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PubMed1966

Changes in some nitrogenous components during the germination of pea seeds.

The changes in major nitrogenous components during the germination of pea seeds have been followed. During the period of rapid axis growth, 3 to 8 days following germination, the nitrogen content of the cotyledons declines rapidly with an accompanying increase of nitrogen in the developing axis. The accumulation of alcohol soluble nitrogen, primarily amino nitrogen, in the cotyledons and axis during germination indicates that the mobilization of nitrogen is facilitated by proteolysis and translocation of the products.Pea cotyledons had an initially high RNA content which declined during germination while axis RNA increased. The increase in axis RNA was greater than the decline in cotyledonary RNA indicating a net nucleic acid synthesis. Thus some of initial nitrogen reserve is interconverted to provide for nucleic acid synthesis.During the depletion of cotyledonary RNA there was no accumulation of nucleotides in the cotyledons, nucleotide content of the axis did, however, increase during the germination period.The DNA content of the axis increased with growth of that organ. There was also an increase in DNA content of the cotyledons during the early stages of germination.

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PubMed1967

Carbohydrate and Lipid Metabolism During Germination of Uredospores of Puccinia graminis tritici.

Uredospores of Puccinia graminis (Pers.) tritici (Eriks. and Henn.) were uniformly labeled with (14)C by permitting the host (Triticum aestivum L.) to carry out photosynthesis in (14)CO(2) during the process of spore production by the obligate parasite. The use of (14)C labeled spores provided advantages in a study of the utilization of endogenous substrates at frequent intervals with small amounts of spores under conditions conducive to germination.Because of previous uncertainties about the nature of the substrates of importance to germination, a detailed study of carbohydrate and lipid components, both in the spores and in the germination medium, was made during the first 7 hours after placing the spores on aqueous media. Diethyl ether and 80% ethanol soluble metabolites each constituted approximately 20% of the total spore carbon. During the first hour nearly 60% of the 80% alcohol solubles disappeared from the spores while the total ether soluble material did not change appreciably. A significant part of the 80% ethanol soluble materials appeared in the germination medium.During germination and germ tube extension, there was rapid utilization of trehalose, arabitol and mannitol even though appreciable amounts of these materials were present as exogenous pools in the germination medium. Although the total amounts of ether soluble components did not change as drastically as the carbohydrate fraction, there was extensive utilization of palmitic, oleic, linolenic and 9,10-epoxyoctadecanoic acids.The results indicate that the germination process in spores of obligate parasites is not based solely on the utilization of lipids and some possible roles of the changes in internal and external pools of soluble carbohydrates are discussed.

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PubMed1967

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

Lettuce seeds were germinated at different hydrostatic pressures and O(2) concentrations. Some combinations of pressure and O(2) concentration stimulated while others retarded germination. The highest germination rates which occurred with pressure application and added O(2) were equalled at atmospheric pressure if the O(2) concentration was increased sufficiently. Increasing pressure generally retarded germination, but the pressure effect was partly offset by added O(2). Sustained pressures affected subsequent germination after pressure was released, but only when O(2) was present. Seeds to which pressure was applied in the absence of O(2) germinated as well after pressure exposure as untreated seeds.

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PubMed1967

Biogenesis of Mitochondria in Germinating Peanut Cotyledons II. Changes in Cytochromes and Mitochondrial DNA.

Biogenesis of mitochondria occurs in the germinating cotyledons of peanuts. This process was demonstrated by measuring both constitutive and enzymatic properties of mitochondria as a function of germination time. Direct counting by phase contrast microscopy of sucrose density gradient preparations showed that the number of mitochondria increased markedly during germination. DNA with a buoyant density distinct from the major cellular DNA was associated with these mitochondrial preparations. During germination the amount of this DNA in mitochondrial pellets increased. This increase closely paralleled the increase in number of mitochondria.Succinoxidase and succinic dehydrogenase increased during germination. Both activities were confined to the mitochondrial fraction. The rate of increase of succinoxidase activity was significantly greater than the rate of increase of succinic dehydrogenase and both increased at least initially at a greater rate than the amount of mitochondrial DNA or numbers of mitochondria.The amounts of cytochromes present in mitochondrial preparations were measured spectrophotometrically. All of the cytochromes increased in amount during germination. The rate of increase of cytochrome a - a(3) very close to the rate of increase in succinoxidase activity.

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PubMed1968

Enzymic mechanism of starch breakdown in germinating rice seeds I. An analytical study.

Time-sequence analyses of carbohydrate breakdown in germinating rice seeds shows that a rapid breakdown of starch reserve in endosperm starts after about 4 days of germination. Although the major soluble carbohydrate in the dry seed is sucrose, a marked increase in the production of glucose and maltooligosaccharides accompanies the breakdown of starch. Maltotriose was found to constitute the greatest portion of the oligosaccharides throughout the germination stage. alpha-Amylase activities were found to parallel the pattern of starch breakdown. Assays for phosphorylase activity showed that this enzyme may account for much smaller amounts of starch breakdown per grain, as compared to the amounts hydrolyzed by alpha-amylase. There was a transient decline in the content of sucrose in the initial 4 days of seed germination, followed by the gradual increase in later germination stages. During the entire germination stage, sucrose synthetase activity was not detected in the endosperm, although appreciable enzyme activity was present in the growing shoot tissues as well as in the frozen rice seeds harvested at the mid-milky stage. We propose the predominant formation of glucose from starch reserves in the endosperm by the action of alpha-amylase and accompanying hydrolytic enzyme(s) and that this sugar is eventually mobilized to the growing tissues, shoots or roots.

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PubMed1971

Phospholipids in the Uredospores of Uromyces phaseoli: II. Metabolism during Germination.

The levels and types of phospholipids changed in distinct phases during the germination of (32)P-labeled uredospores of Uromyces phaseoli. During the first 20 minutes of germination, the phospholipid content dropped to 40% of the pregermination level. Between 2 and 3 hours, phospholipid levels increased to approximately 80% of the pregermination levels, and after germination for 5 hours, catabolism had reduced the (32)P-lipids to about the same level observed prior to the first anabolic phase. A second anabolic phase was observed between 5 and 10 hours of germination. Phosphatidylcholine and phosphatidylethanolamine, the major phospholipids, did not undergo anabolism and catabolism at the same rates during germination. Only small quantities of the more polar phospholipids were released to the germination media.Germinating uredospores were capable of utilizing l-methionine-methyl-(14)C, d,l-(14)C serine, (14)C-choline, and (14)C-ethanolamine for the synthesis of phospholipids. The active one-carbon units from methionine and serine appear to be involved in the methylation of phosphatidylethanolamine to form phosphatidylcholine. Preformed ethanolamine and choline may be incorporated into these two phospholipids also. Evidence for the synthesis of phosphatidylserine, which is subsequently decarboxylated to yield phosphatidylethanolamine, was obtained.

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PubMed1973

Blue Light Interference in the Phytochrome-controlled Germination of the Spores of Cheilanthes farinosa.

Short exposure of the spores of Cheilanthes farinosa to low intensity red light promotes their germination, which is not reversed by a subsequent exposure to far red light. Germination is, however, inhibited by blue light administered before or after red light. Inhibition of germination by blue light is annulled by exposure to a higher intensity of red light, and germination of the repromoted spores is inhibited by far red light. Mutual photoreversibility of germination is also observed in repromoted spores irradiated successively with far red and red light. Although germination appears to be basically under phytochrome control, it is postulated that the presence of a blue light-absorbing pigment interferes with phytochrome transformations in the spores.

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PubMed1973

Control of the formation of amylases and proteases in the cotyledons of germinating peas.

Protease activity increased in attached cotyledons of germinated peas (Pisum sativum L. cv. Alaska) as the stored proteins declined but did not increase in excised cotyledons incubated for the same length of time. Cotyledons of seeds germinated in the presence of a casein hydrolysate solution developed less protease activity than did those germinated on water. These results suggest that accumulation of amino acids regulates the protease level in the cotyledons of germinating peas.In contrast to protease, alpha- and beta-amylase increased during incubation of excised pea cotyledons. Their increase was inhibited by abscisic acid. Abscisic acid did not inhibit (14)C-leucine incorporation into protein or reduce the respiratory rate in the cotyledons; hence, its effect on amylase formation was not the result of a general inhibition of metabolism. An ether-soluble acid fraction, which would contain any abscisic acid present in the material, inhibited amylase formation more when it was obtained from imbibed seeds than when it was obtained from cotyledons of seeds germinated for 10 days. These and other results suggest that amylase formation in germinating peas is regulated by abscisic acid.

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PubMed1973

The Effect of Arsenate and Other Inhibitors on Early Events during the Germination of Lettuce Seeds (Lactuca sativa L.).

The effect of arsenate, arsenite, 2,4-dinitrophenol, and anaerobiosis on early events in seed germination was investigated using both intact and punched seeds of lettuce (Lactuca sativa L.). It was found that punching the seed removes penetration barriers to the entrance of inhibitors without an undue loss of germination or light responses. The kinetics of the action of germination inhibitors were established by 2-hour pulse experiments. Arsenate and 2, 4-dinitrophenol have very different kinetics. The inhibition of germination in punched seeds by arsenate given in conjunction with phosphate compared with the lack of inhibition of arsenate plus phosphate on the growing seedling, suggest a distinct metabolic change in the germinating embryo at some time between the onset of germination and subsequent seedling growth.

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PubMed1974

Actions of gibberellic Acid and phytochrome on the germination of grand rapids lettuce seeds.

Red light and gibberellic acid were about equally effective in promoting germination of Grand Rapids lettuce (Lactuca sativa L.) seeds. With initial far red light treatment more than 80% remained dormant in subsequent dark storage. After 2 days of dark storage, red light effectively promoted germination, while gibberellic acid action was weak. With between 2 and 10 days of dark storage, gibberellic acid had little effect, while promotion by red light decreased slowly and finally disappeared. After 10 days of dark storage, both gibberellic acid and red light were required for germination. The dark storage treatment interferes with phytochrome-independent germination processes and cannot be overcome by added gibberellic acid. However, storage may also decrease the effectiveness of endogenous gibberellins. Phytochrome-dependent germination seems to require only low levels of endogenous gibberellin activity or the addition of gibberellic acid. Gibberellins and red light appear to act on germination by regulation of sequential sites of a branched-looped pathway.

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PubMed1974

The development of isocitric lyase activity in germinating cotton seed.

In cotyledons of germinating cotton (Gossypium hirsutum L. var. Stoneville 213) seedlings, in the dark, isocitric lyase (EC 4.1.3.1) activity peaks after 2 days and thereafter slowly declines to a negligible value after 8 days. The maximum activity of this enzyme in cotyledons of 2-day-old seedlings was 16.2 mumoles of glyoxylate formed/15 min.10 cotyledon pairs. Actinomycin D at a concentration of 10 mug/ml, if added to the imbibing solution, completely prevents the development of isocitric lyase activity in these germinating seed. In cotyledons of germinating cotton seedlings, in the light, isocitric lyase activity peaks after 2 to 3 days and sharply declines to a negligible value after 4 days. The maximum activity of this enzyme in cotyledons of 2- to 3-day-old seedlings was 13.2 mumoles of glyoxylate formed/15 min.10 cotyledon pairs. Actinomycin D at a concentration of 10 mug/ml, if added to the imbibing solution, severely inhibits the development of enzyme activity.In germinating seed, in the light, the synthesis of chlorophyll and glyceraldehyde-3-P dehydrogenase is also limited by the addition of low concentrations of actinomycin D. The new synthesis of fructose-1, 6-diP aldolase, which is detectable after 1 to 2 days of germination, is inhibited by 10 mug/ml of actinomycin D. We, therefore, conclude that the synthetic events leading to the development of chlorophyll, some glyoxysomal and chloroplast enzymes in germinating cotton seedlings depend on newly transcribed mRNA.

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PubMed1975

De Novo Biosynthesis of Deoxyribonucleic Acid Polymerase during Wheat Embryo Germination.

An 8-fold enhancement in the activity of a DNA-dependent DNA polymerase was found in extracts from germinating wheat (Triticum vulgare var. Florence) embryos, as compared to the activity found in extracts from ungerminated embryos. The enhancement of this activity during the first hours of germination is concomitant to the increase of a Dnase activity. The two activities could be separated and the increased level of the DNA polymerase upon germination was observed in an enzymatic fraction which contains very low DNase activity. Addition of the protein synthesis inhibitor, blasticidin S, to germinating wheat embryos, reduced the increase in DNA polymerase. Incorporation of radioactive amino acids into a phosphocellulose preparation, which contains the DNA polymerase starts during the first 6 hours of germination. The amount of radioactivity incorporated is doubled in the next 6 hours, and the incorporation is continued between 12 and 18 hours of germination.

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PubMed1977

Mode of action of gibberellic Acid and light on lettuce seed germination.

The seeds of lettuce (Lactuca sativa L. cv. Grand Rapids) germinate in darkness at 25 C when treated by gibberellic acid (GA(3)) for 1 hour following 2 hours of imbibition. The time of GA(3) application influences the rate and the final percentage of seeds that germinate. In contrast, red light illumination given at different times affects only the rate and not the final germination percentage. The early process(es) of germination initiated by GA(3) or light treatment can be arrested by subjecting the treated seeds to a nongerminative temperature of 35 C. The results suggest differences in the mode of action of light and GA(3) during germination. They indicate that different kinds of processes are involved in the biochemical control of germination.

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PubMed1979

Reversal by Light of Ethylene-induced Inhibition of Spore Germination in the Sensitive Fern Onoclea sensibilis: An Action Spectrum.

Spores of the fern Onoclea sensibilis L. normally germinate to produce two cells of unequal size. The larger cell divides to produce the familiar heart-shaped prothallus. The smaller cell elongates and differentiates into the rhizoid but normally does not divide again. Onoclea spores germinate in complete darkness. Dark germination can be completely inhibited by ethylene gas (10 microliters per liter is saturating). This inhibition can be reversed by light. Broad band colored light studies were designed to determine which area of the spectrum was most effective in overcoming ethylene inhibition. White light treatment resulted in 17% germination. Blue light treatment resulted in 1% germination. Red light treatment resulted in 15% germination. Red light, therefore, was most effective and accounted for most of the effects of white light. A detailed action spectrum was constructed using narrow band interference filters in the wavelength range from 350 to 764 nanometers. The action spectrum has only one major peak at 711 nanometers.

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PubMed1979

Development of Peptide Transport Activity in Barley Scutellum during Germination.

The scutella separated from nongerminated grains of barley (Hordeum vulgare L. cv. Pirkka) took up labeled glycylsarcosine (Gly-Sar), glycylglycine, and leucylleucine at a rate of about 2 nanomoles per scutellum per hour. About 55% of the uptake of Gly-Sar (2 millimolar) was due to nonmediated uptake, but the active component was similar to that operating in the scutella from germinating grains (pH optimum about 4.5, K(m) about 10 millimolar).When the grains were allowed to germinate aseptically on agar, the rate of uptake of Gly-Sar began to increase after a lag of about 5 hours. The increase continued until a maximum value of 70 nanomoles per scutellum per hour (35-fold increase) was attained after 3 days of germination. After this the activity began to decrease slowly. The uptake of glycylglycine and leucylleucine changed in a similar way. When embryo-containing halves of seeds or separated embryos were allowed to germinate for 12 hours on agar containing various inhibitors of RNA and protein synthesis, the increase in the peptide uptake activity was completely or partially inhibited. The increase was not affected by removal of the "embryo proper" or endosperm, or by additions of gibberellic or abscisic acid.Root tips and aleurone layers took up Gly-Sar very slowly; the rate of the mediated component of uptake was less than 1% of the uptake by scutella.The preferential localization of the peptide transport system in the scutellum and its rapid development at the beginning of germination strengthen the earlier conclusion that peptide transport plays an essential role in the mobilization of reserve proteins during germination.

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PubMed1981

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.Jojoba cotyledonary extracts were found to inhibit the enzymic activity of trypsin, chymotrypsin, and pepsin but not the protease from Aspergillus saotoi. The heat-labile trypsin inhibitor substance(s) was found in commercially processed jojoba seed meal and the albumin fraction of seed proteins. Trypsin inhibitor activity decreased with germination.

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PubMed1982

Desiccation of Axes of Phaseolus vulgaris during Development of a Switch from a Development Pattern of Protein Synthesis to a Germination Pattern.

Immature seeds of Phaseolus vulgaris removed from the pod at 32 days of development do not germinate unless first subjected to a desiccation treatment. This change from development to germination caused by premature drying is mirrored in the pattern of protein synthesis by the axes. Rehydrated axes from 32-day-developed seeds cease to synthesize proteins that are uniquely associated with development, but instead synthesize some proteins that are similar to those made in the germinating axes from mature dry seeds. Desiccation of 22-day-developed seeds does not lead to their germination, nor does it cause a switch from a developmental to a germination mode of protein synthesis by the axes. It is proposed that desiccation plays a role in permanently suppressing developmental protein synthesis and in inducing germination protein synthesis.

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PubMed1983

Germination of Echinochloa crus-galli (Barnyard Grass) Seeds under Anaerobic Conditions : Respiration and Response to Metabolic Inhibitors.

Echinochloa crus-galli L. Beauv., a rice-field weed, can germinate and grow for extended periods of time in an anaerobic environment. Compared to pea, which does not germinate under anaerobiosis, the evolution of CO(2) in Echinochloa and rice is lower and the peak rate of CO(2) evolution is delayed when germinated without oxygen. The plants studied also differ with respect to their respiration ratio ([CO(2)] N(2)/[CO(2)] air) and metabolism used during the early stages of germination. Echinochloa does not increase its glycolytic rate under anaerobiosis, whereas pentose phosphate pathway activity appears to increase during the first 40 to 50 hours of germination.Based on its response to metabolic inhibitors (NaF, dinitrophenol, and malonate), anaerobic metabolism in Echinochloa proceeds primarily through glycolysis, with partial operation of the tricarboxylic acid cycle and little or no oxidative phosphorylation. Also, Echinochloa is sensitive to CN during aerobic germination, whereas rice appears to be able to shift to CN-insensitive electron transport. Finally, the effectiveness of cyanide and azide on inhibiting germination of Echinochloa in N(2), but not CO, suggests that cytochrome oxidase is not used to reoxidize pyridine nucleotides in the absence of oxygen. The possible existence of an alternate electron acceptor is discussed.

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