Ribosomal RNA integrity and rate of seed germination

The integrity of ribosomal RNA (the percentage of complete, un-nicked molecules) in seeds was studied by electrophoresis under denaturing conditions. Two batches of carrot seed, harvested at different stages of maturity, and four batches ofNicotiana seed stored for various times were used. Within each species, there was a correlation between the integrity of the rRNA of the dry seed and the rate of germination of that seed. In carrot seed, there was extensive degradation of existing rRNA in both the embryo and endosperm during the first two days of imbibition.     

Inter-nucleosomal DNA fragmentation and loss of RNA integrity during seed ageing

The germination of viable seeds is the basis for new plant growth and development. Seeds lose viability during storage, but the biochemical mechanisms of seed death are not fully understood. This study aimed to investigate degradation patterns of nucleic acids during seed ageing and subsequent water uptake. Seeds of Pisum sativum L. were artificially aged at 50°C and 12% seed water content (WC). Nucleic acids degradation was studied during ageing and during imbibition of four seed lots with differential viability from highly viable to dead. As seeds lost viability during ageing, DNA was gradually degraded into internucleosomal fragments, resulting in ‘DNA laddering’, in conjunction with disintegration of 18S and 28S rRNA bands. During imbibition, non-aged controls had high levels of DNA and RNA integrity through to radicle protrusion. In an aged seed lot with 85% total germination (TG) DNA fragmentation decreased upon imbibition probably due to nucleosome degradation, while rRNA integrity did not improve. In an aged seed lot with 44% TG, neither DNA nor rRNA integrity improved upon imbibition. Dead seeds showed DNA degradation as laddering throughout imbibition along with extensive degradation of rRNA. We present a model in which interlinked programmed and non-programmed events contribute to seed ageing, and suggest that protection of nucleic acids during ageing is key to seed longevity.     

Responses of Vegetable Seeds to Controlled Hydration

Leek, onion and carrot seeds were imbibed in water and in solutionsof polyethylene glycol (PEG) 6000 over the range –0.5to –4.0 MPa osmotic potential, for periods up to 28 dat 15 C. Seeds started to germinate after 7 and 14 d at –0.5MPa and –1.0 MPa PEG, respectively, but in the lattercase, germination did not exceed 5%. No germination occurredin solutions of lower (more negative) osmotic potential. Seedmoisture content increased with osmotic potential in all threespecies and the relationships were unaffected by the durationof imbibition in solutions of –1.0 to –4.0 MPa,though leek seeds had higher moisture contents than the otherspecies for any given osmotic potential. Linear relationships between response to priming (differencebetween mean germination times of primed and untreated seeds)and seed moisture content were obtained for each species, positiveresponses being obtained above 30–35% seed moisture contentwith optima at 46, 44.5 and 44% seed moisture contents in leek,onion and carrot, respectively. Priming had no effect on embryovolume or cell number per embryo in leek and onion. Carrot embryovolume increased by 43% and cell number per embryo doubled inprimed compared with untreated seeds, whereas seeds imbibedin water showed only a slight increase in cell number per embryoat the stage when radicles were beginning to penetrate the seedcoat. Allium cepa L. cv. Rijnsburger Robusta, onion, Allium porrum L. cv. Winterreuzen, leek, Daucus carota L. cv. Nantaise, carrot, germination, priming, polyethylene glycol, seed moisture, cell number, embryo volume     

Germination Inhibitors in some Umbellifer Seeds

Presence of inhibitors of germination in seeds of five umbellifercrops viz. fennel (Foeniculum vulgare Mill.), cumin (Cuminumcyminum L.), carum (Carum copticum Benth. and Hook.), carrot(Daucus carota L.), and coriander (Coriandrum sativum L.) wasdemonstrated. The effect of seed exudates on the pattern ofinhibition of germination of these seeds was also studied. Presence of exudates in general delayed germination althoughthe time of maximum inhibition was different with differentcrop seeds. The inhibitory effect was high at an early stagein the case of carum and carrot and persisted in the latterbut disappeared in the former. The inhibitory effect on cuminand fennel increased up to the seventh day and then began todecrease. Growth of root and hypocotyl of 16-day-old seedlingswas also inhibited by the exudates.     

Biochemical Activities and Compounds in Seeds: Possible Tools for Seed Quality Evaluation

The rate and final percentage of field emergence (seed quality)was measured in 55 seed-lots of carrots (Daucus carota L.),45 seed-lots of onion (Allium cepa L.) and 32 seed-lots of cabbage(Brassica oleracea var. capitata), together with the followingbiochemical indices: rates of ATP-synthesis; reduction of tetrazoliumby seed powder; AMP and malate content. Differences in field performance of seed-lots (seed quality)showed some correlation with particular biochemical indices.Seed quality was most significantly correlated with AMP contentand ATP synthesis in carrot, with malate content, reductionof tetrazolium and ATP synthesis in onion and with malate andAMP content and reduction of tetrazolium in cabbage. Daucus carota L., Allium cepa L., Brassica oleracea var capitata, carrot, onion, seed quality, ATP, biochemical indices     

{beta}-Amylase Activity as an Index for Germination Potential in Rice

Seeds of different vigour also differ in their germination ability.In rice (Oryza sativa), this difference was correlated withthe level of incorporation of ³⁵S-methionine into 25-60% ammoniumsulphate precipitable material that was rich in amylase proteins.This protein fraction, from dry seeds, contained no -amylaseactivity. In contrast, ß-amylase activity was presentin all seed stocks capable of 99% germination, although thelevel was lower in seeds that grew slowly when germinated. Inlow viability low vigour stock (i.e. extensively deterioratedseeds) ß-amylase activity was absent. Alpha-amylaseactivity in all stocks was detected only after 24 h from thestart of imbibition. These results indicate that ß-amylaseactivity is reliable indicator of the germination ability ofrice seed stocks and of their vigour during germination.Copyright1995, 1999 Academic Press Rice (Oryza sativa L.,), germination, ß-amylase, -amylase, seed vigour     

Myo-inositol Biosynthesis and Galactose Utilization by Wild Carrot (Daucus carota L. var. carota) Suspension Cultures

Wild carrot (Daucus carota var. carota) cell suspensions (63–120µm in diameter) were grown on a mineral salt medium containingdifferent carbon sources in the presence (10 mM) and absenceof myo-inositol. The data obtained after 14 and 21 days of growthshow that an external supply of myo-inositol is not essentialfor growth and development of wild carrot embryos. A linearrelationship was found between growth (d. wt) and embryo numberin the presence and absence of myo-inositol. Standard stock cell suspensions never exposed to exogenous myo-inositoland grown in the absence of 2, 4-D with glucose or galactoseas the carbon source synthesized radioactive myo-inositol whenexposed to D-[1–¹⁴C]glucose or D-[1–¹⁴C]galactose.Gas chromatographic analyses revealed the presence of myo-inositolin the bulk tissue grown in the presence of 2.25 µM 2,4-D with glucose, galactose, fructose or mannose as the solecarbohydrate. We could not detect any component indicating anisomer or a methylated derivative of an inositol in the tissueextracts. Stock cultures were maintained (with 2, 4-D) successfully forat least three successive sub-cultures on D-galactose as thesole carbohydrate. The growth achieved over this culture periodshowed that wild carrot cells used by us could quickly adaptto grow on D-galactose as rapidly as they grow on sucrose. Daucus carota L., wild carrot, suspension cultures, myo-inositol, galactose     

Isolation of a Type I Topoisomerase from Carrot Cells

Carbonera, D., Cella, R., Montecucco, A. and Ciarrocchi, G.1988. Isolation of a type I topoisomerase from carrot cells.—J.exp. Bot. 39: 70–78. A DNA topoisomerase activity has been isolated from suspension-culturedcells of Daucus carota, the enzyme has been chromatographedon CM-cellulose, DNA-cellulose and Sephadex G100. Its Mr appearsto be about 100000 by gel filtration. Carrot DNA topoisomeraserelaxes both positively and negatively supercoiled DNA by changingthe linking number of the substrate in steps of one and is notable to unknot the knotted P4 DNA. It does not require ATP orMg²⁺ , has an optimal salt concentration between 40 and 120mol m^–3 KCl and is resistant to nalidixic acid and novobiocine.This carrot enzyme can be designated as a eukaryotic type IDNA topoisomerase. Key words: DNA topoisomerase, Daucus carota     

A Single Equation to Quantify the Hierarchy in Plant Size Induced by Competition Within Monocultures

The following empirical model: Ra(i) = r(1+ln(w(i)/wm)Kn)(1–(w(i)/W))(1–(y/Y)) which is based on the logistic growth equation, is developedto describe the growth of differently sized individuals withinplant communities. The model is tested against extensive setsof carrot (Daucus carota L.) and red beet (Beta vulgaris L.)data and is shown to fit well. The model was used to predictindividual plant weights in independent data. The agreementsbetween observed and predicted weights were often close butsome systematic deviations did occur. Thus, a single equationdescribed most of the complex interactions that occurred withinmonocultures of annual crop plants. Carrot, Daucus carota L., red beet, Beta vulgaris L., model, growth, variation     

4-Hydroxybenzyl alcohol accumulates in flowers and developing fruits of carrot and inhibits seed formation

Somatic embryogenesis in carrot (Daucus carota) is autonomously inhibited by 4-hydroxybenzyl alcohol (4HBA), which is produced by embryogenic cells. Because somatic embryogenesis is used as a model of zygotic embryogenesis, we assayed for 4HBA in carrot seeds and analyzed the effect of 4HBA on seed formation to determine whether 4HBA is also produced during zygotic embryogenesis. HPLC analysis showed that 4HBA accumulated in flowers and immature and mature fruits, but not in vegetative tissues. The concentration of 4HBA was highest after flowering, when the zygote developed into the early globular-stage embryo. 4HBA accumulation then decreased with seed development. Exogenous application of 4HBA to immature carrot fruits inhibited seed formation. Many 4HBA-treated seeds did not include a mature embryo. These results indicate that the production and accumulation of 4HBA occurs during carrot seed development and that 4HBA has an inhibitory effect on carrot seed formation.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.): VII. INFLUENCE OF AFTER-RIPENING AND AGEING ON GERMINATION RESPONSES TO TEMPERATURE AND WATER POTENTIAL

The effects of storage conditions on the germination of developingmuskmelon (Cucumis melo L.) seeds were tested to determine whetherafter-ripening is required to obtain maximum seed vigour. Seedswere harvested at 5 d intervals from 35 (immature) to 60 (fullymature) days after anthesis (DAA), washed, dried, and storedat water contents of 3·3 to 19% (dry weight basis) at6, 20, or 30°C for up to one year. Germination was testedin water and in polyethylene glycol 8000 solutions ( –0·2to –1·2 MPa osmotic potential) at 15, 20, 25 or30°C. Germination percentages and rates (inverse of meantimes to radicle emergence) were compared to those of newlyharvested, washed and dried seeds. For 40 and 60 DAA seeds,one year of storage at 20°C and water contents <6·5%significantly increased germination percentages and rates at20°C, but had little effect on germination at 25 and 30°C.Storage reduced the estimated base temperature (T_b) and meanbase water potential (_b) for germination of both 40 and 60 DAAseeds by approximately 5°C and 0·3 MPa, respectively.Immature 35 DAA seeds showed the greatest benefit from storageat 3 to 5% water content and 30°C, as germination percentagesand rates increased at all water potentials (). Storage underthese same conditions had little effect on the germination ofmature seeds in water, but increased germination percentagesand rates at reduced 's. Accelerated ageing for one month at30°C and water contents from 15 to 19° increased germinationrates and percentages of mature seeds at reduced 's, but longerdurations resulted in sharp declines in both parameters. Immatureseeds lost viability within one month under accelerated ageingconditions. An after-ripening period is required at all stagesof muskmelon seed development to expand the temperature andwater potential ranges allowing germination and to achieve maximumgerminability and vigour. Post-harvest dormancy is deepest atthe point of maximum seed dry weight accumulation and declinesthereafter, both in situ within the ripening fruit and duringdry storage. Key words: Muskmelon, Cucumis melo L., seed, development, dormancy, germination, vigour, after-ripening     

Scutellar Amylase Activity in Naturally Aged and Accelerated Aged Wheat Seeds

Scutellum extracts of dry unaged and differentially aged wheatseeds showed appreciable and ß amylase activities,these being higher in unaged and lower in aged seeds. This indicatesthat scutellar amylase may remain long-lived in seeds. Duringgermination of both unaged and aged seeds an increase in scutellaramylase (particularly ß) occurs. Initiation of thisgermination associated increase in amylase activity, however,was found to occur relatively late during imbibition. Further,while scutellar amylase increased considerably during imbibitionin unaged seeds, in differentially aged seeds the increase wassmall. This also contributes to the reduced scutellar amylaseactivities in aged seeds. Triticum aestivum L., wheat, naturally aged, accelerated aged, unaged, germination, scutellar and ß amylase     

Nucleic acid and protein synthesis and loss of vigour in germinating wheat embryos

A study has been made of the RNA and protein synthesising systems of wheat embryos isolated from seed lots having high viability but differing in vigour. The rate of RNA and protein synthesis in wheat embryos during the early hours of germination is related to the vigour of the seed lot. The imposition of a stress factor, in the nature of a sub-optimal germination temperature, during germination of isolated wheat embryos magnifies the differences in rates of protein and RNA synthesis between high and low vigour seed. Using cell-free protein synthesising systems it has been demonstrated that an important difference between high and low vigour embryos lies in the relative levels of messenger RNA in the embryo. High vigour embryos contain relatively higher levels of poly A⁺-RNA (i.e. potential mRNA species) than lower vigour embryos and furthermore the level of poly A⁺-RNA in high vigour embryos increases during early germination whilst in lower vigour embryos the level decreases. The difference in poly A⁺-RNA levels accounts, at least partially, for the differences in rates of protein synthesis observed between embryos from high and low vigour wheat seed during early germination at both optimal and sub-optimal germination temperatures.Abbreviations HEPES N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid - poly A⁺-RNA polyadenylated RNA - GM germination medium - PMS post-mitochondrial supernatant fraction     

The Germination of Carrot (Daucus carota L.) Seed Harvested on Two Dates: A Physiological and Biochemical Study

The germination of two batches of carrot seed, harvested 44and 104 d after anthesis, was compared at 10 °C. Proteinand nucleic acid contents of the seeds were measured at intervals,together with fresh weight and respiration rate. The matureseed germinated 3.7 d earlier than the immature seed, with nodifference in percentage germination. The dry mature seed containedmore protein and nucleic acid per unit dry matter than did theimmature seed, and proportions of nucleic acid present as rRNAand poly(A)RNA were greater in the mature seed. The sequenceof metabolic reactivation was the same in both batches of seed,as were the relative rates of increase of each nucleic acidcomponent, fresh weight, and respiration rate. Differences inthe composition of the dry seed appear to be responsible forthe observed difference in rate of germination between the twoseed batches.     

Seed vigour and water relations in wheat

The laboratory germination (criterion radicle emergence) of seven seed lots of winter wheat cv. Slejpner was similar. However, they differed in vigour as demonstrated by differences in germination after controlled deterioration carried out at a range of seed moisture contents, at two temperatures and for different times. A vigour assessment for each lot was quantified by calculating a value for the seed lot constant K_i, of the viability equation. Germination in lower water potentials reduced the uptake of water, radicle and coleoptile emergence and radicle and coleoptile extension. There was no difference in the water uptake of seed lots differing in vigour. However, seed lots of lower vigour showed less radicle emergence, less coleoptile emergence and shorter radicles than higher vigour seed lots in low water potentials. Similarly, controlled deterioration resulted in reduced radicle and coleoptile emergence and growth compared to unaged seed, and also to a greater sensitivity to low water potentials. The implications for field establishment are discussed.     

The Effects of Priming and Ageing on Seed Vigour in Tomato

A comparison was made of the effects of seed priming or ageingtreatments on the performance of tomato (Lycopersicon esculentumMill. cv. UC204C) seeds according to a number of indices ofseed vigour. A single lot of tomato seeds was primed in 120mol m^–3 K2HPO4 + 150 mol m^–3 KNO₃ for 5 d at 20?C, or aged at 13% moisture content (dry weight basis) and 50?C for 6 d. Germination percentage (>98%) was unaffectedby priming and reduced to 85% by ageing. X-ray photographs andlongitudinal sections revealed the formation of free space surroundingthe embryo in dry primed seeds, which was not evident in controlor aged seeds. Priming increased the rate of germination atall temperatures above the base temperature (T_b), while ageingdecreased it. T_b was unaffected by priming and only slightlyincreased by ageing. The variation in individual times to germinationwas approximately doubled in both primed and aged seed comparedto the control, based upon the slopes of probit germinationpercentage versus log thermal time curves. Root growth aftergermination tests and seedling growth in both greenhouse andfield tests were not influenced by either priming or ageing.The conductivity test was found to be unreliable as a vigourtest for tomato seeds. The results identify several indiceswhich can be used to quantify seed vigour in tomato. They alsoillustrate that seed priming can enhance seed performance accordingto some criteria, while having no effect or decreasing qualityaccording to other criteria. Seed vigour can apparently be separatedinto various components which can be independently influencedby seed enhancement treatments. Key words: Tomato, seed germination rate, seed priming, seed vigour     

Nucleotide Levels and Loss of Vigour and Viability in Germinating Wheat Embryos

At an optimal germination temperature loss of viability in wheatseed is reflected in reduced levels of nucleoside triphosphatesand nucleotide sugars in the embryo compared to the levels foundin high viability embryos during the early hours of imbibition.These differences are magnified on germination at 10° Cand are accompanied by a greatly reduced rate of protein synthesisin embryos of low viability compared with the correspondingrate in embryos of high viability. Loss of vigour precedes lossof viability in seed but differences in biochemical parametersbetween high and medium vigour seed do not become apparent unlessgermination under stress conditions, e. g. low temperature,occurs. Under these stress conditions, wheat seed of reducedvigour can be distinguished from high vigour seed lots of similarviability by the decreased levels of nucleotides and nucleotidesugars found in embryos from reduced vigour seed lots duringthe early hours of imbibition and by reduced rates of proteinsynthesis in these embryos during this same period. Nucleotideanalysis is a potentially useful method for rapid assessmentof the viability or vigour of a seed lot. Key words: Nucleotides, Vigour, Viability, Wheat     

The Effects of Priming and Accelerated Ageing upon the Nucleic Acid Content of Leek Seeds and their Embryos

Using seed priming and accelerated ageing techniques, a singlelot of leek (Allium porrum) seeds was manipulated to producefour lots of seeds with different germination performance. Changesin content of the major nucleic acid species in whole seedsand embryos of two of these lots (primed and unprimed), weredetermined over the early stages of germination. The major effectof priming was an increased level of RNA species in the seedsand embryos, and this difference was maintained during germination.Comparison of nucleic acid levels in the dry seeds of thesetwo lots and two others (aged and aged then primed) indicatedthat there was no correlation with germination performance.Similar comparisons of the nucleic acid levels in the embryosof seeds imbibed for 1 d showed only a limited correlation betweenrRNA levels and germination performance. Analysis of these datasuggests that accelerated ageing has an adverse effect uponendosperm cells, which results in the degradation of their nucleicacids during priming. Furthermore, the viability of these agedseeds also falls during priming. The data also indicate thatratios of rRNA to DNA correlate with germination performanceof the four lots of seeds studied. It is proposed that sucha relationship is indicative of the efficiency of a primingtreatment, and may be useful in comparisons of naturally varyingseed lots. Key words: Leek, seed, germination, priming, nucleic acids     

Resistance to Azetidine-2-carboxylic Acid and Sodium Chloride Tolerance in Carrot Cell Cultures and Spirulina platensis

Mutants of Spirulina platensis and of Daucus carota resistantto azetidine-2-carboxylic acid were tested for NaCl tolerance.A positive correlation was found between proline overproductionand osmotolerance. In carrot lines proline overproduction wasnot strictly proportional to NaCl tolerance insofar as cellscharacterized by differences in proline overproduction showedsimilar osmotolerance, suggesting that other factors could beinvolved. (Received March 17, 1983; Accepted June 14, 1983)