wheat seeds

Electrophoretic patterns of soluble proteins and enzymes during development from dry seed to first leaf were studied in wheat by means of disc electrophoresis. The patterns considered were those of alcohol dehydrogenase (ADH), peroxidase, and nonspecific esterases. Multiple forms (isoenzymes) were found for all these enzymes in dry seeds. The ADH pattern did not change during development. Esterases and peroxidase patterns changed both qualitatively and quantitatively. Increase in the number of peroxidase bands was marked.Research carried out at Brookhaven National Laboratory under the auspices of the U.S. Atomic Energy Commission.     

徐长卿种子生活力检测及发芽生物学特性观察

对徐长卿[Cynanchum paniculatum(Bge．)Kitag．]种子生活力与发芽生物学特性进行了观察。结果表明：秋收徐长卿种子贮藏至次年3和4月,生活力分别为87％和80％;种子表面用1％H2O2除菌预处理,有利于正常发芽,发芽率达82％,发芽时间提前1d,第3日出现发芽高峰。温度从15℃至30℃,发芽率逐渐升高;25℃时,种子第2天即开始发芽,出现最短发芽持续时间(5d)和最高发芽势(75％),因此,25℃是徐长卿种子发芽的最适温度。     

DNA synthesis and mRNA accumulation during germination of embryos of the orchidSpathoglottis plicata

Summary Germination of embryos of the orchid,Spathoglottis plicata Blume involves the formation of a protocorm in which DNA synthesis and cell divisions are confined to the proximal end whereas cells at the distal end undergo enlargement. Although³H-thymidine was incorporated into the distal cells of the embryo during the early period of germination, DNA synthesis was not followed by mitosis and cytokinesis. Poly(A)-RNA detected by in situ hybridization of sections to³H-poly-(U) was present uniformly in all cells of the embryo of the dry seed. However, coincident with the formation of the apical meristem, there was an increase in the density of auto-radiographic silver grains in the cells of the embryo, with a concentration of silver grains in the proximal part. The results indicate that regulatory events in the embryo prior to seed maturity determine the fate of its proximal and distal parts during germination.Dedicated to the memory of Professor John G. Torrey     

Changes of enzymes and factors involved in DNA synthesis during wheat embryo germination

We have previously purified and characterized wheat germ DNA polymerases A and B. To determine the role played by DNA polymerases A and B in DNA replication, we have measured the level of their activities during wheat embryo germination. The level of cellular proteins known to be associated with DNA synthesis such as PCNA and DNA primase were also investigated. The activity of DNA polymerase A gradually increased reaching a maximal level at 12 h after germination. Three days later, only a residual activity was detected. DNA polymerase B showed the same pattern during germination with very similar changes in activity. Our results indicate a striking correlation between maximal activities of DNA polymerase A, DNA polymerase B and optimal levels of DNA synthesis. These results support a replicative role of these enzymes. The activity of wheat DNA primase that copurifies with DNA polymerase A also increases during wheat germination. Taking together all its properties, and in spite of its behaviour with some inhibitors, DNA polymerase A may be considered as the plant counterpart of animal DNA polymerase . Concerning DNA polymerase B we have previously shown that PCNA stimulates its processivity. Besides studying the changes of DNA polymerases A and B and DNA primase we have also studied changes in PCNA during germination. We show that PCNA is present in wheat embryos at a constant relatively high level during the first 24 h of germination. After 48 h, the absence of PCNA is concomitant with an important decrease in DNA polymerase B activity. In this report we confirm the behaviour of DNA polymerase B as a -like activity.Département de Biologie, Université de Drah-Lmraz,Fez, Maroc     

Underdeveloped embryos and germination in Aristolochia galeata seeds

Aristolochiaceae have been described as having seeds with underdeveloped embryos and morphological or morphophysiological dormancy. Aristolochia galeata is a native climber found in the Cerrado biome, associated with road and gallery forest edges. The aims of this study were to investigate: embryo growth rate, morphology and seed germination parameters under different treatments. Embryos were excised to obtain embryo length at four stages: initial, seeds after coat rupture, radicle tip protrusion and cotyledon emergence from the seed coat. Germination tests were conducted at 30 °C under three nitrate concentrations (1, 10 and 20 mM), fluctuating temperature (27/20 °C) and light and dark conditions. We found that seeds have underdeveloped embryos, which take about 301 ± 178 h (±SD) to achieve seed coat rupture, another 205 ± 126 h to reach radicle protrusion and 176 ± 76 h more to the final stage of cotyledon emergence. Germinability was above 52% in all treatments, except in the dark (15%). For all treatments, average germination time was above 290 ± 123 h. Potassium nitrate increased germinability to >87%. No particular treatment was required for embryo development, but seeds in the population that continued to germinate after 1 month were probably in various states of non-deep, simple morphophysiological dormancy. Increased germinability in nitrate treatments and light requirement for germination could prevent germination under unsuitable environmental conditions and be a strategy to increase seedling establishment in the cerrado.     

Antagonistic effects of high and low temperature pretreatments on the germination and pregermination ethylene synthesis of lettuce seeds

Red light-induced germination of Grand Rapids lettuce seeds (Lactuca sativa L.) incubated at 20 C was inhibited if the seeds were first imbibed at 30 C for 36 hours. This effect was counteracted by exogenous ethylene and associated with a reduction in the rate at which the seeds produced ethylene throughout the pregermination period. A chilling treatment reversed the effect of a prior imbibition at 30 C on both germination and ethylene production. The possibility that the pretreatments influence germination through their effects on ethylene production is discussed.     

三药槟榔种子休眠与萌发的研究

对三药槟榔种子休眠和萌发的基本特性进行研究,结果表明种子的休眠属于综合休眠;种壳对种子 萌发的抑制作用不是由于其对水分透过的限制,而是种皮的机械束缚和透气性差;种子还需要一段低温的生 理后熟过程才能解除休眠。种子经0．2％的高锰酸钾溶液浸泡15 min,0．3％亚硝酸钠和0．2％的硝酸钾溶液 浸种24℃后,发芽速度均显著加快,以0．3％亚硝酸钠处理效果为最佳。种子在15、4℃和室温(昼24～32 ℃／夜18～24℃)三种不同温度下贮藏60 d后,在4℃贮藏的种子发芽情况最好。种子不耐脱水,采用硅胶脱 水,含水量降低至22％以下,种子活力显著降低。