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.     

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.     

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.     

Investigation of biological behavior and colonization ability of Iranian indigenous phosphate solubilizing bacteria

Soil phosphorous (P) uptake mechanisms by the interaction of plants and soil microorganisms benefit plant growth with preparing agriculture inoculants. But it is influenced by soil factors. The present study was performed to identify three effective strains of phosphate solubilizing bacteria (PSB) on the basis of 16S rDNA sequence in vitro. Bacillus lentus strain PS5, Bacillus licheniformis strain PS7 and Pseudomonas putida strain PS13 were isolated from an alkaline soil in Iran and the rhizosphere of Beta vulgaris and Solanum tuberosom. Their high root colonization ability and P solubilizing activity was detected in relation to soil physical and chemical properties influenced by environmental factors including salinity of 800 mM/L and temperature of 42 °C.     

Obtaining and characterizing Pleurotus ostreatus strains for commercial cultivation under warm environmental conditions

Pleurotus ostreatus strains obtained by interbreeding were cultivated under warm conditions (27 °C), with the aim to identify germoplasm adequate to commercial production. Based on the radial growth rate of monokaryotic mycelia isolated from two P. ostreatus strains, a culture isolate was selected from each type of compatibility, and interbred to obtain 16 strains. Pasteurized barley straw was used as the substrate for mushroom production. The mean biological efficiency of the strains was 84.17% and the production rate was 1.76%. In vitro laccase activity was tested for both parental strains and the four most productive strains of progeny. The mean enzyme activity of the strains was 1.21 μg mol/disc/ABTS oxidized. These results show the high potential of these strains for their use in substrate delignification and the production of basidiocarps under natural environmental conditions of warm regions.     

Application method and rate of Trichoderma species as a biological control against Pythium aphanidermatum (Edson) Fitzp. in the production of microgreen table beets (Beta vulgaris L.)

Seed balls of ‘Early Wonder Tall Top’ table beet (Beta vulgaris L.) were incubated for 2 days at 21 °C in moist exfoliated grade 3 vermiculite (3 g seed balls [168 seed balls] and 1.1 g vermiculite) containing equal weights of Trichoderma harzianum Rifai strain KRL-AG2 G41 and T. virens strain G-41 (ThTv) at 0, 0.25, 0.50, 0.75 or 1.00 mg ThTv per seed ball. ThTv also was applied to the peat-lite growth medium 14 days before planting, at the same rates per seed ball as the seed ball treatments. Four days before planting, the peat-lite was inoculated with Pythium aphanidermatum (Edson) Fitzp. (Pa) at 0, 0.5 and 1.0× the rate that resulted in 96% damping-off when non-ThTv-treated dry seed balls were sown in peat-lite containing 1.0 Pa. Increasing ThTv level per seed ball decreased percentage damping-off, with 1.0 Pa giving greater percentage damping-off than 0.5 Pa. At 1 mg ThTv per seed ball, damping-off was 5% and 19% at 0.5 and 1.0 Pa, respectively. Including Agro-Lig UF (mostly humic acids) in the incubating seed-ball-ThTv mixture further decreased damping-off by an average 13 and 10 percentage points in 0.5 and 1.0 Pa, respectively. Increasing ThTv per seed ball in growth media decreased percentage damping-off, but not to the extent achieved with seed ball treatment, with 1 mg ThTv per seed ball giving 20% and 55% damping-off in 0.5 and 1.0 Pa, respectively. Decreasing incidence of damping-off with increasing ThTv application to seed balls or growth media was associated with increasing shoot fresh weight m⁻² at 14 days after planting, a response attributable to increased percentage plant survival and not to a ThTv growth-promoting effect.     

Influences of day and night temperatures on flowering of Fragaria x ananassa Duch., cvs. Korona and Elsanta,at different photoperiods

The effects of photoperiod (12, 13, 14, 15 or 16 h), day temperature (12, 15, 18, 24 or 27 °C) and night temperature (6, 9 or 12 °C) and their interactions on flower and inflorescence emergence were investigated by exposing 4 week old runner plants of strawberry cvs. Korona and Elsanta during a period of 3 weeks. A daily photoperiod of 12 or 13 h resulted in the highest number of plants with emerged flowers. A photoperiod of 14 h or more strongly reduced this number, while no flowers emerged at a photoperiod of 16 h. Plants exposed to photoperiods of 12 or 13 h flowered earlier and had longer flower trusses. A day temperature of 18 °C and/or a night temperature of 12 °C were optimal for plants to emerge flowers and resulted in the shortest time to flowering. A night temperature of 6 °C strongly reduced the number of plants that emerged flowers, especially when combined with lower day temperatures. Photoperiod and temperature had no effect on the number of inflorescences, all flowering plants produced on average one inflorescence. The number of flowers on the inflorescence increased with decreasing day temperature and when photoperiod was raised from 12 to 15 h. In general, ‘Korona’ was more sensitive to photoperiod and temperature as ‘Elsanta’, and had a lower optimal day temperature for flower emergence. Results of this experiment may be used to produce high quality plant material or to define optimal conditions when combining flower induction and fruit production.     

Light quality of continuous illuminating at night to induce floral initiation of Fragaria chiloensis L. CHI-24-1

A wild strawberry strain, Fragaria chiloensis CHI-24-1, produced inflorescences from both parent and asexually propagated daughter plants linked with runners when grown at 23 °C/20 °C (day/night) under a 24 h day-length (DL) of daylight plus nightly lighting by an incandescent lamp, but not under 8 or 16 h DLs. In the present study, the effect of light quality for continuous illuminating at night on floral initiation of CHI-24-1 plants grown under a 24 h DL was examined. The CHI-24-1 plants were grown under a 24 h DL consisting of natural daylight and continuous lighting at night by an incandescent, a blue fluorescent, a red fluorescent or a far-red fluorescent lamp for 40 days in summer and autumn. Also, the CHI-24-1 plants were grown for 40 days in a growth chamber at 25 °C/20 °C (day/night) with natural daylight and continuous lighting at night by red- and four types of far-red light-emitting-diodes (LEDs with peak wavelengths of 660, 700, 735, 780 and 830 nm). In both experiments, floral initiation of the parent and daughter plants was observed under a stereomicroscope. Although more than 50% of the parent and daughter plants initiated flower buds under the incandescent and far-red fluorescent lamps, about 15% and 0% of those initiated flower buds under blue and red fluorescent lamps, respectively. Floral initiation of the parent and daughter plants occurred under the far-red LED light source whose peak wavelength was 735 nm, but not under the red or the other far-red LEDs. From these results, it can be concluded that the effective light wavelength range of nightly continuous illuminating for floral induction in the CHI-24-1 plants is 735 nm in the far-red light region. Hence, the induction of floral initiation by nightly continuous far-red light (735 nm) appeared to be a response mediated by phytochrome.     

Responses of non-primed or primed seeds of ‘Marketmore 76’ cucumber (Cucumis sativus L.) slurry coated with Trichoderma species to planting in growth media infested with Pythium aphanidermatum

Aqueous slurries of commercial preparations of Trichoderma harzianum Rifai strain KRL-AG2 G41 (Th), T. virens Strain G-41 (Tv), or their combination (ThTv, at half rates each of the single application rate) were applied to ‘Marktetmore 76’ cucumber seeds (Cucumis sativus L.) that were non-primed or primed for 3 days at 25 °C either osmotically (−2.5 MPa from 0.337 molal Ca(NO₃)₂) or osmomatrically (−1.0 MPa from 0.135 molal Ca(NO₃)₂ plus −1.5 MPa from 50% water in exfoliated grade 5 vermiculite). Slurries were applied to seeds (1 mg per seed) either before or after priming. Seeds were sown in soilless, peat-based media with or without inoculation with Pythium aphanidermatum (Pa). Protection against damping-off caused by high pressures of Pa (16% emergence in non-coated, non-primed seeds) was increased by slurry coating Th on non-primed (76.4% emergence) or on osmotically primed seeds, with coating either before or after priming having no effect on efficacy (average 62.6% emergence). Slurry coating Th on osmomatrically primed seeds failed to increase final emergence percentage (FEP). Colony forming units per three seeds (CFU, all 10³) was 2.8 for non-primed seeds, and 3.2 and 2.6, respectively, when osmotically and osmomatrically primed seeds were coated after priming. In a second study with lower disease pressure (58.1 FEP from non-coated, non-primed seeds), slurry coating of non-primed or osmotically primed seeds with Th, Tv or ThTv reduced percentage damping-off and increased FEP. The combination coating eliminated damping-off only in non-primed seeds, and tended to reduce percentage damping-off in osmotically or osmomatrically primed seeds compared to coating with Th or Tv alone. In a third study using only non-primed seeds, slurry coatings with mefenoxam fungicide, Th, Tv, or ThTv decreased total damping-off to 2.6%, 7.4%, 2.0%, and 0%, respectively, from the 30.1% occurring in non-coated seeds. Th, Tv or ThTv applied to growth media at the same rate as the seed coating (1 mg per seed) were generally as effective as the seed coatings, and only the ThTv growth medium application eliminated damping-off. A fourth experiment revealed that Th, Tv or ThTv remained viable on non-primed seeds for up to 4 weeks (the longest storage duration) at 21 or 4 °C, but 21 °C storage resulted in faster seed germination by week 3 and higher CFU per three seeds by week 4. In summary, coating of non-primed seeds with Th, Tv or ThTv was more effective than coating primed seeds in reducing percentage damping-off. While priming treatments generally led to faster seedling emergence and greater seedling shoot fresh weight than was achieved with non-primed seeds, only for non-primed seeds was damping-off eliminated by the ThTv seed coating or growth medium application.     

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.     

Different mechanisms to obtain higher fruit growth rate in two cold-tolerant cucumber (Cucumis sativus L.) lines under low night temperature

One cold-sensitive cultivar (Jinyan 4) and two cold-tolerant inbred lines (NY-1 and XC-1) of cucumber (Cucumis sativus L.) were subjected to temperatures of 28 °C/22 °C (day/night, control) or 28 °C/12 °C (day/night, cold treatment) in a 10 h photoperiod (7:00–17:00). Under control conditions, cucumber fruits grew fast during afternoon and early night, and slow during late night and morning. Under 28 °C/12 °C conditions, the two cold-tolerant inbred lines maintained relatively higher fruit growth rates than the cold-sensitive cultivar by different mechanisms. Compared to Jinyan 4, NY-1 fruits had higher growth rates during cold nights while XC-1 fruits grew faster during the next day. Under the 28 °C/12 °C temperature regime, the assimilate accumulation in the fruits of all tested genotypes followed a similar trend with the corresponding fruit growth rates. After a cold night treatment, the net CO₂ assimilation rates of one- and two-fruit plants, which had increased sink demand, were higher than that of plants without fruits in all tested genotypes. This response indicates that feedback inhibition might be an important reason for the reduction of photosynthesis on the next day. In addition, after a cold night treatment, the levels of exportable sugars (sucrose and stachyose) in mature leaves of XC-1 were higher than those measured in Jinyan 4 and NY-1, which might explain why XC-1 fruits had faster assimilate accumulation rates in the next morning. Higher activity of sucrose-phosphate synthase, a key enzyme of sucrose and stachyose biosynthesis, constituted an additional evidence that faster sucrose and stachyose biosynthesis in mature leaves may occur in XC-1 than in Jinyan 4 and NY-1 at that time. In conclusion, our results showed that cucumber genotypes may use different mechanisms to enhance their cold tolerance.     

Effect of hot acetic acid solutions on postharvest decay caused by Penicillium expansum on Red Delicious apples

The objective of this research was improvement of apple hot water treatment efficiency by using acetic acid. The apples (cultivar Red Delicious) were treated using hot acetic acid solutions (1, 2 and 3%) at 50 °C for 1, 2 and 3 min. The results of in vitro study showed that treatment with acetic acid at 50 °C can significantly reduce the growth of Penicillium expansum spores. The treatment of apples with 50 °C acetic acid solutions, in particular 2% acetic acid solution for 3 min or 3% acetic acid solution for 2 min, had significant impacts in reducing the extent of decay of the fruit during the short time storage experiment, while this effect was not significant in the long-time storage.     

Modeling plant morphology and development of petunia in response to temperature and photosynthetic daily light integral

The effects of mean daily temperature (MDT) and mean photosynthetic daily light integral (MDLI) on flowering during the finish stage of two petunia (Petunia × hybrida) cultivars were quantified. Petunia ‘Easy Wave Coral Reef’ and ‘Wave Purple’ were grown in glass-glazed greenhouses at 14–23 °C or 14–26 °C and under 4–19 mol m⁻² d⁻¹ with a 16-h photoperiod. The flower developmental rate was predicted using a model that included a linear MDT function with a base temperature multiplied by an exponential MDLI saturation function. The flower developmental rate increased and time to flower decreased as MDT increased within the temperature range studied. For example, under a MDLI of 12 mol m⁻² d⁻¹, as MDT increased from 14 to 23 °C, time to flower of ‘Easy Wave Coral Reef’ and ‘Wave Purple’ decreased from 51 to 22 d and 62 to 30 d, respectively. Flower developmental rate increased as MDLI increased until saturation at 14.1–14.4 mol m⁻² d⁻¹. Nonlinear models were generated for effects of MDT and MDLI on flower bud number and plant height at flowering. The number of flower buds at flowering increased as MDT decreased and MDLI increased. For example, at an MDT of 14 °C with 18 mol m⁻² d⁻¹, plants had 2.5–2.9 times more flower buds than those grown at 23 °C and 4 mol m⁻² d⁻¹. Models were validated with an independent data set, and the predicted time to flower, flower bud number, and plant height were within ±7 d, ±20 flowers, and ±4 cm, respectively, for 96–100%, 62–87%, and 93–100% of the observations, respectively. The models could be used during greenhouse crop production to improve scheduling and predict plant quality of these petunia cultivars.     

Effect of high temperature stress on the reproductive growth of strawberry cvs. ‘Nyoho’ and ‘Toyonoka’

High temperatures are known to reduce fruit size and fruit weight in strawberry, but cultivar differences in the response to high temperature stress during the reproductive stage up to the second inflorescence have not been sufficiently reported. We examined the effect of two day/night temperature regimes on fruit set and fruit growth in two cultivars, ‘Nyoho’ and ‘Toyonoka’. A high day/night temperature of 30/25 °C reduced the number of inflorescences, flowers, and fruits in both cultivars compared with plants grown at 23/18 °C. The percentage of fruit set in ‘Nyoho’ was not significantly different between the two temperature treatments, while that in ‘Toyonoka’ was much lower at 30/25 °C than at 23/18 °C. Days to ripening was shorter at 30/25 °C than at 23/18 °C, and no cultivar differences were observed. Fresh weight of primary, secondary, and tertiary fruits was greater at 23/18 °C than at 30/25 °C in both cultivars, and no cultivar differences were observed, except in tertiary fruits. The diameter of fruits from all positions was also reduced at 30/25 °C in both cultivars. Relative growth rates of fruits showed two peaks in both cultivars and in both temperature treatments. Both peaks appeared earlier at 30/25 °C than at 23/18 °C. Percentage of fruit set at 30/25 °C in the second inflorescence was also significantly lower in ‘Toyonoka’ than in ‘Nyoho’. These results indicate that high temperature stress negatively affects the reproductive process in strawberry and that plant response to high temperature stress is cultivar-related in such responses.     

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.     

Interactions of photoperiod,temperature, duration of short-day treatment and plant age on flowering of Fragaria x ananassa Duch. cv. Korona

The effects of photoperiod (10, 12, 16, 20 or 24 h), day-temperature (12, 15, 18, 24 or 30 °C), the number of short days (14, 21 or 28 days), plant age (4, 8 or 12 weeks) and their interactions on flower and inflorescence emergence were investigated in strawberry cv. Korona. No flowers emerged in plants exposed to photoperiods of 16, 20 or 24 h or to a short-day treatment for 14 days. All plants exposed to short days at daily photoperiods of 10 or 12 h for 21 days or longer, emerged flowers at temperatures between 12 and 18 °C. A further increase in temperature led to a drastic decrease in the total number of flowers per plant. A short-day treatment (10 or 12 h photoperiod) of 28 days resulted in highest numbers of inflorescences and flowers per plant, while a short-day treatment of 21 days resulted in the highest numbers of flowers per inflorescence. Complete flower induction was observed in only 4-week-old runner plants. The number of inflorescences and the number of flowers per inflorescence increased with plant age. However, the start of flowering was delayed with increasing plant age.     

A protocol to assess heat tolerance in a segregating population of raspberry using chlorophyll fluorescence

A detached leaf protocol was developed to assess heat tolerance in raspberry (Rubus L. subgenus Idaeobatus) by measuring chlorophyll fluorescence. In order to establish the heat screen protocol for field measurements in a segregating population, several baseline experiments were performed on genotypes with known heat tolerance based on visual observations in multiple climates. ‘Mandarin’ and ‘Qualicum’ were used as heat tolerant and heat susceptible controls, respectively. We determined that: optimum collection time was in the morning before ambient temperatures reached 28 °C, critical temperature (Tc) was 45 °C, minimum duration of dark adaptation was 15 min, and optimum heat shock duration was 30 min. Parameters established in baseline experiments were subsequently used for screening a large population (196 genotypes) planted in the field, where warm conditions commonly occur (maximum temperatures ranging between 30 and 35 °C). A significant difference in chlorophyll fluorescence was found between the two controls; ‘Qualicum’, the heat sensitive cultivar, had the lowest Fv/Fm (variable fluorescence/maximum fluorescence) value, while 18.8% of the segregating population had a higher Fv/Fm value than ‘Mandarin’, the heat tolerant cultivar. We have identified 106 individuals with an Fv/Fm above 0.600 (representing 52% of the population), and 7 individuals above 0.680 (4% of the population). The distribution of heat tolerance within the population was essentially normal, and thus quantitative trait analysis is a plausible approach for studying heat tolerance. This protocol has potential application to other Rosaceae species, and is not dependent on visual assessment.     

Photoperiodic reaction of sexual and asexual reproduction in Fragaria chiloensis L. CHI-24-1 plants grown at various temperatures

Floral initiation of a wild strawberry strain, Fragaria chiloensis CHI-24-1, is strongly induced by a 24 h day-length (DL) treatment for 40 days consisting of natural daylight and continuous lighting at night by an incandescent lamp. To use the characteristics of floral initiation in CHI-24-1 as a genetic resource for breeding of cultivated strawberries, the photoperiodic reactions of sexual and asexual reproductive growth under various temperature conditions should be clarified. For that purpose, we examined: (1) floral initiation, inflorescence emergence and runner production seasons of CHI-24-1 plants grown under natural climatic conditions in an open field at the Faculty of Agriculture, Kagawa University and (2) the effects of various DLs and temperatures on floral initiation and runner production of CHI-24-1 plants. When the CHI-24-1 plants were grown under natural conditions, the floral initiation, inflorescence emergence and runner production were observed, respectively, in late autumn, spring, and from spring to autumn. Floral initiation of CHI-24-1 plants was induced strongly by 24 h DL at mean temperatures greater than 20 °C. The maximum floral initiation rates were 90% in the parent plant and 94% in the daughter plants, which were linked by runners to the parent plant. The floral initiation of the daughter plants occurred under 20, 22, and 23 h DL at mean temperatures greater than 20 °C, but not for the parent plants. Floral initiation was induced in 100% of the parent plants by the 8 h DL and the lowest mean-temperature conditions. Results of those experiments indicated that CHI-24-1 was an absolute long day plant having critical DL of about 20 h at mean temperatures greater than 20 °C, even though it was a June-bearing strawberry plant. In addition, CHI-24-1 was a facultative short-day plant at mean temperatures of less than 15 °C.     

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.     

Aneuploid strawberry (2n = 8x + 2 = 58) was developed from homozygous unreduced gamete (8x) produced by second division restitution in pollen

Unreduced gamete formation is significant in the evolutionary development of complex polyploidy series found in wild strawberry, genus Fragaria (Rosaceae). Also, it is important for genetics and breeding in strawberry plants to elucidate the mechanism of unreduced gamete formation. The objective of this study was to search for ploidy anomalies resulting from artificial diploid × octoploid crosses, and examine the mechanism through which these unreduced gametes were produced. Five everbearing cultivars of Fragaria vesca L. diploid (2n = 2x = 14) were crossed with pollen from six June-bearing cultivars of Fragaria × ananassa Duch., octoploid (2n = 8x = 56). A total of 3000 mature seeds, 100 from each of the 30 parental combinations were sown at 23 °C/20 °C (day/night) under artificial lighting with a 16 h day. The seedlings were transplanted to pots and grown in a greenhouse. Reproductive and morphological observations, flow cytometry analyses, chromosome counts and DNA analyses using CAPS markers were performed to identify the genetic background of the offspring. Most of the seed (79%) did not germinate or died soon after germination. Of the seedlings produced, 7% seemed to be pure F. vesca based on morphological characteristics, flow cytometry analyses and chromosome counts; 14% were pentaploids (2n = 5x = 35), 0.1% were hexaploids (2n = 6x = 42), and 0.03% (one individual) was aneuploid (2n = 8x + 2 = 58). Electrophoresis banding patterns obtained by CAPS marker analysis were heterozygotic in the 8x pollen parent but homozygotic in the aneuploid progeny. Judging from the chromosome counts and the CAPS marker analysis, the aneuploid was the result of a homozygous unreduced pollen grain (8x) crossed with an incomplete chromosome compliment from the egg. Because of the homozygosity, the unreduced male gamete must have been derived from second division restitution (SDR) in the octoploid pollen parent.