In vitro germination and viability of buckwheat (Fagopyrum esculentum Moench) pollen

An in vitro method for the germination of common buckwheat pollen was developed. Pollen grains were successfully germinated in an artificial medium consisting of 0.2 g each of MnSO₄, Ca(NO₃)2.4H₂O and KNO₃, 0.04 g H₃BO₃, 15 g sucrose and 30 g polyethylene glycol (molecular weight approximately 20,000) dissolved in 100 ml of double distilled water. The viability of pollen was assessed by in vivo and in vitro germination tests at 20 °C and 25 °C over a 38 h time period. Pollen grains were collected and germinated at 4 h intervals from freshly harvested flowers grown under 16 h day length and a constant temperature. Maximum pollen viability was found 2 h and 6 h after first light when plants were maintained at 25 °C and 20 °C, respectively. Viability, as measured by germination percentage, was similar at both temperature regimes. Some pollen remained viable for approximately 34 to 38 h in intact flowers, but all pollen lost viability in less than an hour when stored at room temperature without humidity control. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cryopreservation of Delphinium pollen at –30 °C

Pollen of garden varieties and species of Delphinium that was cryopreserved at –30 ^°C after 3 hours of air drying and storage on silica gel, had high germination and pollen tube growth in vitroeven after 180-day storage. On the other hand, pollen stored at 25 ^°C showed a marked decrease in the germination rate within 10 days. The best in vitro germination of Delphinium pollen was on a 1% water agar containing 15% sucrose and 0.005 to 0.01% boric acid and at 15 to 20 ^°C. Field pollination with the cryopreserved pollen showed higher fruit and seed set than pollination with pollen stored at 25 ^°C, and was not significantly different from fresh pollen. This revised version was published online in August 2006 with corrections to the Cover Date.     

Storage of avocado pollen

Summary Avocado pollen was stored at a range of temperatures and relative humidities (RH) for up to one year and the pollen was tested for viability in vivo.Pollen stored for one month was capable of germination on the stigma and penetrating the ovule when stored at 4°C with <1,23,55 and 75% r.h. and at -196°C with 0% r.h. Most pollen samples stored at 25 and -15°C at a range of r.h. would germinate on the stigma but none would penetrate the ovule.After one year of storage, pollen at 4°C and <1 and 23% r.h. would germinate on the stigma but would not penetrate the ovule. There was no germination of pollen stored at 4°C and 55 and 75% r.h. Only pollen stored at -196°C and 0% r.h. would penetrate the ovule, but thawing and refreezing once during the year destroyed the viability.     

Cryopreservation of English walnut (Juglans regia L.) pollen

Summary Pollen from eight clones of English walnut (Juglans regia L.) was stored in liquid nitrogen (LN₂) at-196°C for one year. Pollen was monitored for percent germination in vitro before being frozen. Pollen was frozen within two hours of anther dehiscence and subsequently at 24 hr intervals until germination assays indicated that the ability to germinate was lost. Survival after one year was evaluated by determining percent germination in vitro. Freshly collected pollen of only five of the eight clones survived LN₂. By contrast, when the same pollen was held for 24 and 72 hr before freezing, pollen of all eight clones survived. Survival is correlated with moisture content (MC) of the pollen. Pollen samples with MC greater than 7.5% were killed by freezing in LN₂. All pollen samples with MC between 4 and 7.5% survived as did most of the samples with MC between 3.2 and 4%.     

Low UV-C illumination for keeping overall quality of fresh-cut watermelon

The effects of four pre-packaging UV-C illumination doses (1.6, 2.8, 4.8 and 7.2 kJ m^?2) on quality changes of watermelon cubes stored up to 11 days at 5 °C were studied. Non-treated cubes were used as a control. Higher UV-C doses induced slightly higher CO₂ production throughout the storage period, while no changes in C₂H₄ production were monitored. However, UV-C did not significantly affect the final gas partial pressures within modified atmosphere packages where levels of 3–6 kPa O₂ and 13–17 kPa CO₂ were reached for all treatments. UV-C decreased microbial counts just after illumination. After 11 days at 5 °C, mesophilic, psycrophilic and enterobacteria populations were significantly lower in UV-C treated watermelon. Slight changes in CIE colour parameters were observed. According to sensory quality attributes, control and low UV-C treated cubes (1.6 and 2.8 kJ m^?2) can be stored for up to 11 days at 5 °C while the maximum shelf-life of moderate to high UV-C treated fruit was 8 days at 5 °C. Control cubes showed a 16% decrease in lycopene content after 11 days at 5 °C similar to that found for the high UV-C treatment. However low UV-C treated watermelon cubes preserved their initial lycopene content (2.8 kJ m^?2) or it was slightly decreased (1.6 kJ m^?2). UV-C radiation did not significantly affect the vitamin C content while catalase activity and total polyphenols content considerably declined throughout the storage period. However, total antioxidant capacity markedly increased, independently of UV-C doses. As a main conclusion, UV-C radiation can be considered a promising tool for keeping overall quality of fresh-cut watermelon.     

High CO2 effects on postharvest biochemical and textural properties of white asparagus (Asparagus officinalis L.) spears

The effects of high CO₂ concentration (10% CO₂, 17% O₂) on the changes of functional cell wall components (pectic substances, hemicellulose, cellulose, lignin), mechanical properties, content of free soluble sugars (sucrose, glucose, fructose), and respiration activity were studied in harvested white asparagus spears stored at 10 and 20 °C, respectively, for up to 7 d. Spears stored at 2, 10 and 20 °C in air were studied as controls, where the 2 °C condition indicated the effects of cold storage. During storage, respiration activity declined only slightly, irrespective of the CO₂ and temperature regime. Spears stored at 20 °C under both CA and normal air became less stiff and more elastic, however, tissue toughness increased significantly. Changes in toughness were associated primarily with the dynamics of lignin and cellulose, revealing a strong correlation (r² = 0.81). High CO₂ concentration inhibited the synthesis of cellulose and, to some extent, lignin accumulation at 20 °C. Additionally, elevated CO₂ inhibited the degradation of soluble carbohydrates. In contrast, slightly lower temperatures of 10 °C in combination with high CO₂ did not have a pronounced effect on changes in structural carbohydrates (lignin, cellulose, hemicellulose and pectins). The effect low temperature (2 °C) under normal atmosphere conditions resulted in the inhibition of cell wall changes in asparagus spears.     

农杆菌真空渗透法转化棉花花粉的初步研究

 以陆地棉花粉为受体,利用农杆菌真空渗透法将外源Susy基因转入新陆早19,对转化因素作了初步研究。结果表明,花粉在45%蔗糖浓度的花粉悬浮液中能维持适当渗透压,破损率最低;在-80 Pa压力下,菌液OD₆₀₀值在0.6～1.2范围内,真空渗入时间在15～25 min内,能维持较低花粉破损率及较高瞬间转化率;对自然花粉、农杆菌浸染的花粉及农杆菌真空渗透的花粉,分别进行GUS瞬时检测,未抽真空的花粉未被染色,而辅助真空处理的花粉部分变成蓝色,后将其授粉于已去雄的柱头上,观察到花粉管的伸长,说明抽真空后部分花粉仍有活力。     

Viability and storage of bromeliad pollen

Several bromeliad species from two different subfamilies, were used to develop a reliable method to evaluate pollen viability. Pollen germination on a medium containing 20% sucrose, 0.001%H₃BO₃ and 0.5% agar was comparable to germination on a compatible stigma. Maximum germination was reached within 2 to 10 hours depending on the species. Based on this test, six species were considered as being good pollen donors with germination percentages between 49%and 83%. Furthermore, pollen from these species and cultivars could be stored in liquid nitrogen (–196 ^°C) without a considerable loss of viability. For all species, a dehydration period of 4 hours prior to cryopreservation and a rehydration period of 1 hour after cryostorage were essential. Greenhouse humidity influenced anther moisture content and cryostorability. This revised version was published online in July 2006 with corrections to the Cover Date.     

Combined effect of chemical treatment and/or modified atmosphere packaging (MAP) on quality of fresh-cut papaya

The efficacy of chemical dips and modified atmosphere packaging (MAP), alone and in combinations, on the quality of fresh-cut papaya were studied throughout 25 days at 5 °C. Fresh-cut papaya were dipped in a solution of calcium chloride (1% w/v) and citric acid (2% w/v), packed in an atmosphere of 5% O₂, 10% CO₂, 85% N₂ and stored at 5 °C for 25 days. Physico-chemical analysis (package atmosphere, weight loss, pH, total soluble solids, firmness and color) and microbial quality along with a sensory analysis were measured at regular intervals throughout the storage period. Significant differences were found among the chemically treated and non-treated fresh-cut papaya in all the parameters considered. Chemical treatment followed by MAP, showed the best results among the treatment in terms of retaining sensory and quality characteristics and extending the shelf-life of 25 d for fresh-cut papaya.     

Induction of 2n pollen formation in <Emphasis Type="Italic">Begonia</Emphasis> by trifluralin and N<Subscript>2</Subscript>O treatments

In this study, treatments of both trifluralin (at 10, 100 and 1000 μM) and N₂O (in the form of gas under pressure) were applied to Begonia flower buds to induce the formation of 2n pollen. Three male fertile species (B. cucullata, B. subvillosa var. leptotricha and B. fischeri) and two male sterile hybrids (B. schmidtiana × B. cucullata and B. subvillosa var. leptotricha × B. cucullata) were treated. Pollen size, which is related to pollen DNA content, increased after both N₂O and trifluralin treatments, but the induction of large pollen was genotype dependent. Trifluralin induced large pollen only in the male fertile species, while N₂O treatments induced fertile 2n pollen in the male sterile B. schmidtiana × B. cucullata. Cytological studies showed that trifluralin induced multinuclear monads that resulted in 4n gametes in stead of 2n gametes. In general, large pollen obtained after trifluralin treatments showed low germination capability, while large pollen obtained after N₂O treatments retained high germination capability. Seedlings with raised ploidy level could only be obtained after crosses were performed with large pollen obtained from N₂O treatments. Hence, N₂O treatments are preferable to the use of trifluralin to induce 2n gametes in Begonia.     

Storage of sugarbeet pollen

Summary To develop the technology for long-term pollen preservation, sugarbeet pollen was collected from plants grown in the greenhouse and in the field, and was stored 1 day to 1 year at 5, -18, and -196°C. Pollen containing about 12% moisture was successfully stored in liquid nitrogen (LN₂) up to 1 year; this pollen effected fertilization of male-sterile flowers as well as freshly collected pollen. Germination of the resultant seed was good and not different from seed from fresh pollinations. Pollen stored at -18°C for 1 year did not result in as much seed set as fresh pollen, and 1 year at 5°C was essentially lethal. In vitro pollen germination served as a post-storage viability measure, provided the pollen was hydrated before germination. The methods tested in these experiments provided a relatively simple, reliable, and inexpensive means for preservation of sugarbeet pollen for breeding purposes and for preservation of genetic resources.Joint contribution of the Agricultural Research Service, USDA, and the Beet Sugar Development Foundation.     

美国梾木(Cornus spp.)花粉活力及萌发特性

为了了解美国梾木花粉活力及萌发特性,采用扫描电镜技术对13个美国梾木无性系花粉形态进行观测与方差分析,并采用荧光显微技术和TTC染色法对无性系2号花粉萌发特性及花粉活力进行观察。结果显示:(1)不同无性系间花粉大小差异不显著,但外壁纹饰差异较大,极面沟距的变异系数最高且遗传最为稳定。(2)花序开花率为75%时花粉活力最高,整花4℃条件下短期保存花粉活力较高。(3)授粉后,多数花粉可在柱头上正常萌发,72 h后花粉管即可进入子房。研究认为,花粉外壁及极面沟距可作为鉴定美国梾木无性系的形态指标,美国梾木花粉活力及萌发特性是抗逆及丰产杂交组合的重要保证。     

Germination of Brassica pollen and expression of incompatibility in vitro

Summary Pollen derived from cabbage, broccoli, and collards (Brassica oleracea, var. capitata, Italica and acephala, respectively) will germinate in a synthetic medium consisting of sucrose, H₃BO₄, CaCl₂ and purified polyethylene glycol 20,000. The optimum levels of constituents varied from genotype to genotype. With one genotype of cabbage, a germination percentage of 84% and tube lengths of at least 3.5 mm long were obtained. Whereas pollen from broccoli germinated in 1 to 2 hours, cabbage pollen required a 3 to 4 hour incubation period. Optimum germination occurred at pH 5.8 and at a pollen density of 0.5–2.0 mg/ml for cabbage. The effect of flower age on pollen germination varied from genotype to genotype. Of 7 cabbage genotypes tested, pollen germination either increased, decreased or remained the same as flower age increased. Pollen germination in vitro required polyethylene glycol purified by dialysis or passage through Sephadex G-25. Cabbage stigmas release a heat-labile substance capable of inhibiting germination of self-pollen, but stigma extracts from a cross plant have no effect.     

Flesh quality and lycopene stability of fresh-cut watermelon

Red fleshed watermelons are an excellent source of the phytochemical lycopene. However, little is known about the stability of lycopene in cut watermelon. In this study, lycopene stability and other quality factors were evaluated in fresh-cut watermelon. Twenty melons each of a seeded (Summer Flavor 800) and a seedless (Sugar Shack) variety were cut into 5 cm cubes and placed in unvented polystyrene containers, sealed, and stored at 2 °C for 2, 7, or 10 days. At each storage interval, melons were evaluated for juice leakage, changes in carotenoid composition, color, soluble solids content (SSC), and titratable acidity. Headspace carbon dioxide and ethylene were monitored during storage intervals. Juice leakage after 10 days of storage averaged 13 and 11% for the seeded and seedless melons, respectively. Lycopene content decreased 6 and 11% after 7 days of storage for Summer Flavor 800 and Sugar Shack melons, respectively. β-Carotene and cis lycopene contents were 2 and 6 mg kg⁻¹ for Summer Flavor 800 and Sugar Shack, respectively, and did not change with storage. After 10 days of storage, CIE L¹ values increased while chroma values decreased, indicating a lightening in color and loss of color saturation in melon pieces. Symptoms of chilling injury, such as greatly increased juice leakage, or lesions on cubes, were not seen on the fresh-cut cut watermelon after 10 days storage at 2 °C. Puree pH increased and SSC decreased slightly after storage. Carbon dioxide levels increased and oxygen levels decreased linearly during storage, creating a modified atmosphere of 10 kPa each of CO₂ and O₂ after 10 days. Fresh-cut cut watermelon held for 7 or more days at 2 °C had a slight loss of SSC, color saturation, and lycopene, most likely caused by senescence.     

Pollen storage at low temperature as a procedure for the improvement of cold tolerance in spring rape, Brassica napus L.

The possibility of selecting spring rape for cold tolerance at the mature pollen grain stage was studied by investigating the effects of pollen storage at low temperatures on the quality of pollen grains and on the cold tolerance of the plants generated from them. Pollen treatments of F¹ hybrids affected fertilization ability much more than viability and even after 10 days storage at 3 or 10°C the pollen germination percentage was reasonably high. Pollen storage for 7 or 10 days at 3 or 10°C significantly increased the cold tolerance of F² seed germination, with 3°C being more effective. Pollen storage for a shorter time had no effect upon the number of resulting genotypes tolerant to low temperature. This approach may be successfully applied in plant breeding to enrich segregating plant populations with cold-tolerant genotypes.     

Controlled atmosphere storage of guava (Psidium guajava L.) fruit

The effects of controlled atmospheres (CA) on respiration, ethylene production, firmness, weight loss, quality, chilling injury, and decay incidence of three commercially important cultivars of guava fruit were studied during storage in atmospheres containing 2.5, 5, 8, and 10 kPa O₂ with 2.5, 5, and 10 kPa CO₂ (balance N₂) at 8 °C, a temperature normally inducing chilling injury. Mature light green fruit of cultivars, ‘Lucknow-49’, ‘Allahabad Safeda’ and ‘Apple Colour’, were stored for 30 days either in CA or normal air, and transferred to ambient conditions (25–28 °C and 60–70% R.H.) for ripening. CA storage delayed and suppressed respiratory and ethylene peaks during ripening. A greater suppression of respiration and ethylene production was observed in fruit stored in low O₂ (≤5 kPa) atmospheres compared to those stored in CA containing 8 or 10 kPa O₂ levels. High CO₂ (>5 kPa) was not beneficial, causing a reduction in ascorbic acid levels. CA storage was effective in reducing weight loss, and maintaining firmness of fruit. The changes in soluble solids content (SSC), titratable acidity (TA), ascorbic acid, and total phenols were retarded by CA, the extent of which was dependent upon cultivar and atmosphere composition. Higher amounts of fermentative metabolites, ethanol and acetaldehyde, accumulated in fruit held in atmospheres containing 2.5 kPa O₂. Chilling injury and decay incidence were reduced during ripening of fruit stored in optimal atmospheres compared to air-stored fruit. In conclusion, guava cultivars, ‘Lucknow-49’, ‘Allahabad Safeda’, and ‘Apple Colour’ may be stored for 30 days at low temperature (8 °C) supplemented with 5 kPa O₂ + 2.5 kPa CO₂, 5 kPa O₂ + 5 kPa CO₂, and 8 kPa O₂ + 5 kPa CO₂, respectively.     

Influence of the combination of different atmospheres on diphenylamine,folpet and imazalil content in cold-stored ‘Pink Lady®’ apples

‘Pink Lady^®’ apples were harvested at commercial maturity, treated with three different agrochemical products, and stored at 1 °C under either air or controlled atmosphere conditions (2 kPa O₂ + 2 kPa CO₂ and 1 kPa O₂ + 1 kPa CO₂) for 13 and 27 weeks, followed by 4 weeks storage in air at 1 °C. Diphenylamine, folpet and imazalil contents in both the skin and flesh were simultaneously determined after cold storage plus simulated marketing periods of 1 and 7 d at 20 °C. After 27 weeks plus 7 d, diphenylamine and folpet levels in apple skin were lower for fruit stored in low O₂ (2 kPa) or air than for those kept under ultra-low O₂ (1 kPa). An additional storage period of 4 weeks in air reduced diphenylamine and folpet contents in whole apples stored for 13 weeks in the low O₂ controlled atmosphere. For imazalil, the same result was obtained in apple skins stored for 27 weeks under an ultra-low O₂ controlled atmosphere. Differences in diphenylamine and folpet contents were found for skin and flesh samples throughout the simulated marketing period, but there were observable differences in imazalil contents only for flesh samples.     

Establishment of reliable methods of in vitro pollen germination and pollen preservation of Brassica rapa (syn. B. campestris)

A simple and reliable method for evaluating the viability of Brassica pollen was established in which the in vitro germination rate of pollen was adopted as the index of the viability of pollen grains. Pollen grains were preincubated in an atmosphere in which the relative humidity (RH) was fixed to 52% or 66% at 20 °C for 5 hours. They were cultured for 16 hours at 25 °C in a liquid Kwack's medium (1964) supplemented with 20% sucrose, and the pH was adjusted to 8.0. They were then observed under a microscope and the number of germinating and unchanged pollen grains were counted. The germination rate of pollen was improved and stabilized by preincubation and the use of a high pH medium. More than 90% of the freshly harvested pollen grains of Brassica rapa (syn. B. campestris) germinated constantly in these conditions Undehisced anthers were collected from flowers at anthesis and dehydrated by incubation at 20 °C for 16–24 hours in an atmosphere where the RH was fixed to 15% or 32%. They were put into a plastic vial and preserved in a freezer at -20 °C. The germination percentage of the preserved pollen was scored at intervals during preservation. The germination rate of the pollen grains preserved at -20°C for 1 year was higher than 50% and the pollen proved to be efficient for seed set. Most of the seeds germinated normally. This revised version was published online in August 2006 with corrections to the Cover Date.     

Diphenylamine and pre-slicing storage effects on the responses of apple slices to elevated CO2 atmospheres

The effect of treatment with diphenylamine (DPA) and duration of postharvest storage of whole apple fruit on the responses of fresh-cut apple slices to elevated CO₂ storage atmospheres has been investigated. On the day of harvest, ‘McIntosh’, ‘Empire’ and ‘Delicious’ apples were untreated or dipped in DPA, and were held at 0.5 °C overnight or for 6 weeks before slicing. Slices were then stored at 0, 15, 30, 45 or 60% CO₂ in 1% O₂ (balance N₂), atmospheres. Color, firmness and accumulation of acetaldehyde, ethanol and ethyl acetate of the slices were measured. Generally slices were lighter (higher L^* values) when stored in elevated CO₂ atmospheres, but atmosphere and DPA effects varied by cultivar and were affected by pre-slice storage time. Slices prepared from stored fruit were softer compared with slices prepared at harvest. Slice firmness was not affected consistently by CO₂ or DPA concentration, whether they were prepared at harvest or after storage. The effects of increasing CO₂ concentration on acetaldehyde and ethanol accumulations were variable, being affected by cultivar and storage period. DPA treatment did not affect acetaldehyde accumulation of any cultivar, or ethanol accumulation of slices prepared from fruit at harvest. However, DPA-treated ‘Empire’ and ‘Delicious’ apples stored before slicing accumulated less ethanol compared with untreated fruit. Storage of apples before processing increased the accumulation of fermentation volatile compounds by cut apples under storage atmosphere conditions.     

Effects of short-term storage on germinability of pistachio pollen

The effects of short‐term storage under room conditions (constant 25°C and 35% relative humidity), refrigeration (4°C) and freezing (‐20°C) on the germinability of pollen of pistachio, Pistacia vera L., were studied and the effects of desiccation and pollen age on its germinability were reassessed. It was found that: (1) weight loss as a result of desiccation was positively correlated with germinability; (2) pollen grains stored in room conditions only germinated after prehydration, which restored germinability even after 7 days of exposure; (3) refrigerated (4°C) pollen grains retained their germinability for at least a week; (4) frozen pollen grains irreversibly lost most of their germinability; (5) 2‐day‐old pollen grains, within the anthers, retained their germinability under room conditions. These findings will contribute to the improvement of pistachio pollen preservation and help to find a simple and inexpensive method for its short‐term storage, while retaining its germinability.