魔芋试管苗批量生产过程中外植体消毒灭菌技术研究

采用熏气法,可较为彻底地杀死魔芋球茎或根状茎的内生菌,再配合酒精、HgCl2等进行表面灭菌,可使初次接种污染率控制在10%以下,继代培养污染率控制在5%以下,从而建立魔芋试管苗的无菌繁殖体系,为试管苗的批量化生产提供了前提和技术保障。     

Ion transport in roots: measurement of fluxes using ion-selective microelectrodes to characterize transporter function

The transport of mineral ions into and out of tissues and cells is central to the life of plants. Ion transport and the plasma membrane transporters themselves have been studied using a variety of techniques. In the last 15 years, measurement of specific ion fluxes has contributed to the characterization of transport systems. Progress in molecular genetics is allowing gene identification and controlled expression of transporter molecules. However the molecular expression of transporter gene products must be characterized at the functional level. The ion‐selective microelectrode technique to measure specific ion fluxes non‐invasively is ideally suited to this purpose. This technique, its theory, its links with others and its application and prospects in plant science, are discussed. Ions studied include hydrogen, potassium, sodium, ammonium, calcium, chloride and nitrate. Applications discussed include: solute ion uptake by roots; gravitropism and other processes in the root cap, meristematic and elongation zones; Nod factor effect on root hairs; osmotic and salt stresses; oscillations; the effects of light and temperature. Studies have included intact roots, leaf mesophyll and other tissues, protoplasts and bacterial biofilms. A multi‐ion capability of the technique will greatly assist functional genomics, particularly when coupled with imaging techniques, patch clamping and the use of suitable mutants.     

Gliding motility of Peloploca spp., and their distribution in the sediment and water column of a eutrophic lake

Observations indicating gliding motility in the gas-vacuolate, filamentous organism Peloploca were made using microcapillary tubes. Tubes containing semi-solid agar, incubated in sediment cores gave good enrichments of Peloploca spp. The organisms, which had the form of helical bundles of filaments, were seen to corkscrew through the agar at up to 2–3 m s^-1.The vertical distribution of Peloploca spp. in the sediment and water column of a eutrophic lake was examined periodically during summer stratification. The organisms were confined to anoxic conditions in the sediment prior to stratification. With increasing anoxia in the hypolimnion, the population shifted upwards in the sediment, and towards the end of stratification, in the most reducing conditions, appeared in the lower hypolimnion. Anaerobically incubated sediment cores also showed the movement of the Peloploca population from sediment into the overlying water.It is suggested that the gliding motility and helical morphology of Peloploca bundles enable them to migrate through sediment in response to oxygen and Eh gradients, in addition to their use of gas vacuoles to adjust their position in the water column. The taxonomic implications of gliding motility in Peloploca spp. are discussed.     

Aluminum effects on the kinetics of calcium uptake into cells of the wheat root apex

The effects of aluminum on the concentration-dependent kinetics of Ca²⁺ uptake were studied in two winter wheat (Triticum aestivum L.) cultivars, Al-tolerant Atlas 66 and Al-sensitive Scout 66. Seedlings were grown in 100 M CaCl₂ solution (pH 4.5) for 3 d. Subsequently, net Ca²⁺ fluxes in intact roots were measured using a highly sensitive technique, employing a vibrating Ca²⁺-selective microelectrode. The kinetics of Ca²⁺ uptake into cells of the root apex, for external Ca²⁺ concentrations from 20 to 300 M, were found to be quite similar for both cultivars in the absence of external Al; Ca²⁺ transport could be described by Michaelis-Menten kinetics. When roots were exposed to solutions containing levels of Al that were toxic to Al-sensitive Scout 66 but not to Atlas 66 (5 to 20 M total Al), a strong correlation was observed between Al toxicity and Al-induced inhibition of Ca²⁺ absorption by root apices. For Scout 66, exposure to Al immediately and dramatically inhibited Ca²⁺ uptake over the entire Ca²⁺ concentration range used for these experiments. Kinetic analyses of the Al-Ca interactions in Scout 66 roots were consistent with competitive inhibition of Ca²⁺ uptake by Al. For example, exposure of Scout 66 roots to increasing Al levels (from 0 to 10 M) caused the K _m for Ca²⁺ uptake to increase with each rise in Al concentration, from approx. 100 M in the absence of Al to approx. 300 M in the presence of 10 M Al, while having no effect on the V _max. The same Al exposures had little effect on the kinetics of Ca²⁺ uptake into roots of Atlas 66. The results of this study indicate that Al disruption of Ca²⁺ transport at the root apex may play an important role in the mechanisms of Al toxicity in Al-sensitive wheat cultivars, and that differential Al tolerance may be associated with the ability of Ca²⁺-transport systems in cells of the root apex to resist disruption by potentially toxic levels of Al in the soil solution.We would like to thank Dr. Lionel F. Jaffe, Director of the National Vibrating Probe Facility, Marine Biological Laboratory, Woods Hole, Mass., USA, for making his calcium-selective vibrating-mi-croelectrode system available for a portion of this work. The research presented here was supported in part by USDA/NRI Competitive Grant number 91-37100-6630 to Leon Kochian. Contribution from the USDA-ARS, U.S. Plant, Soil and Nutrition Laboratory, Cornell University, Ithaca, N.Y. This research was part of the program of the Center for Root-Soil Research, Cornell University, Ithaca, N.Y. Department of Soil, Crop and Atmosphere Science, paper No. 1741.     

Harvesting Chlorella species using magnetic iron oxide nanoparticles

Microalgae are very useful organisms as it provides many beneficial products for human use. For large scale cultivation and further applications, its harvesting procedure needs to be enhanced to make the production process of the end product highly affordable. Magnetic nanoparticles have great potential to harvest microalgae as it can easily attract and attach to the algal cell surface forming a layer, which can be harvested quickly under the influence of a magnetic field. Our work on Chlorella pyrenoidosa and Chlorella minutissima shows, 500 mg of the synthesized bare iron oxide nanoparticles, harvests 90% of Chlorella pyrenoidosa (1 g L^?1), in 60 s at pH 3 and 600 mg iron oxide nanoparticles, harvests 85% of Chlorella minutissima (1 g L^?1) in 60 s at pH 5, which can decrease the amount of time and energy consumed in the overall production costs.     

Effects of polyoxin D on germination,morphological development and biosynthesis of the cell wall of Trichoderma viride

When polyoxin D is added to a spore suspension of Trichoderma viride at a concentration from 50–100 g/ml, it inhibits from 40–60% of germination. This percentage increases if dimethylsulfoxide (DMSO) is added.Mycelium growing in the presence of polyoxin D becomes irregular and loses its rigidity, showing several bulges along the hypha. Under the electron microscope the features of the cell wall and cytoplasmic content are apparently normal. Nevertheless, after incubation with different lytic systems or with (¹⁴C)glucose, it can be seen that polyoxin D partially inhibits the biosynthesis of -(1-3)glucan and the biosynthesis of chitin to a greater extent attaining inhibition of 83% at 100 g/ml of the antibiotic concentration.Regenerating protoplasts are less affected by polyoxin D. They do regenerate slower but the percentage of regeneration is more than 80%. Aberrant tubes synthesized by these protoplasts are not affected, they manifest their usual morphology and lack of chitin is confirmed in their composition.List of Abbreviations CECT Colecci Española de Cultivos Tipo - GAE Glucose, Asparagine and yeast Extract - DMSO Dimethylsulfoxide - PPO Diphenyloxazole - POPOP 1,4-bis-(4-methyl-5-phenyloxazol-2-yl)     

Lifetime and temporal occurrence of ectomycorrhizae on ponderosa pine (Pinus ponderosa Laws.) seedlings grown under varied atmospheric CO2 and nitrogen levels

Climate change (elevated atmospheric CO2, and altered air temperatures, precipitation amounts and seasonal patterns) may affect ecosystem processes by altering carbon allocation in plants, and carbon flux from plants to soil. Mycorrhizal fungi, as carbon sinks, are among the first soil biota to receive carbon from plants, and thereby influence carbon release from plants to soil. One step in this carbon release is via fine root and mycorrhizal turnover. It is necessary to know the lifetime and temporal occurrence of roots and mycorrhizae to determine the capacity of the soil ecosystem to sequester carbon assimilated aboveground. In this study, ponderosa pine (Pinus ponderosa Laws) seedlings were grown under three levels of atmospheric CO2 (ambient, 525 and 700 mol CO2 mol-1) and three levels of annual nitrogen additions (0,100 and 200 kg N ha-1) in open-top chambers. At a two-month frequency during 18 months, we observed ectomycorrhizal root tips observed using minirhizotron tubes and camera. The numbers of new mycorrhizal root tips, the numbers of tips that disappeared between two consecutive recording events, and the standing crop of tips at each event were determined. There were more mycorrhizal tips of all three types seen during the summer compared with other times of the year. When only the standing crop of mycorrhizal tips was considered, effects of the CO2 and N addition treatments on carbon allocation to mycorrhizal tips was weakly evident. However, when the three types of tips were considered collectively, tips numbers flux of carbon through mycorrhizae was greatest in the: (1) high CO2 treatment compared with the other CO2 treatments, and (2) intermediate N addition treatment compared with the other N addition treatments. A survival analysis on the entire 18 month cohort of tips was done to calculate the median lifetime of the mycorrhizal root tips. Average median lifetime of the mycorrhizal tips was 139 days and was not affected by nitrogen and CO2 treatments.     

Evolution and speciation of cave-dwelling Fulgoroidea in the Canary Islands (Homoptera: Gixiidae and Meenoplidae)

Ten new cavernicolous Fulgoroidea species of the families Cixiidae and Meenoplidae are described from the Canary Islands: Cixius palmeros, C. pinarcoladus, C. ratonicus, C. tacandus, Memoplus claustrophilus from La Palma, C. ariadne, C. nycticolus, M. charon from El Hierro, and Tachycixius crypticus, T. retrusus from Tenerife. Notes on their ecology and distribution are given, and where possible, phylogenetic affinities to existing epigean species of the corresponding families are discussed. Despite the present-day relict status of the majority of cavernicolous species, it cannot be concluded with certainty whether their evolution has followed allopatric or parapatric modes of speciation.