Onset Of Germination Research Articles

A comparative study of the seed germination biology of a narrow endemic and two geographically-widespread species ofSolidago(Asteraceae). 2. Germination responses of buried seeds in relation to seasonal temperature cycles

AbstractSeeds of the narrow-endemicSolidago shortiiand of the geographically-widespreadS. altissimaandS. nemoraliscollected and buried in 1992 and 1993 were incubated in light and in darkness at 15/6°, 20/10°, 25/15°, 30/15° or 35/20°C following various periods of burial in soil in a non-temperature-controlled glasshouse. At maturity in November, seeds of the three species germinated to 0–1% in light at 15/6°C and to 10–77% at 20/10°, 25/15°, 30/15° and 35/20°C. Seeds exhumed each April from 1993 to 1996 and incubated in light at 15/6° and 20/10°C germinated to ≥83% and ≥90%, respectively, whereas those exhumed each September of 1993–96 germinated to ≤2%and ≤40%, respectively. At 25/15°, 30/15° and 35/20°C in light, seeds of S.altissimaandS. shortiigerminated to ≥52% and those of S.nemoralisto ≥19/, regardless of when they were exhumed. Timson's index, which integrates percentages, rates and times for onset of germination, was higher at all temperature regimes for seeds exhumed in April 1995 than for those exhumed in September 1995. Freshly-matured seeds of the three species germinated to 0–11% in darkness. Furthermore, regardless of when they were exhumed, seeds ofS. altissimaandS. nemoralisincubated in darkness germinated mostly to only 0–9% over the range of temperature regimes. In contrast, ≤88% and ≤6% of the seeds ofS. shortiiexhumed and incubated in darkness each April and September of 1993–96, respectively, germinated, ≤1% of them germinating while buried in soil. Thus, although buried seeds of all three species exhibited an annual conditional dormancy/non-dormancy cycle, only those ofS. shortiiexhibited cyclic changes in their germination response in darkness.

A comparative study of the seed germination biology of a narrow endemic and two geographically-widespread species ofSolidago(Asteraceae). 1. Germination phenology and effect of cold stratification on germination

AbstractSolidago shortiiis endemic to a small area in northcentral Kentucky (USA), whereas two of its sympatric congeners,S. altissimaandS. nemoralis, are geographically widespread. Seeds (achenes) ofS. shortii(0.370 mg) are significantly larger (PLSD,P=0.05) than those ofS. altissima(0.070 mg) andS. nemoralis(0.068 mg). Germination percentages of freshly-matured seeds of the threeSolidagospecies collected in November 1991, 1992 and 1994 were 0–2% in light at 15/6°C, 1–37% at 20/10°C, 9–56% at 25/15°C and 10–85% at 30/15 and 35/20°C. Stratification increased the percentage and rate of germination and decreased the time to the onset of germination (measured by Timson's index only at 20/10°C in light) in the three species. Following 12 weeks of cold stratification in light, seeds of the three species germinated to 72–100% in the light and to 22–100% in darkness over the range of thermoperiods; those cold-stratified in darkness germinated to 39–100% in light. Freshly-matured seeds ofS. altissimaand ofS. nemoralisgerminated to 0–4% in darkness, whereas those cold-stratified for 12 weeks in darkness germinated to 0–28% in darkness. On the other hand, freshly-matured and cold-stratified (in darkness) seeds ofS. shortiigerminated to 0–13 and 13–73%, respectively, in darkness. Under near-natural temperatures in a glasshouse without temperature control, germination of the three species peaked in March. Thus, the primary difference in dormancy-breaking and germination requirements of the three species is that the endemic germinates to a much higher percentage in darkness than its two congeners. Seeds ofS. shortiido not have any special dormancy-breaking or germination requirements that could not be fulfilled outside its present-day geographic range.

Metabolism of inositol 1,4,5-trisphosphate in Candida albicans: significance as a precursor of inositol polyphosphates and in signal transduction during the dimorphic transition from yeast cells to germ tubes.

The metabolism of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] was examined in yeast cells and germ tubes of Candida albicans. Methods have been developed for analysis of the two key metabolic enzymes, Ins(1,4,5)P3, kinase and phosphatase. ATP-dependent Ins(1,4,5)P3 kinase activity was detected predominantly in the soluble fraction of cell extracts and exhibited a Km of approximately 9 microM. The apparent Km of Ins(1,4,5)P3 phosphatase for Ins(1,4,5)P3 was approximately 480 microM. The slow rate of dephosphorylation of Ins(1,4,5_P3 to inositol bisphosphate suggests a lower importance of the phosphatase within cells compared to the kinase. Since both yeast cells and germ tubes of C. albicans rapidly phosphorylated Ins(1,4,5)P3 to inositol tetrakisphosphate and inositol penta/hexakisphosphate, it is suggested that Ins(1,4,5)P3 has an important role as a precursor for production of these compounds. A sustained increase in cellular Ins(1,4,5)P3 levels was observed during germ tube formation and, prior to the onset of germination between 1 and 2 incubation, the Ins(1,4,5)P3 content increased up to eightfold. Transient increases in the level of Ins(1,4,5)P3 were also observed during yeast-like growth of C. albicans. The possible role and relative importance of Ins(1,4,5)P3 as a precursor for inositol polyphosphates and in signal transduction involving Ca2+ release from internal stores is discussed.

Improvement of Citrus somatic embryo development by temporary immersion

Liquid medium improves and facilitates somatic embryo development from Citrus deliciosa Ten. suspension cultures. Three different culture conditions were compared to determine a means of overcoming poor somatic embryo development. Somatic embryos derived from suspension cultures were plated on solid medium, maintained in suspension culture or temporarily immersed. About 60% of somatic embryos plated on solid medium developed to the cotyledonary stage, but were hyperhydric. Continuous growth in suspension culture at 100 rpm hindered cotyledon and protoderm formation, and somatic embryos were unable to develop beyond the globular stage. Temporary immersion promoted somatic embryo development, i.e. 66% of the somatic embryos produced were cotyledonary, and were morphologically similar to nucellar embryos. This latter culture system also improved regeneration synchronization by hampering secondary embryogenesis at the onset of germination. Irrespective of the culture system used, most cotyledonary somatic embryos studied had no caulinary meristem or starch and protein reserves, thus explaining the low germination rates obtained.

Germination of Alternaria linicola conidia on linseed: effects of temperature, incubation time, leaf wetness and light regime

Conidia of Alternaria linicola germinated on both water agar and linseed leaves (detached or attached) over a wide range of temperatures (5–25°C) by producing one to several germ tubes. At temperatures between 10°C and 25°C and under continuous wetness in darkness, germination started within 2 h after inoculation and reached a maximum (100%) by 8 to 24 h, depending on temperature. At 5°C, the onset of germination was later and the rate of germ tube elongation was slower than that at 10–25°C. During germination, conidia of A. linicola were sensitive to dry interruptions of wet periods and to light. Short (2 h) or long (12 h) dry interruptions occurring at any time between 2 and 6 h after inoculation stopped conidial germination and germ tube elongation. With continuous wetness, light periods 2 to 12 h long immediately after inoculation inhibited conidial germination, which was resumed only when a dark period followed subsequently. However, germination and germ tube elongation of A. linicola conidia stopped and the viability of the conidia was lost during exposure to dry light periods immediately after inoculation with spore suspensions. Penetration of leaves by A. linicola was evident after 12 h and occurred mainly through epidermal cells (direct) with or without the formation of appressoria.

Gemmules of sponges from a warm lake

1. Two types of gemmules were found, each in a different species of sponge, from the warm monomictic Lake Kinneret: (i) clustered gemmules, sharing the pneumatic layer of the gemmular capsule and resembling gemmules of Eunapius; (ii) gemmules that develop non‐synchronously, containing amphidisc spicules within the gemmular capsule and resembling those of Ephydatia. Algal cells were not detected within either type of gemmule although they exist in the developed sponges.2. Sponges began producing gemmules in the lake with the onset of lake drawdown and ceased when lake level was minimal. The gemmules hatched when the lake level began to rise.3. Under experimental conditions gemmules hatched between 13 and 35 °C. Germination was optimal at 20–25 °C. Chilling of young gemmules prior to incubation at 25 °C improved germination rate.4. The percentage of germinating dry gemmules diminished 4–6 months after their collection from the lake. None germinated after 10 months. Submerged gemmules maintained high viability with ageing (up to 100% germination 18 months after collection). Desiccation influenced gemmule viability over time, by both decreasing the percentage of germinating gemmules and increasing the lag time before onset of germination.5. Gemmules kept in the dark germinated significantly less than those illuminated for 12 h day

Tissue-specific expression of the rolA gene mediates morphological changes in transgenic tobacco.

The spatial and temporal activity of the entire and individual promoter domains of the rolA gene of Agrobacterium rhizogenes was investigated and correlated with the distinctive features of the phenotypes of transgenic tobacco plants. The GUS assay was performed in the presence of an oxidative catalyst during the development of transgenic plants expressing chimeric genes containing the beta-glucuronidase coding sequence under the control of the different promoter domains. In situ hybridization was also used on transgenic plants harbouring rolA under the control of the entire or deleted promoter. This paper demonstrates for the first time that the entire rolA promoter, composed of domains, A, B and C, is silent in seeds, then activated at the onset of germination in the cotyledons and in the elongation zone of the radicle and is finally expressed throughout the vegetative and floral phases. Domains B + C, which were sufficient to induce wrinkled leaves and short internodes, were active in all the stem tissues, but only in the companion cells of the phloem strands of the leaves. Domain C, which specified a dwarf phenotype with normal leaves, was weakly expressed in the stem vascular bundles and in the leaf internal phloem. These results indicate that the vascular bundles are the primary targets for the generation of the short internode phenotype. Furthermore, the local expression of rolA in the stem vascular bundles induced a size reduction of the surrounding parenchyma cells, suggesting the existence of some diffusible factor(s) associated with the expression of the rolA gene.

RELEASE AND ACTIVATION OF BARLEY β-AMYLASE

The aim of this study was to determine the role of low molecular weight thiols both in the release and activation of β-amylase during grain germination. In quiescent barley grains (Hordeum vulgare L. cv. Torrent) about 55% of the β-amylase was extracted with buffer, the remaining 45% was in the bound form. During micromalting the bound form was progressively solubilised between germination days 1 and 4. When free β-amylase, extracted from ungerminated grains, was incubated with dithiothreitol the enzymic activity increased by 15%-20%. This activation did not occur when free β-amylase, from grain germinated for 3 days or more, was incubated with DTT. The release of bound β-amylase with thiols was pH dependant, occurring most rapidly at and above pH 8.0. At the onset of germination the embryo released soluble thiol (approximately 5 nmol per embryo) into the endosperm. Degermed grains were dosed with reduced glutathione and incubated for 72 h. The addition of 60 nmol glutathione caused the release of about 80% of the bound β-amylase. When less glutathione was used, 5 nmol (an amount similar to that released by the embryo in vivo) no significant release of the bound enzyme was detected. When degermed grains were dosed with oxidised glutathione (60 nmol), no bound β-amylase was released. However, addition of the disulphide bis-hydroxyethyldisulphide (60 nmol) did cause the release of about 90% of the bound enzyme. The aleurone layer reduced the bis-hydroxyethyldisulphide to a thiol, presumably 2-mercaptoethanol. Oxidised glutathione and cystine were not significantly reduced to thiols by isolated aleurone layers. The aleurone layer did cause the disappearance of cysteine from solution. When preparations of bound β-amylase were incubated with extracts from the endosperms of grains germinated for three days, the bound enzyme was released. This release was due to the high molecu lar weight material (>5 kDa) in the extract and not to low molecular weight thiols. It seems unlikely that simple thiols, such as glutathione, are solely responsible for the release of bound β-amylase.

Isolation and characterization of two distinct cDNA clones encoding corn seed cysteine proteinases

We obtained two cDNA clones encoding corn seed cysteine proteinases (CCP1 and CCP2). Sequence analysis showed that CCP1 consists of 371 amino acid residues, in a prepro-protein form, with two unique short insertions in the mature protein region that are not found in papain or other common CPs. CCP2 consists of 360 amino acid residues with a vacuole sorting signal in the pro-sequence region. An amino acid sequence similarity of 42% was found between the mature protein region of CCP1 and that of CCP2. Although CCP1 is highly homologous to pea 15a CP (72%) and Arabidopsis thaliana RD19 CP (79%), both of which are known to be induced only when the plant is exposed to a dehydrated environment, it showed very low homologies to other known cysteine proteinases (38–43%). CCP2 showed as much as 87% and 89% identity to rice oryzain γ and barley aleurain, respectively. We also observed that the CCP1 mRNA is expressed in ripened corn seeds, although its expression reaches a maximum 5 days after the onset of germination. On the other hand, the CCP2 mRNA is expressed only during germination, with maximum expression at the 3-day stage. These results suggest the presence of at least two cysteine proteinases playing differential roles in the corn seeds.

Catabolism of the Seed Storage Proteins From Lupinus Albus: Fate of Globulins During Germination and Seedling Growth.

The degradation of the major seed storage globulins of Lupinus albus L. was studied during the first 11 days after the onset of germination. The collagenolytic activity present in cotyledonary extracts was found to closely follow the rate and pattern of total globulin degradation. Little or no degradation of the three main globulins was observed during the first days of germination and seedling growth. The rate of proteolysis of the �-conglutin main subunit, composed of one heavy glycopolypeptide chain and one light non-glycosylated polypeptide chain, occurs gradually between days 3 and 11, with no apparent change in the globulin structure being detected during this time. Between days 3 and 5, �-conglutin, with at least 14 of its heavier polypeptide chains glycosylated, suffers a dramatic change in its structure and concentration, involving the appearance of a new set of polypeptides including a higher molecular mass group, whose concentration steadily declines until complete disappearance after 11 days, and a lighter molecular mass group, whose concentration surprisingly increases from 5 to 11 days. Finally, �-conglutin, with each of its four main subunits composed of a heavy glycopolypeptide chain and a 19 kDa non-glycosylated polypeptide chain, undergoes a pattern of degradation intermediate to those described for �- and �-conglutins in the sense that the new set of polypeptides coexist with polypeptides that were already present in the dry seed. The degradation of a-conglutin subunits starts at the level of their heavier polypeptide chains, releasing the intact 19 kDa polypeptide which gradually disappears after day 7. Our results indicate that �-, �- and �-conglutins are subjected to differential proteolysis during germination and seedling growth. Furthermore, the different polypeptide chains of each conglutin are also degraded at significantly different rates.

Ecological Studies of the Endangered Rutidosis leptorrhynchoides. II. Patterns of Seedling Emergence and Survival in a Native Grassland

The emergence, survival and growth of seedlings of the endangered Rutidosis leptorrhynchoides F.Muell. were followed in a Themeda triandra grassland during 1991 and 1992. The effect of summer irrigation on seedling survival was also investigated. Seedling emergence occurred in both years within 2 weeks of the 'autumn break' when soil moisture rose above 20%. Ninety percent of emergence was observed within 4 weeks of the onset of germination and 87% of seedlings were within 20 cm of an established plant. No emergence was observed after 8 weeks. Thirteen percent of the 1991 cohort survived for 14 months. Mortality of most seedlings (63%) was attributed to soil moisture stress in summer. Small seedlings (Ͱ4 3 leaves) were no more susceptible to drought than larger seedlings. Seventy two percent of the 1991 cohort produced four leaves before subsequently dying. In 1992, however, most early seedling mortality was amongst cotyledonary seedlings. No seedlings flowered in their first year. Above-ground growth was slow and by 14 months, 60% of surviving seedlings had seven or fewer live leaves. Irrigation in a year of below-average rainfall had no significant effect on the survival and growth of seedlings. This suggests that seedling recruitment is not restricted to climatically favourable years (i.e. is not episodic) but rather, is potentially on-going provided suitable microsites are available for seedling survival.

Adhesion of ungerminated conidia of Colletotrichum graminicola to artificial hydrophobic surfaces

Conidia of Colletotrichum graminicola , the causal agent of corn anthracnose, begin to adhere to hydrophobic surfaces immediately after contact with the surface and hours before the onset of germination. Conidia rarely adhered to glass, a hydrophilic surface. The maximum percentage adhesion of ungerminated conidia was approx. 30–40% on polystyrene Petri dishes and 20–25% on dimethyldichlorosilane (DMS)-coated glass slides. The maximum adhesion on both surfaces was attained within 30 min of the time of contact. Thus, conidia appear to require hydrophobic surfaces for adhesion. The extracellular mucilage that is produced in association with conidia was shown to not be involved in adhesion. Adhesion was not influenced by conidium concentration but was influenced by conidium age. Exposure of conidia to different temperatures in adhesion assays delayed the ability of conidia to adhere. The respiration inhibitor, sodium azide and the transcription inhibitor, actinomycin D, had no effect on adhesion. In contrast, inhibitors of glycoprotein transport (brefeldin A) and protein synthesis (cycloheximide) reduced conidial adhesion by 30 and 50%, respectively. Treatment of conidia with pronase E prevented adhesion completely. If conidia were pre-treated with pronase E and then washed free of the enzyme, approx. 20 % of the conidial population recovered the ability to adhere. Conidia treated with Concanavalin A at a concentration of 1 mg ml −1 also failed to adhere. This effect was partially reversed by the haptens glucose and mannose. Together, these results suggest that glycoprotein synthesis is involved in the adhesion of ungerminated conidia.

Wheat seed ageing and embryo protein degradation

AbstractWheat seed lots of different viability were obtained by an accelerated ageing technique in which seeds were hydrated up to 12.5–14.5% on a fresh weight basis and stored at 35°C. Ageing was accompanied by a progressive delay of the onset of germination and a decrease of [35S]-methionine uptake and incorporation into soluble proteins of the imbibing embryos. A detailed analysis of newly synthesized proteins by two-dimensional polyacrylamide gel electrophoresis showed that normal protein patterns during the germination process were impaired by ageing. Only two groups of polypeptides with molecular masses of 84.9–69.5 and 66.6–60.2 kDa, respectively, were actively produced while most proteins decreased or were definitely degraded. Our results suggest that the biochemical changes required for germination are gradually opposed by a deterioration process and that some newly-synthesized polypeptides can be taken as markers in assessing seed survival of ageing stress.

Regulation of protein kinase A subunits during germination of Mucor rouxii sporangiospores.

Levels of protein kinase A (PKA) subunits and of cAMP have been measured during aerobic germination of the sporangiospores of the dimorphic fungus Mucor rouxii; further, the holoenzyme and its catalytic (C) and regulatory (R) subunits have been visualized through sucrose gradient centrifugation. Sporangiospores contain around 0.06 microM of a dimeric holoenzyme species of 5.5 S and a sixfold excess of a free R subunit of 2.7 S. Both these species are proposed to be derived by proteolysis from the native forms. Enzymic activity at this stage is highly inhibited, as demonstrated with permeabilized cells. Immediately upon germination, and after a transient increase in cAMP concentration from 10 microM to 90 microM, C-subunit levels fall to 30%. After the onset of germination, the specific activity and concentration of both the 5.5 S holoenzyme species and the 2.7 S species of free R subunit decrease in parallel to the increase in total protein and volume. Net synthesis of C and R subunits to form a native holoenzyme species of 8.8 S is apparent 4 h onwards after germination. A significant increase in cellular concentration is observed at 6 h. At 7 h of growth, when germ-tube emission is complete, the holoenzyme concentration is around 0.23 microM; there is almost no free R subunit and the intracellular concentration of cAMP is around 3 microM. A role for PKA during germination and morphogenesis is discussed.

A freeze-fracture study of mesophyll cell membranes in the embryonic Phaseolus leaf, in the dry seed and during the initial stages of germination

Using rapid-freezing and freeze-fracturing techniques, we have examined cellular organisation within mesophyll cells in primary leaves of Phaseolus, in the dry seed and over a period of 48 hr from the onset of germination. The rapidly-frozen, unetched image reveals a dynamic cytoplasm with membrane shape and role able to change and fluctuate with the increase in cell metabolism. The ER in this tissue is highly mobile, becoming progressively associated with protein and lipid bodies, then plastids, mitochondria and nuclei as cytoplasmic requirements change. The ER thus provides an efficient transport of substrates to metabolic sites within the cytoplasm. Nuclei and plastics change shape in early germination and the protein body membrane changes in role and character to that of a vacuolar tonoplast. The freeze-fractured image emphasizes the considerable movement that occurs within the cytoplasm during germination, with flexible and mobile membranes of the various organelles able to associate and interact with others. We have also measured the density of intramembrane particles on most of the membranes within the mesophyll parenchyma cell, during this period of early germination. Using our method of rapid-freezing, we find consistently higher IMP density on the protoplasmic face of each membrane. The counts provide a set of comparative values for membranes of a specific tissue, and for a specific cryofixation technique.

Confocal image analysis of spatial variations in immunocytochemically identified calmodulin during pollen hydration, germination and pollen tube tip growth inNicotiana tabacumL.

Using monoclonal anti-calmodulin antibodies in conjunction with confocal scanning laser microscopy we have analysed the spatial variations in the distribution pattern of calmodulin (CaM) during the sequential events of pollen hydration, germination and tube growth in Nicotiana tabacum. These immunocytochemical observations have been complemented by immunochemical studies wherein the anti-calmodulin antibody raised against pea CaM recognises a polypeptide of c. 18 kDa in the pollen extracts. Digitisation of confocally acquired optical sections of immunofluorescence images reveals that in hydrated pollen a high level of CaM is consistently present in the region of the germinal apertures. Subsequently, with the onset of germination a high CaM concentration was found associated with the plasma membrane of the germination bubble and in the cytoplasm in its vicinity, while in the vegetative cytoplasm a weak diffuse and intense punctate signal was registered. CaM immunostain was also detected in association with the plasma membrane of the tube tips in both short and long pollen tubes. Furthermore, the cytosol of the tubes invariably manifested an apically focused CaM gradient. We were, however, unable to detect any vacuolar association of CaM in the older regions of the pollen tubes. Although punctate immunostain was obvious across the pollen tube numerous punctate structures were invariably present in the extreme tip. The possible implications of these findings in development of cell polarity, polarised growth, maintenance of calcium homeostasis and CaM interactions with other mechanochemical motor proteins in effecting propulsion of organelles during pollen hydration, germination and pollen tube growth are discussed.

Characterization and genetic control of germination-emergence responses of grain sorghum to salinity

When grain sorghum is grown in saline soils, one cause of low yield is poor crop establishment. The objectives of this study were to assess the response of grain sorghum to salinity in the germination-emergence stages, study the inheritance of salt tolerance at this stage, and determine the relative contribution to final emergence of salt effects during imbibition, and after onset of germination. Twelve inbred lines and 18 F1 hybrids, resulting from an incomplete 6×6 factorial mating design, were tested for germination and emergence in folded paper at 10 salt concentrations, from 1.8 to 36 dSm-1. The mean EC50 (the electrical conductivity at which the variable score declines by 50%) for emerged seedlings production was 21.2 dSm-1. Large genotypic differences were observed for salt tolerance at germination and emergence stages, which were not related to the viability of seeds, and poorly related to seed weight (considered as an estimate of intrinsic seed vigor). In the hybrids, these differences were due to SCA and female GCA for emergence, and female GCA for germination, though the male GCA was also significant for both characters. Line per se performance was significantly correlated to individual GCA estimates for emergence, but not for germination. Heterosis was only detected in three crosses for final emergence and in one cross for germination. The genetic differences in final emergence were mainly due to effects occurring after the onset of germination rather than a consequence of effects during imbibition.

Effect of oxidative stress during imbibition of soybean embryonic axes

SynopsisBoth respiration and generation by soybean embryonic axes showed a sharp increase upon germination, leading to a significant increase in the steady-state concentration of and H2O2 after 6 h of imbibition. An assay was developed to assess in vivo generation of reactive oxygen species, based upon DCFH-DA oxidation. Fluorescence of the external medium was dependent on reaction time and axes number and was inhibited by catalase.α-Tocopherol content declined significantly after 24 h of incubation, as compared to the content at the onset of germination. Incubation in the presence of redox cycling agent paraquat (4 mM) for 24 h increased α-tocopherol content to 1.9±0.2 nmol per axis from 1.0 ± 0.1 nmol per axis in the absence of paraquat. Supplementation of the incubation medium with 500 μM Fe-EDTA increased α-tocopherol content to 1.8±0.1 nmol/axis and DCFH-DA oxidation by two-fold.The data presented here showed that active metabolism at the onset of germination increased steady-state concentration of oxygen active species and suggest that cellular content of α-tocopherol is physiologically adjusted as a response to conditions of oxidative stress.

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 conditions during storage of infested soil on infection of bait plants by Polymyxa betae and beet necrotic yellow vein virus

Infectivity of resting spores ofPolymyxa betae in soil stored air-dry or moist was determined by assessing infection of bait plants that were exposed to the soil. Storage of soil under air-dry conditions at room temperature resulted in a delayed onset of germination of resting spores compared to germination in soil stored under moist and cool conditions, as inferred from the infection of the bait plants. Bait plants had to be exposed for more than 12 h to flooded infested soil before germination and infection had occurred. However, when soil was prewetted for 24 h before exposing bait plants, germination, infection and transmission of beet necrotic yellow vein virus (BNYVV) were accomplished within 12 h, but only with the moistly stored soil. When resting spores isolated from roots were stored for 4 and 8 weeks under dry conditions at 22°C, germination of viruliferous spores, as measured by detection of BNYVV in bait plants exposed for 48 h to the spores, was less than that of spores stored in moist soil at 22°C. Approximately 100% of bait plants were infected after exposure to resting spores that were frozen in demineralized water or stored cool (5°C) in water or moist soil for 42 weeks. Air-dry cool storage for 42 weeks resulted in a low percentage of infection. Storage conditions of soil influence the results of bioassays for detection of rhizomania when short baiting periods are applied, whereas differences in infectivity were not detected using a bioassay with long duration.