Dormancy and Impotency of Cocklebur Seeds: VII. Inability of Dormant Cotyledons to Form Chlorophyll

Dormant seeds of cocklebur (Xanthium pennsylvanicum Wallr.) were characterized by the lack of ability to form chlorophyll. Such an inability of cotyledons of the dormant seeds was improved by the application of various factors and reagents which were capable of breaking the dormancy and of increasing cotyledon enlargement. Of these, ethylene, benzyladenine, and high temperature treatments were particularly effective, and, in turn, oxygen enrichment, gibberellic acid, thiourea, carbon doxide, and potassium nitrate were also promotive to the greening of the dormant cotyledons. The effects of benzyladenine, oxygen enrichment, and high temperature were reduced in the presence of absorbents for endogenously evolve carbon dioxide and ethylene. δ-Aminolevulinic acid could not restore their greening ability.     

Ethylene Production, 1-Aminocyclopropane-l-Carboxylic Acid Content and Its Conversion to Ethylene in Axial Segments of Dormant and Nondormant Cocklebur Seeds

During the imbibition of water, the change in the ethylene productionof axial segments of nondormant (ND) cocklebur (Xanthium pennsylvanicumWallr.) seeds paralleled the change in the content of free 1-aminocyclopropane-l-carboxylicacid (ACC), but not the change in conjugated hydrolysable ACCin the axes. Aminoethoxyvinylglycine, anoxia and a-aminoisobutyricacid inhibited ethylene production, the lattertwo compoundscausing the accumulation of free ACC. Administration of ACCgreatly enhanced ethylene production in the axes. Thus, freeACC seems to be a direct precursor of ethylene production inthe axial tissue of cocklebur seeds. Imbibed dormant (D) axes characterized by inferior ethyleneproduction had less ability to convert exogenously applied ACCto ethylene as compared to ND axes. But, there was little differencebetween the D and ND axes in the endogenous contents of freeand conjugated ACC. This suggests that the inferior ethyleneproduction found in detached D axes is associated with the lowactivity of an ACC-ethylene converting system. (Received December 17, 1982; Accepted April 30, 1983)     

Dormancy and Impotency of Cocklebur Seeds VIII. Lack of Germination Responsiveness in Primarily Dormant Seeds to Cyanide, Azide, Anoxia and Chilling

Germination responsiveness to KCN, NaN₃, chilling or anoxiaand respiration activity was compared between non-after-ripenedand after-ripened upper cocklebur (Xanthium pennsylvanicum Wallr.)seeds. The latter, coat-imposed dormant seeds, could germinatein response to the above chemicals and conditions, whereas theformer, primarily dormant seeds, could not respond. There waslittle difference in the respiratory properties of both typesof seeds. (Received June 22, 1981; Accepted October 30, 1981)     

Failure of Chlorophyll Formation in the Cotyledons from Secondarily Dormant Cocklebur Seeds

The cotyledons of secondarily dormant cocklebur (Xanthium pennsylvankumWallr.) seeds lacked not only growing potential but also chlorophyll-formingability. These properties developed as the seeds establishedsecondary dormancy during a soaking period. The inability toform chlorophyll in cotyledonary segments was not improved inthe presence of S-amino levulinic acid. But this ability wasgreatly restored by benzyladenine or ethylene treatment, whichis effective in increasing cotyledonary growth. Similarly, theapplication of ethylene together with enriched oxygen and carbondioxide, or subsequent to a KCN treatment, which were the mosteffective means for breaking secondary dormancy, completelyrestored both chlorophyll formation and growing ability of thecotyledons. The depression of the greening-ability in cotyledonsmay be involved in development of secondary dormancy of cockleburseeds in concert with the decline of their growth pontential. (Received September 24, 1982; Accepted December 27, 1982)     

Metabolic Changes in Partially Dormant Wheat Seeds during Storage

Changes in germination, seedling growth, respiration, response to applied gibberellic acid, and glucose-U-¹⁴C utilization were investigated in partially dormant wheat (Triticum aestivum L., Pa 151 × 107) seeds which were stored under various conditions for periods up to 1 year. Only seeds stored at −20 C and 12.4% moisture maintained partial dormancy, which was overcome by germinating in 10⁻³m gibberellic acid. Germination and seedling growth of seeds stored at 25 C and 15.1% moisture declined within 12 weeks and the percentage of seeds infected with storage fungi increased. Gibberellic acid produced faster growing seedlings, particularly from those seeds with partial dormancy, but did not overcome growth reduction which was caused by deterioration. Seeds kept under laboratory conditions (B), 25 C and 12.1% moisture (C), and 25 C and 15.1% moisture (D) for 12 weeks utilized 35, 55, and 80% less glucose, respectively, than those stored at −20 C and 12.4% moisture (A). Seeds stored under B and C consistently had higher germination, growth, and respiratory rates than seeds from A and D. The respiratory rate declined as deterioration advanced under D. Respiratory quotients ranged from 1.0 for seeds stored under A to 1.6 for seeds stored under D.     

Changes in Ethylene Production and in 1-Aminocyclopropane-1-carboxylic Acid (ACC) and Malonyl-ACC Contents of Cocklebur Seeds during Their Development

Ethylene production in developing cocklebur (Xanthium pennsyluanicumWallr.) seeds peaked when the dry weight of the seeds beganto increase in the early period of development. The productionthen began to decrease and stopped when the dry weight increasewas completed. The upsurge of ethylene production in the earlydevelopmental period paralleled increases in ACC synthase activityand the 1-aminocyclopropane-1-carboxylic acid (ACC) contentof the seeds, both of which rapidly decreased later. Malonyl-ACC (MACC) accumulated in developing cocklebur seedsduring the early period of development, before the ACC contentand ethylene production increased. Although the ACC synthaseactivity, ACC content and ethylene production showed markeddecreases, the MACC content remained almost unchanged duringthe middle period of seed development, with a pronounced decreaseoccurring in the late period. Exogenous application of MACCdid not promote ethylene production of seeds collected at thelate developmental stage. Aminoethoxyvinylglycine, an inhibitorof ACC synthase, strongly inhibited the ethylene productionof the same lot of seeds. Therefore, the decrease in the MACCcontent in developing cocklebur seeds was not due to reuse ofMACC for ethylene production. (Received May 24, 1984; Accepted August 15, 1984)     

Changes in Ribonuclease Activity and Content of RNA during Dormancy and Growth of Bird-cherry Buds

Ribonuclease has been extracted from buds of bird-cherry (Prunus padus L.) by a three-step method. In step 1 ribonuclease was extracted from the homogenized tissue at p^H 6. In step 2 the tissue residue from (1) was extracted at p^H 7. In step 3 the tissue residue from (2) was extracted with a buffer of p^H 8.5 and containing 0.7 M KCl. The ribonuclease from step 1 and 2 is named soluble and that from step 3 bound ribonuclease. The activity of the soluble ribonuclease was very low in October and increased slowly until March. When the buds sprouted in April the activity of this fraction rose rapidly. The activity of the bound ribonuclease was rather high in October and increased continuously from autumn to spring. RNA accumulated slowly between autumn and spring. The change in activity of the two ribonuclease fractions is discussed in relation to development.     

The Metabolism of Hormones during Seed Germination and Release from Dormancy: III. The Effects and Metabolism of Zeatin in Dormant and Nondormant Ash Embryos

Zeatin and zeatin-9, β-ribonucleoside enhance the germination of dormant ash embryos. While the first macroscopic signs of germination appear only after about 72 hours, 12 hours of exposure to 50 μm zeatin is as effective as continuous incubation. There must be barriers against transport out of the embryos since 8-¹⁴C-zeatin and its metabolites, zeatin-9, β-ribonucleoside, the 5′-mono and the suspected di- and triphosphates, accumulate against a concentration gradient. Zeatin ribonucleoside is about as effective as zeatin in enhancing embryo germination, yet the internal 8-¹⁴C-zeatin level is lower by a factor of about 50 when the ribonucleoside is fed. The physiological effects of zeatin and abscisic acid on the germination of ash embryos are antagonistic. There is, however, no evidence that abscisic acid has a significant effect on 8-¹⁴C-zeatin uptake or conversions.     

Dormancy and Impotency of Cocklebur Seeds: IV. Effects of Gibberellic Acid, Benzyladenine, Thiourea, and Potassium Nitrate on the Growth of Embryonic Axis and Cotyledon Segments

Germination of nondormant but impotent small cocklebur seeds (Xanthium pennsylvanicum Wallr.) was promoted profoundly with thiourea or benzyladenine, and slightly with gibberellic acid. Gibberellic acid was ineffective in causing the germination of dormant cocklebur seeds, although thiourea and benzyladenine were effective. Experiments with excised seed pieces showed that the promotive effects of thiourea, benzyladenine, and gibberellic acid on cocklebur seed germination were associated with the enhancement of growth of seed parts; thiourea stimulated predominantly the axial growth, whereas benzyladenine stimulated predominantly the cotyledonary growth.     

Identification and Content of 1-Malonylaminocyclopropanecarboxylic Acid in Germinating Cocklebur Seeds

A major conjugate of 1-aminocyclopropanecarboxylic acid in germinatingcocklebur (Xanthium pennsylvanicum Wallr.) seeds was isolatedand identified as 1-malonylaminocyclopropanecarboxylic acid(MACC). The change in MACC content during the germination periodof this seed also was examined. (Received November 4, 1983; Accepted March 15, 1984)     

Dormancy in Dioscorea: Rapid Germination of Detached Embryos from Dormant Seeds of D. tokoro

The intact dormant seeds of Dioscorea tokoro germinate slowlyif at all between 11-23°C; for full and rapid germinationthey require prior chilling treatment [Okagami and Kawai (1982)Bot. Mag. Tokyo 95: 155]. The germination abilities of zygoticembryos detached from dormant seeds of this species were studiedunder various nutritional and temperature regimes. For germinationof embryos, the minimum nutritional components in Murashigeand Skoog's (1962) medium that were required were sucrose andNO^–₃ or SO^2–₄. As the source of carbohydrate forgermination of detached embryos, sucrose, mannose and maltosewere effective; glucose and fructose were less effective; andrhamnose was entirely unable to support germination. Embryos detached from dormant seeds, incubated with the sucroseplus KNO₃, germinated more rapidly with increasing temperatureup to 35°C. However, application of sucrose and KNO₃ didnot induce germination of intact seeds above 26°C. Therefore,it is very possible that the endosperm exerts an inhibitoryfunction on germination at such high temperatures. When seeds were incubated after a cut was made over a smallpart of the edge of the endosperm in which the radicle of theembryo is encased, germination occurred rapidly but the increasein germination percentage was slight. This result suggests thatthe endosperm suppots part of the germination inhibition bymeans of a mechanical barrier or its impermeability to wateror gases. Physiological features of the endosperm alone or interactionsbetween the embryo and endosperm may contribute significantlyto the characteristics of dormancy of intact seeds of this species. (Received May 30, 1988; Accepted January 11, 1989)     

Changes in Respiratory Potential of Dormant and Non-dormant Galium aparine L. (cleavers) Seeds during Dry Storage

Hilton, J. R. and Thomas, J. A. 1987. Changes in respiratorypotential of dormant and non-dormant Galium aparine L. (cleavers)seeds during dry storage.—J. exp. Bot. 38: 1484–1490. Pre-germinative rates of O₂ uptake of two collections of Galiumaparine L. seed were compared throughout a 9 month period ofdry storage at 4 °C, 23 °C and at ambient (frost-protected)temperatures. Uptake of O₂ by the dormant seeds was generallyhigher than that of the less dormant seeds except when freshly-harvested.Moreover, changes in the O₂ consumption of seeds stored at ambienttemperatures could be associated with periods of germinationand seedling emergence in the field. The results are discussedin relation to changes in respiratory metabolism during dormancybreakage.     

Rhythmic Behavior in Germination of Cocklebur Seeds

Non-dormant, lower seeds of cocklebur (Xanthium pensylvanicum Wallr.) germinated with unimodal flush after 20 and 36 h from the start of water imbibition at 33 and 23°C, respectively. At 28°C, however, germination occurred bimodally, the time of each peak coinciding with that at 23 and 33°C. This type of germination behavior was induced even at 33°C, when the seeds were contacted with some osmotica. Further, the application of different osmotica at 28°C caused a rhythmic multimodal germination with a period of about 16 h. It was suggested that an endogenous rhythmicity may be involved in the control of cocklebur seed germination.     

Activity of the Pentose Phosphate Pathway and Changes in Nicotinamide Nucleotide Content during Germination of Seeds of Cicer arietinum L.

Changes in the level of nicotinamide nucleotides, rate of ¹⁴CO₂output from [1^–14C] or [6¹⁴ C₆/C₁ ratios, glucose-6-phosphatedehydrogenase, 6-phosphogluconate dehydrogenase, and NAD kinaseactivities were determined during the first 72 h of germinationof seeds of Cicer arietinum L. The level of oxidized and reducedforms of nicotinamide nucleotides, together with the activityof glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase,NAD kinase, and C₆/C₁ ratios, suggest that the pentose phosphatepathway is activated during early germination in cotyledonsof chick pea seeds. The results obtained in embryonic axes seemsto indicate a lower participation of the PP pathway, probablydue to the development of the activity of the glycolytic-TCApathway.     

Changes in Phosphorus Fractions and Phytase Activity of Rice Seeds during Germination

The present experiment provides information on the phosphorus compounds in rice seeds and elucidates the changes they undergo during germination. In ungerminated seeds, the bulk of total-P appears in phytin (about 76 per cent). It is then dephosphorylated in course of germination with a simultaneous accumulation of large amounts of inorganic-P. Lipid-P increases very rapidly from 0 to 24 hours. The increase up to 72 hours is gradual, after which it drops at 96 hours and again rises to a maximum after 120 hours. The ester-P and RNA-P, fractions increase in concentration with time of germination (except 120 hours). Protein-P begins to fall after 48 hours, while DNA-P remains more or less constant throughout the experiment. The two pH optima recorded for phytase activity at 4.0 and 9.0, suggests that there exist two phytases, one acidic and the other alkaline. Both behave similarly during germination with a continuous increase throughout the course of the experiment. The enzyme with an optimal pH at 4 hydrolyses phytin more actively than the other with the pH optimum at 9.0. Phytase shows maximum activity at a stage when most of the phytin has disappeared; the metabolic significance is uncertain.     

The Activity of Principal Antioxidant Enzymes and the Content of Metabolites in Dormancy Breaking and Germination of Davidia involucrate Seeds

The principal antioxidant enzymes and metabolites were reported to play a very important role in eliminating reactive oxygen species and maintaining vigour in dormancy breaking and germination of Davidia involucrate seeds. However, the researches on the change laws of content and activity are seldom. In present study, we chose Davidia involucrate seeds which collected from the Mabian Nature Reserve as experimental materials, and then investigated the activity of principal antioxidant enzymes and the content of metabolites in dormancy breaking and germination of the seeds. The results showed that the activity of peroxidase (POD), catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR) exhibited an increasing trend but the activity of total antioxidant capacity (T AOC) decreased greatly in early stage of stratification. Comparing with the decrease of protein (Pr), the content of ascorbic acid (Vc) and free amino acid (FAA) significantly increased. In middle stage of stratification, the activity of SOD and T AOC increased firstly and then decreased at last. The activity of POD always maintained a higher level although the activity of CAT and GR decreased greatly. In terminal stage of stratification, the activities of POD, GR, T AOC and the content of FAA and Vc exhibited an increasing trend, while the activity of CAT and SOD maintained a low level. In germination stage, the activity of all antioxidant enzymes and the content of Vc and FAA were increased obviously (P < 005), but the content of the protein was decreased. These results suggested that Davidia involucrata seeds may have a good ability to efficiently remove the effects on cell structure caused by reactive oxygen species (ROS) by keeping the interaction of antioxidant enzymes and metabolites successively during the period of dormancy breaking and germination. This adjustment helps to ensure the germination of seeds by acceleration the post ripening in morphological and physiological of progresses.     

Water Content and the Conversion of Phytochrome Regulation of Lettuce Dormancy

In an effort to determine which biological reactions can occur in relation to the water content of seeds, the regulation of lettuce seed dormancy by red and far red light was determined at various hydration levels. Far red light had an inhibiting effect on germination for seeds at all moisture contents from 4 to 32% water. Germination was progressively stimulated by red light as seed hydration increased from 8 to 15%, and reached a maximum at moisture contents above 18%. Red light was ineffective at moisture contents below 8%. Seeds that had been stimulated by red light and subsequently dried lost the enhanced germinability if stored at moisture contents above 8%. The contrast between the presumed photoconversion of phytochrome far red-absorbing (Pfr) to (Pr) occurring at any moisture content and the reverse reaction occurring only if the seed moisture content is greater than 8% may be explained on the basis of the existence of unstable intermediates in the Pr to Pfr conversion. Our results suggest that the initial photoreaction involved in phytochrome conversion is relatively independent of water content, while the subsequent partial reactions become increasingly facilitated as water content increases from 8 to 18%.     

Phytochrome Transformation and Action in Seeds of Rumex crispus L. during Secondary Dormancy

Promotion of germination by red light fails after prolonged dark imbibition of Rumex crispus L. seeds, indicative of a secondary dormancy. The degree and rate of inception of the dormancy increases with increasing temperature. Following establishment of the dormancy, germination response to red light can be restored by either prolonged cold treatment or brief high temperature shifts. Loss of phytochrome was not a factor in the initial establishment of the dormancy. When the seeds are in secondary dormancy, the chromophore of phytochrome can be transformed to the far red-absorbing form, but the far red-absorbing form cannot induce germination. The responses to changes in temperature suggested dependence of germination on order disorder transitions in components of the seeds.