A modified thermal convection drying technique for herbarium preservation

A simple and efficient installation for drying plants that is both rapid and preserves colour, and which works on the principle of autonomous thermal convection, is presented. The described apparatus offers significant advantages including: uniform heat distribution and short drying times; a system for applying pressure to prevent shrinkage, which uses polyurethane foam sheets and elastic straps; a compact, space-saving system suitable for field or laboratory applications; absorbent materials which can be re-used immediately; ability to run off differing power supplies (110/220 V) and to use infra-red lamps of varying specifications; low cost and ease of use.     

A modified thermal convection drying technique for herbarium preservation

A simple and efficient installation for drying plants that is both rapid and preserves colour, and which works on the principle of autonomous thermal convection, is presented. The described apparatus offers significant advantages including: uniform heat distribution and short drying times; a system for applying pressure to prevent shrinkage, which uses polyurethane foam sheets and elastic straps; a compact, space-saving system suitable for field or laboratory applications; absorbent materials which can be re-used immediately; ability to run off differing power supplies (110/220 V) and to use infra-red lamps of varying specifications; low cost and ease of use.     

A new method for the preservation of fungus stock cultures by deep-freezing

Recovery of 66 fungus stock cultures including Oomycota, Zygomycota, Ascomycota, Basidiomycota, and mitosporic mycetes were examined after cryopreservation. Almost all the stock cultures remained viable when the mycelia that had grown over the sawdust medium containing 10% glycerol as the cryoprotectant (65% moisture content, W/W) were frozen rapidly at −85°C and then allow to thaw naturally at room temperature. Test stock cultures were preserved for more than 10 years by this preservation method without any programmed precooling and rapid thawing for their cryopreservation. Most of the test fungi could survive for 5 years in medium containing 10% glycerol even after alternate freezing and thawing at intervals of 6 months. When a strain of Flammulina velutipes was tested for mycelial growth rate and productivity of fruit-bodies after cryopreservation for 3 years, the fungus reproduced with its initial capability. These results demonstrate that the sawdust-freezing method using a cryoprotectant is expected to be a reliable and easy preservation method for fungus stock cultures. Received: December 7, 2000 / Accepted: December 19, 2001     

A new method for the preparation of uniformly 14C-labelled compounds by using Anacystis nidulans

1. Growth conditions were devised which enabled Anacystis nidulans to be grown on high specific radioactivity (14)CO(2) (at over 98% isotopic abundance of (14)C) in a closed system with greater than 85% substrate utilization. 2. The DNA and RNA content of A. nidulans was estimated by various methods and compared with that of Chlorella pyrenoidosa. 3. A procedure was developed for the quantitative extraction and separation of the DNA and RNA from A. nidulans. 4. The fractionation and analysis of the DNA and RNA includes methods for the preparation of the uniformly (14)C-labelled deoxyribonucleoside monophosphates and the uniformly (14)C-labelled ribonucleoside monophosphates. 5. The base 6-methylaminopurine was identified as a component of the DNA. 6. The preparation and total yield of uniformly (14)C-labelled amino acids from A. nidulans are also briefly described. The results are discussed in relation to the use of other micro-organisms for the preparation of compounds labelled with (14)C.     

Acetone preservation: a practical technique for molecular analysis

In attempts to establish a convenient and reliable method for field collection and archival preservation of insects and their endosymbiotic microorganisms for molecular analysis, acetone, ethanol, and other organic solvents were tested for DNA preservability of the pea aphid Acyrthosiphon pisum and its intracellular symbiotic bacterium Buchnera sp. After 6 months' storage, not only the band of high-molecular-size DNA but also the bands of rRNA were well preserved in acetone, ethanol, 2-propanol, diethyl ether and ethyl acetate. Polymerase chain reaction (PCR) assays confirmed that the DNA of both the insects and their symbionts was well preserved in these solvents. In contrast, methanol and chloroform showed poor DNA preservability. When water-containing series of acetone and ethanol were examined for DNA preservability, acetone was apparently more robust against water contamination than ethanol. Considering that most biological materials contain high amounts of water, acetone may be a more recommendable preservative for DNA analysis than ethanol which has been widely used for this purpose. The DNA of various insects could be preserved in acetone at room temperature in good condition for several years. In addition to the DNA of the host insects, the DNA of their endosymbionts, including Buchnera and other mycetocyte symbionts, Wolbachia, and gut bacteria, was amplified by PCR after several years of acetone storage. The RNA and protein of the pea aphid and its endosymbiont were also preserved for several years in acetone. After 2 years' storage in acetone, proteins of A. pisum could be analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, and the endosymbiotic bacteria were successfully detected by immunohistochemistry and in situ hybridization on the tissue sections.     

草菇采后生理及保鲜技术

概述了草茹采集后的生理现象,并就草茹的采集和储藏过程中应采取的措施及其保鲜技术的应用提出了积极的建议。     

枇杷果实采后生理与保鲜技术研究进展(综述)

本文综述枇杷果实采后生理与保鲜技术的研究现状,并提出枇杷保鲜技术发展方向。     

A new mode of ammonite preservation – implications for dating of condensed phosphorite deposits

Unusual phosphatic casts of the ammonites Mortoniceras (Subschloenbachia) sp. and Stoliczkaia sp. from the upper Albian condensed phosphorite bed at Annopol, Poland, are discussed in terms of their taphonomic history. These specimens are interpreted as ‘secondary’ external casts of ammonite replicas preserved originally as attachment scars on oyster shells. The following genetic history is suggested for this previously undocumented mode of ammonite preservation: (1) settling of shells of dead ammonites on the seafloor; (2) colonization of these shells by oysters and formation of ammonite replicas on left valves of oysters; (3) dissolution of ammonite shells; (4) reworking and fragmentation of oyster shells; (5) casting of ammonite replicas by phosphatic material; and (6) separation of ammonite casts from oyster shells, either through mechanical disintegration or dissolution of the latter. The specimens studied were formed after dissolution of the ammonite conchs, not prior to this event as in the case of typical ammonite steinkerns (internal moulds). Therefore, they are here referred to as ‘pseudo‐steinkerns’. The time interval between loss of the original ammonite shells and the formation of oyster‐mediated pseudo‐steinkerns may be very extensive. Therefore, the pseudo‐steinkerns may potentially mislead in biostratigraphic dating of condensed phosphorite deposits.     

A simple technique for producing 13C or 14C-labelled fronds of Lemna gibba and their use in soil incubation investigations

The duckweed Lemna gibba required light and a suitable energy source such as sucrose, glucose or fructose, for maximum growth in culture. The requirement for light was relatively unimportant and the plants grew well in a photon flux density of only 52 μmol m^-2s^-1 PAR. The uptake and incorporation of uniformly labelled ¹⁴C-glucose into fronds was related only to the concentration of the sugar. When incubated with soil, labelled L. gibba behaved in a manner similar to that of labelled ryegrass roots which had been produced by a more elaborate technique using a ¹⁴CO₂ labelled atmosphere. During incubation with soil for 224 days the L. gibba material (specific activity 6133 Bq mg^-1 d. wt) lost 64% of its radioactivity as ¹⁴CO₂ and ryegrass (specific activity 6634 Bq mg^-1 d. wt) lost 49%. Alkaline extracted humic and fulvic acids from soil had specific activities for the L. gibba incubation of 3409 and 407 Bq mg^-1 solid and for ryegrass roots of 4609 and 546 Bq mg^-1 solid respectively. The production of ¹³C or ¹⁴C-labelled L. gibba can be undertaken using only simple equipment producing material the specific radioactivity of which can be controlled by adjusting the activity of the sugar energy source.     

Screening for lignin degrading bacteria by means of 14C-labelled lignins

Several Nocardia and Pseudomonas spp., as well as some unidentified bacteria, isolated from lake water containing high loads of waste lignin, were tested for their capacity to release ¹⁴CO₂ from specifically ¹⁴C-labelled dehydropolymer of coniferyl alcohol (DHP) or corn stalk lignins. The bacteria were selected according to their ability to degrade phenolic compounds. However, only some of them could release significant amounts of ¹⁴CO₂ from the labelled lignin. The tested Nocardia spp. were more active than the Pseudomonas spp. and the unidentified bacteria. The most active strains belonged to N. autotrophica. These strains released CO₂ significantly from the methoxyl group and transformed the other carbons from the phenylpropane skeleton of lignin also into CO₂. Other less demethylating strains also released little CO₂ from the other carbons of the lignin molecule. From corn stalk materials which were specifically labelled in the lignin part only small amounts of labelled CO₂ were released.Non-Common-Abbreviation Used DHP dehydropolymers of coniferyl alcohol     

鲜切花的衰老与保鲜(综述)

本文概述鲜切花在瓶插过程中水分代谢、物质代谢、植物激素,细胞膜变化与切花衰老之间的关系,以及鲜切花保鲜技术的研究进展。     

A new technique for selective identification and mapping of enhancers within long genomic sequences

We report a new experimental method of direct selection, identification, and mapping of potential enhancer sequences within extended stretches of genomic DNA. The method allows simultaneous cloning of a quantity of sequences instead of tedious screening of the separate ones, thus providing a robust and high-throughput approach to the mapping of enhancers. The selection procedure is based on the ability of such sequences to activate a minimal promoter that drives expression of a selective gene. To this end a mixture of short DNA fragments derived from the segment of interest was cloned in a retroviral vector containing the neomycin phosphotransferase II gene under control of a cytomegalovirus (CMV) minimal promoter. The pool of retroviruses obtained was used to infect HeLa cells and then to select neomycin-resistant colonies containing constructs with enhancer-like sequences. The pool of the genomic fragments was rescued by PCR and cloned, forming a library of the potential enhancers. Fifteen enhancer-like fragments were selected from 1-Mb human genome locus, and enhancer activity of 13 of them was verified in a transient transfection reporter gene assay. The sequences selected were found to be predominantly located near 5' regions of genes or within gene introns.     

果实热处理保鲜技术研究(综述)

应用热处理技术控制果实采后病虫害和延缓果实衰老是近年来果实保鲜领域的研究热点.本文概述热处理对果实形态、生理生化和品质的影响,以及热处理的应用现状与应用前景.     

A successful technique for the preservation of rabbit embryos

A technique for successfully freezing, thawing and transferring rabbit embryos has been developed. Morula stage embryos were collected from super-ovulated female rabbits by flushing both oviducts and uterine horns with a tissue culture medium. Well developed, viable embryos were then transferred to freezing vials and a cryoprotectant, dimethyl sulfoxide (DMSO) was added in several steps to bring its final concentration to 1.6 molar. To freeze the embryos the temperature was lowered slowly (either 0.5 degrees C/min or 1.0 degrees C/min) to -80 degrees C at which point the vials were transferred directly to liquid nitrogen (-196 degrees C). Thawing was done at 8 degrees C/min. After thawing, phosphate buffered saline was added in a stepwise manner to dilute the DMSO. The thawed embryos were then cultured at 37 degrees C. Transfer of the embryos was accomplished by laparotomizing a pseudopregnant doe and introducing the embryos into the fimbriated ends of the oviducts. The 101 positively transferred embryos resulted in 45 implantations and 34 live born young.     

A role for microwave processing in the dry preservation of mammalian cells

Dry preservation involves removing water from samples so that degradative biochemical processes are slowed and extended storage is possible. Recently this approach has been explored as a method for preserving living mammalian cells. The current work explores the use of microwave processing to enhance evaporation rates and to improve drying uniformity, thereby overcoming some of the challenges in this field. Mouse macrophage cells (J774) were pre-incubated in full complement media containing 50 mM trehalose, for 18-h, to allow for endocytosis of trehalose. Droplets of experimental and control (no intracellular trehalose) cell suspensions were placed on coverslips in a microwave cavity. Water was evaporated using intermittent microwave heating (600 W, 30 s intervals). Samples were dried to various moisture levels, rehydrated, and then survival was assessed after a 45-min recovery period using Calcein-AM/PI fluorescence and Trypan Blue exclusion assays. The metabolic activity of dried cells (4.3 gH(2)O/gdw) was assessed after rehydration using a resazurin reduction assay. Apoptosis levels were also measured. Post- rehydration survival correlated with the final moisture content achieved, consistent with other drying methods. Intracellular trehalose provided protection against injury associated with moisture loss. Metabolic assays revealed normal growth in surviving cells, and these survival levels were consistent with results from apoptosis assays (P > 0.05). Brightfield and fluorescence images of microwave-dried samples revealed a uniform distribution of cells within the dried matrix and profilometry analysis demonstrated that solids were uniformly distributed throughout the sample. Microwave-processing successfully facilitated rapid and uniform dehydration of cell-based samples.     

Mineralization of 14C-labelled maize in alkaline saline soils

The turnover of organic material determines the availability of plant nutrients in unfertilized soils, and this applies particularly to the alkaline saline soil of the former Lake Texcoco in Mexico. Uniformly labelled [¹⁴C] maize and its neutral detergent fibre (NDF) fraction, mainly containing cellulose and hemi-cellulose, were added to these soils to investigate dynamics of C and N and the importance of the NDF fraction. Soil with electrolytic conductivity (EC) of 1.2, 3.2, 24.6 and 32.7 dS m^–1 was incubated aerobically, while CO₂ and ¹⁴CO₂ production, and inorganic N dynamics (NH₄ ⁺, NO₂ ^–, NO₃ ^–) were monitored. The amount of ¹⁴C-labelled maize mineralized after 97 days was >500 mg C kg^–1 dry soil (D.S.) of the 1000 mg C kg^–1 D.S. added in soils with EC 24.6 dS m^–1, but only 257 mg C kg^–1 D.S. in soil with EC 32.7 dS m^–1. The decomposition of the NDF fraction showed a lag, greatest in the soil with the largest EC and the amount of ¹⁴C-labelled NDF fraction mineralized after 97 days was > 300 mg C kg^–1 D.S. in soils with EC 3.2 dS m^–1, but in the soil with EC 32.7 dS m^–1 it was only 118 mg C kg^–1D.S. Application of ¹⁴C-labelled maize and the NDF fraction induced a priming effect, most accentuated at the onset of the incubation. The ratio between the amount of CO₂ produced due to the priming effect and the ¹⁴CO₂ produced was 16-times larger when 250 mg maize-C kg^–1 D.S. was added and only 3-times when 2000 mg maize-C kg^–1 D.S. was added. Oxidation of NO₂ ^– occurred in soil with EC 32.7 dS m^–1 as witnessed by decreases in concentration of NO₂ ^– and increases in concentration of NO₃ ^–. It was found that EC affected the decomposition of maize, the NDF fraction and the priming effect. Decomposition of cellulose and oxidation of NO₂ ^– occurred in soil with EC 32.7 dS m^–1 although cellulolytic micro-organisms and autotrophic NO₂ ^– oxidizers could previously not be isolated from this soil.     

植物乳杆菌高活力保存方法的研究及其应用

本研究旨在探讨植物乳杆菌的高活力保存方法,为植物乳杆菌饲料添加剂规模化、工业化生产奠定基础。采用4℃低温保存法、36℃烘干后常温保存法、阳离子活性载体保存法等3种方法对植物乳杆菌进行活力保存比较试验。以保存后活菌数不低于原值50%为参照标准,结果显示,对照组可保存15 d,4℃低温保存法可保存30 d,36℃烘干后常温保存法只能保存一周,而阳离子活性载体保存法则可保存2个月以上。结果表明,阳离子活性载体保存法可应用于植物乳杆菌饲料添加剂的规模化、工业化生产实践中。