Lipid composition and food quality of some freshwater phytoplankton for cladoceran zooplankters

The nutritional value of several planktonic algae was testedby means of feeding trials with three cladoceran zooplankters.The algae were monocultures and included two blue-greens, fourgreens and four flagellates with a size range of 5–48µm. The specific growth rates of the zooplankters werechosen as the measure of the nutritional value of the algae.The three cladocerans showed large differences in growth ratein the different algae, but the two cryptomonads were withoutdoubt best suited as food for all. The fatty acid compositionfor the cryptomonads were different from the other algae. Theycontained high percentages of the polyunsaturated fatty acids20:5æ3 (EPA) and 22:6æ3 (DHA), which also are commonin fish. It is suggested that the lipid composition is a probablefactor determining the nutritional quality of the algae.     

Induction and activity of NO synthase in bone-marrow-derived macrophages are independent of Ca2+.

The aim of the present study was to analyse whether an increase in the intracellular free Ca2+ concentration ([Ca2+]i) plays a role as a signal mediating synthesis of nitric oxide (NO) in bone-marrow-derived macrophages, either by stimulating induction of NO synthase or by regulating the activity of the enzyme. Therefore we compared the effects of various synthetic analogues of bacterial lipopeptide and of lipopolysaccharide (LPS) on NO production (assessed as nitrite formation during an incubation for 24 h) and on [Ca2+]i [measured with the fluorescent probe indo-1 (1-[2-amino-5-(6-carboxyindol-2-yl)phenoxy]-2- 2-(2'-amino-5'-methylphenoxy)ethane-NNN'N'-tetra-acetic acid)]. Strongly dissociating effects were evoked on nitrite formation and on [Ca2+]i by the stimuli. LPS was preferentially effective on nitrite formation, whereas the Ca2+ ionophore ionomycin and AlF3 induced increases only in [Ca2+]i. The lipopeptides N-palmitoyl-(S)-[2,3-bis(palmitoyloxy)-(2RS)- propyl]-(R)-cysteinylalanylglycine, N-palmitoyl-(S)-[2,3-bis(palmitoyloxy)- (2RS)-propyl]-(R)-cysteinylseryl-lysyl-lysyl-lysine and (S)-(1,2- dicarboxyhexadecyl)ethyl-N-palmitoylcysteinylseryl-lysyl-lys yl-lysine stimulated both parameters, but the maximal effects on nitrite formation and the shape of the dose-response curves did not parallel the effects on [Ca2+]i. Reduction of extracellular Ca2+ with EGTA significantly inhibited increases in [Ca2+]i, but did not change nitrite formation. Furthermore, NO synthesis in the cytosolic fraction of stimulated macrophages was not affected by Ca2+ over the concentration range 10 nM-2 microM. We conclude that increases in [Ca2+]i are not required for NO production in bone-marrow-derived macrophages. Thus the cellular regulation of NO production strikingly differs from that in the vascular endothelium, brain and adrenal gland.     

Thidiazuron-induced adventitious shoot regeneration of sweetpotato (Ipomoea batatas)

Adventitious shoots of sweetpotato (Ipomoea batatas L. Lam.) were produced in vitro using a two-stage culture method. Petiole explants were incubated on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (0.2 mg·liter^−1) for 3 d, and transferred to MS medium with thidiazuron (0 to 0.4 mg·liter^−1). Shoot regeneration was observed in most explants (78.2%) of genotype PI 318846-3 within 28 days when cultured on thidiazuron at 0.2 mg·liter^−1. Histological studies of cultured petiole explants showed meristematic activity within cells of vascular bundles and throughout the ground tissue. Explants isolated from apical leaves exhibited higher shoot regeneration frequency than those isolated from the basal portion of the shoot. Leaf lamina explants exhibited lower frequency of regeneration than petiole explants. In contrast to thidiazuron, the use of zeatin riboside, and kinetin resulted in a lower frequency of shoot regeneration although more sweetpotato genotypes could be regenerated using either of these two cytokinins. The sweetpotato plants regenerated using thidiazuron grew vigorously and rooted easily when transferred to the greenhouse.     

Stimulation of Pod and Ovule Growth of Soybean, Glycine max (L.) Merr. by 6-Benzylaminopurine

Flowers at distal nodes on soybean racemes usually fail to setpods and subsequently abscise. Physiological and histologicalstudies were performed to determine the influence of 6-benzylaminopurine(BAP) on distal pod development. The pedicels of fully openedflowers on terminal racemes of field-grown IX93-100 soybeanplants were treated three times with 200 mg kg^-1 BAP in lanolinover a 6-d period. Racemes were then excised and ³²P uptakewas recorded for each flower position within a raceme; histologicalfeatures of pedicels and ovules also were determined. Applicationof BAP increased pod and ovule length, width and weight at allfour distal nodes (D, D-1, D-2, D-3) relative to controls treatedwith lanolin. Length and width of parietal endosperm cells weresmaller in BAP-treated ovules at the most proximal node beingstudied (D-3), and greater numbers of parietal endosperm cellswere observed at D-1 and D-3 nodes when compared to lanolincontrols. Smaller amounts of starch were found in suspensorcells, endosperm, and integuments of lanolin-treated ovules,and starch depletion over time was observed within starch sheathsof pedicels from lanolin-treated pods when compared to BAP-treatedtissues. BAP-treated racemes had more ³²P uptake at the fourmost distal nodes. A higher rate of uptake (cpm mg^-1 f. wt)was evident in ovules than in ovary tissues. These results suggestthat for racemes otherwise destined to abscise, applicationof BAP promotes pod set and growth by stimulating ovule development.Copyright1993, 1999 Academic Press Pod, ovule soybean, abscission, 6-benzylaminopurine     

A rapid method for extraction of cotton (Gossypium spp.) genomic DNA suitable for RFLP or PCR analysis

Extraction of high-quality genomic DNA fromGossypium (cotton) species is difficult due to high levels of polysaccharide, oxidizable quinones, and other interfering substances. We describe a procedure that consistently permits isolation of cotton genomic DNA of satisfactory size and quality for RFLP and PCR analysis, as well as for most routine cloning applications. Several antioxidants, phenol-binding reagents, and phenol oxidase inhibitors are used throughout the procedure, and most polysaccharides are eliminated early in the procedure by isolation of nuclei.     

Coevolution of Bacteriophage PP01 and Escherichia coli O157:H7 in Continuous Culture

The interaction between Escherichia coli O157:H7 and its specific bacteriophage PP01 was investigated in chemostat continuous culture. Following the addition of bacteriophage PP01, E. coli O157:H7 cell lysis was observed by over 4 orders of magnitude at a dilution rate of 0.876 h⁻¹ and by 3 orders of magnitude at a lower dilution rate (0.327 h⁻¹). However, the appearance of a series of phage-resistant E. coli isolates, which showed a low efficiency of plating against bacteriophage PP01, led to an increase in the cell concentration in the culture. The colony shape, outer membrane protein expression, and lipopolysaccharide production of each escape mutant were compared. Cessation of major outer membrane protein OmpC production and alteration of lipopolysaccharide composition enabled E. coli O157:H7 to escape PP01 infection. One of the escape mutants of E. coli O157:H7 which formed a mucoid colony (Mu) on Luria-Bertani agar appeared 56 h postincubation at a dilution rate of 0.867 h⁻¹ and persisted until the end of the experiment (~200 h). Mu mutant cells could coexist with bacteriophage PP01 in batch culture. Concentrations of the Mu cells and bacteriophage PP01 increased together. The appearance of mutant phage, which showed a different host range among the O157:H7 escape mutants than wild-type PP01, was also detected in the chemostat culture. Thus, coevolution of phage and E. coli O157:H7 proceeded as a mutual arms race in chemostat continuous culture.