Storage of Pollen Grains in Organic Solvents: Effect of Organic Solvents on Leaching of Phospholipids and its Relationship to Pollen Viability

In vitro germinability and membrane integrity (as revealed bythe fluorochromatic reaction (FCR) test) of pollen grains ofCrotalaria retusa L. stored in various organic solvents forsix months at –20±2 °C were studied and correlatedwith leaching of lipids, phospholipids, sugars and free aminoacids from pollen grains into organic solvents during storage.Pollen grains stored in organic solvents with low dielectricconstants (a measure of their non-polar nature), such as hexane,cyclohexane and diethyl ether, showed high scores for germinationand FCR and very little leaching of phospholipids, sugars andamino acids. Pollen grains stored in solvents with high dielectricconstants (a measure of their polar nature) such as isopropanoland methanol did not show germination or positive FCR scores,but showed extensive leaching of phospholipids, sugars and freeamino acids. The viability of pollen grains stored in organicsolvents seems to be determined largely by the effect of theorganic solvents on pollen phospholipid composition, which inturn affects membrane integrity and consequently pollen viability. Crotalaria retusa, organic solvents, pollen storage, viability, phospholipids     

Acceleration of the Growth of Camellia sasanqua Pollen by Soaking in Organic Solvent

Camellia sasanqua pollen that had been soaked in acetone or diethyl ether for only 5 minutes grew three to four times longer pollen tubes than unsoaked pollen. Although the acceleration of pollen tube growth was observed when the pollen had been soaked in cold solvents (5 C, −15 C) for 100 days, soaking in warm solvents (30 C, 24 C) caused it to decrease gradually, and the pollen eventually lost the ability to germinate. The acceleration may be caused by removing inhibitor from the pollen grain by the organic solvents.     

钝裂银莲花花粉活力测定方法的研究

以3种花色（黄色、浅黄色和白色）钝裂银莲花为研究对象,分别采用TTC染色、I2-I染色、无机酸测定和蔗糖萌发4种方法,测定了钝裂银莲花（Anemone obtusiloba）的花粉活力,同时比较了不种方法对花粉活力的测定效果,并了解了花粉活力在一天内的动态变化情况。结果表明：（1）钝裂银莲花花粉可育率很高,达80%左右。花粉直径为28.75~33.75 m 。（2）TTC浓度影响花粉活力的测定,当浓度为0.5%时,所测花粉活力的效果较好。（3）I2-I法的染色效果不理想,染色率仅为8.33%~22.2%,且3种花色花粉活力测定数据间差异不显著。（4）3种花色钝裂银莲花花粉活力有差异,黄色最高,浅黄色次之,白色最低。TTC染色法是简单快速测定钝裂银莲花花粉活力的最适方法。一天内花粉活力在12:00~14:00最大。     

Sputtering,a New Method for Coating Pollen Grains in Scanning Electron Microscopy

Sputtering is an easy, rapid and effective method for metal coating of pollen grains for examination in the scanning electron microscope. A very thin, regular and stable metal layer is obtained by bombarding a metal target with ions under a low vacuum, so that it ejects atoms on to the specimen. This allows of the observation of exine sculpturing which would be completely masked by a coating resulting from evaporation of carbon and metal. The sputtering method was tested on pollen of Deschampsia flexuosa, Molinia caerulea and Betula pubescens. Very narrow perforations could be discerned in the exine, similar to those seen in the images of carbon replicas obtained by many authors with the transmission electron microscope.     

Embryoid Formation in Pollen Grains of Nicotiana tabacum

Anthers of Nicotiana tabacum (n = 24) were cultured on nutrientagar and examined at intervals for pollen embryoids. Embryoidswere formed in anthers of varying developmental stage, the youngestof which coincided with the liberation of free microspores fromtetrads, and the oldest with the formation of bicellular grains.This period in the development of the anther occupied 4–5days. Older anthers within this range were more successful thanyounger anthers. The first mitosis of the pollen was typicallyasymmetric and resulted in the formation of unequal generativeand vegetative cells. Some of the grains then went into a lagphase for at least 5–6 days, after which the mitotic conditionwas restored. Embryoids were formed by repeated division ofthe vegetative cell. If the generative cell divided, it didso only once or twice. Occasionally the first mitosis was symmetricand gave rise to equal cells, and in these instances both cellsprobably participated in embryoid formation. The youngest anthersexamined were probably less successful because fewer grainssurvived to enter mitosis. The number of embryoids produced varied considerably from oneanther to another both within the same bud and between differentbuds: values ranging from less than 400 to 10 000 per antherwere encountered. Most of these degenerated after the firstfew divisions, partly because they burst prematurely from thepollen grain wall. Embryoids which continued to develop formedplantlets and/or callus. The largest number of plantlets obtainedfrom one anther was 32. Haploid plantlets were also regeneratedfrom callus by transferring it to a low-sugar medium withoutauxin. The behaviour of grains not forming embryoids was also noted.     

Mesoscale Transport of Corylus Pollen Grains in Winter Atmosphere

Measurements of atmospheric pollen concentration were made at different sites of the Po Valley, Italy, during the winter of 1978. The concentration was determined with volumetric samplers both near the ground and at different heights up to 500 m. Only Corylus and Alnus pollen grains were found during the experimental period. Due to the large quantity of grains released and the detailed information available about the source areas, the Corylus pollen was used to investigate the features of atmospheric transport. It was found that the three-dimensional patterns of pollen concentration are strongly influenced by meteorological conditions. Moreover, it appears that pollen is present in the atmosphere for a longer time with respect to the emission period, due to the relevant effect of the resuspension of the deposited pollen grains. This effect is detectable at all the sampling points, and not only near the sources.     

Kinetics of Encapsulated Yeast Alcohol Dehydrogenase Dispersed in an Organic Solvent

Microcapsules dispersed in organic solvents provide a suitable environment for conducting enzyme reactions involving cofactors and hydrophobic substrates. Encapsulated YADH is active and stable in cyclohexane provided the pH is adjusted appropriately. Mass transfer does not influence batch reaction rates. Conversion in a fluidized-bed reactor containing encapsulated YADH/NAD⁺ and employing cyclohexane as the continuous phase depends strongly on residence time and inlet cinnamyl alcohol concentration. However, interpretation of these results is complicated by enzyme inactivation by the product, cinnamaldehyde, and interference from residual encapsulating agents.     

Kinetics of Encapsulated Yeast Alcohol Dehydrogenase Dispersed in an Organic Solvent

Microcapsules dispersed in organic solvents provide a suitable environment for conducting enzyme reactions involving cofactors and hydrophobic substrates. Encapsulated YADH is active and stable in cyclohexane provided the pH is adjusted appropriately. Mass transfer does not influence batch reaction rates. Conversion in a fluidized-bed reactor containing encapsulated YADH/NAD⁺ and employing cyclohexane as the continuous phase depends strongly on residence time and inlet cinnamyl alcohol concentration. However, interpretation of these results is complicated by enzyme inactivation by the product, cinnamaldehyde, and interference from residual encapsulating agents.     

Biodegradation by an Arthrobacter Species of Hydrocarbons Partitioned into an Organic Solvent

An Arthrobacter strain mineralized naphthalene and n-hexadecane dissolved in 2,2,4,4,6,8,8-heptamethylnonane. The extent of mineralization increased with greater volumes of solvent. Measurements under aseptic conditions of the partitioning of naphthalene into the aqueous phase from the solid phase or from heptamethylnonane showed that the rates were rapid and did not limit mineralization. The rate of mineralization of hexadecane was rapid, although partitioning of the compound into aqueous solution was not detected. The Arthrobacter sp. grown in media with or without heptamethylnonane did not excrete products that increased the aqueous solubility of naphthalene and hexadecane. Measurements of the number of cells in the aqueous phase showed that the Arthrobacter sp. attached to the heptamethylnonane-water interface, but attachment was evident even without a substrate in the heptamethylnonane. Tests with small inocula of the Arthrobacter sp. demonstrated that at least a portion of naphthalene or hexadecane dissolved in heptamethylnonane was degraded by cells attached to the solvent-water interface. The cells did not adhere in the presence of 0.1% Triton X-100. The surfactant prevented mineralization of the hexadecane initially dissolved in heptamethylnonane, but it increased the rate and extent of mineralization of naphthalene initially dissolved in heptamethylnonane. The data show that organic solvents into which hydrophobic compounds partition affect the biodegradation of those compounds and that attachment of microorganisms to the organic solvent-water interface may be important in the transformation.     

Discharge in a Subthreshold Microwave Beam as an Effective Means for Mercaptan Decomposition

Plasma Physics Reports - A subthreshold microwave discharge in atmospheric-pressure air is used to process mixtures of mercaptans (thiols) with air and with air and methane. It is found that, at...     

Osmoconditioning as a Means of Counteracting the Ageing of Pepper Seeds during High-temperature Storage

Sweet pepper seeds were osmotically conditioned in 0.4 M mannitolsolution for 4 d (at 25 °C, in darkness) before or afterstorage at 35 °C for up to six months, and their germinationand viability was compared with that of untreated seeds storedunder the same conditions. Seeds that had been osmoconditionedprior to storage retained a high rate of germination and germinatedto a high final percentage (from 80 to 50 per cent) at both15 and 25 °C throughout the storage period. By contrast,both the rate and total level of germination of untreated pepperseeds declined rapidly at both germination temperatures, andby three months of storage the total level of seed viabilitywas already less than 10 per cent. Seeds that were first storedat 35 °C, and then osmoconditioned just prior to germination,showed a decline in germinability which when tested at 25 °Cwas the same as for untreated seeds, while tested at 15 °Coccurred at a slightly slower rate than for untreated seeds. It is evident that osmoconditioning prior to storage, in additionto the acceleration of germination, resulted in a dramatic delayof the ageing rate, thus increasing considerably the longevityof seeds. On the other hand, osmoconditioning after storagedid not seem to have any significant effect on seed viability,though it enhanced the germination rate. Capsicum annuum, sweet pepper, seed, germination, osmoconditioning, priming, storage, viability, ageing, longevity     

Irradiation Can Selectively Kill Tumor Cells while Preserving Erythrocyte Viability in a Co-Culture System

An understanding of how to safely apply intraoperative blood salvage (IBS) in cancer surgery has not yet been obtained. Here, we investigated the optimal dose of ¹³⁷Cs gamma-ray irradiation for killing human hepatocarcinoma (HepG2), gastrocarcinoma (SGC7901), and colonic carcinoma (SW620) tumor cells while preserving co-cultured erythrocytes obtained from 14 healthy adult volunteers. HepG2, SGC7901, or SW620 cells were mixed into the aliquots of erythrocytes. After the mixed cells were treated with ¹³⁷Cs gamma-ray irradiation (30, 50, and 100 Gy), tumor cells and erythrocytes were separated by density gradient centrifugation in Percoll with a density of 1.063 g/ml. The viability, clonogenicity, DNA synthesis, tumorigenicity, and apoptosis of the tumor cells were determined by MTT assay, plate colony formation, 5-ethynyl-2''-deoxyuridine (EdU) incorporation, subcutaneous xenograft implantation into immunocompromised mice, and annexin V/7-AAD staining, respectively. The ATP concentration, 2,3-DPG level, free Hb concentration, osmotic fragility, membrane phosphatidylserine externalization, blood gas variables, reactive oxygen species levels, and superoxide dismutase levels in erythrocytes were analyzed. We found that ¹³⁷Cs gamma-ray irradiation at 50 Gy effectively inhibited the viability, proliferation, and tumorigenicity of HepG2, SGC7901, and SW620 cells without markedly damaging the oxygen-carrying ability or membrane integrity or increasing the oxidative stress of erythrocytes in vitro. These results demonstrated that 50 Gy irradiation in a standard ¹³⁷Cs blood irradiator might be a safe and effective method of inactivating HepG2, SGC7901, and SW620 cells mixed with erythrocytes, which might help to safely allow IBS in cancer surgery.     

西瓜花粉壁超微结构与花粉ATP酶细胞化学定位

研究了西瓜花粉壁超微结构以及单核花粉液泡化时期ATP酶活性超微细胞化学定位。花粉壁的外壁分为外层和内层 ,外层包括覆盖层、基粒棒和基足层等三层 ,内层只包含一层。外层电子密度相对较小 ,内层电子密度相对较大 ;外层与内层之间有缝隙。ATP酶活性反应产物主要分布在细胞质基质、质体、内质网和花粉内壁中     

Chemical Inactivation of Lipase in Organic Solvent: A Lipase from Pseudomonas aeruginosa TE3285 is More Like a Typical Serine Enzyme in an Organic Solvent than in Aqueous Media

A microbial lipase from Pseudomonas aeruginosa TE3285 was treated in anhydrous diisopropyl ether with three kinds of serine-reactive reagents, ethyl p-nitrophenyl methylphosphonate (ENMP), diisopropyl fluorophosphate (DFP), and phenylmethylsulfonyl fluoride (PMSF) to lose its catalytic activity for both transesterification in an organic solvent and ester hydrolysis in aqueous system. In contrast with the facile inactivation in an organic solvent, no or very slow inactivation was observed in an aqueous solution. The lipase was shown to behave more like a typical serine enzyme in an organic solvent than in aqueous solution with regard to the chemical inactivation by serine-reactive reagents. The unique behavior of the lipase in an organic solvent may be associated with inferfacial activation of the lipase, which is one of the most distinct characteristics of the lipase family, and the activiation of lipase could be induced by a hydrophobic interaction with an organic solvent.