Protein Synthesis in Dormant and Non-Dormant Cocklebur Seed Segments

Using the axial and cotyledonary segments of lower cocklebur (Xanthium pensylvanicum Wallr.) seeds, protein synthesis as shown by incorporation of radioactive leucine was examined in relation to their dormant status. During the first 9 h of water imbibition, the protein synthesis was higher in the dormant axes than in the non-dormant, after- ripened ones. When imbibed for more than 12 h non-dormant axes had a higher activity than dormant ones. This was also the case with the cotyledonary segments. Cyctoheximide, an inhibitor of protein synthesis, blocked protein synthesis in the axial tissue regardless of its dormant status, and thereby inhibited germination of the non-dormant seeds. In the dormant seeds, however, cycloheximide at 3 mM slightly stimulated germination without stimulating the C₂H₄ production. Based on these results, it is suggested that in cocklebur seeds there may be some proteinaceous system which is involved in the maintenance of dormancy.     

Inhibitory Effects of Methyl Jasmonate on the Germination and Ethylene Production in Cocklebur Seeds

Methyl jasmonate (JA-Me) inhibited the germination of cocklebur (Xanthium pennsylvanicum Wallr.) seeds. The inhibition of the germination of cocklebur seeds treated with JA-Me at concentrations less than 300 μm was nullified by ethylene applied exogenously, although the inhibitory effect of 1,000 μm JA-Me was not recovered completely even by high concentrations of ethylene (10,000 μL/liter). JA-Me inhibited ethylene production before seed germination. The level of 1-aminocyclopropane-1-carboxylic acid (ACC) in the cotyledonary tissues treated with JA-Me decreased but not the level of 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC). JA-Me inhibited the conversion of ACC to ethylene in the tissues. These results suggested that JA-Me inhibits ethylene production by prevention of ACC oxidation in addition to ACC synthesis. We believe that the inhibition of ethylene production by JA-Me results in the retardation of the germination of cocklebur seeds. Received June 4, 1997; accepted October 23, 1997     

Stimulation of ethylene production by hydroxamic acid, in particular, by benzohydroxamic acid

The effects on the ethylene production of known inhibitors ofa cyanide-insensitive, alternative respiration in plants wereinvestigated using cotyledonary segments of cocklebur (Xanthiumpennsylvanicum Wallr.) seeds. Benzohydroxamic acid (BHAM) at3 mM stimulated ethylene production 4- to 8-fold over the control,but respiration of the segments was hardly affected at thatconcentration. The stimulatory effects of 3-chlorobenzohydroxamicacid (CLAM) and salicylhydroxamic acid were far smaller thanthat of BHAM. BHAM at 3 mM also markedly stimulated the ethyleneformation in the epicotyl or hypocotyl sections of etiolatedpea (Pisum sativum L.) and mung bean (Vigna radiata [L.] Wilczek)seedlings. Moreover, 3 mM BHAM further promoted the increasedethylene formation which was caused by administration of 1-aminocyclopro-pane-1-carboxylicacid (ACC) to the cotyledonary segments. The promoting effectsby BHAM and CLAM were also found in the conversion of ACC intoethylene in pea stem homogenates. (Received July 26, 1980; )     

Stimulation of ethylene production by hydroxamic acid, in particular, by benzohydroxamic acid

The effects on the ethylene production of known inhibitors ofa cyanide-insensitive, alternative respiration in plants wereinvestigated using cotyledonary segments of cocklebur (Xanthiumpennsylvanicum Wallr.) seeds. Benzohydroxamic acid (BHAM) at3 mM stimulated ethylene production 4- to 8-fold over the control,but respiration of the segments was hardly affected at thatconcentration. The stimulatory effects of 3-chlorobenzohydroxamicacid (CLAM) and salicylhydroxamic acid were far smaller thanthat of BHAM. BHAM at 3 mM also markedly stimulated the ethyleneformation in the epicotyl or hypocotyl sections of etiolatedpea (Pisum sativum L.) and mung bean (Vigna radiata [L.] Wilczek)seedlings. Moreover, 3 mM BHAM further promoted the increasedethylene formation which was caused by administration of 1-aminocyclopro-pane-1-carboxylicacid (ACC) to the cotyledonary segments. The promoting effectsby BHAM and CLAM were also found in the conversion of ACC intoethylene in pea stem homogenates. (Received July 26, 1980; )     

Transition from primary dormancy to secondary dormancy in cocklebur seeds

Abstract The transition from primary dormancy to secondary dormancy was examined using upper cocklebur (Xanthium pennsylvanicum Wallr.) seeds. The non-after-ripened seeds with primary dormancy responded to chilling, anoxia, KCN, and NaN₃ with an increase in germination. However, their maximal responses to these treatments only occurred after a period of water imbibition, probably a reflection of the increasing growth potential of the axial tissue which was accompanied by the increase in the capacities of respiration and ethylene production. On the other hand, the establishment of secondary dormancy was accompanied by a decrease in respiration and ethylene production of seeds, and in the growth potential of both axial and cotyledonary tissues. The decrease in growth potential of these tissues occurred regardless of whether they were excised from after-ripened seeds or non-after-ripened seeds. It is inferred that the primary dormancy of cocklebur seeds is a state maintained in un-germinated seeds for a long time through a spontaneous transition to secondary dormancy.     

Endogenous ethylene and reversing methyl jasmonate inhibition of Amaranthus caudatus seed germination by benzyladenine or gibberellin

BA at 10^–5 M, GA₃ at 3×10^–4 M or GA₄₊₇ at 3×10^–5 M partially or largely reversed the inhibition of Amaranthus caudatus seed germination due to JA-Me. BA or GA₃ did not affect ethylene production and ACC oxidase activity in vivo in the presence of JA-Me before radicle protrusion. However, both increased ethylene production after 72 h of incubation, when the reversal of the JA-Me inhibition of seed germination was observed. AVG at 3×10^–4 M decreased ethylene production when it was applied simultaneously with BA and JA-Me or GA₃ and JA-Me, but it had no effect on seed germination. NBD almost completely reversed the stimulatory effect of BA, GA₃ or GA₄₊₇ on the germination of seeds in the presence of JA-Me. Exogenous ethylene reversed the inhibitory effect of NBD. The results indicate that action of endogenous ethylene is involved in the response of JA-Me inhibited seeds to BA or GA_s.     

Induction of secondary somatic embryogenesis in hybrid larch (Larix x leptoeuropaea) as related to ethylene

Induction of secondary somatic embryogenesis was studied with hybridlarch (Larix x leptoeuropaea)cotyledonary somatic embryos obtained after 3, 4, 5 and 6 weeks of culture on amaturation medium supplemented with abscisic acid. Almost all 3-week maturedcotyledonary somatic embryos can develop embryonal masses whereas only 78, 27and 12% of them are able to do so after 4, 5 and 6 weeks of maturation,respectively. During the first week of culture on the induction medium, somaticembryos with high embryogenic potential (i.e. 3-weekmatured) release little ethylene (less than 1.5 nL h^–1g^–1 FW), whereas those which have almost completelylosttheir ability to induce embryonal masses (i.e. 6-weekmatured) produce much more ethylene. Thereafter, ethylene production by bothtypes of embryos is very similar at around 5–6 nLh^–1 g^–1 FW. Enrichment of theatmosphere with ethylene, or addition of 2-chloroethylphosphonic acid(ethephon)or ACC in the induction medium strongly reduced the induction of secondarysomatic embryogenesis. Moreover, inhibitors of ethylene action(AgNO₃and 2,5-norbornadiene) improved the development of embryonal masses fromsomaticembryos, particularly from the 6-week maturated ones. The results obtainedclearly suggest that ethylene is involved in the regulation of somaticembryogenesis in hybrid larch. The possible relationship between somaticembryogenic potential and ethylene biosynthesis by the explants or sensitivityof the latter to ethylene is discussed.     

Aspects of variation in a neotropical seed dispersal system

This paper explores the causes and consequences of seed and seedling mortality of the tree Virola nobilis (Myristicaceae) Central Panama in order to understand the advantage to local seed dispersal by birds and monkeys. Post-dispersal mortality due to insects (primarily Conotrachelus spp., Curculionidae) accounts for 30–35% of seed and seedling death during the first 12 weeks after seed fall. Because more seeds and seedlings are killed under and near fruiting trees than 15–45 m away, seed dispersal confers a 20–40 fold advantage on seeds carried 45 m from fruiting adults. In contrast, >60% of seed and seedling death during the first year is due to seed predation by mammals, with >90% due to mammals among the <2% seeds that survive until maternal endosperm is exhausted ±12 weeks after seed fall. Mortaliy due to mammals is independent of distance from parent trees, confering no advantage to seed dispersal. Insects account for variation in mortality attributable to distance effects, mammals to between site effects.Early weevil infestations put a premium on seed removal by large birds (Ramphastos swainsonii, R. sulfuratus, Penelope purpurascens), which carry >50% of the seeds that they eat >40 m, as compared with smaller birds (Baryphthengus martii, Tityra semifasciata, Trogon massena) and monkeys (Ateles geoffroyi), which leave most or all of the seeds that they eat under or near the tree crown.     

Germination of cocklebur seed and growth of their axial and cotyledonary tissues in response to C2H4, CO2 and/or O2 under water stress

Abstract. Germination modes of lower seeds of cocklebur (Xanthium pennsylvanicum Wallr.) under different water stresses, prepared with mannitol solution, were examined in relation to gaseous factors. As the concentration of mannitol increased, germination was increasingly inhibited at a mode which was drawn by two straight lines having different slopes and meeting at an angle. One is a sharp line occurring at the lower concentrations of mannitol; the other is a gentle line occurring at higher concentrations of mannitol. The former reflected the growth response of axial tissues to mild water stress, whereas the latter reflected the growth response of cotyledonary tissues to severe water stress. The germination potential of cocklebur seeds increased with increasing temperature. Thus, the seeds were more resistant to water stress at higher than al lower temperatures. This increased germination potential under water stress resulted from the greater growth potential of axial tissues, but not cotyledonary tissues, at higher temperature. Increased O₂ levels improved both the reduced axial and cotyledonary growth under water stress. Carbon dioxide predominantly enhanced axial growth under water stress, whereas C₂H₄ exclusively enhanced cotyledonary growth. Thus, these gases were effective in potentiating germination under water stress. When combined with each other, these gases caused more pronounced growth of the axial and cotyledonary tissues, leading to germination under more severe water stresses. Maximal axial and cotyledonary growth under water stress occurred in the simultaneous presence of CO₂, C₂H₄ and O₂, which allowed the germination at higher mannitol concentrations above 0.6 kmol m^?3 From these results, it was suggested that cocklebur seeds would override water stress by depending upon both the Corresponding axial growth and the C₂H₄-responding cotyledonary growth.     

Dispersal and predation in alien Acacia

Summary I investigated seed removal in the litter layer of alien Acacia stands at bimonthly intervals throughout one year. Both ants (dispersers) and rodents (predators) removed significant quantities of seeds and may compete for seeds in low density Acacia stands. Seed removal from depots was greatest prior to seed-fall (Sept.–Nov.) and lowest during seed-fall (Jan.–Mar.). As rodents may consume a large proportion of the annual seed production at low Acacia densities, I propose that ants have played a critical role in accumulating Acacia seed banks.     

Soil seed banks of woodland, heathland, grassland, mire and montane communities, Cairngorm Mountains, Scotland

The size and species composition of soil seed banks were assessed at 111 altitudinally diverse sites in the Cairngorm Mountains. Mean densities of germinable seeds varied from 83 000 m^–2 in Scots pine (Pinus sylvestris L.) woodland at 230–490 m to 200 m^–2 in moss (Racomitrium lanuginosum (Hedw.) Brid.) heath at 1000–1120 m. Seed banks were dominated by Calluna vulgaris (L.) Hull, not only wherever it was prominent in the vegetation, but also at some sites with less than 5% cover of parent plants in the ground vegetation. Many species conspicuous in the vegetation were under-represented in or absent from the seed bank and surface vegetation generally was more species rich than was the underlying seed bank, especially in high montane communities. Multiple regression was used to examine the relationship between the density of buried Calluna seeds and the abundance of parent plants in the vegetation, site altitude and the organic matter content of the soil. The model fitted to woodland communities accounted for 95% of the variation in seed density. The heathland model was less predictive but still explained 52% of the variation in seed bank size. In mire communities there was no relationship, collective or individual, between buried seed density and the measured environmental variables, possibly due to variations in the duration and frequency of waterlogging at these sites. The potential role of seed banks for initiating the recolonisation of disturbed ground is discussed. Densities of buried seeds at most Calluna-dominant sites were probably sufficient to generate successful recolonisation but the prospects for recovery were poor at other sites, particularly in graminaceous communities at 800 m or higher.     

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.     

Stimulation of ethylene production and gas-space (aerenchyma) formation in adventitious roots of Zea mays L. by small partial pressures of oxygen

Adventitious roots of two to four-weekold intact plants of Zea mays L. (cv. LG11) were shorter but less dense after extending into stagnant, non-aerated nutrient solution than into solution continuously aerated with air. Dissolved oxygen in the non-aerated solutions decreased from 21 kPa to 3–9 kPa within 24 h. When oxygen partial pressures similar to those found in non-aerated solutions (3, 5 and 12 kPa) were applied for 7 d to root systems growing in vigorously bubbled solutions, the volume of gas-space in the cortex (aerenchyma) was increased several fold. This stimulation of aerenchyma was associated with faster ethylene production by 45-mm-long apical root segments. When ethylene production by roots exposed to 5 kPa oxygen was inhibited by aminoethoxyvinylglycine (AVG) dissolved in the nutrient solution, aerenchyma formation was also retarded. The effect of AVG was reversible by concomitant applications of 1-aminocyclopropane-1-carboxylic acid, an immediate precursor of ethylene. Addition of silver nitrate, an inhibitor of ethylene action, to the nutrient solution also prevented the development of aerenchyma in roots given 5 kPa oxygen. Treating roots with only 1 kPa oxygen stimulated ethylene production but failed to promote gas-space formation. These severely oxygen-deficient roots seemed insensitive to the ethylene produced since a supplement of exogeneous ethylene that promoted aerenchyma development in nutrient solution aerated with air (21 kPa oxygen) failed to do so in nutrient solution supplied with 1 kPa oxygen. Both ethylene production and aerenchyma formation were almost completely halted when roots were exposed to nutrient solutions devoid of oxygen. Thus both processes require oxygen and are stimulated by oxygen-deficient surroundings in the 3-to 12-kPa range of oxygen partial pressures when compared with rates observed in air (21 kPa oxygen).Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine     

Fruit dimorphism inPhaseolus sublobatus (Fabaceae) and its evolutionary significance

Populations of the annualPhaseolus sublobatus from different ecogeographical zones are genetically differentiated. In twelve populations from the western ghats (Maharashtra range) chasmogamous flowers are arranged in peduncled capitate racemes borne in leaf axils of higher nodes. One population (Poona Race S₄) has additional inflorescences, also with chasmogamous flowers, on the main axis between the cotyledonary node and the ground. When the pods of these flowers ripen, the inflorescence gradually coils and, eventually, gets bury the fruits in the soil. This phenomenon is not known in any other plant.—The seed-coat patterns as revealed by SEM, and germination behaviour of both aerial and subterranean seeds are similar, and both types of seeds lack dormancy.—Heavy seed predation selection pressure is probably the principal cause for the evolution of fruit dimorphism inP. sublobatus. The origin of geocarpy in relation to fruit dimorphism and seed predation is discussed.     

Selective inhibitors of germination of legume seeds in activated sludge compost

Water extracts of the compost produced from activated sludge and coffee residue were found to be selectively inhibitory to seed germination of some legumes. Germination rate of white clover (Trifolium repens L.), red clover (Trifolium pratense L.) and alfalfa (Medicago sativa L.) seeds were reduced to 2, 29 and 73% of the control, respectively, by water extracts of the compost (20 g l^–1). However, the extracts did not show any inhibition to seed germination of sorghum (Sorghum bicolor Moench), African millet (Eleusine coracana Gaertn.), and Komatsuna (Brassica rapa L.) at the same concentration. The inhibitors in the compost extracts were separated by ion-exchange chromatography and reverse-phase high performance liquid chromatography (HPLC) and the inhibitory activities of seed germination were tested with white clover seeds. Five inhibitors were isolated and identified as 3,4-dichlorophenylacetic acid (3,4-DCP), 3,4-dichlorobenzoic acid (3,4-DCB), 3,4,5-trichlorophenylacetic acid, 3,4,5-trichlorobenzoic acid and mono-2-ethylhexylphthalate by ¹H-, ¹³C-NMR spectroscopy and mass spectrometry. The inhibitory activities of some authentic chemicals of the inhibitors and the related compounds were compared. The results indicated that the main inhibitor in the compost could be 3,4-DCB, which was contained at the concentration of 6.58 mg kg^–1 compost and showed the strongest inhibitory effect on seed germination of white clover among the tested compounds.     

Biométrie des graines et nombres chromosomiques dans la sous-sectionHeterophylla du genreCampanula (Campanulaceae)

Length measurements of seeds belonging to 12 taxa of theCampanula rotundifolia group, including six Pyrenean endemics, prove a certain relation between seed length and ploidy level (2x–4x–6x).

     

Effects of 6-methoxy-2-benzoxazolinone on the germination and α-amylase activity in lettuce seeds

Germination of lettuce seeds was inhibited by 6-methoxy-2-benzoxazolinone (MBOA) at concentrations greater than 0.03 mmol/L. MBOA also inhibited the induction of α-amylase activity in the lettuce seeds at concentrations greater than 0.03 mmol/L. These two concentration–response curves for the germination and α-amylase indicate that the percentage of the germination was positively correlated with the activity of α-amylase in the seeds. Lettuce seeds germinated around 18 h after incubation and inhibition of α-amylase by MBOA occurred within 6 h after seed incubation. These results show that MBOA may inhibit the germination of lettuce seeds by inhibiting the induction of α-amylase activity.     

Efficient<Emphasis Type="Italic"> Agrobacterium tumefaciens</Emphasis>-mediated transformation of soybeans using an embryonic tip regeneration system

Here, we report the establishment of an efficient, in vitro, shoot organogenesis, regeneration system for soybeans [Glycine max (L.) Merr.]. Mature soybean seeds were soaked for 24 h, the embryonic tips were collected and cultured on MSB5 medium supplemented with 3.5 mg l^–1 N⁶-benzylaminopurine (BAP) for 24 h, and explants were transferred to MSB5 medium supplemented with 0.2 mg l^–1 BAP and 0.2 mg l^–1 indolebutyric acid. Use of embryonic tips yielded a higher regeneration frequency (87.7%) than regeneration systems using cotyledonary nodes (40.3%) and hypocotyl segments (56.4%) as starting materials. Regenerated embryonic tips were inoculated with Agrobacterium tumefaciens strain EHA105, which contains the binary vector pCAMBIA2301, and cultured for 20 h. Our results showed that the T-DNA transfer efficiency reached up to 78.2% and the transformation efficiency reached up to 15.8%. These data indicate that the embryonic tip regeneration system can be used for efficient, effective Agrobacterium-mediated transformation.Abbreviations GUS -Glucuronidase - T-DNA Transferred DNA - BAP N⁶-Benzylaminopurine - IBA Indolebutyric acid     

Seed viability and seed dormancy of non-native Phragmites australis in suburbanized and forested watersheds of the Chesapeake Bay, USA

The non-native, invasive haplotype of Phragmites australis is rapidly invading tidal and non-tidal wetlands across North America. Phragmites has the potential to spread by seeds and rhizomes. Seed viability and dormancy differences were quantified among 18 patches of non-native Phragmites in subestuarine wetlands in developed (i.e., suburbanized) vs. forested watersheds of the Chesapeake Bay. We used tetrazolium and germination assays to assess seed viability and compared germination percentages and rate of germination among fresh seeds, cold–moist treated seeds, and warm–dry treated seeds to evaluate seed dormancy. Seed viability was <1% in most patches but a few patches produced abundant viable seeds (5–21%). Seed viability, however, did not differ significantly between wetlands in forested vs. developed watersheds. Contrary to studies of Phragmites seed dormancy in European populations, some Phragmites seeds were dormant at maturity; cold–moist treated seeds germinated faster and to higher percentages than fresh seeds or warm–dry treated seeds.