Effect of potassium nutrition on frost damage and yield of potato plants on alluvial soils of the Punjab (India)

Summary Field experiments were conducted during 1974 and 1975 on 14 alluvial soils, varying in available K from 56 to 158 ppm, at Jullundur-Punjab (India) to study the effect of K nutrition on frost damage to potato crops. Frost damage was inversely related to the available K content of the soils and the K concentration of the potato leaves. K fertilization significantly reduced the frost damage and increased tuber yields in soils containing less than 114 ppm available K. Frost damage was significantly related to potato response to K application and negatively correlated to tuber yield.     

Structural analysis of telechelic polymer solution using dissipative particle dynamics simulations

A telechelic polymer is an amphiphilic polymer that can form micellar structures when dissolved in water. A telechelic polymer solution shows viscoelastic behaviour owing to the formation of characteristic networks, i.e. loops, bridges and dangling chains. For industrial purposes, telechelic polymers have many applications as thickening agents, such as in paints and cosmetics. Thus, it is desirable to predict and control the rheological properties of telechelic polymers. However, detailed studies at the molecular level have not yet been performed. In this study, I use the dissipative particle dynamics (DPD) method to investigate the relationship between the characteristic structural properties and the molecular structure in telechelic polymer solutions. I show that the morphology of telechelic polymer solutions depends on the concentration and chain length, the distribution of the end-to-end distance, the mean square end-to-end distance, the mean square radius of gyration and the time-averaged mean square displacement. Although an effect of entanglement is important for properties of polymer melts, the polymer chain composed of DPD particles cannot reproduce it. Therefore, I compare telechelic polymer solutions with and without the segmental repulsive potential (SRP), which can simulate the effect of entanglement in DPD simulations. The results indicate that it is necessary to include the SRP in DPD simulations to correctly analyse the behaviour of telechelic polymer solutions.     

Recovery of the bilberry (Vaccinium Myrtillus L.) from artificial spring and summer frost

Vegetative and sexual recovery of bilberry (Vaccinium myrtillus L.) was monitored in the field for three growing seasons after artificially applied spring and early summer frost. Bilberry recovered vegetatively (density and biomass) by vigorous production of new ramets and by production of large shoots in the damaged ramets. Recovery did not occur sexually (production of flowers), however. Summer frost was slightly more detrimental than the spring frost, but the difference was significant only in the percentage of flowering ramets. The results showed that bilberry is buffered against moderate frost injury. The rapid vegetative recovery guarantees survival of the plants after frost damage, whereas sexual reproduction may need accumulation of resources for many seasons. Consequently, the ability of bilberry to recover from sporadic frosts increases its potentiality to persist in a changing environment, where increasing temperature leads to earlier snowmelt in the spring, and increases the risk of frost injuries in plants that overwinter under snow.     

Modelling frost resistance of Scots pine seedlings using temperature,daylength and pH of cell effusate

The annual course of frost resistance (LT₅₀) and the pH of the cell effusate in needles of two-year-old Scots pine seedlings were monitored in a field experiment in Oulu, Northern Finland (65° N, 25° E) during 1995. The aim of the work was to to develop model to predict the annual variation in frost resistance by pH of the cell effusate and meteorological data. The seedlings were covered with a fibre cloth shelter which transmitted sufficient light for them to experience the photoperiod, but prevented the accumulation of snow over them. The shelter above the seedlings was removed at the beginning of May and erected again at the end of September. The seedlings were watered only for the time when the shelter was removed, and received fertilizer only during the previous summer (1994).Frost resistance was only -5° C during the growing season but more than -100° C during the winter rest period. It was about -10° C at the end of August, increased to -55° C in the next three weeks, and reached -100° C at the beginning of October. The pH of the cell effusate was lowest during the growing season and highest in winter, the difference being about one and half pH unit. Needles exposed to -196° C showed pH from 4.0 in summer to 5.5 in winter, while pH of the non-frozen needles varied from 5.0 to 6.5, respectively. Seasonal variation in frost resistance was explained by a regression model well (R² = 0.9) when day length, minimum air temperature and pH were entered as variables.     

Effects of a kaolin-based particle film on oviposition and feeding of gypsy moth (Lep., Lymantriidae) and forest tent caterpillar (Lep., Lasiocampidae) in the laboratory

Abstract:  In the laboratory we investigated a kaolin-based crop protectant, Surround^® WP, that was mixed in a spray carrier (94% water, 2% methanol, 4% Triton X100) and applied as particle film against gypsy moth Lymantria dispar (L.) and forest tent caterpillar Malacosoma disstria Hubner. Paired-choice feeding tests showed that gypsy moth larvae were significantly (P < 0.05) less disposed to feed on red oak Quercus rubrus L. treated with high concentrations (60 and 120 g/l carrier) of kaolin than on untreated foliage. There was no significant difference (P > 0.05) between gypsy moth's consumption of kaolin-treated and untreated red oak when given a single food choice, but the adults were less inclined to oviposit on the kaolin-treated surfaces than on controls. However, no kaolin treatment completely deterred gypsy moth feeding or oviposition. Kaolin particle film did not influence forest tent caterpillar feeding and there were insufficient data to evaluate the effect of kaolin on the species' oviposition. The spray carrier did not increase the efficacy of the kaolin, but as an independent, albeit impractical treatment it constrained feeding by both insect species but did not affect gypsy moth oviposition. These initial laboratory findings suggest that applying kaolin-based particle film to forests would present significant challenges; but if these could be surmounted, the technique might assist in the management of gypsy moth but not likely in that of forest tent caterpillar.     

Production of fertile transgenic lentil (Lens culinaris Medik) plants using particle bombardment

Summary A reproducible system for gene transfer in lentil through particle bombardment is presented. Lentil cotyledonary nodes excised from germinated seedlings were bombarded with a plasmid containing a mutant acetolactate synthase gene (ALS) from tobacco conferring resistance to sulfonylurea herbicides. Putative transgenic shoots regenerated on Murashige and Skoog medium supplemented with 6-benzylaminopurine (BA) and chlorsulfuron (5 nM for first 4 wk followed by 2.5 nM for the remainder of the culture period) were micrografted and successfully transferred to soil. T₀ and selfed progeny plants were screened using metsulfuron herbicide leaflet painting. The non-transformed escapes died and transformed plants survived the test. The surviving plants were phenotypically normal and produced viable seeds. The presence and stable transmission of the transgene into genomic DNA of screened T₁ transformants was confirmed by PCR and Southern hybridization. This method for producing transformed plants will allow new opportunities for lentil breeding to produce improved cultivars.     

Production of transgenic cassava (Manihot esculenta Crantz) plants by particle bombardment using luciferase activity as selection marker

Cassava embroids derived from friable embryogenic callus of the genotype TMS60444 were bombarded with DNA of the constructs pJIT100 or pJIT64. Both constructs contain the non-invasive reporter gene luciferase from firefly driven by the CaMV 35S promoter. The influence of several particle gun machine parameters and pretreatment of plant material on transient luciferase activity were studied to determine the most essential conditions for stable transformation. Two weeks after bombardment pieces of friable calli with luciferase activity were selected. In total, 67 independent selected calli with luciferase activity (spots), derived from five different experiments, were further cultured either in liquid or on solid medium. Per plate or flask one spot was cultured. In subsequent selection rounds all spots of one individual plate or flask were cultured as one individual group. In this way different transformation events were separated and multiplied.Eight weeks after bombardment 34 cultures still contained luciferase activity. The mean number of luciferase spots per culture had increased from 1 to 4.6 spots in liquid and to 2.5 spots on solid medium. After two more months of subsequent culture and luciferase selection presence of the construct in these cultures was confirmed at the molecular level using the polymerase chain reaction assay and Southern analysis.Friable embryos derived from four transformation events were cultured for maturation. Between 3% and 21% of the mature embryos of the different transformation events were luciferase-positive. After multiplication of the luciferase-positive mature embryos by secondary somatic embryogenesis they were germinated. The plantlets analysed contained one to several copies of the inserted DNA. The method presented enables the transformation of this particular cassava genotype, thus allowing the genetic improvement of this important tropical crop by transgenesis.     

A kaolin‐based particle film for suppression of the olive fruit fly Bactrocera oleae Gmelin (Dip., Tephritidae) in olive groves

The efficacy of a kaolin‐based particle film formulation M‐99‐099 to control olive fruit fly, Bactrocera oleae Gmelin, field infestations was investigated in north‐western Syria. The results showed that fruit infestation levels were significantly reduced on kaolin‐treated trees compared with untreated trees. Kaolin particle film successfully suppressed B. oleae populations and provided season‐long insect control (>14 weeks) whereas the insecticide dimethoate failed to protect olives for as long a period after the last spray. Consistent with previous findings, the M‐99‐099 kaolin particle film proved to be a promising alternative method to synthetic insecticides and could be used to control B. oleae in olive groves.     

Modelling and simulation of the adsorption of the lipase from Geotrichum sp. on hydrophobic interaction columns

The Langmuir model fitted well the adsorption isotherms of lipase on the hydrophobic resin. The model parameters, Q _m and k _d, were affected by NaCl concentration: Q _m increased from 31 to 80 U g^–1 resin, and k _d changed from 9.4 to 3 U ml^–1. Column modelling and the simulation data were compared with the experimental data with good agreement. The highest achieved column efficiency was 71%.     

Dissipative particle dynamics simulation of the soft micro actuator using polymer chain displacement in electro-osmotic flow

ABSTRACT

In this research, the numerical simulation of a soft polymer micro actuator performance has been investigated using the dissipative particle dynamics method in electro-osmotic flow. Effective factors including electro-osmotic flow and polymer chain parameters have been studied. First of all, considering a wide range of electro-osmotic parameters, the validation of analytical results is carried out in a simple micro channel. The electric field and zeta potential changes are linearly related to the flow rate, and the kh parameter behaves nonlinearly to around the kh?=?10. In the following, a convergent–divergent channel is used for the soft micro actuator simulation in which a polymer chain as a heart of actuation is embedded in the middle. As the main control parameter, the direction of the electric field is changed every 4?s, and it leads to a reciprocating motion. The numerical results indicate that the displacement of the soft polymer chain will be increased by enhancing the electric field, the number of beads, decreasing the harmonic bond coefficient and also exposing more length of a polymer chain in front of fluid flow. The results of this study may be useful for some future applications such as artificial fibres and muscles.     

Insert engineering and solubility screening improves recovery of virus‐like particle subunits displaying hydrophobic epitopes

The Polyomavirus coat protein, VP1 has been developed as an epitope presentation system able to provoke humoral immunity against a variety of pathogens, such as Influenza and Group A Streptococcus. The ability of the system to carry cytotoxic T cell epitopes on a surface‐exposed loop and the impact on protein solubility has not been examined. Four variations of three selected epitopes were cloned into surface‐exposed loops of VP1, and expressed in Escherichia coli. VP1 pentamers, also known as capsomeres, were purified via a glutathione‐S‐transferase tag. Size exclusion chromatography indicated severe aggregation of the recombinant VP1 during enzymatic tag removal resulting from the introduction the hydrophobic epitopes. Inserts were modified to possess double aspartic acid residues at each end of the hydrophobic epitopes and a high‐throughput buffer condition screen was implemented with protein aggregation monitored during tag removal by spectrophotometry and dynamic light scattering. These analyses showed that the insertion of charged residues at the extremities of epitopes could improve solubility of capsomeres and revealed multiple windows of opportunity for further condition optimization. A combination of epitope design, pH optimization, and the additive l ‐arginine permitted the recovery of soluble VP1 pentamers presenting hydrophobic epitopes and their subsequent assembly into virus‐like particles.     

A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants

Introduction –  RNA quality and integrity are critical for many studies in plant molecular biology. High‐quality RNA extraction from grapevine and other woody plants is problematic due to the presence of polysaccharides, polyphenolics and other compounds that bind or co‐precipitate with the RNA. Objective  – To develop an optimised cetyltrimethylammonium bromide (CTAB)‐based protocol, to reduce the time and cost of extraction without reducing quality and yield of RNA extracted from polysaccharide‐rich tissues of several plants. Methodology  – Several changes were introduced to the original CTAB protocol. All centrifugation steps were carried out at 4°C, the sample weight was decreased and the concentrations of PVP‐40 and LiCl were increased reducing incubation time prior to RNA precipitation. This rapid CTAB protocol was compared with six different RNA extraction methods from three grapevine tissues, namely, in vitro plantlets, and leaves and mature canes from actively growing field vines. Results –  The rapid CTAB method gave high‐quality RNA in only 3 h at low cost with efficiency equal to or higher than that obtained with other time‐consuming and expensive protocols. The procedure was applied to RNA extraction from other grapevine tissues and other woody species including olive, lemon, poplar, chestnut, apple, pear, peach, cherry, apricot, plum and kiwi fruit. RNA of high quality could be isolated from all tissues and from all species. Conclusion –  The study has shown that the improvement of a CTAB‐based protocol allows the rapid isolation of high‐quality RNA from grapevine and many woody species. Copyright © 2008 John Wiley & Sons, Ltd.     

Improving the performance of Rosetta using multiple sequence alignment information and global measures of hydrophobic core formation

This study explores the use of multiple sequence alignment (MSA) information and global measures of hydrophobic core formation for improving the Rosetta ab initio protein structure prediction method. The most effective use of the MSA information is achieved by carrying out independent folding simulations for a subset of the homologous sequences in the MSA and then identifying the free energy minima common to all folded sequences via simultaneous clustering of the independent folding runs. Global measures of hydrophobic core formation, using ellipsoidal rather than spherical representations of the hydrophobic core, are found to be useful in removing non-native conformations before cluster analysis. Through this combination of MSA information and global measures of protein core formation, we significantly increase the performance of Rosetta on a challenging test set. Proteins 2001;43:1-11.     

甘肃国家重点保护野生植物资源保护利用研究

甘肃省分布27种国家重点保护野生植物资源,其中有国家一级保护植物苏铁银杏、红豆杉、水杉、珙桐、光叶珙桐、独叶草等7种,占国家51种的13．73％,国家二级保护植物岷江柏木、秦岭冷杉、大果云杉、巴山榧树、连香树、油樟、红豆树、野大豆、红椿、鹅掌楸、厚朴、凹叶厚朴、西康玉兰、水曲柳、香果树、瓣鳞花、榉树、水青树、喜树、发菜、虫草等21种,占国家202种的10．4％,研究甘肃本地分布的国家级保护植物的分布特点、生物多样性保护的战略对策、利用价值,为保护和利用提供科学依据。     

Influence of tryptophan tags on the purification of cutinase, secreted by a recombinant Saccharomyces cerevisiae, using cationic expanded bed adsorption and hydrophobic interaction chromatography

During cationic bed adsorption (EBA), with cutinase with varying length tryptophan tags (WP)(2)and (WP)(4), 33% and 10% of adsorption capacity and 80% and 32% eluted specific activity were observed in relation to wild type (wt)-cutinase in the conventional process. Therefore, as the hydrophobicity of the protein increases, it is important to integrate the EBA step with a hydrophobic interaction chromatography (HIC) process. As the length of the hydrophobic tag-(WP) increases from n = 2 to n = 4, the purification factor obtained by HIC was 1.8 and 2.2-fold higher than wt-cutinase. However, the recovery yield obtained in HIC decreases substantially as the length of hydrophobic tag increases (97%, 84% and 70% for wt-cutinase, cutinase-(WP)(2) and cutinase-(WP)(4)). The integration of two purification steps, EBA followed by HIC, resulted in the highest overall purity level for cutinase-(WP)(2), and the highest overall recovery yield for wt-cutinase. When optimizing the design of a hydrophobic tag fused to a protein secreted by Saccharomyces cerevisiae it must be considered that the cultivation parameters could impair the downstream process, and consequently the optimum tag is not necessarily the one that presents the highest purification factor in HIC.     

Improvement of the enantioselectivity and activity of lipase from Pseudomonas sp. via adsorption on a hydrophobic support: kinetic resolution of 2-octanol

Pseudomonas sp. lipase (PSL) was successfully immobilized on a novel hydrophobic polymer support through physical adsorption and the immobilized PSL was used for resolution of (R,S)-2-octanol with vinyl acetate as acyl donor. Enhanced activity and enantioselectivity were observed from the immobilized PSL compared with free PSL. The effects of reaction conditions such as temperature, water activity, substrate molar ratio and the amount of immobilized lipase were investigated. Under optimum conditions, the residual (S)-2-octanol was recovered with 99.5% enantiomeric excess at 52.9% conversion. The results also indicated that the immobilized PSL could maintain 94% of its initial activity even after reusing it five times.     

Dissipative particle dynamics simulation on the properties of the oil/water/surfactant system in the absence and presence of polymer

Dissipative particle dynamics is used to simulate the oil/water/surfactant system in the absence and presence of polymer. Structural properties, interfacial properties, and their dependence on the surfactant concentration, polymer concentration and oil/water ratio were investigated. The snapshots illustrate the variation of the structure of oil/water/surfactant system. In the presence of polymer, the interface is supersaturated at a lower surfactant concentration. The end-to-end distance increases with surfactant concentration and polymer chains but shows weak dependence on the oil/water ratio. The peak of density grows higher with surfactant concentration, but it is not affected by oil/water ratio. The density profiles of polymer grow higher with polymer chains, indicating that most of the polymer chains stay at the interface for stability. Interfacial thickness shows an adsorption of polymer/surfactant complexes at the interface, where the polymer is in an extended conformation at the interface. The formation of polymer/surfactant complexes is favourable for the decrease of oil/water interfacial tension.