Preparation of chloroplast DNA from pea plastids isolated in a medium of high ionic strength

A simple, rapid, and inexpensive method for the preparation and purification of chloroplast DNA (cpDNA) from pea has been developed. The crucial step is the isolation of chloroplasts in a medium of high ionic strength (I congruent equal to 1.40 M). CpDNA from pea prepared according to this method has successfully been used for restriction enzyme mapping, Southern transfers, and cloning.     

Molecular phylogeny of Heritiera Ait on (Sterculiaceae), a tree mangrove: variations in RAPD markers and nuclear DNA content

Random amplified polymorphic DNA (RAPD) markers are used to estimate interspecific variation among mangrove and non-mangrove Heritiera fomes, H. littoralis and H. macrophylla. All the species have 2n = 38 chromosomes, with minute structural changes distinguishing the karyotype of each species. Significant variation of 4C DNA content occurs at the interspecific level. Interspecific polymorphism ranged from 14.09% between H. fomes and H. littoralis to 52.73% between H. fomes and H. macrophylla. H. macrophylla showed wide polymorphism in the RAPD marker with H. littoralis (51.23%) and H. fomes (52.73%). Two distinct RAPD products obtained from OPA-10 (1000 bp) and OPD-15 (900 bp) found characteristic molecular markers in H. macrophylla , a species from a non-mangrove habitat. H. macrophylla was more distantly related to H. fomes [genetic distance (1-F) = 0.305] than to H. littoralis [genetic distance (1-F) = 0.273]. H. littoralis was of a closer affinity to H. fomes [genetic distance (1-F) = 0.218] than to H. macrophylla.     

DNA content of spermatozoa in different genetic lines of turkeys

The variability of DNA content in turkey spermatozoa of four different lines and its correlation with body weight and sperm concentration were studied. In lines selected for lower body weight the DNA content was 2.034 and 2.036 pg per spermatozoon. In lines selected for higher body weight the DNA content was 2.267 and 2.370 pg per spermatozoon. Sperm concentration in 1 mm³ of semen, however, was higher in lines with a lower body weight (6.08–6.21 million) in comparison with lines selected for higher body weight (5.46–5.67 million). The correlations between the DNA content and sperm concentration were negative (r ranged from ?.457 to ?.860).     

Selectivity in plant food consumption in the lizard Liolaemus lutzae from southeastern Brazil

Selectivity in the consumption of plant matter from the natural habitat by the tropidurid lizard Liolaemus lutzae, endemic to the beach habitats of restingas of southeastern Brazil, and the differences in the qualitative properties of the plants consumed were studied in the Barra de MaricÝ restinga, Rio de Janeiro State. The diets of 180 lizards were analysed and the plant species present in the stomachs and their frequencies were recorded. Only four of the 19 species which occur on the beach (Phylloxerus portulacoides, Althernantera maritima, Ipomoea littoralis and I. pes-caprae) were consumed by the lizard and their frequencies in the stomachs differed from that of occurrence. Analysis of the composition of the leaves of the 13 most abundant plant species indicated that the plants consumed by the lizards had the highest contents of water, total nitrogen, total sugar and the lowest amount of gross fibres. Thus, it appears that L. lutzae is not a generalist herbivore, but feeds selectively on those plants in its environment that are most easily digested and assimilated. A seletividade no consumo de algumas entre as espécies vegetais ocorrentes no habitat de praia pelo lagarto tropidur¡deo Liolaemus lutzae e, as diferenças nas propriedades qualitativas presentes nas plantas consumidas em relação às demais plantas mais abundantes do habitat foram estudada na restinga da Barra de Maricá, Sudeste do Brasil. A dieta de 180 lagartos foi analisada tendo sido anotadas as espécies de plantas presentes no estômago e suas respectivas frequências. Apenas quatro entre as 19 espécies registradas na praia (Phylloxerus portulacoides, Althernantera maritima, Ipomoea littoralis and I. pes-caprae) foram consumidas por L. lutzae. As frequências destas na dieta do lagarto diferiram da frequência com que as plantas ocorrem no habitat. A análise da composição das folhas de 13 entre as espécies de plantas mais frequentes indicou que as plantas consumidas pelo lagarto possuem as mais elevadas proporçôes de água, nitrogênio total, açúcar total e a menor proporção de fibras. Aparentemente L. lutzae não é um herbivoro generalista mas seleciona no seu ambiente as plantas as quais são mais facilmente digeridas e assimiladas.     

Estimating recent growth in the cuttlefish Sepia officinalis: are nucleic acid-based indicators for growth and condition the method of choice?

A laboratory calibration study was undertaken with juvenile Sepia officinalis (80-85 g initial wet weight) to investigate the effects of different food rations and different starving intervals on RNA/dry weight (DW) ratios and RNA/DNA ratios in cephalopod mantle muscle at two different temperatures. The digestive gland index was also used as an additional indicator of recent growth. High food rations and low temperature went along with high RNA/DW ratios and high RNA/DNA ratios. Starving resulted in a linear decline in growth performance and a concomitant decrease in RNA/DW and RNA/DNA ratio, with RNA/DNA ratios representing the growth data better. RNA/DNA ratios decreased faster at higher temperatures. A fluorimetric assay for nucleic acid analysis was optimized for cephalopod mantle tissues and yielded reproducible RNA/DNA ratios with a relative variance below 10%. Thus, it may be possible to use this estimator of recently encountered feeding regime for the evaluation of mortality rates of early teuthid paralarvae to eventually support stock management. Also, log relative digestive gland weight showed a strong relationship with starving time, but, surprisingly, not with temperature. Data from the two temperatures analyzed could be combined to form a common regression line of relative digestive gland index with starving time. This indicator for recent growth might be especially suitable for large specimens with a well-developed digestive gland.     

Rad52 recruitment is DNA replication independent and regulated by Cdc28 and the Mec1 kinase

Recruitment of the homologous recombination machinery to sites of double‐strand breaks is a cell cycle‐regulated event requiring entry into S phase and CDK1 activity. Here, we demonstrate that the central recombination protein, Rad52, forms foci independent of DNA replication, and its recruitment requires B‐type cyclin/CDK1 activity. Induction of the intra‐S‐phase checkpoint by hydroxyurea (HU) inhibits Rad52 focus formation in response to ionizing radiation. This inhibition is dependent upon Mec1/Tel1 kinase activity, as HU‐treated cells form Rad52 foci in the presence of the PI3 kinase inhibitor caffeine. These Rad52 foci colocalize with foci formed by the replication clamp PCNA. These results indicate that Mec1 activity inhibits the recruitment of Rad52 to both sites of DNA damage and stalled replication forks during the intra‐S‐phase checkpoint. We propose that B‐type cyclins promote the recruitment of Rad52 to sites of DNA damage, whereas Mec1 inhibits spurious recombination at stalled replication forks.     

Fluid replacement following dehydration reduces oxidative stress during recovery

To investigate the effects of hydration status on oxidative DNA damage and exercise performance, 10 subjects ran on a treadmill until exhaustion at 80% VO_2max during four different trials [control (C), 3% dehydration (D), 3% dehydration + water (W) or 3% dehydration + sports drink (S)]. Dehydration significantly decreased exercise time to exhaustion (D < C and S). Plasma MDA levels were significantly higher at pre-exercise in D than C. Plasma TAS was significantly lower at pre-exercise in C and S than in D, and was significantly lower in S than D at 60 min of recovery. Dehydration significantly increased oxidative DNA damage during exercise, but fluid replacement with water or sports drink alleviated it equally. These results suggest that (1) dehydration impairs exercise performance and increases DNA damage during exercise to exhaustion; and (2) fluid replacement prolongs exercise endurance and attenuates DNA damage.