PCR-based markers to differentiate the mitochondrial genomes of petaloid and male fertile carrot (Daucus carota L.)

Cytoplasmic male sterility (CMS) is essential for the development of highly adapted and uniform hybrid varieties of carrot and other crops. The most widely used type of CMS in carrot is petaloidy, in which the stamens are replaced by petals or bract-like structures. We have developed a series of mitochondria-specific PCR markers to distinguish cytoplasms inducing petaloidy (Sp) and male-fertility (N). The markers target the atp1,atp6, atp9, orfB (atp8), nad6 and cob loci from the mitochondrial genomes of a diverse collection of male fertile and petaloid carrots. We report 14 primer pairs that amplify marker fragments from either Sp or N cytoplasms and three primer pairs that amplify fragments with length polymorphism. The amplification products span sites of insertions, deletions or recombinations adjacent to or within the coding regions of the targeted genes. The markers reported here are useful tools to identify the type of cytoplasm in cultivated carrot and to evaluate variation in the mitochondrial genomes within the genus Daucus. This revised version was published online in August 2006 with corrections to the Cover Date.     

Crop nitrogen demand and canopy area expansion in winter wheat during vegetative growth

The effect of nitrogen (N) on crop growth and productivity is mediated through effects on both light interception (green crop area) and radiation use efficiency (RUE). The effects of N nutrition on these factors were studied using data on green area index (GAI), above-ground dry matter and N uptake from growth analysis measurements in winter wheat from a number of experiments in Denmark with different application rates of N. Only measurements taken prior to anthesis were used in the statistical analyses. The N uptake was found to be proportional to GAI, and to have an additional curvilinear response to dry matter implying decreasing N concentration with increasing dry matter. This supports the hypothesis that nitrogen is associated with both the green surfaces of the crop canopy and with the dry matter component. A model of GAI expansion is presented incorporating three limiting factors: an exponential increase in GAI in thermal time, a minimum leaf area ratio, and a minimum ratio of GAI to N content in above-ground dry matter. This simple function has potential to be used as a tool for targeting timing and rates of N fertilisation in relation to a desired development of GAI. Such N application strategies should also consider the nitrogen nutrition index (NNI), which was defined based on the relationship between N uptake and both GAI and dry matter. The response of RUE to NNI showed a curvilinear response with a tendency for saturation at high NNI. The design of N application strategies should therefore ensure that low NNI is avoided during the most productive periods in the growing season.     

Genotypes of Perennial Ryegrass (Lolium perenne L.) with High Anther-Culture Response through Hybridization

Anther-culture response in Lolium perenne was studied in the hybrid populations from three responsive diploid parent clones to obtain genotypes with superior anther-culture response through hybridization. Genetic variation for anther-culture response was significant both within and between the four hybrid families and their parents. A reciprocal effect was indicated for embryo formation and regeneration percentages for one of the cross combinations. Among 55 hybrid clones tested in two environments, 6 clones were identified with an anther-culture response level superior to their parents. While the parents produced one green plant or less per 100 anthers, these superior clones produced from 11 to 59 green plants/100 anthers, mainly as the result of improved embryo formation and improved percentages of regenerated plants being green. A significant correlation (r = 0.397^**) was observed between embryo formation and percentage of green plants. The results are discussed with regard to the possible transfer of anther-culture response to non-responsive breeding material through hybridization.     

A polycross pattern formula

Summary A formula defining a polycross pattern is introduced. The pattern appears to be a special latin square in which every clone has any one other clone as a neighbour and just once in each of the four directions North, South, East, and West, presenting an ideal polycross pattern. The formula holds good for n clones when n+1 is a prime.     

A completely balanced polycross design

Summary A polycross design for n² replicates of n distinct clones is introduced. The design consists of n distinct n x n polycross patterns, provided n+1 is a prime number.The design is completely balanced in that it has the following properties with respect to nearest neighbours: In anyone of the four main directions, North, South, East, and West, every clone has any other clone as a nearest neighbour exactly n times, while no clone has itself as a nearest neighbour. In any of the four intermediate directions, NorthEast, SouthEast, SouthWest, and NorthWest, every clone has any other clone as a nearest neighbour exactly n–2 times, and itself as a nearest neighbour exactly n–1 times.     

Segregation of Isozyme Marker Loci Amongst Androgenetic Plants of Lolium perenne L.

The segregation of isozyme markers has been examined in the green and albino androgentic progenies of two plants of Lolium perenne. Overall the pooled data for the loci PG1/2 and GOT/2 does not deviate from expectation, but there is considerable heterogeneity between families and between green and albino plants. Specific alleles predominate in the different progeny sets. The results may be accounted for by possible linked loci affecting anther culture response or of linked lethal genes.     

Dephosphorylation of the calcium pump coupled to counterion occlusion

P-type ATPases extract energy by hydrolysis of adenosine triphosphate (ATP) in two steps, formation and breakdown of a covalent phosphoenzyme intermediate. This process drives active transport and countertransport of the cation pumps. We have determined the crystal structure of rabbit sarcoplasmic reticulum Ca2+ adenosine triphosphatase in complex with aluminum fluoride, which mimics the transition state of hydrolysis of the counterion-bound (protonated) phosphoenzyme. On the basis of structural analysis and biochemical data, we find this form to represent an occluded state of the proton counterions. Hydrolysis is catalyzed by the conserved Thr-Gly-Glu-Ser motif, and it exploits an associative nucleophilic reaction mechanism of the same type as phosphoryl transfer from ATP. On this basis, we propose a general mechanism of occluded transition states of Ca2+ transport and H+ countertransport coupled to phosphorylation and dephosphorylation, respectively.     

Risk factors for nuisance barking in dogs

Objective   To determine the key variables related to the environment, animal and owner that influence nuisance barking by dogs in a city with a subtropical climate.
Design and Population   A case–control survey of dog owners in Brisbane, Queensland, was conducted using a questionnaire investigating key variables connected to nuisance barking. Owners of dogs exhibiting nuisance barking were obtained from a list of dogs being treated in a Brisbane behaviour clinic, and those of control dogs were selected from a telephone directory.
Results   Univariate analysis showed that animal, owner and environmental factors all potentially influence the occurrence of nuisance barking. Multivariate analysis identified the following factors, with the relevant odds ratios (OR) as significant: age of the dog (young dog vs old dog, OR 11.2); multiple dogs in the household vs single (OR 5.6); origin of the dog (home bred vs obtained from breeder or friend, OR 4.0); type of dog, (herding vs other types, OR 3.2) and dog with access to the home vs dog without access (OR 2.5).
Conclusion   The greatest risk for nuisance barking occurs with a young dog of the herding type that is home bred and with access to the house in a multiple dog household.