The Effect of Sodium Hypochlorite on Germination of Lettuce Seed at High Temperature

Investigations into the effects of treatment with sodium hypochloritesolution on high-temperature germination of lettuce seed aredescribed. A 2 h treatment in a solution with 10% availablechlorine enabled up to c. 70% of seeds to germinate at 35 °C,without impairing germination and normal seedling developmentat 20 °C. A 10 min rinse in 0.01 N HCI following such treatmentenhanced its effect and treated seeds retained their abilityto germinate at high temperature for at least 18 months. Seedlingdevelopment at 20 °C following treatment was influencedby treatment temperature and batch of hypochlorite solution.Emergence from compost in a cycling (30/15 °C) temperatureregime was improved in three out of five cultivars tested, andthere were indications that hypochlorite treatment enabled oneof the other cultivars to escape induced thermodormancy. Key words: Lactuca sativa L., Sodium hypochlorite, Thermo-inhibition     

The effects of thermotherapy and sodium hypochlorite treatments on pepino seed germination, a crucial step in breeding programmes

We evaluated the effects of treatments with thermotherapy (80°C for 24 h) and dipping in sodium hypochlorite (NaOCl; 0.8% for 10 min) on the germination percentage, germination rate, vigour index and ToMV inactivation of seeds from three pepino (Solarium muricatum) accessions (96–5, B–2 and OV–8) and two wild relatives (S. tabanoense EC–26 and S. caripense EC–40). Thermotherapy decreased the germination percentage (especially in the wild species), germination rate in wild species but not in cultivated pepino, and vigour index in all cases. Sodium hypochlorite increased germination, except for nonthermotreated seeds of OV–8, improved the germination rate in wild species and the vigour index in all cases except for non thermotreated OV–8. Seed coats were altered by both treatments, especially by NaOCl. Thermotherapy was totally effective in the inactivation of seed-borne ToMV, while NaOCl was not. Thermotherapy combined with NaOCl allows effective seed disinfection of S. muricatum seeds without negatively affecting seed germination. For the wild species, although thermotherapy reduces germination, the higher number of seeds per fruit in these species reduces the magnitude of the problem. Even when no disinfection is necessary, treatments with NaOCl are always advisable as they improve germination.     

不同处理对海甘蓝种子发芽和幼苗生长的影响

由于受种子生理休眠作用的影响和硬而厚的种皮所产生的抑制作用,使海甘蓝（ＣｒａｍｂｅｍａｒｉｔｉｍａＬ．）种子发芽慢,发芽率低。为探索加快海甘蓝种子发芽和提高种子发芽率的有效方法,我们先后进行了８种不同的种子预处理试验和６种不同次氯酸钠溶液浸种的发芽试验。结果发现：（１）种子剥皮处理可以很大程度地促进发芽和提高发芽率;（２）用浓度为０．０５％的赤霉酸溶液浸种１８ｈ对海甘蓝种子发芽也有很好的促进作用;（３）用０．２０％的代森锰锌４５Ｍ（Ｄｉｉｔｈａｎｅ４５Ｍ）溶液浸种２０ｍｉｎ的消毒处理对海甘蓝种子发芽产生一定程度的抑制作用,但可减少海甘蓝幼苗死亡率;（４）适宜浓度的次氯酸钠漂白水（法文名１＇ＥａｕｄｅＪａｖｅｌ）的溶液（１０％）浸种５ｍｉｎ对促进海甘蓝种子发芽和减少幼苗死亡均有良好效果;浓度低于１０％时．不足以腐蚀种子硬而厚的种皮而促进种子发芽,也不足以杀死种子携带的病菌而减少幼苗死亡率;浓度大于１０％时,对种子的胚和种子内的酶活性产生不良影响,从而抑制种子发芽和影响幼苗的正常生长。     

THE SURFACE DECONTAMINATION OF SEEDS TO PRODUCE AXENIC SEEDLINGS

A variety of sterilizing agents were tested to develop a standard procedure for surface decontaminating seeds to produce axenic seedlings. The use of calcium hypochlorite (0.5% phosphate buffer, pH 6) for 10 min followed by three sterile water rinses was among the most effective agents, and it did not injure some species as did sodium hypochlorite, formaldehyde, ethylene oxide and mercuric chloride. Some species contained internal microbes requiring severe treatments which killed or injured the seedling, while other species were “decontaminated” with a sterile water rinse. The percentage of seeds with internal microbes varied considerably among plant species, seed lot, and the length of seed storage. Thus, with seeds not easily decontaminated, screening of additional seed lots would be more profitable than testing additional decontamination agents. Release of microbes from the seed's interior is associated with germination, and microbial testing must last at least 11 days. Nutrient agar permitted growth, although the seedlings outgrew petri plates too quickly for adequate certification. These seedlings were transferred to nutrient agar in quart jars in which an internal pool of broth was periodically agitated to permit microbial sampling of the leaves while the plant grew.     

Improving germination and vigour of aged and stored onion seeds by matriconditioning

Germination and vigour of accelerated aged (AA) and naturally stored onion seeds were examined. Accelerated ageing was conducted at 40 °C and 100 % relative humidity (RH). Non aged seeds were stored for 34 months at 3 or 15 °C and 40, 60 or 90 % RH. To restore seed viability, stored and aged seeds were matriconditioned with Micro-Cel E. A distinct loss of germination was observed after 5 days of accelerated ageing. Naturally stored seeds maintained high viability for 34 months, when stored at 3 °C and 40, 60 and 90 % RH or at 15 °C and 40 %. An increase of RH to 60 and 90 % at 15 °C caused loss of germination and vigour. Matriconditioning improved germination and increased endogenic ethylene release and in vivo ACC oxidase activity of both aged and stored seeds.     

Effects of matriconditioning on onion seed germination, seedling emergence and associated physical and metabolic events

The effect of matriconditioning, the physiological presowing seed technique, using Micro-Cel E on Allium cepa L. cv. Czerniakowska seed quality was studied. Several ratios of seeds, carrier, water and time of priming were tested. The most effective treatment for improving onion seed germination at most tested temperatures was priming to a ratio of 2 g seed:1 g Micro-Cel:3 g water for 5 days in light at 15 °C. Matriconditioning greatly improved the germination and emergence percentage, seedling fresh and dry weight and reduced electrolyte leakage compared to that of untreated seeds; this beneficial effect was especially evident at suboptimal temperatures. Matriconditioning improved the germinability of aged seeds, the effect being more pronounced in the more aged seeds. No significant differences in ethylene production by primed and non-primed seeds were observed in the absence of its precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), but its presence during imbibition caused an increase in ethylene production; an enhanced activity of in vivo ACC oxidase in Allium cepa matriconditioned seeds in comparison to untreated seeds, indicates that the endogenous level of ACC is a limiting factor of ethylene production. Likewise, the activity of ACC oxidase isolated from matriconditioned seeds was higher than that from untreated seeds. Higher endo--mannanase and total dehydrogenase activities were observed in primed air-dried seeds in comparing to non-primed seeds.     

Stimulatory effects of sodium and calcium hypochlorite, pre‐chilling and cytokinins on the germination of Cypripedium macranthos seed in vitro

Cypripedium macranthos is a wild orchid that is becoming endangered. Efficient methods for its propagation from seed, which is indispensable for conservation, production and breeding, have not been reported. The effects of sodium and calcium hypochlorite, pre‐chilling and cytokinins on the germination of seeds of Cypripedium macranthos Swartz were examined. The duration of treatment with a solution of hypochlorite prior to sowing was one of the critical factors that affected germination. Approximately 70% of seeds that had been treated with either a solution of NaClO that contained 0.5% available chlorine for 60 min or with one of Ca(ClO)₂ with 3.2% available chlorine for 7 h, germinated after 3 months of culture at 20°C, subsequent to 2 months chilling at 4°C. Chilling seeds at 4°C prior to culture at 20°C was another factor that stimulated germination. Even chilling for 2 weeks had a promotive effect on germination, and chilling for 2 months enhanced it most effectively: the frequency of germination was 67% after 3 months of culture at 20°C. However, the promotive effects of chilling on germination were reduced by holding seeds at 20°C for 3 and 6 weeks prior to chilling treatment. Germination of 58‐70% was achieved by the addition of 1 µ M cytokinin to the medium, while the frequency was only 17% in cytokinin‐free medium. We report a reproducible and efficient method for enhancing seed germination of C. macranthos , which involves treatment with hypochlorite prior to sowing, and the combination of chilling at 4°C prior to germination and exposure to a cytokinin.     

Effect of water stress and gibberellic acid on germination of flax,sesame and onion seeds

The effects of different osmotic stresses (from 0 to –8× 10⁵ Pa) obtained with NaC or polyethylene glycol 6000 solutions on the germination of flax, sesame and onion seeds were investigated. The effect of presoaking with gibberellic acid (GA³) on the germination of the above mentioned seeds was also studied. It was found that the rate of seed germination and the final germination percentages as well as the amount of water absorbed by the seeds were considerably lowered with the rise of osmotic stress levels whatever the stress agent used, more considerable reduction was obtained under polyethylene glycol 6000 than under NaCl. Presoaking with gibberellic acid increased the rate and the final germination percentage of osmotically stressed flax and sesame seeds, while those of stressed onion seeds were slightly retarded.     

Acceleration of germination of super-hard rice cultivar EM10 by soaking with red onion

Red onion accelerated the germination of rice seeds and inhibited microbial infestation during germination. After germination with red onion for 16 h at 35 °C, super-hard rice EM10 showed a higher germination ratio (2.3 times), and it contained more GABA (2.3 times) and glucose (2.9 times) than that soaked without onion. Due to soaking with red onion, germinated EM10 was fortified with quercetin (18 mg/100 g).     

Physiology of Oil Seeds: II. Dormancy Release in Virginia-type Peanut Seeds by Plant Growth Regulators

Germination, ethylene production, and carbon dioxide production by dormant Virginia-type peanuts were determined during treatments with plant growth regulators. Kinetin, benzylaminopurine, and 2-chloroethylphosphonic acid induced extensive germination above the water controls. Benzylaminopurine and 2-chloroethylphosphonic acid increased the germination of the more dormant basal seeds to a larger extent above the controls than the less dormant apical seeds. Coumarin induced a slight stimulation of germination while abscisic acid, 2,4-dichlorophenoxyacetic acid, and succinic acid 2,2-dimethylhydrazide did not stimulate germination above the controls. In addition to stimulating germination, the cytokinins also stimulated ethylene production by the seeds. In the case of benzylaminopurine, where the more dormant basal seeds were stimulated to germinate above the control to a larger extent than the less dormant apical seeds, correspondingly more ethylene production was induced in the basal seeds. However, the opposite was true of kinetin for both germination and ethylene production. When germination was extensively stimulated by the cytokinins, maximal ethylene and carbon dioxide evolution occurred at 24 and 72 hours, respectively. Abscisic acid inhibited ethylene production and germinaton of the seeds while carbon dioxide evolution was comparatively high. The crucial physiological event for germination of dormant peanut seeds was enhancement of ethylene production by the seeds.     

Acceleration of Germination of Super-Hard Rice Cultivar EM10 by Soaking with Red Onion

Red onion accelerated the germination of rice seeds and inhibited microbial infestation during germination. After germination with red onion for 16 h at 35 °C, super-hard rice EM10 showed a higher germination ratio (2.3 times), and it contained more GABA (2.3 times) and glucose (2.9 times) than that soaked without onion. Due to soaking with red onion, germinated EM10 was fortified with quercetin (18 mg/100 g).     

Effect of Ash Pond Soil and Amendments on the Growth and Arbuscular Mycorrhizal Colonization of Allium cepa and Germination of Arachis hypogaea,Lycopersicon esculentum,and Vigna mungo Seeds

The effect of fly ash pond soil on the growth and yield of onion (Allium cepa var. microaggregatum) plants grown in pots was investigated. The fly ash pond soil was amended with combinations of red soil and press mud, a waste product from sugar mills. Water-holding capacity of ash pond soil amended with press mud increased; however, addition of press mud delayed onion bulb development. Onion bulb germination took place rapidly in ash pond soil; however, subsequent bulb development declined. The addition of red soil and press mud increased the growth and yield of onion plants. In all the soils and amendments, onion plant roots were colonized by native arbuscular mycorrhizal species. Both vesicles and arbuscules were present in the roots. Colonization was low in ash pond soil but increased with the addition of red soil. Effect of fly ash pond soil on germination of groundnut (Arachis hypogaea), tomato (Lycopersicon esculentum), and black gram (Vigna mungo) seeds was evaluated and compared with red soil in the laboratory. Ash pond soil increased the germination of tomato seeds but did not affect the germination of groundnut and black gram seeds.     

Bleach-induced Germination and Breakage of Dormancy of Seeds of Alectra vogelii

The effects of solutions of sodium hypochlorite and calcium hypochlorite (bleach) on dormancy and germination of Alectra vogelii seeds were investigated. Dry (non-pretreated) and 10-day water-pretreated seeds were exposed to various bleach concentrations, as well as to the host (Vigna unguiculata) root exudate. The 15-month-old Alectra seeds used were partially dormant in that only 40% or less of the 10-day pretreated seeds could be stimulated to germinate by the standard host root exudate. Comparable percentage germination of nonpretreated seeds was achieved with certain concentrations of bleach and the halogens, chlorine and bromine. Bleach and the halogens served not only as germination stimulant for pretreated Alectra seeds but appropriate concentrations induced also high percentage (70–90%) germination indicating a breakage of seed dormancy as well. The activity of the bleach in stimulating high percentage germination could be significantly reduced when the optimally bleach-treated seeds were rinsed daily with water during the germination period. Vigna root exudate, but not bleach, was shown to act as root stimulant for Alectra seedlings. The possible mechanism of the bleach effect on germination and dormancy of Alectra seeds is discussed.     

Dormancy and germination responses of halophyte seeds to the application of ethylene

Ethylene is invariably produced during seed germination but its role in regulating seed dormancy and germination is poorly understood. Seeds of 22 halophytic species having different life forms – salt secreting dicots, salt secreting monocots, stem succulents and leaf succulents were germinated in Petri dishes kept in a growth chamber set at 20/30 °C (night/day) temperature and a 12 hr light period. Sodium chloride and ethephon were added to the medium from the beginning of the experiment. Seed germination was recorded every other day for twenty days. Application of ethylene did not have any significant effect on releasing seeds from innate dormancy. However, it appeared to have a role in alleviation of salinity effects which varied from negative in certain species to almost complete alleviation of high salinity effects in others.Our data indicates that ethylene appears to have little role in breaking innate seed dormancy however, in most halophyte seeds studied, application of ethylene alleviate the salinity effect to various degrees. Halophyte seeds which could germinate under saline conditions approaching twice the salinity of seawater may offer clues to understand management of seed germination under highly saline conditions. To cite this article: M.A. Khan et al., C. R. Biologies 332 (2009).     

Requirement for the action of endogenous ethylene during germination of non-dormant seeds of Amaranthus caudatus

The role of endogenous ethylene during germination of non-dormant seeds of Amaranthus caudatus L. was investigated. The seeds readily germinated in water and darkness at 24°C. Application of ethylene or of its precursor I-aminocyclopropane-I-carboxylic acid (ACC) slightly increased the rate of germination. Both compounds effectively antagonized osmotic inhibition by polyethyleneglycol. Application of aminoethoxyvinylglycine (AVG) reduced ethylene production by 90% but did not inhibit germination. However, germination was inhibited by 2,5-norbornadiene, a competitive inhibitor of ethylene action. This inhibition was counteracted by ethylene, ethephon or ACC and enforced by AVG. It is concluded that the action of endogenous ethylene is an indispensable factor during germination of non-dormant seeds of A. caudatus. Ethylene action is required from the start of imbibition on. In water, low levels of endogenous ethylene are sufficient for this action. PEG increased the ethylene requirement considerably.     

Ethylene as a Component of the Emanations From Germinating Peanut Seeds and Its Effect on Dormant Virginia-type Seeds

The embryonic axes of Spanish-type peanut seeds that do not exhibit dormancy to any extent were found to produce ethylene during germination. Virginia-type peanut seeds of the extremely dormant variety NC-13 produced low levels of ethylene when imbibed but not germinating. Treatments that released dormancy of NC-13 peanut seeds resulted in increased ethylene production by the embryonic axis. The estimated internal concentration of ethylene in Virginia-type peanut seeds was 0.4 ppm at 24 hr of germination. Fumigation with an external concentration of 3.0 to 3.5 ppm for 6 hr was sufficient to break dormancy of Virginia-type peanut seeds. These results suggest that ethylene is associated with the germination processes of non-dormant seeds and participates in the breaking of seed dormancy of dormant peanut varieties.     

Enhancement of germination rate of aged seeds by ethylene

Naturally and artificially aged seeds of rape, Brassica napus L., produced less ethylene than freshly harvested seed during the early stage of germination. With freshly harvested seeds one peak of ethylene production was observed during germination, which coincided with the emergence and elongation of root and cotyledon, accompanied by splitting of the seed coat. Application of exogenous ethylene was effective in accelerating germination in aged seeds but did not significantly improve the percentage of germination. Ethylene as a hormone was considered to serve as a stimulator of germination and growth. One of the factors causing seed aging might be the degeneration of an ethylene-producing system in the seed. Exogenous ethylene may be effective only for the seeds in which the ethylene-producing system is weakened but the following responding systems are still functional.     

Action of ferric and aluminium ions on the dormancy breakage of <Emphasis Type="Italic">Stylosanthes humilis</Emphasis> seeds

Dormancy of scarified seeds of Stylosanthes humilis was broken by acidic Al³⁺ and Fe³⁺ solutions. Fe⁺³-stimulated seeds exhibited a high activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and produced great amounts of ethylene, which showed correlated with the germination process. In addition, specific inhibitors of ethylene biosynthesis and action largely depressed the Fe³⁺-stimulated germination, leading to the conclusion that the ion broke dormancy by triggering ethylene production by the seeds. By contrast, inhibitors of ethylene biosynthesis and action did not impair germination of Al³⁺-stimulated dormant seeds. Moreover, ethylene production and activity of ACC oxidase of Al³⁺-treated seeds was substantially decreased by inhibitors of ethylene biosynthesis, but germination kept large. Together these data suggest that ethylene biosynthesis was not required in the chain of events triggered by Al³⁺ leading to dormancy breakage. Methyl viologen (MV), a reactive oxygen species-generating compound, broke dormancy of seeds to the same extent as Al³⁺ did. Germination of both Al³⁺- and MV-stimulated dormant seeds was inhibited by sodium selenate, an antioxidant compound; selenate, however had no effect on germination of Fe³⁺-stimulated seeds. Together these data indicate that the mechanisms underlying the germination of Al³⁺- and Fe³⁺-treated seeds are not the same.     

Physiology of oil seeds: I. Regulation of dormancy in virginia-type peanut seeds

The germination and ethylene production by dormant Virginia-type peanut seeds were observed in relation to phytohormone treatments that could conceivably release the dormancy of these seeds. A comparison was made between the effects of these treatments on the less dormant apical seeds and the more dormant basal seeds. Indole-3-acetic acid did not stimulate ethylene production by, or germination of, the dormant seeds to any extent. Gibberellic acid at 5 × 10⁻⁴ M stimulated ethylene production by apical seeds to 17 millimicroliters per hour and germination to only 40% above the control. The more dormant basal seeds were affected even less by gibberellic acid than the seeds. Ethylene gas at 8 microliters per liter stimulated germination to 85% above the control for both apical and basal seeds. At this ethylene concentration the physiology of the more dormant basal seeds was altered, so that they behaved in a manner similar to the inherently less dormant apical seeds. 2-Chloroethylphosphonic acid at 10⁻³ and 5 × 10⁻⁴ M provided results similar to ethylene gas. Both apical and basal seeds germinated 100% at 48 hours. Among the phytohormones tested in this study, ethylene gas produced the greatest germination at low concentrations, and it appears must directly related to initiating the reactions required for converting the quiescent cells to an active state of growth.     

Antagonistic effects of high and low temperature pretreatments on the germination and pregermination ethylene synthesis of lettuce seeds

Red light-induced germination of Grand Rapids lettuce seeds (Lactuca sativa L.) incubated at 20 C was inhibited if the seeds were first imbibed at 30 C for 36 hours. This effect was counteracted by exogenous ethylene and associated with a reduction in the rate at which the seeds produced ethylene throughout the pregermination period. A chilling treatment reversed the effect of a prior imbibition at 30 C on both germination and ethylene production. The possibility that the pretreatments influence germination through their effects on ethylene production is discussed.