Calcium transport and ATPase activity in a microsomal vesicle fraction from corn roots

Abstract. An investigation has been made of methods for isolating membrane vesicles from corn ( Zea mays L.) roots active in calcium transport and K⁺-stimulated ATPase. Pretreating and grinding the roots at room temperature with EGTA and fusicoccin increases basal ATPase activity. Improvement in Ca²⁺ uptake requires isolation of a scaled vesicle fraction by the method of Sze(1980). Sorbitol is superior to sucrose as an osmoticant. The pH optimum for Ca²⁺ uptake is 7.5. whereas that for associated ATPase activity is 6.5. Calmodulin strongly stimulates Ca²⁺ uptake in a process little affected by uncouplers and ATPase inhibitors, but blocked by chlorpromazine. Fusicoccin gives less stimulation of Ca²⁺ uptake which is sensitive to uncouplers, and is dependent upon isolation with fusicoccin present. It appears that the sealed vesicle fraction may possess two Ca²⁺ transport systems: a calmodulin-activated Ca²⁺-transporting ATPase, and a Ca²⁺/H⁺ antiport coupled through the protonmotive force to a fusicoccin-stimulated H⁺-ATPase.     

Density of Parasites in Various Organs and the Relation to Numbers of Trypomastigotes in the Blood during Acute Infections of Trypanosoma cruzi in Mice*

Quantitative methods were used to (a) determine the density of Trypanosoma cruzi in organs of CF₁ mice following intraperitoneal inoculation of 50,000 trypomastigotes of a Brazil strain of T. cruzi and (b) study the relation of the numbers of these intracellular stages to the numbers of trypomastigotes in the blood. Tissue stages (predominantly amastigotes) in heart, skeletal muscle (triceps), diaphragm, cerebrum, cerebellum, and musculature of stomach, duodenum, esophagus, jejunum, cecum, and rectum increased in numbers during the 1st 3 weeks of infection, reached maximum density 21–28 days after inoculation and subsequently declined in numbers until mice were histologically negative for intracellular parasites by 30–40 days. The density of tissue stages in the urinary bladder, uterine body, and ileum was similar with the exception that maximum numbers of parasites were observed slightly earlier at 15 days. The greatest density of intracellular stages was seen in heart, urinary bladder, diaphragm, and triceps muscle where mean counts of 44.6–60.0 × 10⁶ parasites/cc of muscle were recorded while maximum density of parasites in the uterine body, cerebrum, stomach, cerebellum, duodenum, esophagus, jejunum, ileum, cecum, and rectum was 13.0 × 10⁶/cc of muscle or less. Amastigotes were not observed in sections of lymph node, thymus, salivary glands, liver, spleen, or kidney and only a single pseudocyst containing 5 amastigotes was seen in lung. With the exception of the brain and lung, intracellular parasites were located exclusively in the musculature. Trypomastigotes in the blood increased during the 1st 3 weeks of infection, reached maximum numbers 21–28 days after initiation of infection, and subsequently decreased until by 30–40 days parasites were observed only rarely in the blood of a few animals. Thus generally close correlation was noted between the numbers of intracellular stages of T. cruzi in the organs and trypomastigotes in the blood throughout acute Chagas’ disease in mice as evidenced by the concomitant increase in numbers of both stages, the coincidence of days of maximum parasite levels, and the simultaneous decline in numbers of both stages. The mean number of parasites/pseudocyst section varied in the organs studied. Of the 15 positive organs studied, the pseudocyst sections in skeletal muscle contained the highest mean number of parasites (64.3 parasites/pseudocyst section) and those pseudocyst sections seen in the musculature of the small intestine contained the lowest mean number (5.5–6.8 parasites/pseudocyst section respectively in ileum and jejunum). Serial sections of skeletal muscle, heart, urinary bladder, and stomach revealed the largest pseudocysts in skeletal muscle while those in the musculature of the urinary bladder were the smallest.     

Macroinvertebrate size-distributions of two contrasting freshwater macrophyte communities

SUMMARY.

• 1 Macroinvertebrates (>0.1 mg, fresh weight) were collected every 2 weeks for 18 weeks from weedbeds dominated by either the macroalga Chara or rooted plants (mostly Isoetes sp., Potamogeton filiformis, Nuphar variegatum and Myriophyllum exalbescens) in Narrow Lake, Alberta, Canada. Significant differences in total biomass, taxonomic composition, and size-structure of the macroinvertebrate community were found between the two weedbed types.
• 2 Total biomass of macroinvertebrates in the Chara beds (seasonal mean 26.4 g m^?2) was consistently higher than in the rooted-plant weedbeds (seasonal mean 7.6 g m^?2).
• 3 The macroinvertebrate community of the Chara beds was dominated by chironomids, anisopterans, gastropods and sphaerid clams whereas amphipods dominated the community in the rooted-plant weedbeds.
• 4 On average, the proportion of total biomass in the various size-classes (biomass size-spectrum) of the macroinvertebrate community in Chara beds varied very little between 1 and 512 mg. In rooted plant weedbeds, the spectrum showed a strong peak in the 4–8 mg size-class, a weak peak in the 32–64 mg size-class, and proportionately low biomass in the 128–256 mg and 256–512 mg size-classes. The slope of the normalized size-spectrum for the macroinvertebrate community of the Chara beds (-0.89) was significantly different from that of the rooted plant community (-1.11). The biomass of large organisms (>64 mg) was 10 times greater in the Chara beds than in the rooted-plant weedbeds.
• 5 The species composition of aquatic macrophyte beds can greatly influence the abundance, taxonomic composition, and size-structure of the littoral zone macroinvertebrate community. Therefore, any changes in plant species composition of weedbeds, through introduction of exotic plant species or introduction (or loss) of an efficient grazer (e.g. crayfish), have the potential to alter greatly the structure of the macroinvertebrate community in a lake, with unknown consequences to fish and waterfowl dependent upon these prey.

     

Wild and agronomically important Agave species (Asparagaceae) show proportional increases in chromosome number,genome size,and genetic markers with increasing ploidy

Agave (Asparagaceae) includes cultivated and wild varieties of henequen used for hard fibre production. As part of a breeding programme to improve Agave production, species with different ploidy levels were genetically characterized: two diploids [A. tequiliana Weber and the hybrid H11648 ((A. amaniensis Trel. & Nowell × A. angustifolia Haw.) × A. amaniensis)], a triploid (A fourcroydes Lem. var. kitam ki), a tetraploid (A. angustifolia var. letona), three pentaploids (A. fourcroydes var. sac ki, A. fourcroydes var. yaax ki, and A. sisalana Perrine), and two hexaploids (A. angustifolia var. chelem ki from two locations). Chromosome spreading was used to determine the chromosome number, flow cytometry was employed to measure the genome size, and fluorescent in situ hybridization was performed using 45S and 5S ribosomal DNA (rDNA) and the telomeric sequences (TTAGGG)_n and (TTTAGGG)_n as genetic markers. There were proportional increases with ploidy level of the following: (1) chromosome number (from diploid 2n = 2x = 60 to hexaploid 2n = 6x = 180), including the number of large and small chromosomes in the bimodal karyotype of Agave; (2) genome size, with a mean monoploid genome size (1Cx) of 7.5 pg (range, 7.36–7.61 pg); and (3) the number and distribution of 45S and 5S rDNA loci, with one locus of each per basic, monoploid genome. Thus there was complete additivity in genome structure with increasing ploidy, as reported in some angiosperm polyploids. However, as other analyses of polyploids have revealed a decrease in 1Cx values with increased ploidy, possible explanations for the observed genomic stability were considered. With the (TTAGGG)_n probe, the signal was localized at the telomeres, consistent with published data showing that many species in the order Asparagales have this type of telomere sequence. It is speculated that sporadic telomeric signals using the (TTTAGGG)_n probe are probably derived from either errors in telomerase activity or relic ancestral‐type telomeric sequences. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 215–222.