Isolation and fusion studies on protoplasts from pollen tetrads

Summary different enzymes were tested for isolation of intact protoplasts from pollen tetrads. About 80% isolation was achieved from pollen tetrads of Cajanus cajan and Zea mays and about 60% from Luffa cylindrica and Lycopersicon esculentum after 4 h of treatment with 5% cellulase. When these mononucleate protoplasts were incubated in presence of 0.05 M CaCl₂ in 0.3 M glucose at pH 10.5, 70–80% fusion was achieved. Fusion was rare in sodium nitrate solutions.This work was supported by a grant from the Indian Council of Agricultural Research, New Delhi (No. 1-32/73, Ed. I) in the form of a Senior Fellowship awarded to P. C. D.Work carried out at Molecular Cytogenetics Research Unit, Dept. of Genetics and Plant Breeding, Banaras Hindu University, Varanasi, India.     

The effect of actinomycin D on the production of viruses by the protoplasts of Chinese cabbage infected in vitro by the yellow mosaic virus of turnips]

Chinese Cabbage (Brassica sinensis L. var. Cantonner) protoplasts were infected by Turnip yellow mosaic virus (TYMV) and inoculated in the presence or absence of actinomycin D. Virus production was determined 40 hrs. after inoculation, the time required for the virus replication cycle to be terminated. While actinomycin D had no effect on TYMV production when present at a concentration of 1 microng/ml, a 50 to 80% inhibition of virus production was noticed at concentrations of the order of 5 to 10 microng/ml, and the inhibition reached 90% with 25 microng/ml.     

Reversion of Lactobacillus lactis protoplasts

More than 99% of L. lactis cells have been converted to protoplasts upon digestion of cell walls with mutanolysin (N-(acetyl)muramidase). Functional protoplasts were obtained even with the lowest level of the enzyme that was used (0.1 U.ml-1 of the cell suspension) and after incubation at 37 degrees C for 2 min. The regeneration of the polymerized cell wall appears to be induced by a cell homogenate of the same organism.     

Factors affecting high-frequency fungal protoplast fusion

Summary Factors influencing the fusion frequency of protoplasts have been examined with auxotrophic mutants ofAspergillus nidulans. The optimum conditions were a total of 5 to 15 million protoplasts per ml, and 25% polyethylene glycol (PEG) 4000 or 6000 as fusogenic agent in 10 to 100 mM CaCl₂ solution.     

Transfer of isolated nuclei into protoplasts of Aspergillus nidulans.

Nuclei were isolated from protoplasts of a haploid auxotrophic Aspergillus nidulans strain. Transformation of protoplasts prepared from a complementary haploid auxotrophic strain with these purified nuclei resulted in both heterokaryotic and diploid colonies. The nutritionally-complementing colonies appeared at a frequency of 5 x 10(-7) to 10(-8).     

Airborne pollen content in the atmosphere of central Italy (1982–1986)

Summary This report describes qualitatively and quantitatively the content of pollen in the atmosphere of central Italy during the five years 1982–1986. Total production in this period showed fluctuations depending on the flowering seasons of the anemophilous taxa. The season of maximum pollen concentration was from April to July, with a prevalence of arboreal pollen in thefirst months, and of pollen from herbaceous plants in the last months of the year. During the five years of research more than 81 different types of pollen grains were recorded and identified. In both the cities investigated Curpessaceae/Taxaceae, Fagaceae, Oleaceae Gramineae and Urticaceae were responsible for the greatest amounts of pollen.     

Transfer of isolated nuclei into protoplasts ofAspergillus nidulans

Nuclei were isolated from protoplasts of a haploid auxotrophicAspergillus nidulans strain. Transformation of protoplasts prepared from a complementary haploid auxotrophic strain with these purified nuclei resulted in both heterokaryotic and diploid colonies. The nutritionally-complementing colonies appeared at a frequency of 5×10^–7 to 10^–8.     

Inhibition of insect larval growth by phenolics in glandular trichomes of tomato leaves

Summary We have found that the foliar tetracellular glandular trichomes (tetrads) of the tomato plant,Lycopersicon esculentum Mill., contribute significantly to the antibiotic effect of the leaf against the fruitwormHeliothis zea (Boddie), as measured by reduction in larval growth. This effect is attributable to phenolic compounds localized within the tetrads. We have found that the cellular fluid of the tetrads is particularly rich in the flavonol glycoside rutin, accompanied by lesser amounts of other phenolics.Acknowledgments. Supported by CFMTAB grant No. 21, and USDA-SEA grant No. 5901-0269. We thank Shell Development Company of Modesto, California, for supplying eggs ofH. zea, J. S. Rolston for preparation of the figures, and B. C. Campbell for the SEM photograph in fig. 1.     

Are rat alveolar macrophages capable of phagocytizing pollen grains?]

Plane-tree pollen grains were incubated in vitro with alveolar macrophages from inbred Rats, and the possibility of phagocytosis was investigated. No phagocytosis was observed even after 48 hrs incubation. But alveolar cells were bound to pollen grains generally at the apertures. This binding did not require the Fc receptor on the macrophage membrane.     

A comparative study of proteoglycans from bovine lung,trachea, tracheal mucosa,and aorta

Summary Proteoglycans were isolated from bovine lung, trachea, tracheal mucosa, and aorta by dissociative extraction with 4M guanidinium hydrochloride. Fractionation of these tissue extracts by cesium chloride density centrifugation and gel chromatography allowed the isolation from each extract of a high molecular weight fraction consisting mainly of proteochondroitin sulfate and hyaluronic acid.Acknowledgment. This work was supported by a National Heart, Lung, and Blood Institute grant to K. Ehrlich (HL21617).     

Turnover of 5-hydroxydopamine in adrenergic nerves

Summary In adrenergic nerve endings of the guinea-pig vas deferens the population of small granular vesicles increases from 19% in control animals to 80–90% 1–3 h after the administration of 5-hydroxydopamine, and gradually declines to control values in 10 days. Large granular vesicles were also loaded but the loss of enhanced granulation was more rapid than in the small granular vesicles.M. R. C. Scholar. I am greatly indebted to Prof. G. Burnstock, Prof. G. Gray and Dr G. Gabella for helpful advice and criticism. The technical assistance of J. Barron and G. Barrett is gratefully acknowledged.     

The use of cell free extracts derived from fungal protoplasts in the study of aflatoxin biosynthesis.

A supernatant fraction derived from protoplasts of Aspergillus flavus was shown to be capable of converting both sterigmatocystin and versiconal hemiacetal acetate to aflatoxin B1. Versicolorin A was not converted under the same conditions.     

Voltage variation inLilium longiflorum pistils induced by pollination

Summary Voltage variations inLilium longiflorum pistils arose 120 min after pollination with pollen of the same flower. No action potential was registered if pollen ofPetunia hybrida was placed on the stigma. Apparently the voltage variations were evoked by the germination of the pollen grains, and/or the penetration of the style by the pollen tubes.     

Pollen carryover and neighbourhood inRanunculus bulbosus

The pattern of gene flow has a major influence on the spatial scale of evolutionary processes. In plant populations pollen carryover will influence the pattern of gene flow. InRanunculus bulbosus L. pollen carryover was found to occur over a maximum of 5 flowers, with seed set per flower decreasing in a curvilinear fashion with increasing visit number in a sequence of visits by a bee after pollen pick-up from a donor plant. The effect of pollen carryover is to increase both neighbourhood area and neighbourhood size by two-fold.     

New scanning electron microscopy contribution to the study of yeast protoplasts]

Study by scanning electron microscopy enables us to describe with precision the morphology of protoplasts. We can confirm that the wrinkles more or less perceptible on the protoplasts studied, do belong to the morphology of those just mentioned, that the presence or the absence of globulous component is connected with the physiological activity of the cell and that it is possible to distinguish the different protoplasts thanks to their morphological aspect.     

The use of cell free extracts derived from fungal protoplasts in the study of aflatoxin biosynthesis

Summary A supernatant fraction derived from protoplasts ofAspergillus flavus was shown to be capable of converting both sterigmatocystin and versiconal hemiacetal acetate to aflatoxin B₁. Versicolorin A was not converted under the same conditions.     

Selective killing of smooth muscle cells in culture by the ricin A-chain conjugated with monoclonal antibodies to a cell surface antigen via a dextran bridge

Monoclonal antibodies to a surface antigen of the modulated smooth muscle cells originally isolated from the rat aorta media were conjugated with ricin A-chain via an oxidized dextran bridge. The interaction of cultured cells with the conjugates obtained and with control substances was monitored following incorporation of 14C-leucine radioactivity. It was found that 14C-leucine incorporation was suppressed by 80-90% at a conjugate concentration of 10(-6)-10(-7) M. Antigen-negative cells (line IAR; rat hepatocytes) were insensitive to the conjugate at any concentration used. Control use of purified ricin A-chain, native or oxidized dextran, specific and nonspecific IgG did not affect normal 14C-leucine incorporation. The data obtained may be useful for designing targeted drug transport systems and for selective screening of modulated smooth cells in vascular pathology models in vivo.     

Identification of 2 phosphorylated amino acids in chicken bone phosphoproteins]

Phosphoproteins of Chicken bone have been extracted by 0.5 M EDTA pH 7.5. Their amino acid composition was similar to that of phosphoproteins of other calcified tissues. The crude EDTA extract contained 80 to 90% proteins, only 70% of the total organic phosphorus was bound to the proteins. We have studied and identified two phosphorylated components. o-phosphoserine and o-phosphothreonine have been identified by amino acid analysis at pH 1.7 from a partial acid hydrolysate, and confirmed by the liberation of the parent amino acids after total hydrolysis. In addition, phosphorus was found equimolecular to both of them. The presence of these phosphorylated groups is important for an understanding of the role of these proteins in the mechanism of mineralization.     

The isolation,culture and organ differentiation from mesophyll protoplasts ofMollugo nudicaulis Lam., a C4 species

Summary Isolation and culture of mesophyll protoplasts fromMollugo nudicaulis, a C₄ species, is reported. Protoplasts developed into callus with root formation. However, no shoot was differentiated. This work was carried out as a part of our overall objective of inducing somatic fusion between the protoplasts of C₃ and C₄ species ofMollugo.     

Genotypic variation in foraging responses to environmental stimuli by honey bees,Apis mellifera

We experimentally tested a model predicting that colony-level genotypic diversity contributes to colonylevel foraging flexibility in honey bees. We established a colony into which we placed individually marked workers from three genetically distinct groups. The colony was placed in an enclosure that contained feeding stations with pollen and sugar syrup. Foraging resources, stores of pollen and the quantity of brood within the colony were varied temporally. Individual foragers switched between resource types in response to changes in relative resource quality and colony need, demonstrating flexibility in resource choice at the individual level. However, genetic groups within the colony varied in their tendency to collect pollen versus nectar, and in lability of response to changes in foraging stimuli. Our data suggest that within-colony genotypic diversity contributes to a resilient foraging response to environmental variation.