Effect of drying temperature on carrageenan yield and quality of Kappaphycus alvarezii (Rhodophyta,Solieriaceae) cultivated in Brazil

This study evaluates the effect of different drying temperatures on the properties of semi-refined (SR) and refined (R) carrageenan extracted from Kappaphycus alvarezii cultivated in Brazil. Drying kinetics was studied in seaweeds under the following treatments: sun drying (control) and drying at 40, 60, and 90 °C in convective air dryer. Drying was carried out until the moisture content of seaweeds reached values below of 30 % on wet basis. Significant reductions in drying time were observed with the increase of temperature. At 90 °C, 30 % moisture content was reached in 100 min, as compared with the 1,440 min required by the sun-drying treatment. SR yields showed no significant differences when compared to the control, varying from 40 to 44 %, while R had a significantly higher yield (30 %) at 90 °C in relation to control (26 %). Gel strength of SR was significantly higher in the sun-dried samples (1,685.1 g cm^?2) and 60 °C samples (1,727.2 g cm^?2), but no significant differences were observed in R gel strengths. Lowest syneresis was observed in both SR (9.8 %) and R (10.3 %) after the treatment at 90 °C. Significantly lower viscosity values were observed for SR at 60 °C (233 mPa s) and at 90 °C (175 mPa s), as for R, the lowest value was observed at 90 °C (205 mPa s). Based on these results, it was concluded that best results for both types of carrageenan are obtained drying at 60 °C.     

Denaturation and in Vitro Gastric Digestion of Heat-Treated Quinoa Protein Isolates Obtained at Various Extraction pH

The aim of this study was to determine the influence of heat processing on denaturation and digestibility properties of protein isolates obtained from sweet quinoa (Chenopodium quinoa Willd) at various extraction pH values (8, 9, 10 and 11). Pretreatment of suspensions of protein isolates at 60, 90 and 120 °C for 30 min led to protein denaturation and aggregation, which was enhanced at higher treatment temperatures. The in vitro gastric digestibility measured during 6 h was lower for protein extracts pre-treated at 90 and 120 °C compared to 60 °C. The digestibility decreased with increasing extraction pH, which could be ascribed to protein aggregation. Protein digestibility of the quinoa protein isolates was higher compared to wholemeal quinoa flour. We conclude that an interactive effect of processing temperature and extraction pH on in vitro gastric digestibility of quinoa protein isolates obtained at various extraction pH is observed. This gives a first indication of how the nutritional value of quinoa protein could be influenced by heat processing, protein extraction conditions and other grain components.     

Changeability in Retama raetam essential oils chemical composition,antioxidant and antimicrobial properties as affected by the physiological stage

Abstract

Retama raetam essential oils (EOs) composition and biological activities were assessed during three developmental periods. The essential oil yield varied significantly among the developmental stages and the optimal was detected at the fresh fruiting stage (0.34%). In addition, EOs composition varied significantly (p < 0.05) according to the different developmental stages. In fact, 2-Methoxy-4-vinylphenol, linalool, and 1-octen-3-ol were the main compounds in the vegetative, flowering, and the fresh fruiting stages, respectively. Developmental stage had also a strong effect on EOs antioxidant and antimicrobial activities. In fact, during the fresh fruiting stage, IC₅₀ and EC₅₀ values of the antioxidant assays were 2 to 3 times inferior to all others stages. Concerning the determination of the diameter of inhibition, a slight to high antimicrobial activity was revealed against 12 bacteria and 4 yeasts. Once again, EOs from the fresh fruiting stage had higher bactericidal effect than those from the flowering and vegetative ones (IZ varied from 10 to 13 mm). The results of this investigation showed for the first time the high accumulation of EOs at the early stages of fruit development making the fresh fruiting optimal stage for the extraction of powerful antioxidant and antimicrobial EOs.     

Subcritical Water Extraction of Phenolic Compounds from Vaccinium Dunalianum Wight Leaves and Their Antioxidant and Tyrosinase Inhibitory Activities in Vitro

Subcritical water extraction was used to extract bioactive phenolic compounds from Vaccinium dunalianum Wight leaves. The optimal extraction conditions were determined as an extraction temperature of 150 °C, an extraction time of 40 min, and a liquid-solid ratio of 35 : 1 mL/g. The total phenolic content reached 21.35 mg gallic acid /g, which was 16 % higher than that by hot water extraction. The subcritical water extraction extract exhibited strong scavenging activity of DPPH free radical and ABTS⁺ free radical, as well as significant tyrosinase inhibitory activity. The study suggests that subcritical water extraction can alter the composition of the extracts, leading to the production of various phenolic compounds, effective antioxidants, and tyrosinase inhibitors from Vaccinium dulciana Wight leaves. These findings confirm the potential of Vaccinium dunalianum Wight as a natural antioxidant molecule source for the medicine and food industries, and for the therapy of skin pigmentation disorders.     

Combined action of antioxidant defense system and osmolytes in chilling shock-induced chilling tolerance in Jatropha curcas seedlings

Low non-freezing temperature is one of the major environmental factors affecting growth, development and geographical distribution of chilling-sensitive plants, Jatropha curcas is considered as a sustainable energy plants with great potential for biodiesel production. In this study, chilling shock at 5 °C followed by recovery at 26 °C for 4 h significantly improved survival percentage of J. curcas seedlings under chilling stress at 1 °C. In addition, chilling shock could obviously enhance the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR), and the levels of antioxidants ascorbic acid (AsA) and glutathione (GSH), as well as the contents of osmolytes proline and betaine in leaves of seedlings of J. curcas compared with the control without chilling shock. During the process of recovery, GR activity, AsA, GSH, proline and betaine contents sequentially increased, whereas SOD, APX and CAT activities gradually decreased, but they markedly maintained higher activities than those of control. Under chilling stress, activities of SOD, APX, CAT, GR and GPX, and contents of AsA, GSH, proline and betaine, as well as the ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)] in the shocked and non-shock seedlings all dropped, but shocked seedlings sustained significantly higher antioxidant enzyme activity, antioxidant and osmolyte contents, as well as ratio of reduced antioxidants to total antioxidants from beginning to end compared with control. These results indicated that the chilling shock followed by recovery could improve chilling tolerance of seedlings in J. curcas, and antioxidant enzymes and osmolytes play important role in the acquisition of chilling tolerance.     

Differential Sensitivity of Telomerase from Human Hematopoietic Stem Cells and Leukemic Cell Lines to Mild Hyperthermia

We have investigated the effects of hyperthermia (HT) on cell proliferation and telomerase activity of human hematopoietic stem cells (HSCs) and compared with human leukemic cell lines (TF-1, K562 and HL-60). The cells were exposed to HT at 42 and 43 °C up to 120 min. The cells were incubated at 37 °C for 96 h. Then the cells were collected and assayed for cell proliferation, viability, telomerase activity, and terminal restriction fragment (TRF) lengths. The enzyme activity from HSCs was decreased up to 68.6 at 42 and 85.1 % at 43 °C for 120 min. This inhibition in leukemic cells was up to 28.9 and 53.6 % in TF-1; 53 and 63.9 % in K562; 45.2 and 61.1 % in HL-60 cells. The treated cells showed TRF lengths about 5.3 kb for control HL-60 cells, 5.0 kb for HL-60 cells treated at 42 and 4.5 kb at 43 °C for 120 min. In HSCs, the TRF length was about 4.5 kb for untreated cells and 4.0–4.5 kb for treated cells at 42 and 43 °C for 120 min. The time response curves indicated that, inhibition of the enzyme activity in leukemic cells was dependent to the time of exposure to HT. But in HSCs, the inhibition was reached to steady state at 15 min exposure to 43 °C heat stress. TRF length was constant at treated two types of cells, which implies that in cells subjected to mild HT no telomere shortening was observed.     

Changes in quality of Phellinus gilvus mushroom by different drying methods

This study was conducted to investigate the changes in characteristics of the Phellinus gilvus mushroom as influenced by drying methods after harvest. The lowest weight loss rate of P. gilvus mushroom was 75.8% with drying in the shade and 80% by dryer (60°C). The size loss rate of pileus was 19.3% of that in a hot air dryer (60°C). The hardness of dried material context using a hot air dryer (60°C) was the lowest (20 kg/cm²), and that by a dry oven (60°C) was the highest (457 kg/m²). For ΔE value, 4.9 of context and 2.6 of tubes using drying in the shade (20°C) were found to be the lowest. The survival rate of sarcoma 180 treated with P. gilvus dried in the sun was the lowest (51.8%), and this was considered the most effective method for antitumor activity against sarcoma 180.     

Effect of hot water and ultra violet radiation on the incidence of seedborne fungi of okra

Seed samples of okra (Abelmoschus esculentus (L.) Moench) variety Arka anamika were subjected to hot water treatment at 42, 52 and 62°C for a period of 30 min and UV light treatment for 10, 20, 30 min at 28 ± 2°C. Their efficacy was tested against some seedborne fungal species. Among them, seeds under hot water treatment at 52°C for 30 min and UV light at 20 min were found to be more effective in the improvement of crop, both in greenhouse and field conditions. Ultimately, there was increase in the total number of leaves, fruits, length of the fruit, girth and biomass of the plants. Apart from these the total number of seeds per fruit, 1000 seed weight and ascorbic acid content were also found to be enhanced. These treatments also reduced the incidence of mycoflora in the seeds and thereby enhanced the seed germination percentage and vigour index of the seedlings.     

Time-of-day effects of exposure to solar radiation on thermoregulation during outdoor exercise in the heat

High solar radiation has been recognised as a contributing factor to exertional heat-related illness in individuals exercising outdoors in the heat. Although solar radiation intensity has been known to have similar time-of-day variation as body temperature, the relationship between fluctuations in solar radiation associated with diurnal change in the angle of sunlight and thermoregulatory responses in individuals exercising outdoors in a hot environment remains largely unknown. The present study therefore investigated the time-of-day effects of variations in solar radiation associated with changing solar elevation angle on thermoregulatory responses during moderate-intensity outdoor exercise in the heat of summer. Eight healthy, high school baseball players, heat-acclimatised male volunteers completed a 3-h outdoor baseball trainings under the clear sky in the heat. The trainings were commenced at 0900 h in AM trial and at 1600 h in PM trial each on a separate day. Solar radiation and solar elevation angle during exercise continued to increase in AM (672–1107 W/m² and 44–69°) and decrease in PM (717–0 W/m² and 34–0°) and were higher on AM than on PM (both P < 0.001). Although ambient temperature (AM 32–36°C, PM 36–30°C) and wet-bulb globe temperature (AM 31–33°C, PM 34–27°C) also continued to increase in AM and decrease in PM, there were no differences between trials in these (both P > 0.05). Tympanic temperature measured by an infrared tympanic thermometer and mean skin temperature were higher in AM than PM at 120 and 180 min (P < 0.05). Skin temperature was higher in AM than PM at the upper arm and thigh at 120 min (P < 0.05) and at the calf at 120 and 180 min (both P < 0.05). Body heat gain from the sun was greater during exercise in AM than PM (P < 0.0001), at 0–60 min in PM than AM (P < 0.0001) and at 120–180 min in AM than PM (P < 0.0001). Dry heat loss during exercise was greater at 0–60 min (P < 0.0001), and lower at 60–120 min (P < 0.05) and 120–180 min (P < 0.0001) in AM than PM. Evaporative heat loss during exercise was greater in PM than AM at 120–180 min (P < 0.0001). Total (dry + evaporation) heat loss at the skin was greater during exercise in PM than AM (P < 0.0001), at 0–60 min in AM than PM (P < 0.0001) and at 60–120 and 120–180 min in PM than AM (P < 0.05 and 0.0001). Heart rate at 120–150 min was also higher in AM than PM (P < 0.05). Neither perceived thermal sensation nor rating of perceived exertion was different between trials (both P > 0.05). The current study demonstrates a greater thermoregulatory strain in the morning than in the afternoon resulting from a higher body temperature and heart rate in relation to an increase in environmental heat stress with rising solar radiation and solar elevation angle during moderate-intensity outdoor exercise in the heat. This response is associated with a lesser net heat loss at the skin and a greater body heat gain from the sun in the morning compared with the afternoon.     

Involvement of antioxidant defense system in chill hardening-induced chilling tolerance in Jatropha curcas seedlings

Jatropha curcas L. is a sustainable energy plant with great potential for biodiesel production, and low temperature is an important limiting factor for its distribution and production. In this present work, chill hardening-induced chilling tolerance and involvement of antioxidant defense system were investigated in J. curcas seedlings. The results showed that chill hardening at 10 or 12 °C for 1 and 2 days greatly lowered death rate and alleviated electrolyte leakage as well as accumulation of the lipid peroxidation product malondialdehyde (MDA) of J. curcas seedlings under severe chilling stress at 1 °C for 1–7 days, indicating that the chill hardening significantly improved chilling tolerance of J. curcas seedlings. Measurement of activities of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), and glutathione reductase (GR), and the levels of the antioxidants ascorbic acid (AsA) and glutathione (GSH) showed the chill hardening at 12 °C for 2 days could obviously increase the activities of these antioxidant enzymes and AsA and GSH contents in the hardened seedlings. When the hardened and non-hardening (control) seedlings were subjected to severe chilling stress at 1 °C for 1–7 days, the chill-hardened seedlings generally maintained significantly higher activities of the antioxidant enzymes SOD, APX, CAT, POD, and GR, and content of the antioxidants AsA and GSH as well as ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)], when compared with the control without chill hardening. All above-mentioned results indicated that the chill hardening could enhance the chilling tolerance, and the antioxidant defense system plays an important role in the chill hardening-induced chilling tolerance in J. curcas seedlings.     

Effect of treatment of cow’s urine “Gomutra” and antioxidants in alleviating the lindane-induced oxidative stress in kidney of Swiss mice (Mus musculus)

The study aimed to evaluate the effect of cow urine and combination of antioxidants against lindane induced oxidative stress in Swiss mice. Male healthy mice, 8–10 weeks old, weighing 30 ± 5 g were randomly selected and divided into eight groups, namely, control (C); lindane (L); antioxidant (A), antioxidant+lindane (A+L), cow urine (U), cow urine+lindane (U+L), cow urine+antioxidants (U+A) and cow urine+antioxidants+lindane (U+A+L). Group C animals were administered only the vehicle (olive oil); doses selected for other treatments were: lindane: 40 mg/kg b.w.; antioxidants: 125 mg/kg b.w. (vitamin C: 50 mg/kg b.w., vitamin E: 50 mg/kg b.w., α-lipoic acid: 25 mg/kg b.w.) and cow urine: 0.25 ml/kg b.w. In group A+L and U+L antioxidants and cow urine were administered 1 h prior to lindane administration and in group U+A and U+A+L cow urine was administered 10 min before antioxidants. All treatments were administered orally continuously for 60 days. Lindane treated group showed increased lipid peroxidation, whereas glutathione, glutathione peroxidase, superoxide dismutase, catalase, protein and endogenous levels of vitamin C and E were significantly decreased compared to control. Administration of cow urine and antioxidants alleviated the levels of these biochemical parameters.     

Are winter-active species vulnerable to climate warming? A case study with the wintergreen terrestrial orchid,Tipularia discolor

In the eastern United States, winter temperature has been increasing nearly twice as fast as summer temperature, but studies of warming effects on plants have focused on species that are photosynthetically active in summer. The terrestrial orchid Tipularia discolor is leafless in summer and acquires C primarily in winter. The optimum temperature for photosynthesis in T. discolor is higher than the maximum temperature throughout most of its growing season, and therefore growth can be expected to increase with warming. Contrary to this hypothesis, experimental warming negatively affected reproductive fitness (number of flowering stalks, flowers, fruits) and growth (change in leaf area from 2010 to 2012) in T. discolor. Temperature in June–July was critical for flowering, and mean July temperature greater than 29 °C (i.e., 2.5 °C above ambient) eliminated reproduction. Warming of 1.2 °C delayed flowering by an average of 10 days and fruiting by an average of 5 days. Warming of 4.4 °C reduced relative growth rates by about 60 %, which may have been partially caused by the direct effects of temperature on photosynthesis and respiration. Warming indirectly increased vapor pressure deficit (VPD) by 0.2–0.5 kPa, and leaf-to-air VPD over 1.3 kPa restricted stomatal conductance of T. discolor to 10–40 % of maximum conductance. These results highlight the need to account for changes in VPD when estimating temperature responses of plant species under future warming scenarios. Increasing temperature in the future will likely be an important limiting factor to the distribution of T. discolor, especially along the southern edge of its range.     

Evaluation of drying methods on nutritional constituents and antioxidant activities of Chlorella vulgaris cultivated in an outdoor open raceway pond

Chlorella vulgaris is known for its protein, growth factor, and nutritional constituents. Chlorella vulgaris was cultivated in a 1000-L outdoor open raceway pond with a maximum volumetric productivity of 130 mg L^-1 day^-1. The harvested biomass was dried through different methods, viz., sun drying (30 °C), oven drying (60 °C), lyophilization (?110 °C), drum drying (120 °C), and spray drying (100–150 °C). The effect of the drying method on proximate composition, pigments (chlorophyll, carotenoids), bioactive compounds (total phenolic content, flavonoid content), vitamin B₁₂, antioxidant properties (ferric reducing antioxidant power, DPPH, and total antioxidant activity), and the color quality of C. vulgaris biomass was evaluated. Surface characterization by scanning electron microscopy (SEM) and functional group characterization through Fourier transform infrared spectroscopy were also performed. The biomass dried through lyophilization and sun drying retained maximum bioactive compounds and antioxidant activities. In contrast, drum drying resulted in a loss of nutrients, viz., protein (up to 44%), lipid (up to 41%), vitamin B₁₂ (up to 40%), total phenolic content (> 50%), total flavonoid content (> 50%), and antioxidant activity (> 50%). Oven drying led to a loss of 30% in total flavonoid content and 17% in ferric reducing antioxidant power. SEM showed the destruction of cell wall integrity in the drum-dried sample and porous structure in the spray-dried sample. This study suggests that drying methods affect the nutrients and bioactive compounds of C. vulgaris biomass, and therefore a drying method should be selected carefully depending on the end use of the biomass.

     

Optimization of Ultrasonic-Assisted Extraction and Radical-Scavenging Capacity of Phenols and Flavonoids from Clerodendrum cyrtophyllum Turcz Leaves

Ultrasonic-assisted extraction (UAE) was developed to extract phenolic and flavonoid antioxidants from Clerodendrum cyrtophyllum Turcz leaves. The optimal experimental parameters for antioxidant extraction from C. cyrtophyllum leaves were measured using single-factor experimentation combined with response surface methodology (RSM). Total phenolic content (TPC) and total flavonoid content (TFC) assays were used to quantify antioxidant compounds. Next, antioxidant radical scavenging capacity was measured using 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′ -azino-bis(3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) radicals. Optimized extraction conditions for UAE from C. cyrtophyllum leaves were as follows: 60.9% ethanol, 85.4 min, and 63.3°C for maximal TPC extraction (16.8±0.2 mg GAE/g DW); 67.7% ethanol, 82.9 min, and 63.0°C for maximal TFC extraction (49.3±0.4 mg RT/g DW); 48.8% ethanol, 85.1 min, and 63.9°C for maximal DPPH radical-scavenging capacity (86.8±0.2%); and 50.6% ethanol, 81.3 min, and 63.4°C for maximal ABTS radical-scavenging capacity (92.9±0.5%). Ethanol concentration was the most important factor in the extraction process. Our work offers optimal extraction conditions for C. cyrtophyllum as a potential source of natural antioxidants.     

Increased phenolic content and antioxidant capacity of the heated leaves of yacon (Smallanthus sonchifolius)

ABSTRACT

We investigated the content of phenolic compounds and antioxidant capacity of two batches of non-heated and heated leaves of the yacon cultivar “Andes no yuki”, grown in Japan. Lyophilized yacon leaves heated at 160°C for 20 min and 100°C for 60 min had a 1.96 to 9.69-times higher total phenolic content than that of the non-heated leaves. Heated leaves exhibited a 1.98 to 4.07-times higher antioxidant capacity than that of the non-heated leaves in three different free radical scavenging assays. Heated leaves were more efficient at attenuating the superoxide anion radical production in human granulocytic cells than the non-heated leaves. High-performance liquid chromatography analysis revealed that, in the heated leaves, the caffeic acid content was 2.13 to 3.64-times higher and the chlorogenic acid content was slightly lower than those in the non-heated leaves. Hence, heat processing may affect the active constituent contents in yacon leaves, potentiating its antioxidant capacity.

Abbreviations: ABTS⁺: 2,2′-azinobis(2-ethylbenzothiazoline-6-sulfonic acid) cation; DPPH: 1,1-diphenyl-2-picrylhydrazyl, HPLC: high-performance liquid chromatography; NBT: nitroblue tetrazolium; O₂^?: superoxide anion; PMA: phorbol 12-myristate 13-acetate; PMS: phenazine methosulfate; TEAC: Trolox equivalent antioxidant capacity     

Investigations into the effects of processing on the retention of the carotenoid fractions of Leucaena leucocephala during storage,and the effects of processing on mimosine concentration

Leucaena leucocephala leaves were subjected to several drying treatments followed by pelleting and/or inclusion of ethoxyquin as antioxidant and analysed monthly to determine rates of loss of carotenes and xanthophylls. Rapid sun drying yielded leucaena leaf meal with carotene and xanthophyll concentrations of 446 and 865 mg/kg, respectively, but considerable losses occurred during drying at 60 and 145°C, and as a result of blanching in hot water or ferrous sulphate solution. Loss rates of carotenes during storage were in the range 19–40 mg/kg per month, and of xanthophylls, 29–53 mg/kg per month. Carotenoids in sun-dried leaf meal were most stable during storage. Pelleting or inclusion of ethoxyquin did not reduce carotenoid loss rates below those of unpelleted leaf meal.The mimosine content of untreated sun-dried leucaena leaf meal was 3.2%, but the concentration decreased to 2.5 and 1.8% on drying at 60 and 145°C, respectively. The concentrations of mimosine in sun-dried leafmeals of water-blanched and ferrous sulphate-blanched leucaena were 2.0 and 2.1%, respectively. Treatments causing maximum losses of mimosine were those which brought about maximum losses of carotenoids.     

Induction of trehalose in spores of the biocontrol agent Trichoderma harzianum

Spores of the biocontrol agent Trichoderma harzianum P1 produced in liquid media and harvested in the stationary sporulation stage SSS (after 60?h), had higher viability after slow (>4×) and fast drying (>12×) than their counterparts harvested in the exponential sporulation stage, ESS (after 30?h). The trehalose content of SSS spores was almost 20× higher than that of ESS spores (0.16 vs. 3.4?mg/100?mg, respectively). Heat shock (40?°C?×?90?min) effectively increased the trehalose content 2.5× with respect to untreated SSS spores. The trehalose content achieved in heat-treated SSS spores was almost 60% higher than the maximum reached by holding the spores under water-stress at 97% relative humidity prior to drying.     

Effects of heat treatment on oil-binding ability of rice flour

Heat-treated (120 °C for 120 min) rice flour showed high affinity to oil (oil-binding ability). This oil-binding ability could be observed by shaking the heat-treated rice flour (2.0 g), oil (4.0 mL), and water (20 mL) vigorously in a test tube, and the oil bound to the rice flour sank into the water. To examine the time-dependent levels of the oil-binding ability, rice flour was heat-treated at 120 °C for 10, 20, 40, 60, and 120 min, and the precipitated volume of oil/rice flour complex increased with an increase of the heating time. The oil-binding ability of the rice flour was not affected by the treatments with diethyl ether or boiled chloroform/methanol (2:1) solutions, which suggested no relationship to the oil in the rice flour, but was lost upon alkali (0.2% NaOH solution) or pepsin treatment, which suggested its relationship to the rice proteins.     

Salicylic acid-induced antioxidant protection against low temperature in cold-hardy winter wheat

“Dongnongdongmai 1” is a cultivated winter wheat which can endure cold temperature as low as ?30 °C with a reviving rate of 85 %. We aimed to explore the involvement of antioxidant protection system in salicylic acid (SA)-enhanced cold resistance of winter wheat. Seedlings were prayed with 0.1 mM SA at three-leaf stage, followed by cold acclimation at tillering stage (4 °C for 5 days) prior to cold treatment at 4, 0, ?10 or ?20 °C for 2 days. Under low temperature, the relative electrical conductivity (REC) of rhizomes and H₂O₂ content in rhizomes were lower compared with leaves, while in the reactive oxygen species (ROS) removal system, only the POD activity was higher. Foliar spray with SA significantly inhibited the cold-increased REC of rhizomes at ?20 °C and REC of leaves at ?10 and ?20 °C. In addition, application of SA prior to ?10 or ?20 °C treatment suppressed the increase in H₂O₂ content both in rhizomes and leaves. SA enhanced the activities of SOD, POD, and CAT in wheat following low-temperature treatment, especially at ?10 and ?20 °C. In addition, spray with SA resulted in 1.1-to-4.9-fold enhanced activities of the key enzymes in AsA–GSH cycle, including APX, DHAR, and MDHAR. Our results suggested that SA could improve the resistance of winter wheat against extreme low temperature by enhancing the activities of antioxidases to eliminate ROS and maintain the redox homeostasis. In addition, the less damage to rhizomes in comparison with leaves may be attributed to enhanced POD activity.     

Cryopreservation,early seedling development,and genetic stability of <Emphasis Type="Italic">Oncidium flexuosum</Emphasis> Sims

An efficient cryopreservation protocol was developed for mature seeds of Oncidium flexuosum Sims. Seed morphology, protocorm formation, and early seedling development were also assessed. The effects of phloroglucinol and Supercool X-1000^® as cryoprotectant additives in the vitrification solution were investigated. Dehydration using the plant vitrification solution 2 (PVS2) for 60 and 120 min prior to immersion in liquid nitrogen promoted the highest frequency of in vitro seed germination 6 weeks following culture on half-strength Murashige and Skoog (½ MS) medium. Mature seeds submitted to vitrification for 120 min in PVS2 and 1 % phloroglucinol at 0 °C enhanced germination by 68 %, whereas in PVS2 and 1 % Supercool X-1000^® germination was just moderately enhanced (26 %). In vitro-germinating seedlings developed healthy shoots and roots without the use of plant growth regulators. After 6 months of growth, there were no differences between in vitro- and ex vitro-grown seedlings for various phenotypic characteristics, including shoot length, number of leaves, number and length of roots, and fresh and dry weight. Seedlings were transferred to greenhouse conditions and successfully acclimatized, further developing into normal plants with over 90 % survival. Comparative analysis of seedlings from control and vitrified seeds using flow cytometry indicated that no change in ploidy levels occurred as a result of cryopreservation, therefore maintaining seedlings genetic stability. In this study, vitrification with PVS2 for 120 min with the addition of 1 % phloroglucinol offers a simple, safe, and feasible protocol for cryopreservation of O. flexuosum mature seeds.