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.     

Seed treatments with Vitavax for the control of loose smut of wheat and barley

Seed dressings with Vitavax—75 % w.p.—eliminated Ustilago nuda in spring barley seeds and greatly reduced infection due to U. tritici in winter wheat. Emergence and yield of these crops were not adversely affected. Seed soak treatments including 0·2 % aqueous Vitavax for 6 h at 30 °C (wheat and barley), 0·2% thiram for 24 h at 30 °C (barley) and 0·2% Vitavax for 1 h at 30 °C (barley) also rid the seeds of infection. In other tests with barley 2 h soaks in 0·2 % aqueous suspensions of Vitavax at 30 °C gave equivalent control to 12 h soaks in 0·2% thiram at 30 °C.     

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     

Morphological and functional seed traits of the wild medicinal plant Dioscorea strydomiana,the most threatened yam in the world

• Morphological and functional seed traits have important roles in characterising the species regeneration niche and help to understand the reproductive biology of rare and threatened plants, which can thus support appropriate plant conservation measures.
• Seed morphometric and dispersal kinetics of the critically endangered Dioscorea strydomiana were measured and compared with those of four other Dioscorea species, and seed germination response under constant temperatures (5–35 °C) was compared with that of the congeneric and widespread D. sylvatica.
• Seed mass of D. strydomiana (ca. 14 mg) was twice that of D. sylvatica, but similar to or smaller than the other species examined. Seeds of D. strydomiana have the lowest speed of descent and lowest variability in most of the morphological traits considered, suggesting lower phenotypic plasticity but higher variance in the wing‐loading value. Seeds of D. strydomiana reached maximum germination at 15 °C (ca. 47%), which decreased slightly to ca. 37% at 25 °C and was completely inhibited at 35 °C. D. sylvatica seeds started to germinate at 10 °C (ca. 3%), reached 75–80% germination at 15–20 °C and maximum (ca. 90%) at 25–30 °C. Base temperatures for germination (T_b) were 9.3 and 5.7 °C, for D. strydomiana and D. sylvatica, respectively. Due to the higher germination percentages of D. sylvatica, ceiling and optimum temperatures could also be modelled for this species, suggesting higher sensitivity to high temperature for seeds of D. strydomiana.
• The detected poor seed lot quality of D. strydomiana suggests difficulties in reproduction from seed, highlighting the need for further investigation and conservation actions for this threatened yam species.

     

The effects of some halogen-containing compounds on Bacillus subtilis endospores

Sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) were more active against Bacillus subtilis 8236 spores in both viability and in germination and outgrowth studies than were polyvinylpyrrolidone-iodine (PVP-I) and Lugol's solution. Of the two chlorine compounds studied NaOCl proved to be the more active. The two iodine-containing compounds gave contrasting results with the Lugol's solution demonstrating increased antibacterial activity with increasing available iodine concentration. The antibacterial behaviour of PVP-I, however, reflected the more complex nature of aqueous iodine—surfactant mixtures. Initially, non-complexed iodine concentration (the active species) increased with increasing total available iodine concentration, resulting in increasing antibacterial activity. However, due to changes in the physical properties of the mixture, a maximum concentration of non-complexed iodine was reached so that a further increase in total available iodine resulted in a decrease in non-complexed iodine concentration and consequently a decrease in the antibacterial activity of the solution was observed.
A greater inhibitory effect was observed in subsequent germination and outgrowth studies when spores were pre-treated with respective biocide than when untreated spores were added to germination media containing biocide at t = 0. This may reflect a combination of different contact times plus the neutralizing effect of the germination media on such halogen compounds.     

The effects of some halogen-containing compounds on Bacillus subtilis endospores

Sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) were more active against Bacillus subtilis 8236 spores in both viability and in germination and outgrowth studies than were polyvinylpyrrolidone-iodine (PVP-I) and Lugol's solution. Of the two chlorine compounds studied NaOCl proved to be the more active. The two iodine-containing compounds gave contrasting results with the Lugol's solution demonstrating increased antibacterial activity with increasing available iodine concentration. The antibacterial behaviour of PVP-I, however, reflected the more complex nature of aqueous iodine-surfactant mixtures. Initially, non-complexed iodine concentration (the active species) increased with increasing total available iodine concentration, resulting in increasing antibacterial activity. However, due to changes in the physical properties of the mixture, a maximum concentration of non-complexed iodine was reached so that a further increase in total available iodine resulted in a decrease in non-complexed iodine concentration and consequently a decrease in the antibacterial activity of the solution was observed. A greater inhibitory effect was observed in subsequent germination and outgrowth studies when spores were pre-treated with respective biocide than when untreated spores were added to germination media containing biocide at t = 0. This may reflect a combination of different contact times plus the neutralizing effect of the germination media on such halogen compounds.     

Growth-altering effects of sodium hypochlorite in cultured human dermal fibroblasts

Sodium hypochlorite, the most widely used antimicrobial active chlorine compound in chemical disinfection, is little used as an antiseptic in clinical practice. This study aimed to assess the capacity of hypochlorite to alter human dermal fibroblast growth in vitro in relation to the concentration and exposure time. Effects of decreasing concentrations of hypochlorite (0.5%-0.00025%) on fibroblast adherence capacity and proliferation, according to varying exposure times and fetal calf serum (FCS) concentrations were investigated combining XTT assay, which provides cytochemical quantification of metabolically-active cell number, and total cell protein content, an indirect method for assessing substrate-adhered cell number. Initial cytotoxicity was produced at 0.0075% hypochlorite within contact time of two hours, provoking concentration-dependent cell detachment. From 0.1% upwards, NaOCl exerted a profound cytotoxic effect on fibroblasts. At later stages (4 h) and concentrations > or = 0.01% hypochlorite produced dose-dependent mitochondrial dysfunction: cell survival progressively diminished from 71% to 10%. Cytotoxic effects were not significantly affected by exposure-time periods, probably because maximum chlorine is released within the first four hours. Hypochlorite concentrations from 0.005% to 0.00025% were found to have no inhibitory effects on cell growth; in fact, they appear to exhibit the opposite effect. Increments in protein content found after 24 h exposure ranged from 30% to 120% above control values. Hypochlorite is highly cytotoxic for fibroblasts at concentrations > or = 0.01% provoking concentration-dependent loss of cell adherence capacity and mitochondrial dysfunction. In contrast, a mitogenic effect was observed with concentrations < or = 0.005% which supports NaOCl as a source growth-promoting activity in cultured human fibroblasts. Hypochlorite proved to be a highly reactive molecule which inhibits or stimulates cell division according to the concentration.     

Comparative assessment of different biocides in swimming pool water

For disinfection of swimming pool water chlorine of chlorine-based products are normally used. In practice, these products have proven their worth regarding killing of pathogenic micro-organisms. Detailed values of their biocidal activity in swimming pool water were not found in literature. In the given study the efficacy of sodium hypochlorite (NaOCl) versus five micro-organisms was investigated.It is known that chlorination of swimming pool water may lead to formation of specific unwanted products like haloform. Nowadays, the concentration of those by-products in swimming pool water is limited and specific measures exist to minimize their formation. Nevertheless, there is increasing interest in alternative methods without by-product formation like e.g. hydrogen peroxide (H₂O₂) treatment.In the given study the antimicrobial activity of sodium hypochlorite was compared with that of different hydrogen peroxide-based products. The test procedure used was specifically designed to simulate practical conditions in a swimming pool but at the same time to lead to adequate reproducibility. Five test organisms were selected being relevant for the swimming pool area: Pseudomonas aeruginosa, Escherichia coli, Legionella pneumophila, Staphylococcus aureus and Candida albicans.The swimming pool water for the test was artificially prepared. Water hardness, temperature and pH value were adjusted to a defined level. Regarding simulation of organic load it was found that a mixture of urea, creatinine and several amino acids was most appropriate.Addition of the test organisms was done in three portions: one big in the beginning and two smaller after 10 and 20 min to simulate recontamination by bathers. Total test period was 30 min. The number of surviving cells was determined after 30 s as well as after 10, 20 and 30 min.Sodium hypochlorite was tested at a concentration of 1 ppm active chlorine. Compared to that three products based on hydrogen peroxide were investigated: pure hydrogen peroxide, hydrogen peroxide + silver nitrate and a trade product based on hydrogen peroxide.Sodium hypochlorite resulted in total kill of the inoculated organisms after 10, 20 and 30 min corresponding to a log 4 reduction. In contrast to that the biocidal effect achieved by the hydrogen peroxide-based products was significantly lower than one log cycle notwithstanding a very high concentration of up to 150 ppm.The test results confirm the very good killing activity of sodium hypochlorite versus micro-organisms relevant for the swimming pool area. Products based on hydrogen peroxide, with or without silver ions, are from a microbiological point of view no real alternative to chlorine disinfection in swimming pools.     

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.     

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。     

Pitfalls in using sodium hypochlorite as a seed disinfectant in C incorporation studies

Seeds sterilized with sodium hypochlorite (NaOCl) retained sufficient amounts to interfere with studies of amino acid metabolism of the sterilized seeds during germination. Repeated washing in water did not remove NaOCl completely. However, soaking the seeds for 10 min in 0.01 n HCl removed NaOCl completely, without reducing germinability.Residual NaOCl reacted with the amino acids and reduced their concentrations in the incubation media. This reaction resulted in high production of CO(2) and low uptake of amino acids by the seeds. Decarboxylation of the amino acids occurred in the incubation medium outside the seed, was independent of the presence of seeds in the reaction, and therefore was not related to amino acid metabolism by the seeds. Effects of NaOCl on uptake, incorporation, and CO(2) production from indoleacetic acid were similar to those of the amino acids studied.     

Sporicidal Properties of Some Halogens

S ummary . Sodium hypochlorite, sodium dichloro iso cyanurate, dichlorodimethyl hydantoin, dibromodimethyl hydantoin and an iodophor have been examined for disinfectant activity against spores of Bacillus cereus. Under the test conditions used sodium hypochlorite was the most effective compound and dibromodimethyl hydantoin was least affected by increase in pH from 6.5 to 8; the activity of the iodophor was unaffected in the pH range 2.3–6.5. Bacillus subtilis spores were much more resistant to the disinfectants than were B. cereus spores. The addition of KBr to solutions of sodium dichloro iso yanurate enhanced its activity at pH 9 but not at pH 7 or 8.
Mixtures of 1.5–4% of NaOH with NaOCl (200 p of available chlorine/m) were much more rapidly sporicidal than either NaOH or NaOCl (pH 9 and above) alone.     

风箱果种子催芽技术研究

以风箱果自然种群成熟的种子为研究对象,对种子的生活力和催芽方法进行了研究,结果表明：TTC染色法测得风箱果种子的生活力达到94.44%;清水浸种处理的发芽势和发芽率均较低,以浸种48 h的效果较好,发芽率为28.33%;不同化学药剂处理均能不同程度的提高发芽率,其中50 mg·L^-1 GA₃+4%KNO₃的混合药剂处理72 h的萌发效果最好,发芽率达到90%;100和200 mg·L^-1 GA₃能极显著提高发芽势（P<0.01）;低温处理28 d能显著提高发芽势和发芽率（P<0.05）。     

Inactivation of Bacillus anthracis Spores by Liquid Biocides in the Presence of Food Residue

Biocide inactivation of Bacillus anthracis spores in the presence of food residues after a 10-min treatment time was investigated. Spores of nonvirulent Bacillus anthracis strains 7702, ANR-1, and 9131 were mixed with water, flour paste, whole milk, or egg yolk emulsion and dried onto stainless-steel carriers. The carriers were exposed to various concentrations of peroxyacetic acid, sodium hypochlorite (NaOCl), or hydrogen peroxide (H₂O₂) for 10 min at 10, 20, or 30°C, after which time the survivors were quantified. The relationship between peroxyacetic acid concentration, H₂O₂ concentration, and spore inactivation followed a sigmoid curve that was accurately described using a four-parameter logistic model. At 20°C, the minimum concentrations of peroxyacetic acid, H₂O₂, and NaOCl (as total available chlorine) predicted to inactivate 6 log₁₀ CFU of B. anthracis spores with no food residue present were 1.05, 23.0, and 0.78%, respectively. At 10°C, sodium hypochlorite at 5% total available chlorine did not inactivate more than 4 log₁₀ CFU. The presence of the food residues had only a minimal effect on peroxyacetic acid and H₂O₂ sporicidal efficacy, but the efficacy of sodium hypochlorite was markedly inhibited by whole-milk and egg yolk residues. Sodium hypochlorite at 5% total available chlorine provided no greater than a 2-log₁₀ CFU reduction when spores were in the presence of egg yolk residue. This research provides new information regarding the usefulness of peroxygen biocides for B. anthracis spore inactivation when food residue is present. This work also provides guidance for adjusting decontamination procedures for food-soiled and cold surfaces.     

Pea seed infection by Mycosphaerella pinodes and Ascochyta pisi and its control by seed soaks in thiram and captan suspensions

Apparently normal pea seeds from pods bearing lesions of Mycosphaerella pinodes were often internally infected with the fungus. When infected seeds were sown in sterile grit almost all the seedlings showed disease lesions, at or below soil level, 4–6 weeks after sowing. Seed infected with Ascochyta pisi gave only 40% infection of seedlings: these showed lesions on the stem and first two leaves within 4 weeks of sowing. Infection of seeds by both pathogens could be eradicated by soaking the seeds for 24 hr. in 0.2% suspensions of thiram or captan at 30d?C. In laboratory or greenhouse tests these treatments did not check germination, but in the field the captan treatment reduced emergence. The treated seeds became fully imbibed but could be dried and stored: the thiram treatment was used for semi-commercial treatment of quantities of seed up to 3 cwt.     

Inactivation of Bacillus anthracis spores by liquid biocides in the presence of food residue

Biocide inactivation of Bacillus anthracis spores in the presence of food residues after a 10-min treatment time was investigated. Spores of nonvirulent Bacillus anthracis strains 7702, ANR-1, and 9131 were mixed with water, flour paste, whole milk, or egg yolk emulsion and dried onto stainless-steel carriers. The carriers were exposed to various concentrations of peroxyacetic acid, sodium hypochlorite (NaOCl), or hydrogen peroxide (H(2)O(2)) for 10 min at 10, 20, or 30 degrees C, after which time the survivors were quantified. The relationship between peroxyacetic acid concentration, H(2)O(2) concentration, and spore inactivation followed a sigmoid curve that was accurately described using a four-parameter logistic model. At 20 degrees C, the minimum concentrations of peroxyacetic acid, H(2)O(2), and NaOCl (as total available chlorine) predicted to inactivate 6 log(10) CFU of B. anthracis spores with no food residue present were 1.05, 23.0, and 0.78%, respectively. At 10 degrees C, sodium hypochlorite at 5% total available chlorine did not inactivate more than 4 log(10) CFU. The presence of the food residues had only a minimal effect on peroxyacetic acid and H(2)O(2) sporicidal efficacy, but the efficacy of sodium hypochlorite was markedly inhibited by whole-milk and egg yolk residues. Sodium hypochlorite at 5% total available chlorine provided no greater than a 2-log(10) CFU reduction when spores were in the presence of egg yolk residue. This research provides new information regarding the usefulness of peroxygen biocides for B. anthracis spore inactivation when food residue is present. This work also provides guidance for adjusting decontamination procedures for food-soiled and cold surfaces.     

Effect of temperature and cold stratification on seed germination of the Mediterranean wild aromatic Clinopodium sandalioticum (Lamiaceae)

A germination study was carried out on seeds of Clinopodium sandalioticum (Bacch. & Brullo) Bacch. & Brullo ex Peruzzi & Conti (Lamiaceae), a wild aromatic plant endemic to Sardinia. Seeds were incubated at a range of constant (5–25°C) and an alternating temperatures regime (25/10°C), with 12 hours of irradiance per day. The results achieved at 10°C were also compared with those obtained after a period of cold stratification at 5°C for three months. Final seed germination ranged from ca. 28% (5°C) to ca. 72% (25/10°C). A base temperature for germination (T_b) of ca. 5°C and a thermal constant for 50% germination (S) of 89.3°Cd were identified and an optimal temperature for germination (T_o) was estimated to be comprised between 20 and 25°C. Cold stratification negatively affected seed viability and germination at 10°C. Although a typical “Mediterranean germination syndrome”, could not be detected for C. sandalioticum seeds, these results were coherent with those previously reported for other Mediterranean Lamiaceae species.     

Evaluation of chemical seed coat sterilants

Summary Three seed coat sterilants were evaluated for effectiveness as surface sterilants and for effects on seed germination. Several exposure times in sodium hypochlorite, hydrogen peroxide and peracetic acid were evaluated. Seeds of the following crop plants were used: wheat (Triticum aestivum L.); sorghum [Sorghum bicolor (L.) Moench]; soybean [Glycine max (L.) Merrill]. All treatments reduced germination of wheat and soybean seed, but only the most severe peracetic acid treatments substantially reduced germination of sorghum seed. Considering both sterilization effectiveness and seed germination, a treatment with 5% sodium hypochlorite for 45 min appeared to be the most satisfactory of the treatments used.     

In vitro reproduction of three Limodorum species (Orchidaceae): impacts of scarification methods and nitrogen sources on mature seed germination and seedling development

Limodorum species are hard-to-cultivate, germinating with difficulty or not germinating at all on a number of media. The present work was focused on the effects of seed sterilization-scarification methods and of different nitrogen sources on in vitro seed germination and seedling development of Limodorum abortivum, L. brulloi and L. trabutianum. Our results showed that sodium hypochlorite solution with a 1% concentration of available chlorine was ineffective for both sterilization and scarification of Limodorum seeds, while a 5% concentration was found to be an effective sterilizing and scarifying solution for L. trabutianum. The positive effect of organic nitrogen in promoting asymbiotic germination in many terrestrial orchids was confirmed by this study for L. trabutianum and L. brulloi, which showed significantly higher germination percentages on Basal Medium modified than on Knudson C, which contains inorganic nitrogen. This is the first report of in vitro asymbiotic germination of mature seeds of L. brulloi and L. trabutianum, especially successful for L. trabutianum that showed high germination percentages (>63%) and an active seedling development. These results may be useful for conservation purposes, since L. brulloi is a narrow Italian endemics and L. trabutianum is threatened with extinction both in Italy and in France.     

Understanding biological and ecological factors affecting seed germination of the multipurpose tree Anogeissus leiocarpa

• Anogeissus leiocarpa (DC.) Guill. & Perr. (Combretaceae) has important economic and cultural value in West Africa as source of wood, dye and medicine. Although this tree is in high demand by local communities, its planting remains limited due to its very low propagation via seed.
• In this study, X‐rays were used to select filled fruits in order to characterise their morphology and seed germination responses to treatment with sulphuric acid and different incubation temperatures.
• Morphological observations highlighted a straight orthotropous seed structure. The increase in mass detected for both intact and scarified fruits through imbibition tests, as well as morphological observations of fruits soaked in methylene blue solution, confirmed that they are water‐permeable, although acid‐scarified fruits reached significantly higher mass increment values than intact ones. Acid scarification (10 min soaking in 98% H₂SO₄) positively affected seed germination rate but not final germination proportions. When intact fruits where incubated at a range of temperatures, no seeds germinated at 10 °C, while maximum seed germination (ca. 80%) was reached at 20 °C. T₅₀ values ranged from a minimum of ca. 12 days at 25 °C to a maximum of ca. 34 days at 15 and 35 °C. A theoretical base temperature for germination (T_b) of ca. 10 °C and a thermal requirement for 50% germination (S) of ca. 195 °Cd were also identified for intact fruits.
• The results of this study revealed the seed germination characteristics driven by fruit and seed morphology of this species, which will help in its wider propagation in plantations.