Chromosome observation method at metaphase and pro-metaphase stages in diploid and octoploid strawberries

Chromosome observation is necessary to elucidate the structure, function, organization, and evolution of octoploid strawberry plants’ genes and genomes. However, distinguishing strawberries’ chromosomes from one another using light microscopy is extremely difficult, not only because of their small size and large number, but also because current chromosome observation methods are insufficient. Chromosome preparation and staining using maceration enzymes, acetic acid, and DAPI (4′,6-diamidino-2-phenylindole) were improved for this study to obtain clear images of somatic chromosomes in Fragaria vesca (2n = 14) and Fragaria×ananassa (2n = 56). Collected root tips of octoploid plants were placed in 0.002 M 8-hydroxyquinoline solution for 1 h and stored at 4 °C for 16 h. Subsequently, they were fixed using 3:1 absolute alcohol:glacial acetic acid for 40 min, hydrolyzed in the 1N HCl solution at room temperature for 2 h, macerated using an enzyme solution for 25 min at 42 °C, and stained in 1.5% lacto-propionic orcein solution. On the other hand, in case of DAPI staining, the macerated root tips of octoploid plants were soaked in 60% acetic acid for 5 min before staining. Clear digital images of F. vesca and F.×ananassa were obtained using light and fluorescent microscopy. Their respective 14 and 56 chromosomes were counted. Fluorescent microscopy yielded clear chromosome images at the pro-metaphase in F. vesca and F.×ananassa. This chromosome observation method alleviates the difficulties that have heretofore hindered chromosome analyses of strawberry plants.     

Improvement of staining method for observation of mitotic chromosomes in octoploid strawberry plants

An important strategy to conduct intentional breeding of octoploid strawberry plants is to recognize the functions of every chromosome. To do so, a methodology must be developed to distinguish chromosomes one by one. We reported the possibility of distinguishing chromosomes using light microscopy when somatic cells of octoploid strawberry plants were stained using ordinary methods with lacto-propionic orcein (LPO). However, karyotype analysis of octoploid strawberry plants required clearer chromosome images. This study obtained clearer chromosome images of octoploid Fragaria × ananassa and F. chiloensis plants. Three staining methods were examined: 60% acetic acid (AA) alone, 1.5% LPO alone, and two-step treatments with 60% AA and 1.5% LPO. Collected root tips of the plants were placed in 2 mM 8-hydroxyquinoline solution for 1 h and were subsequently stored at 4 °C for 15 h. The samples were then fixed in a 3:1 absolute alcohol: glacial acetic acid solution for 40 min, followed by mixture with 1N HCl solutions at room temperature for 2 h and then at 60 °C for 10 min. For separate staining using 60% AA and 1.5% LPO, the root tip was expelled on the glass slide with a drop of each solution for a few minutes to stain the chromosomes. For the two-step staining method, the samples stained with 60% AA were frozen at −80 °C for at least 5 min. The cover slip was removed using a razor blade. Subsequently, the specimens were air-dried and stained with the 1.5% LPO for 3 min. Digital images of chromosomes were obtained using light microscopy. Samples of the two-step staining method produced the clearest chromosome images in both F. × ananassa and F. chiloensis. Furthermore, the greatest color difference between the chromosomes and the cytoplasm was obtained from images of the two-step staining method among the three staining methods. These results demonstrate that the two-step staining method is useful for chromosome counting and karyotype analysis in strawberry plants.     

Changes in antioxidants and fruit quality in hot water-treated ‘Hom Thong’ banana fruit during storage

The effects of hot water treatment on antioxidants and fruit quality were investigated in banana fruit of cv. Gros Michel (Musa acuminata, AAA Group, locally called cv. Hom Thong) by immersing fruits in hot water (50 °C) for 10 min, before storage at 25 °C for 10 days or 14 °C for the first 8 days followed by storage at 25 °C for the second 8 days until ripening. Quality parameters including peel color and pulp firmness indicated that hot water treatment helped to delay banana fruit ripening at both storage conditions. Hot water treatment decreased the levels of hydrogen peroxide (H₂O₂) and malonydialdehyde (MDA) during storage at 25 °C. Glutathione (GSH and GSSG) contents and the ratio of GSH/GSSG during fruit approaching ripening were significantly induced in hot water-treated fruits while ascorbic acid (AA) contents were slightly increased. In addition, the combined treatment increased free phenolics and flavonoids during storage. Results suggest that hot water treatment has led to an induction of antioxidants in banana fruits as indicated by an increase of antioxidants and a decrease of H₂O₂ during ripening, and all of which result in a delayed ripening of banana fruit.     

Physicochemical changes in Mazafati date fruits incubated in hot acetic acid for accelerated ripening to prevent diseases and decay

A method was developed and optimized for the accelerated ripening of date fruits of cultivar ‘Mazafati’ to prevent diseases and decay. The date fruits were incubated in hot acetic acid solution 0.5% at 40 + 1 °C for 72 h. During the process some physicochemical changes in the fruits were studied and were found to be comparable with the changes in the fruits that naturally ripened on the tree. Fruit firmness, water insoluble solid (WIS), protein, pH, L*a*b* and E decreased during accelerated ripening whereas in control samples at 4 °C increased. Total solid (TS), total soluble solid (TSS) and acidity were slightly higher in treated fruits compared to control fruits. The greatest loss of fruit firmness occurred during the first 12 h of incubation. Organoleptic tests also showed little difference between the naturally ripened fruits on trees and accelerated ripened fruits in hot acetic acid. Overall there was no difference between the fruits and were readily acceptable to consumers.     

Promotion by 5-aminolevulenic acid of pepper seed germination and seedling emergence under low-temperature stress

The effects of incorporating 5-aminolevulenic acid (ALA) into the priming solution on low-temperature germination and emergence percentage performance of red pepper (Capsicum annuum cv. Sena) seeds before and after seed storage were investigated. Seeds were primed in 3% KNO₃ solution for 6 days at 25 °C in darkness containing 0 ppm, 1 ppm, 10 ppm, 25 ppm, 50 ppm or 100 ppm ALA. Following priming, seeds were either immediately subjected to germination and emergence tests at 15 °C or stored at 4 °C or 25 °C for 1 month after which they were subjected to germination and emergence tests at 15 °C. Priming pepper seeds in the presence of ALA improved final germination percentage (FGP) and germination rate (MGT) at 15 °C compared to non-primed seeds. The highest FGP was obtained from seeds primed in the presence of 25 ppm and higher ALA concentrations while the highest MGT was obtained from seeds primed in KNO₃ supplemented with 10 ppm ALA. Emergence percentages were the highest for the seeds primed in the presence of 25 ppm ALA and 50 ppm ALA while non-primed seeds had the lowest emergence percentage. Highest emergence rates (MET) and heaviest seedlings were also obtained from seeds primed in KNO₃ supplemented with 50 ppm ALA. Although all priming treatments improved germination and emergence performance of pepper seeds at 15 °C following 1 month of storage under two different temperatures, inclusion of 25 ppm and 50 ppm ALA into the priming solution resulted in higher germination and emergence percentages and faster germination and emergence compared to seeds primed in KNO₃ only and non-primed seeds. These results indicate that priming seeds in 25 ppm and 50 ppm ALA incorporated into the KNO₃ solution can be used as an effective method to improve low-temperature performance of red pepper seeds and that these seeds can be stored for 1 month at 4 °C or 25 °C and still exhibit improved germination and emergence performance at 15 °C.     

Biocontrol of mildew with Bacillus subtilis in bitter gourd (Momordica charantia L.) seeds during germination

The bitter gourd seed has a thick, hard seed coat. Mildew often occurs during germination and causes uneven and low rates of seed germination. However, the problems caused by mildew can be overcome by treating seeds with hot water, by soaking in water, or by using microorganisms. Seeds of the ‘Ching Pi’ bitter gourd were treated in a water bath at 60 °C for 10 min and then soaked in tap water at 25 °C for 24 h. The resulting germination percentage was 86.7%, and the resulting percentage of mildewed seeds was 10%. The biocontrol potential of three commercially available Bacillus subtilis solutions was examined. For seeds primed with Huodijun B. subtilis solution, the germination percentage was 73.3% and the mildewed percentage 6%. In dual cultures, the antibiotic content in the Huodijun B. subtilis solution was significantly greater than in Yunghsing and Huolibao, the other B. subtilis solutions examined. B. subtilis effectively inhibited the growth of Rhizoctonia solani mycelium and caused abnormal mycelial growth.     

Response of gladiolus (Gladiolus spp) plants after exposure corms to chitosan and hot water treatments

The state of Morelos, Mexico has gradually become an important producer of gladiolus. Some preconditioning treatments of corms are empirically done causing uneven emergence and low quality of flowers. In this investigation, before planting, gladiolus corms var. ‘Blanca Borrego’ were dipped in chitosan (chitosan reagent and commercial chitosan Biorend^®), in hot water at various temperatures and in treatments combined with Biorend^® at 1.5% and hot water. Results indicated that the most influenced variables were corm germination, number of flowers per spike, number of cormlets and vase life. Overall, the commercial product Biorend^® at 1.5% accelerated corm emergence in approximately 4 days, the number of flowers increased by 2–7 and the vase life extended for 3 days. The number of cormlets was also duplicated. Corms dipped in the commercial chitosan Biorend^® at 1.5% at different intervals of time were not greatly affected except for the emergence and number of cormlets. However, for this experiment there were significant effects on the number of leaves and flowers because of the interactions between chitosan and the immersion time. The temperature of 55 °C affected plant development because emergence was delayed by 6 days; and there were less number of leaves, flowers and cormlets. On the other hand, the incidence of Fusarium oxysporum in naturally infected corms was 0% at temperatures of 55 °C and 50 °C. Immersion times (0, 10, 15 and 20 min) in hot water at 50 °C did not show significant effects on plant development and vase life. Corms dipped in Biorend^® at 1.5% and hot water at 50 °C accelerated their emergence for about 1–7 days, the number of flowers increased by two, extended the storage life for 1–3 days and increased the number of cormlets. The integration of these two treatments -Biorend^® and hot water- might be a good option for increasing the gladiolus plant quality and vase life.     

Droplet-vitrification of apical shoot tips of Rubus fruticosus L. and Prunus cerasifera Ehrh.

Apical shoot tips excised from in vitro plantlets of blackberry (Rubus fruticosus L. ‘?a?anska Bestrna’) and cherry plum (Prunus cerasifera Ehrh.) were tested for recovery after cryopreservation using the droplet-vitrification technique. Following treatment for 30 min with a loading solution comprising 1.9 M glycerol and 0.5 M sucrose, explants were dehydrated with a highly concentrated cryoprotectant solution, so called vitrification solution. Shoot tips were dehydrated for 10, 20 and 30 min at room temperature with a solution derived from the original PVS2 solution (containing 37.5% (w/v) glycerol, 15% (w/v) dimethylsulfoxide, 15% (w/v) ethylene glycol and 22.5% (w/v) sucrose) and for 60, 90 and 120 min using the PVS3 solution (containing 50% (w/v) glycerol and 50% (w/v) sucrose). Explants were cooled by direct immersion in LN in 10 μl droplets of vitrification solution placed on aluminium foil strips. Rewarming was done by direct plunging of foil strips in a preheated (37 °C) unloading solution (0.8 M sucrose) for 30 s, after which an equal volume of unloading solution (at room temperature) was added for further incubation for 30 min. As for regrowth of blackberry, PVS3 proved more effective than the modified PVS2, but the difference was significant (P < 0.05) only for the shortest treatment duration. The duration of PVS3 treatment had no significant effect on regrowth of cryopreserved shoot tips (45.8–70%). By contrast, a 30-min treatment with modified PVS2 solution resulted in a significant increase in regeneration percentage (30%), as compared with a 10-min treatment with the same solution (5%). Cherry plum shoot tips were very sensitive to both vitrification solutions and growth recovery of cryopreserved samples was generally lower (5–20%) than that of blackberry explants. No significant influence of PVS treatment (both type of solution and treatment duration) on regrowth of cryopreserved shoot tips was observed with cherry plum shoot tips. Experiments performed in France and in Serbia produced similar results, thereby showing the robustness and reproducibility of the protocols developed.     

An effective storage protocol for improving the postharvest performance in cut spikes of Consolida ajacis Nieuwl cv. Violet blue

The synergistic effect of cold treatment and protein synthesis inhibition was investigated in cut spikes of Consolida ajacis cv. Violet blue to determine the effective postharvest storage and transportation protocol, and to improve the postharvest performance. The 35 cm cut spikes were cool wet stored at 5 °C for 72 h before or after pulse treatment with protein synthesis inhibitor, cycloheximide, at 0.01 mM concentration. A set of unpulsed spikes wet stored at 5 °C represented control. The treatment effects were evaluated by placing the spikes in holding solutions containing distilled water (DW) or sucrose 0.2 M + 8-HQS 100 mg/l (SUC + HQS) at room temperature under laboratory conditions. The spikes pretreated with CHI before 72 h wet storage at 5 °C showed an enhancement of vase life besides maintaining higher fresh and dry mass of flowers, soluble protein content in sepals and sustained rate of blooming as compared to controls. The present study recommends that pulse treating the spikes of C. ajacis with 0.01 mM CHI before wet storage at 5 °C for 72 h, followed by transfer to holding solution containing SUC + HQS can be used as an effective postharvest storage treatment to bring out their transportation within 72 h without affecting their vase life.     

Addition of salicylic acid to nutrient solution combined with postharvest treatments (hot water,salicylic acid,and calcium dipping) improved postharvest fruit quality of strawberry

This experiment was conducted to study the effect of salicylic acid addition to nutrient solution and different postharvest treatments on fruit quality of strawberry cv. Camarosa after 7 days at 2 °C. Plants were irrigated with two complete nutrient solutions, with salicylic acid (0.03 mM) or without salicylic acid as the control. Fruits were then treated with eight different postharvest treatments (25 °C water, 45 °C water, 25 °C or 45 °C water containing CaCl₂ (1%), 25 °C or 45 °C water containing salicylic acid (2 mM) and 25 °C or 45 °C water containing both CaCl₂ (1%) and salicylic acid (2 mM)). Fruits which received SA in their nutrient solution had less weight loss and decay and higher firmness. All of the postharvest treatments improved fruit quality characteristics. Fruits dipped in salicylic acid solution had less weight loss, decay and a* (redness) and higher firmness and hue angle than control. Heat treated fruits had less decay and a* and higher hue angle than control. Fruits dipped in CaCl₂ solution had less weight loss, decay and a* and higher firmness than control. Combination of the three postharvest treatments improved firmness, decay, weight loss and vitamin C.     

Interspecific variations in seed germination of Corylopsis

This study was initiated to investigate the differences in germination percentages and rates between Corylopsis coreana Uyeki and Corylopsis sinensis var. calvescens Rehder & E.H. Wilson following a warm stratification (WS) and cold stratification (CS), and to study the effect of different WS temperatures interacting with different durations of CS. Warm stratification at 10 °C, 15 °C, 20 °C, and 25 °C was given for 1 month (1 M 10 °C, 15 °C, 20 °C, and 25 °C WS) followed by 0 M, 1 M, 2 M, and 3 M of CS at 5 °C (0 M, 1 M, 2 M, 3 M CS) and seeds were germinated in an air conditioned greenhouse maintained at 18.5 °C/18 °C. On average, less than 1% of C. coreana seeds germinated when sown without any WS and CS or with 1 M 15 °C, 20 °C, and 25 °C WS without CS treatment. However, 26% C. coreana seeds germinated after 1 M 10 °C WS without any CS treatment. Germination was not affected by WS temperatures when followed by 2 M 5 °C CS. It is concluded that C. coreana exhibited low seed germination at 10 °C and that this temperature could be considered the upper limit of CS for C. coreana. Only 2 M CS was required for more than 90% seeds to germinate. However, C. sinensis var. calvescens required longer than 3 M CS for more than 29% seeds to germinate. This clearly shows that there is an interspecific variation in optimum dormancy-breaking requirements.     

Can a post-harvest ripening treatment extend the longevity of Rhododendron L. seeds?

Pre-dehiscent capsules were collected from two Rhododendron griersonianum (Balf.f. & Forrest) trees and either immediately dried in a dry-room (15% relative humidity, 15 °C) or placed in a high humidity room (80% relative humidity, 15 °C) for 30, 60, or 90 d. Further capsules were also collected from the trees at 30 and 60 d, but seeds had been dispersed by 90 d. Seed ageing experiments (60% relative humidity, 45 °C) carried out on these seed-lots and on seeds from a further 10 Rhododendron (L.) species confirmed that short seed lifespans is a trait of the genus, with a mean P₅₀ value of ca. 20 d for this storage environment.     

Heat treatment and harvest date interact in their effect on superficial scald of ‘Granny Smith’ apple

A study was made of the effect of hot water dips (HWD) at temperatures of 42, 44, 46 and 48 °C (HWD 42 °C, HWD 44 °C, HWD 46 °C and HWD 48 °C, respectively) for 3 min on development of superficial scald and the concentration of α-farnesene and conjugated trienols (CT), CT₂₅₉, CT₂₆₉, CT₂₈₁, as well as OD₂₀₀ on Granny Smith apple fruits harvested on three dates and stored 125 days in air at 2 °C. HWD 48 °C efficiently decreased surface scald in the second and third harvest. α-Farnesene and CT were measured spectrophotometrically and by HPLC. No clear relationship of OD₂₀₀ and scald development was observed. Correlation of scald index and OD₂₀₀ at the end of storage was negative for the second harvest date. There was no significant correlation between the scald index and CT₂₅₉. Scald index was positively correlated with CT₂₆₉ after 80 days for the second and third harvest and at the end of storage for the second harvest. CT₂₈₁ was spectrophotometrically detectable only at the end of the storage, for the third harvest date, in control, HWD 42 °C, and HWD 44 °C. HWD 42 °C had significantly higher CT₂₈₁ compared to HWD 44 °C and control. HPLC analysis of control samples revealed presence of CT₂₈₁ in all three harvest dates, and presence of at least two components, as was the case of CT₂₅₉ and CT₂₆₉. The ratio of these two components was different for all three CT species. Fruit maturity was an important factor determining the response of fruit to heat and occurrence of superficial scald. The results indicate that a successful treatment using HWD to control superficial scald may be obtained after further research and that there are still some questions on the role of different CT's in scald biochemistry that should be addressed in future research.     

Cryopreservation of carnation (Dianthus caryophyllus L.) shoot tips by encapsulation-vitrification

Shoot tips excised from in vitro cultured plants of Dianthus caryophyllus L. (cv. Pallas, cv. Pink Candy and cv. Wanessa) were successfully cryopreserved using an encapsulation-vitrification method. Shoot tips (2–3 mm in length) were encapsulated in sodium alginate, precultured on liquid Murashige and Skoog (1962) medium supplemented with various sucrose concentrations (0.25, 0.5, 0.75, 1.0 M) for 24 h or 48 h and dehydrated with the vitrification solution PVS2 (up to 4 h) at 24 °C or 0 °C prior to direct immersion in liquid nitrogen (−196 °C). A maximum of shoot regeneration from cryopreserved shoot tips was obtained with the following combinations: preculture in 0.5 M sucrose and 180 min dehydration treatment at 0 °C for cv. Pallas (60% shoot formation), or preculture in 0.75 M and 200 min dehydration at the same temperature for cv. Pink Candy (66.6% shoot formation) and cv. Wanessa (73% shoot formation).     

Induction of disease resistance and ROS metabolism in navel oranges by chitosan

The objective of this work was to evaluate how disease resistance and reactive oxygen species (ROS) metabolism in harvested navel oranges (Citrus sinensis L. Osbeck) may be affected by chitosan. Fresh navel oranges were treated with 2% chitosan or 0.5% glacial acetic acid (control) solution for 1 min, and some were inoculated with Penicillium italicum and Penicillium digitatum. Then, the fruit were stored at 20 °C and 85–95% RH. Treatment with 2% chitosan significantly reduced the disease incidence and the lesion diameter compared with control fruit. This treatment effectively enhanced the activities of peroxidase (POD) and superoxide dismutase (SOD), and levels of glutathione (GSH) and hydrogen peroxide (H₂O₂), inhibited the activities of catalase (CAT) and the decreases of ascorbate (AsA) content during navel orange fruits storage. Ascorbate peroxidase (APX) activity in the navel orange fruit was induced slightly by the chitosan treatment during 14–21 days storage. However, glutathione reductase (GR) activity in the fruit was not enhanced by the chitosan treatment. These results indicated that chitosan treatment could induce the navel orange fruit disease resistance by regulating the H₂O₂ levels, antioxidant enzyme and ascorbate–glutathione cycle.     

Effect of synthetic auxins on fruit development of ‘Bing’ cherry (Prunus avium L.)

The main cherry cultivar grown in the warm climate of Israel, ‘Bing’, produces relatively small fruit. Over three consecutive years (2003–2005), application of 50 mg l⁻¹ 2,4-dichlorophenoxypropionic acid [2,4-DP; as its butoxyethyl ester (Power™)], 10 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid [3,5,6-TPA; as the free acid (Maxim^®)], or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) plus 30 mg l⁻¹ naphthaleneacetic acid (NAA; 0.3% Amigo™), at the beginning of pit-hardening when fruitlet diameter was ca. 13 mm caused appreciable and significant increases in fruit size and total yield, except when the crop load was heavy. Anatomical studies revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect on fruit quality, either at harvest or after 1 month of storage at 0 °C, or on return yield in the following year.     

Production protocol development for greenhouse cut Linaria, Lupinus, and Papaver flowers

Linaria maroccana Hook. f. Ann., ‘Lace Violet’, Lupinus hartwegii ssp. cruikshankii Lindl. ‘Sunrise’ and Papaver nudicaule L. ‘Meadow Pastels’ seeds were directly sown into 105 cell plug trays and received either ambient light or supplemental high intensity discharge (HID) lighting. For each species, a 2 × 3 × 3 factorial was used with two light intensities during propagation, three transplant stages, and three night temperatures. Seedlings were transplanted at the appearance of 2–3, 5–6, or 8–9 true leaves. Transplanted Linaria and Papaver seedlings were placed at 5/11, 10/16, or 15/21 ± 1 °C night/day temperatures and Lupinus seedlings were placed at 15/24, 18/25, or 20/26 ± 2 °C night/day temperatures. For this study, the optimum production temperature for Linaria was 10/16 °C as the cut stems produced at 15/21 °C were unmarketable and production time was excessively long at 5/11 °C. At 10/16 °C, Linaria seedlings should be transplanted at the 2–3 leaf stage to maximize stem number, stem length and profitability. For Lupinus the optimum temperature was 15/24 °C due to long stems and high profitability per plant. Lupinus seedlings should be transplanted at the 2–3 leaf stage when grown at 15/24 °C to obtain the longest and thickest stems; however, $/m² week was higher for plants transplanted at the 8–9 leaf stage due to less time in finishing production space. For Papaver, the 15/21 °C temperature was optimal as that temperature produced the longest stems in the shortest duration, resulting in the highest $/m² week. At 15/21 °C Papaver plants should be transplanted at the 2–3 leaf stage. Supplemental HID lighting had no effect on any of the species.     

Effects of temperature and hydrogen peroxide on mycelial growth of eight Pleurotus strains

Effective use of hydrogen peroxide as a chemical sterilant in mushroom production and selection of cultivable mushroom strains for tropical conditions require knowledge of the genetic diversity in the tolerance of the strains to hydrogen peroxide and to high temperatures. Therefore, three experiments were conducted to examine the sensitivity of Pleurotus mycelium to temperature and hydrogen peroxide. In Experiment 1, eight Pleurotus strains, which included two Pleurotus sajor-caju strains, three Pleurotus ostreatus strains, Pleurotus salmoneo stramineus, Pleurotus cornicopae and Pleurotus eryngii were cultured aseptically on agar at 25, 30 and or 35 °C. In Experiment 2, the eight strains were cultured aseptically on agar at six hydrogen peroxide concentrations (0–0.1%, v/v) at 27 °C. In Experiment 3, P. sajor-caju strain 1, a fast growing strain, was cultured non-asceptically at six hydrogen peroxide concentrations (0–0.1%, v/v) at 27 °C. In Experiment 1, mycelial growth was maximal at 25–30 °C, whereas a temperature of 35 °C was detrimental to mycelial growth except in one strain. At the highest temperature tested (35 °C), the relative mycelial growth rate (% of maximum) ranged from 6 to 91%, indicating marked differences in tolerance of the strains to high temperature. In Experiment 2, the mycelial growth rate in all strains increased when hydrogen peroxide was increased from 0 to 0.001% (v/v), and then decreased with further increments in hydrogen peroxide concentration. The strains differed markedly in sensitivity to hydrogen peroxide. The hydrogen peroxide concentration associated with 50% reduction in maximum mycelial growth rate due to toxicity (EC₅₀) ranged from 0.009 to 0.045% (v/v). It was noted that P. sajor-caju strain 1 which was the most tolerant strain to high temperature was also the most tolerant to high hydrogen peroxide concentration. In Experiment 3, involving non-aseptic culture of P. sajor-caju strain 1, bacterial growth was observed at concentrations ≤0.016%, whilst the upper hydrogen peroxide concentration limit for fungal growth was 0.025% (v/v). The highest hydrogen peroxide concentrations 0.016% (v/v) and 0.025% (v/v) in which bacteria and fungi, respectively, were observed to grow were within the concentration range 0.009–0.028% (v/v) that was found in Experiment 2 to cause a 50% reduction in mycelia growth in six of the eight Pleurotus strains tested. Use of hydrogen peroxide as a chemical sterilant in conjunction with strains highly tolerant of its toxicity offers a very cheap method of producing spawn as well as the mushrooms, and opens up opportunities for poor rural people.     

Nutritional quality of greenhouse lettuce at harvest and after storage in relation to N application and cultivation season

The effect of five levels of nitrogen fertilization on the growth and nutritional quality of Cos lettuce (Lactuca sativa L. cv. Parris Island) at harvest and after storage was studied during autumn and winter in South-West Greece. Plants were cultivated hydroponically in a greenhouse and the nitrate, chlorophyll and ascorbic acid (vitamin C) concentrations within the plant tissues were measured at harvest and following storage at 5 or 10 °C for 10 days. Nitrate accumulated in the leaves with increasing amounts of N within the nutrient solution and was higher in the winter than in the autumn. At the lowest N level (20 mg L⁻¹), the inner leaves accumulated more nitrate than the outer leaves, whereas at higher N levels (140, 200 or 260 mg L⁻¹) nitrate accumulation was higher in the outer leaves. Overall, the highest nitrate concentrations were detected in the petiole and the proximal end of the leaf, but at the lowest N application rate (20 mg L⁻¹) nitrate accumulated in the distal region of the leaf too. Although the nitrate concentrations within the leaves did not change significantly during 10 days storage at 5 or 10 °C, the chlorophyll and vitamin C concentrations decreased. Chlorophyll loss was higher in lettuce that was grown under low N levels and was higher at 10 °C than at 5 °C, but was reduced by enclosure of the lettuce in polyethylene film. It is concluded that the optimum N application rate for Cos lettuce grown hydroponically under cover during autumn and winter in South-West Greece, and in other areas with a similar climate, is 200 mg N L⁻¹ because at this N rate yield is satisfactory and leaf nitrate concentrations are below the maximum acceptable level for human consumption. Nutritional value (vitamin C concentration) and market quality (chlorophyll content) are highest at harvest and decrease during storage, but quality in terms of nitrate concentration does not change.     

Effect of nitrogen form and radiation on growth and mineral concentration of two Brassica species

Three greenhouse experiments were carried out to determine the growth, yield, nitrate, total N and S concentration in shoots, and water uptake of hydroponically grown Brassica rapa L. subsp. nipposinica var. chinoleifera and Brassica juncea L. In each experiment, daily photosynthetically active radiation (PAR) level was 5.0 mol m⁻² (low), 6.8 mol m⁻² (medium) or 9.0 mol m⁻² (high). Plants were supplied with nutrient solutions having equal N concentrations of 11 mM in different forms: 100% NH₄, 50% NH₄ + 50% NO₃, and 100% NO₃. Nitrogen supplied as 100% NH₄ reduced fresh and dry shoot biomass, leaf area, and leaf number in both Brassica species, especially at low and medium PAR levels. In both Brassica species, S concentrations were highest, while nitrate concentrations were lowest in leaves of plants grown at N supplied as 100% NH₄. No differences in leaf nitrate concentrations were observed between 50% NH₄ + 50% NO₃ and 100% NO₃ treatments. Low and high PAR levels increased the nitrate concentrations and decreased the N/S ratio in leaves of both crops compared to medium PAR level. Fresh shoot biomass was maximized in Brassica rapa when PAR level was above the medium value and nitrate was supplied in the nutrient solution as NO₃ or as a mixture of 50% NO₃ and 50% NH₄. The highest fresh shoot biomass of Brassica juncea was observed in all nutrient solution treatments at high PAR level.