Genomic analysis of genetic markers associated with inherited cardiomyopathy (round heart disease) in the turkey (Meleagris gallopavo)

In turkeys, spontaneous cardiomyopathy or round heart (RH) disease is characterised by dilated ventricles and cardiac muscle hypertrophy. Although the aetiology of RH is still unknown, the disease can have a significant economic impact on turkey producers. In an initial attempt to identify genomic regions associated with RH, we utilised the chicken genome sequence to target short DNA sequences (sequence-characterised amplified regions, SCARs) identified in previous studies that had significant differences in frequency distribution between RH+ and RH- turkeys. SCARs were comparatively aligned with the chicken whole-genome sequence to identify flanking regions for primer design. Primers from 32 alignments were tested and target sequences were successfully amplified for 30 loci (94%). Comparative re-sequencing identified putative SNPs in 20 of the 30 loci (67%). Genetically informative SNPs at 16 loci were genotyped in the UMN/NTBF turkey mapping population. As a result of this study, 34 markers were placed on the turkey/chicken comparative map and 15 markers were added to the turkey genetic linkage map. The position of these markers relative to cardiac-related genes is presented. In addition, analysis of genotypes at 109 microsatellite loci presumed to flank the SCAR sequences in the turkey genome identified four significant associations with RH.     

Construction of a quinoa (Chenopodium quinoa Willd.) BAC library and its use in identifying genes encoding seed storage proteins

Quinoa (Chenopodium quinoa Willd.) is adapted to the harsh environments of the Andean Altiplano region. Its seeds have a well-balanced amino acid composition and exceptionally high protein content with respect to human nutrition. Quinoa grain is a staple in the diet of some of the most impoverished people in the world. The plant is an allotetraploid displaying disomic inheritance (2n=4x=36) with a di-haploid genome of 967 Mbp (megabase pair), or 2C=2.01 pg. We constructed two quinoa BAC libraries using BamHI (26,880 clones) and EcoRI (48,000 clones) restriction endonucleases. Cloned inserts in the BamHI library average 113 kb (kilobase) with approximately 2% of the clones lacking inserts, whereas cloned inserts in the EcoRI library average 130 kb and approximately 1% lack inserts. Three plastid genes used as probes of high-density arrayed blots of 73,728 BACs identified approximately 2.8% of the clones as containing plastid DNA inserts. We estimate that the combined quinoa libraries represent at least 9.0 di-haploid nuclear genome equivalents. An average of 12.2 positive clones per probe were identified with 13 quinoa single-copy ESTs as probes of the high-density arrayed blots, suggesting that the estimate of 9.0× coverage of the genome is conservative. Utility of the BAC libraries for gene identification was demonstrated by probing the library with a partial sequence of the 11S globulin seed storage protein gene and identifying multiple positive clones. The presence of the 11S globulin gene in four of the clones was verified by direct comparison with quinoa genomic DNA on a Southern blot. Besides serving as a useful tool for gene identification, the quinoa BAC libraries will be an important resource for physical mapping of the quinoa genome.     

Interaction between Paranosema locustae and Metarhizium anisopliae var. acridum, two pathogens of the desert locust, Schistocerca gregaria under laboratory conditions

The interaction between two pathogens, the microsporidian Paranosema locustae Canning and the fungus Metarhizium anisopliae var. acridum Driver and Milner was studied under laboratory conditions in an attempt to develop an improved method of microbial control for the desert locust, Schistocerca gregaria Forsk?l. Fifth-instar locust nymphs, reared in the laboratory, were treated with various concentrations of one of the two pathogens or with both pathogens. The numbers of locusts killed were recorded each day and the production of pathogen spores within the dead locusts was assessed at the end (day 21) of each experiment. Locust nymphs treated with both P. locustae and M. anisopliae died sooner than nymphs infected with only one of the pathogens. At the lower concentrations of pathogen tested, the effects of the two pathogens were additive. At the higher concentrations the combined effects were synergistic. In terms of locust mortality, there was no evidence of any antagonistic effects between the two pathogens. However, the production of spores by P. locustae was reduced considerably when the host insects were infected also with M. anisopliae. For example, nymphs treated initially with P. locustae and then treated 3 and 10 days later with M. anisopliae produced 3-20 times and 2.5-8 times fewer spores, respectively, than nymphs treated only with P. locustae. Hence, in areas where M. anisopliae is applied, the natural persistence of P. locustae in the local grasshopper and locust populations may be diminished.     

PHYSIOLOGY WITHOUT RESTRAINT IN DIVING MAMMALS

Forced submersions and natural dives are compared with regard to heart rate, blood distribution and O₂ consumption. Heart rate has been used as a primary indicator of blood distribution. However, often the dive heart rate has been compared to inappropriate surface conditions, and it is suggested that it should be compared to resting apneusis in the arrhythmic breathing animal. Furthermore, natural dives are frequently short relative to the breathhold capacity of a diver, and often during these dives there is little change in heart rate or apparent kidney function from the resting state. Calculations are made to determine the breathhold limit without O₂ conservation measures. Further calculations are made to estimate the expected metabolism and metabolite production during an exceptionally long dive in a seal when O₂ conservation must be invoked. These results are compared to known values obtained from a Weddell seal that made a 61 min dive. Based on the results of these observations and calculations, three possible models of blood distribution during natural dives are discussed. This discussion shows the limitation of present data and the type of data needed to resolve the question of how aquatic vertebrates manage O₂ stores during dives of varying durations.     

Concerted transpositions of mobile genetic elements coupled with fitness changes in Drosophila melanogaster

In an inbred low-activity (LA) strain of Drosophila melanogaster with a low level of fitness and a complex of inadaptive characters, in situ hybridization reveals an invariant pattern of distribution of three copia-like elements (mdg-1, mdg-3, and copia). Rare, spontaneous, multiple transpositions of mobile elements in the LA strain were shown to be coupled with a drastic increase of fitness. A changed pattern of various types of mobile elements was also observed on selecting the LA strain for higher fitness. High-fitness strains show transpositions of mobile elements to definite chromosomal sites ("hot spots"). Concerted changes in the location of three different mobile elements were found to be coupled with an increase of fitness. The mdg-1 distribution patterns were also examined in two low-fitness strains independently selected from the high-fitness ones. Fitness decrease was accompanied by mdg-1 excision from the hot spots of their location usually detected in the high-fitness strains. The results suggest the existence of a system of adaptive transpositions of mobile elements that takes part in fitness control.