Identification of defence proteins from the seed exudates of Cicer arietinum L. and its effect on the growth of Fusarium oxysporum f.sp. ciceri

Germinating seeds tend to release a variety of proteins into their surrounding surfaces; some of which have an inhibitory action against plant pathogens. The aim of this study was to investigate and identify defence proteins present in the exudates from water-imbibed and chitosan-imbibed (0.1% w/v) seeds of chickpea (Cicer arietinum L). Chickpea seeds imbibed in chitosan released a higher amount of proteins in the exudate when compared to the seeds imbibed in water. The obtained exudates were analysed in regard to specific protein activities by enzymatic assays and SDS-PAGE analysis. Results showed that the exude obtained from chickpea seeds imbibed in chitosan solution exhibited a new isoform of chitinase, chitosanase and protease inhibitors. These exudates also have an “in vitro” inhibitory effect on the growth of the fungus, Fusarium oxysporum f.sp. ciceri. Our results suggest that seed exudates protect seeds during their germination from soil pathogens.     

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     

Sugars in Natural and Artificial Pollen Germination Substrates

The stigmatic exudates and pollen grains of five unrelated specieswere tested for sugars. Glucose, fructose, and sucrose werefound in the stigmatic fluid of Yucca aloifolia L. and glucoseand fructose in that of Oenothera adrummondii Hook. In in vitroexperiments with Y. aloifolia pollen, high germination percentageswere obtained in artificial media containing glucose or sucrose.Fructose, which is present in the stigmatic fluid of the Yuccasp. resulted in high in vitro pollen germination only when borateand calcium were added to the medium. Presence of bound sugarsis indicated in the stigmatic secretion of Citrus aurantiumL. and pollen of the single plant tested germinated at a lowpercentage in artificial sugar media. No sugars were detectedin the stigmatic fluids of Hemerocallis fulva L. and Zea maysL. and in these two species in vitro pollen germination in sugarymedia was negligible or absent. The pollen grains of all five species contain sucrose and thoseof Oenothera and Citrus also reducing sugars.     

黄瓜种子及其萌发期的化感作用研究

以大白菜、萝卜、番茄和黄瓜种子为受体,采用实验室培养皿种子发芽生物测试法研究了黄瓜种子浸提液、种子萌发、胚根和芽苗分泌物、芽苗腐解物和芽苗浸提液的化感效应。结果表明:(1)黄瓜种子浸提液对大白菜、萝卜、番茄和黄瓜种子萌发均有化感抑制作用,即黄瓜种子内含有某些化感抑制物质。(2)在水浸提过的黄瓜种子萌发过程中,它不仅对其近邻套种的大白菜、萝卜和番茄种子萌发产生化感抑制作用,而且其胚根和芽苗分泌物对后茬播种的4种蔬菜种子发芽也表现出不同程度的化感抑制作用;黄瓜芽苗腐解物和芽苗水浸提液也对各受体蔬菜种子发芽与生长产生不同程度的化感抑制作用,且随着腐解芽苗量的增加或浸提液浓度的升高,各受体蔬菜种子的发芽指标值、化感效应指数值和综合效应值随之降低。(3)黄瓜种子浸提液及芽苗各器官的化感物质对黄瓜种子的萌发与生长产生了自毒作用,且黄瓜芽苗腐解物、芽苗浸提液、胚根及芽苗分泌物对受体黄瓜的自毒作用均为最大。研究发现,黄瓜种子浸提液、种子萌发时期以及芽苗各器官的化感物质主要是通过抑制受体胚根的生长而起化感抑制作用,即受体蔬菜种子胚根对化感效应最为敏感;因黄瓜种子及萌发期释放化感物质的途径有所不同,导致受体大白菜、萝卜、黄瓜和番茄的化感响应也不相同;在黄瓜种子萌发和芽苗生长的早期,化感物质即开始在芽苗体内进行合成与积累,一部分可通过胚根和芽苗分泌途径释放到环境中,另一部分可通过芽苗腐解途径释放化感物质,并对受体蔬菜种子萌发与生长表现出较强的化感抑制作用。     

REQUIREMENT OF LOW TEMPERATURE TREATMENT FOLLOWING ILLUMINATION IN THE GERMINATION OF SEED OF ELSHOLTZIA

Effect of light and temperature on the germination of seedsof Elsholtzia Argyi L?V. var. nipponica (OHWI) MURATA was investigated. When kept at a constant temperature, the seeds did not germinateeven after photoperiodical treatments. High germination percentageswere obtained only when the seeds were transferred to 5? immediatelyafter light exposure and kept at this temperature for a certainperiod. When the illuminated seeds were exposed to 25? beforebeing transferred to the low temperature, the germination wasinhibited. There was some relation between the durations ofillumination and of the cold treatment. The inhibitory effect of far-red radiation was not reversedby red radiation. (Received March 15, 1960; )     

Germination Inhibitors in Fruit Extracts of Red Beet (Beta vulgaris cv. rubra

Germination inhibitors in methanol and water extracts of redbeet fruits (Beta vulgaris cv. rubra L.) have been studied usinglettuce and red beet seed germination as bioassays. The methanolextracts contained substances which inhibited lettuce seed germination,but had no effect on the germination of red beet seeds. Germinationof both lettuce seeds and of water-leached or sulphuric acid-treatedred beet seed balls were inhibited by the water extracts. Theconcentrations of ammonia, ferulic acid, and oxalate in thewater extracts were much lower than required for inhibitionof red beet seed germination. The water extracts contained,however, large amounts of inorganic ions, and the results clearlydemonstrated that the inhibitory effect of the water extractson red beet seed germination was mainly due to the content ofsuch inorganic ions.     

Response of Seed Germination in Three Genera of Compositae to White Light of Varying Photon Flux Density and Photoperiod

Various photon doses (net number of photons per unit area perday), provided by varying both photon flux density and photoperiod,were applied to imbibing seeds of seven lots of four speciesof Compositae in various germination test regimes. In all fourspecies germination was dependent upon photon dose, the productof photon flux density and daily duration of exposure. The responsewas quantified by linear relations between the probit of percentagegermination and the logarithm of photon dose. In general, photonflux density and photoperiod only influenced the stimulationof germination by the low energy reaction indirectly (as factorsof daily photon dose), whereas there was a tendency for photoperiodto have a direct influence on the inhibition of germinationby the high irradiance reaction. Reducing the germination testtemperature from 25?C to 20?C and 15?C not only increased thedark germination of L. sativa L., but also broadened the photondose range at which full germination occurred by reducing theminimum value necessary for the germination of the most dormantseeds, and increasing the maximum value which failed to inhibitthe germination of any seeds. Differences between L. sativaand L. serriola L. in the response of germination to white lightwere only quantitative, rather than qualitative. The singlemost promotory dose for all four species was 3 ? 10^–3mol m^–2 d^–1, although the inhibitory action of dosesup to 10– mol m– d– was generally only slight. Key words: Light, seed germination, seed dormancy, Compositae     

The effect of volatile and gaseous metabolites of swelling seeds on germination of fungal spores

Effects of volatile and gaseous metabolites of swelling seeds of pea, bean, wheat, corn, cucumber, tomato, lentil, carrot, red pepper and lettuce on germination of spores of five genera of fungi were found to depend rather on the fungal than on the plant genus. Germination of spores ofBotrytis cinerea, Mucor racemosus andTrichoderma viride was most severely inhibited. Spores ofVerticillium dahliae were less sensitive and germination of spores ofFusarium oxysporum was inhibited only in two cases. On the other hand, exudates of pea and bean stimulated germination of spores ofFusarium oxysporum. Also spores ofTrichoderma viride germinated better in an atmosphere enriched with exuded metabolites of swelling lettuce seeds. When carbon dioxide produced by the swelling seeds was absorbed in potassium hydroxide, spores ofTrichoderma viride andVerticillium dahliae did not germinate at all, the inhibitory effects of volatile and gaseous exudates on germination of spores ofMucor racemosus were accentuated, and also the percentage of germinated spores ofFusarium oxysporum decreased. Germination of spores ofBotrytis cinerea was not influenced. Absorption of volatile and gaseous metabolites in a solution of potassium permanganate decreased in most cases their inhibitory effects, particularly inBotrytis cinerea.     

Prehydration and Priming Treatments that Advance Germination also Increase the Rate of Deterioration of Lettuce Seeds

The influence of prehydration in water or priming in –1.5 MPa polyethylene glycol 8000 solution for various periods,followed by redrying, on germination rate and longevity of lettuce(Lactuca sativa L.) seeds (achenes) was determined during controlleddeterioration at 10% moisture content (fresh weight basis) and40°C. Short prehydration treatments (up to 1 h) had littleeffect on either germination rate or longevity, but significantlyimproved root growth rates. Increasing durations of prehydrationor priming reduced the mean time to germination by up to 61%relative to untreated seeds, but also reduced mean seed longevityby as much as 84% Prehydration and priming altered the relationshipsbetween germination rate and viability and between normal andabnormal seedlings during ageing. Prehydration in abscisic acidor at a temperature inhibitory to germination did not preventthe reduction in longevity under controlled deterioration conditions.While prehydration or priming treatments effectively acceleratesubsequent germination rates of lettuce seeds, the redried seedsare nonetheless highly susceptible to deterioration in storage. Key words: Lettuce, Lactuca sativa L., seed priming, seed deterioration, germination rate     

Bioassay and Identification of Root Exudates of Three Fruit Tree Species

A laboratory bioassay was designed to determine the aileiopathic potential of root exudates of three fruit tree species on apple germination. The results showed that root exudates of apple （Malus pumila L.） and peach （Prunus persica L.）, each at concentrations of 0.02 and 0.2 mg/L, inhibited germination and radicle growth of apple seeds by 56.7%, 60.7%, 51.5%, and 59.3%, respectively. The corresponding shoot growth inhibition rate was 49.5%, 46.7%, 36. 4%, and 44%, respectively. Root exudates of jujube （Ziziphus jujuba Mill.） had no significant effect on apple seeds. Qualitative determination of root exudates of apple, peach, and jujube tree was developed with gas chromatography-mass spectrometry. The root exudates of apple seedlings mainly contain organic acids, glycol, esters, and benzenphenol derivatives. Peach root exudates contained phenolic acids and benzenphenoi derivatives in addition to two unidentified compounds. The root exudates of jujube did not contain any phenolic acids.     

Effect of High Temperature on the Development of Cyanide-Sensitive Respiration in the Germination Process of Spinach Seeds

Germination of spinach (Spinacia oleracea L. var. grabra cv.Nobel) seeds was inhibited at a high temperature (35?C). Effectsof KCN on the respiration of seeds incubated at 20 and 35?Cwere compared in order to investigate the mechanism of inhibitionof seed germination by high temperature. Respiration of germinatingseeds incubated at 20?C was inhibited about 50% by 5 mM. KCNsolution, whereas it hardly inhibited the weak respiration ofthe seeds at 35?C. Germination of seeds was delayed by exogenousKCN. When the KCN solution was renewed daily, germination wascompletely inhibited. Pericarp removal promoted germinationat 35?C, but atypical germination (cotyledons emerging earlierthan a radicle) took up more than half of the total germination.The inhibitory action of KCN on the respiration of seeds wasnot altered by pericarp removal. A KCN addition, even at 20?C,elicited atypical germination in the pericarp-less seeds. Theseresults show that cyanide-sensitive respiration is needed toinduce typical spinach seed germination (root emergence), butis rendered inoperative by high temperatures thus bringing aboutpoor germination and atypical germination. (Received December 1, 1984; Accepted February 8, 1985)     

Effect of Salinity on Germination and Seedling Growth of twoAtriplexspecies (Chenopodiaceae)

Salinity is one of the environmental factors that has a criticalinfluence on the germination of halophyte seeds and plant establishment.Salinity affects imbibition, germination and root elongation.However, the way in which NaCl exerts its influence on thesevital processes, whether it is through an osmotic effect ora specific ion toxicity, is still not resolved. Dimorphic seedsof the halophytesAtriplex prostrataandA. patulawere treatedwith various iso-osmotic solutions of NaCl and polyethyleneglycol (PEG). For each treatment, imbibition, germination rate,percent germination, germination recovery and nuclear area ofroot tip cells were compared. Higher concentrations of NaCl(-1.0 MPa) were more inhibitory to imbibition, germination andseedling root elongation than iso-osmotic PEG solutions. Allseeds recovered from a pre-treatment with -2.0 MPa NaCl andPEG solutions, except large seeds ofA. prostratawhich failedto germinate following transfer from -2.0 MPa NaCl. NaCl causeda greater increase in nuclear volume than iso-osmotic PEG solutions.These data suggest that the influence of NaCl is a combinationof an osmotic effect and a specific ion effect.Copyright 1998Annals of Botany Company Atriplex patula,Atriplex prostrata,cytophotometry, osmotic potential, salinity, seed germination.     

Defense Proteins from Vigna Unguiculata Seed Exudates: Characterization and Inhibitory Activity Against Fusarium Oxysporum

Plants exude a variety of substances through their roots, germinating seeds and aerial parts. Some of these released compounds seem to have an inhibitory effect against pathogens. The aim of this work was to investigate and identify antifungal proteins present in exudates from imbibed cowpea seeds (Vigna unguiculata (L.) Walp). The obtained exudation was analyzed in regard to specific protein activities by enzymatic or immunological assays for plant defense proteins, from 4 h to 48 h of seed imbibition. Our results show that cowpea seeds exudates present several defense related proteins characterized as β-1,3-glucanases, cystatins, vicilins and lipid transfer proteins (LTPs), as well as a storage vacuole membrane α-TIP protein, since the very first hours of imbibition. These exudates also have an “in vitro” inhibitory effect on the growth of the fungus Fusarium oxysporum f. sp. phaseoli. Our results suggest that seed exudates should promote seed protection from soil pathogens.     

Effects of Ageing on Amylase Activity and Scutellar Cell Structure during Imbibition in Wheat Seed

In wheat seed the scutellum plays an important role in the hydrolysisof stored substrate during germination. This layer is activatedfirst, whilst the aleurone becomes activated later. A good correlationexists between the initiation of visible germination and theappearance of enzyme activity in the scutellum. Enzyme activityin the aleurone becomes apparent only when the germinating seedlingreaches the rapid growth phase. Electron microscopic observationsshow that during the later stages of germination the scutellarcells develop finger like projections. These may serve to absorbendospermic reserves hydrolysed by aleurone amylase. The scutellumof aged non-germinating seeds showed no amylase activity andno finger like projections were produced even after prolongedimbibition.Copyright 1993, 1999 Academic Press Wheat (Triticum aestivum L.), deteriorated, germination, scutellum, scanning electron microscopy, aleurone     

Seed Dormancy in Acer: The Role of Testa-imposed and Embryo Dormancy in Acer velutinum

The embryo dormancy shown in freshly harvested samples of Acervelutinum seeds is weakly established and very short-lived.Loss of this embryo dormancy occurred during post-harvest fruitstorage at either 5 or 17 C. In contrast, the dormancy of intactfruits and seeds was overcome only during storage at the lowertemperature. Removal of the cotyledons from embryos of freshlyharvested fruits allowed more rapid germination of the embryonicaxes, indicating that the cotyledons exert an inhibitory effect,although the axes still retained a measure of innate dormancy.The inhibitory effect of the cotyledons became less marked withincreasing duration of fruit storage, this loss of inhibitoryeffect occurring at both storage temperatures. Applied ABA stronglysuppressed germinative capacity in intact embryos and isolatedembryonic axes from freshly harvested fruits, but when ABA wasapplied to embryos of fruits that had been stored for variousperiods at 5 or 17 C, the inhibitory effect was first weakenedand then lost with increased storage. Although dormancy in the seeds of A. velutinum may be describedas intermediate between testa-imposed dormancy and true dormancy,it is perhaps more properly included in the former category. Acer velutinum Boiss. var. vanvolxemii, abscisic acid, embryo dormancy, germination, seed storage, testa-imposed dormancy, tissue sensitivity     

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.     

SUCCESSIVE PROCESSES INVOLVED IN THE GERMINATION RESPONSE OF NASTURTIUM SEEDS

1. The seeds ofNasturtium palustreDC. do not germinate, eitherin the light or darkness, at various constant temperatures,but require for their full germination a certain period of alow temperature (5°) applied immediately after light irradiation.These results indicate the existance of at least two processes,a light-dependent process and a low temperature-requiring process,in the initiation of germination ofNasturtiumseeds. Experimentalevidence indicated further that the light exposure causes twodifferent processes in the seed germination. 2. When a dark period at 23° was inserted between the lightirradiation and the low temperature treatment the germinationwas suppressed. The inhibitory effect of the inserted dark periodat 23° was eliminated by a short irradiation during thedarkness (light-break). 3. Prolonged exposure ofNasturtium seeds to any concentrationof gibberellin brought about no germination when exposure wasgiven in complete darkness. The germination was promoted onlywhen light irradiation was applied to the seeds. A short applicationof gibberellin at a fairly high concentration was, however,remarkably effective for the germination even in the darkness,and the germination was inhibited as the gibberellin applicationwas lengthened. It was considered that gibberellin could substitutefor the combined effect of light irradiation and low temperaturetreatment to induce the germination of Nasturtium seeds, andthat gibberellin was inhibitive toward the reactions followingthe above treatments which induced the germination (Received October 31, 1996; )     

Studies on the Germination of the Seeds of Striga hermonthica: III. On the Nature of Pretreatment and after-ripening

An attempt has been made to elucidate some of the changes whichtake place in seeds of Striga hermonthica (immature, ripe andoverripe), (a) during moisture-treatment at three temperatures(15°C., 22°C., and 32°C.), and (b) during after-ripeningin dry storage. Observations on the drift of respiration of immature seed throughoutpretreatment at 15°C. show that an inverse relationshipexists between germination and respirational activity. Thissuggests that germination depends on the accumulation of somemetabolite during pretreatment. It would seem that this metabolitemay also be used as a substrate for respiration during pretreatment. After-ripening of the immature seeds has been shown to changethe nature of the seeds in some way so that their peak respirationon exposure to moisture is at first progressively reduced. Thisreduction has been ascribed to the presence of some barriercapable of restricting gaseous diffusion. The data presented, togetherwith those of the previous paperof this series, suggest that the specific metabolite requiredfor germination might be proteinaceous. Certain other data appearto rule out the suggestion that germination capacity is conditionedby the sugar content of the seeds. Reasons are given which supportthe view that one of the main effects of the stimulating solutionon the seed is to increase the permeability of the seeds togaseous diffusion.     

Inhibition of germination and α-amylase induction by 6-methoxy-2-benzoxazolinone in twelve plant species

6-Methoxy-2-benzoxazolinone (MBOA) inhibited germination of rice (Oryza sativa L.), wheat (Triticum aethiopicum Jakubz), rye (Secale cereale L.), onion (Allium cepa L.), wild oat (Avena fatua L.), barnyard grass [Echinochloa crus-galli (L.) Beauv.], ryegrass (Lolium rigidum Gaudin), cress (Lepidium sativum L.), lettuce (Lactuca sativa L.), tomato (Lycopersicum esculentum Mill.), carrot (Daucus carota L.) and amaranth (Amaranthus retroflexus L) and the inhibition increased with increasing MBOA concentrations. MBOA also inhibited the induction of α-amylase in these plant seeds and the inhibition increased with increasing MBOA concentrations. There were variations in sensitivity of these plant species to MBOA, and species of family Poaceae (barnyard grass, wild oat, rice, rye, ryegrass, and wheat) were less sensitive to MBOA than the other plant species.     

Analysis of Variation in the Rates of Germination and Early Seedling Growth in Tomato

Studies have been made on the time to germination and earlyseedling growth of the tomato Lycopersicon esculentum var. Potentateand Lycopersicon pimpinellifolium var. Red Currant, as wellas on derivative generations from their hybridization. Althoughtime to germination showed a genetic component, the relationshipsbetween different genotypes was much influenced by environmentalfactors. A marked maternal effect on time to germination dueto the varying seed sizes was noted while variation betweendifferent genotypes of similar seeds size was ascribed to anendospermic effect. The F¹ hybrid embryo with L. Pimpinellifoliumas maternal parent contained more cells than that of the parentbut the hybrid cmbryo with L. esculentum as maternal parentcontained a similar number of cells to that of the parent itself.It is suggested that the results for embryo size both supportAshby‘s assertion that embryo size may be important indetermining heterosis, and also Hatcher’s findings in1940, that heterosis for hypocotyl extension is found in hybridsfrom parcnts of different sized seeds provided the small seededvariety is the maternal parent.