Germination Of Dormant Embryos Research Articles

The Mechanisms Involved in Seed Dormancy Alleviation by Hydrogen Cyanide Unravel the Role of Reactive Oxygen Species as Key Factors of Cellular Signaling during Germination

The physiological dormancy of sunflower (Helianthus annuus) embryos can be overcome during dry storage (after-ripening) or by applying exogenous ethylene or hydrogen cyanide (HCN) during imbibition. The aim of this work was to provide a comprehensive model, based on oxidative signaling by reactive oxygen species (ROS), for explaining the cellular mode of action of HCN in dormancy alleviation. Beneficial HCN effect on germination of dormant embryos is associated with a marked increase in hydrogen peroxide and superoxide anion generation in the embryonic axes. It is mimicked by the ROS-generating compounds methylviologen and menadione but suppressed by ROS scavengers. This increase results from an inhibition of catalase and superoxide dismutase activities and also involves activation of NADPH oxidase. However, it is not related to lipid reserve degradation or gluconeogenesis and not associated with marked changes in the cellular redox status controlled by the glutathione/glutathione disulfide couple. The expression of genes related to ROS production (NADPHox, POX, AO1, and AO2) and signaling (MAPK6, Ser/ThrPK, CaM, and PTP) is differentially affected by dormancy alleviation either during after-ripening or by HCN treatment, and the effect of cyanide on gene expression is likely to be mediated by ROS. It is also demonstrated that HCN and ROS both activate similarly ERF1, a component of the ethylene signaling pathway. We propose that ROS play a key role in the control of sunflower seed germination and are second messengers of cyanide in seed dormancy release.

Carbohydrate concentrations and interactions in afterripening-responsive dormantAvena fatuacaryopses induced to germinate by gibberellic acid

AbstractIt has been proposed that gibberellic acid (GA3) promotes germination by overcoming restrictions in sugar production and utilization in afterripening-responsive dormant caryopses. While their germination rates were similar, germination commenced sooner in afterripened wild oat (Avena fatuaL.) caryopses than in dormant caryopses treated with GA3and dormant excised embryos treated with GA3plus fructose (Fru). Limited germination occurred in dormant excised embryos cultured with GA3alone. Carbohydrate concentrations were measured over time in dormant caryopses and excised embryos whose germination was induced with GA3and GA3plus Fru. The concentration of sucrose (Suc) in the endosperm declined prior to germination of dormant GA3-treated caryopses. Raffinose (Raf) family oligosaccharides in the embryos of dormant GA-treated caryopses remained relatively constant prior to and shortly after the onset of germination. In contrast, Raf family oligosaccharides in the embryos of afterripened caryopses declined prior to germination. Together this suggests Raf family oligosaccharide utilization is not associated with germinationper se.Increased starch levels, which occurred in dormant excised embryos treated with Fru and GA3plus Fru, were associated with dormancy because similar effects were not apparent in afterripened embryos cultured with Fru. An initial decline in the concentration of Raf family oligosaccharides in dormant embryos cultured with GA3or GA3plus Fru seems to be a result of the excision process. GA3appears to stimulate the germination of dormant embryos by enhancing the uptake or utilization of Fru. It appears that GA3and afterripening-induced changes in carbohydrate utilization in dormant caryopses are different.

Effect of Soluble Sugars and Gibberellic Acid in Breaking Dormancy of Excised Wild Oat (Avena fatua) Embryos

Dormant line M73 wild oat caryopses were utilized to develop a system for the culture of excised embryos, to evaluate whether embryo dormancy exists, and to investigate the physiological basis for breaking dormancy. Dormant embryos cultured on N6 medium solidified with 0.25% Bacto agar displayed 70% germination in 2 d compared with approximately 20% for the other gelling agents. The non-plant-based gelling agent gellum at a concentration of 0.25% was selected for further experiments on breaking dormancy after it was determined that concentrations ≥ 0.5% decreased the rate of germination. Amending N6 medium with concentrations of 0.1 to 10 μM gibberellic acid (GA) increased the rate and extent of germination. Embryos treated with 0 to 0.01 μM GA required 6 d to attain 90% germination. Germination of dormant embryos on N6 medium without GA suggested that either true embryo dormancy did not exist in M73 or some constituent of the N6 medium promoted breaking of dormancy. Subsequent experiments indicated that the 88 mM sucrose was the constituent in the N6 medium responsible for breaking dormancy. Concentrations of sucrose from 40 to 200 mM were effective in breaking dormancy. Ten μM GA increased the rate and extent of germination of embryos cultured with 88 to 200 mM sucrose. At 88 mM, fructose, maltose, glucose, and sucrose all broke embryo dormancy. Fructose was the most active soluble sugar for breaking embryo dormancy, promoting nearly 100% germination in 4 d. As with sucrose, there was an interaction between GA and the soluble sugars in breaking dormancy. Ten μM GA with 88 mM fructose provided nearly 100% germination in 1 d. Amylose, but not amylopectin or pullulan, may substitute for soluble sugars. However, with 10 μM GA amylose, amylopectin and pullulan were equally effective in breaking dormancy. Breaking dormancy of embryos on N6 medium was independent of temperatures from 12 to 24 C in the presence of GA, but in its absence the optimum was 12 C. Application of GA to dormant caryopses significantly increased and decreased the level of glucose and sucrose, respectively, in the embryo. Gibberellic acid had a similar effect on glucose and sucrose in the endosperm tissue, except the differences were not significant at all times after treatment. The change in carbohydrate metabolism, especially in embryo tissue, may be important when considered in context with the observation that soluble sugars and GA act independently in breaking dormancy in excised M73 embryos. Breaking wild oat embryo dormancy with GA may be mainly a substitution for sugar requirement.

The role of lipases in the removal of dormancy in apple seeds

It was found that the temperature optimum for apple (Malus domestica Borb.) seed acid lipase is the same as that for seed after-ripening process. The activity of the enzyme occurs between the 40th and 70th days of stratification, whereas the activity of alkaline lipase very low at that time appears about 20 days later. The changes of both enzyme activities were also studied during dark and light culture of embryos isolated from seeds after different times of stratification. Only the alkaline enzyme activity is under the control of light. It was concluded that essentially the same process, i.e. the hydrolysis of reserve fats is catalysed by two different enzymes: acid lipase acting during the cold-mediated breaking of embryo dormancy and alkaline lipase acting during the germination of dormant embryos, thus being under light control.

Regulation of the Krebs cycle and pentose phosphate pathway activities in the control of dormancy of Avena fatua

Malonic acid (10−2 M) promoted germination of excised dormant embryos of Avena fatua L. but inhibited germination of non-dormant embryos. Malonate inhibited oxygen consumption and carbon dioxide evolution of dormant embryos which resulted in an increased respiratory quotient and a decrease in the glucose C6/C1 ratio. Germination is oxygen-dependent, and malonate-sensitive oxygen consumption decreased during afterripening. Succinic acid (5 × 10−2 M) promoted germination of dormant embryos. Gibberellic acid (GA) (1.0 ppm) and succinate (5 × 10−2 M) interacted synergistically in promoting germination. Adenosine diphosphate also promoted germination but only after a considerable lag period. Germination of non-dormant embryos was inhibited by 2,4-dinitrophenol indicating the importance of adenosine triphosphate for germination. Storage of caryopses in various gaseous environments indicated that the response of dormant embryos to GA treatment was dependent on the operation of oxidative processes during afterripening. A model is proposed, indicating the nature of the regulation of the oxidative systems involved in the control of dormancy. The model accounts for the known effects of oxygen treatments on seeds, provides an explanation of the mechanism of afterripening, and explains the survival of imbibed dormant caryopses during adverse conditions. Such a model may be of widespread significance and is discussed in relation to its interaction with GA in developmental processes.

Increased participation of pentose phosphate pathway in response to afterripening and gibberellic acid treatment in caryopses of Avena fatua

The rates of oxygen consumption of dormant and non-dormant excised embryos of Avena fatua L. before germination are similar. Gibberellic acid (GA) treatment stimulates germination of dormant embryos without affecting oxygen consumption. Thus dormancy is not the result of restricted oxygen uptake. The fat content of dormant and non-dormant caryopses remains constant during germination. Dormant and non-dormant embryos have respiratory quotients near unity supporting the hypothesis that starch degradation occurs before germination. 6-Phosphogluconate dehydrogenase, a key enzyme of the pentose phosphate pathway, is present in dormant and non-dormant dry embryos but the pre-germination C6/C1 ratio of non-dormant embryos is markedly lower than that of dormant embryos, indicating a greater participation of the pentose phosphate pathway in the respiratory metabolism of non-dormant embryos. Release from dormancy is associated with a shift in metabolism from the glycolytic pathway to the pentose phosphate pathway. GA treatment, which stimulates germination of dormant embryos, causes a similar qualitative change in the oxidative metabolism of dormant embryos. Thus the action of GA is to cause the increased degradation of glucose via the pentose phosphate pathway, which is an essential step in the preparation for germination.

Metabolically Active Glucosides in Oleaceae Seeds

The seeds of six woody species of Oleaceae representing three genera, contain high concentrations of water-soluble glucosides, with major absorption maxima below 240 nanometers. In Fraxinus americana seeds three of these compounds, designated GL-3, GL-5, and GL-6, account for almost 10% of the dry weight. They are found in the endosperm and embryo but not in the pericarp. While the level of GL-5 is not particularly influenced by the physiological state of the embryo, that of GL-3 and GL-6 decreases as a result of germination and growth during a 10-day period. As the concentrations of GL-3 and GL-6 decrease, new ultraviolet-absorbing compounds are formed. The changes in the concentration of the ultraviolet-absorbing glucosides during cold temperature after-ripening, prior to germination, are small. When germination of dormant embryos is induced with gibberellic acid, the concentrations of GL-3 and GL-6 decrease in a manner similar to that observed with nondormant embryos. In the presence of abscisic acid no losses of GL-3 and GL-6 were observed. It is suggested that GL-3 and GL-6 fulfill some definite functions in the germination and growth of F. americana embryos, and that gibberellic acid and abscisic acid can exert a regulatory effect on the metabolism of these glucosides.

Dormancy studies in seed of Avena fatua. 6. Germinability of the immature caryopsis

Caryopses of a dormant strain of Avena fatua L. fail to germinate on moist filter paper when immature whereas non-dormant caryopses germinate at 10 days after fertilization and at later stages of development. Embryos of the dormant strain excised at any stage of development fail to germinate in a complex nutrient medium which supports germination and growth of non-dormant embryos at, and after, 10 days following fertilization. Gibberellicacid (GA) promotes germination of dormant caryopses and excised dormant embryos before and at maturity in the presence of either endosperm or the nutrient medium. This requirement for gibberellin is not absolute, since germination of dormant embryos in mid-development occurs when the nutrient medium is solidified with agar in the absence of gibberellin. The stimulatory effect of solidifying the medium is probably due to differential leaching action upon embryos affecting levels of endogenous gibberellins. The genetic source of endosperm does not influence germinability of the embryo, and endosperms of both dormant and non-dormant strains show an increasing capability of supporting postgerminal growth of non-dormant embryos. A block to the use of these nutrients by the embryo, reversible by gibberellin, is present at maturity in endosperm of the dormant strain.