Rapid and efficient screening of a Representational Difference Analysis library using reverse Southern hybridisation: Identification of genetic differences between Haemophilus parasuis isolates

Representational Difference Analysis (RDA) is an established technique used for isolation of specific genetic differences between or within bacterial species. This method was used to investigate the genetic basis of serovar-specificity and the relationship between serovar and virulence in Haemophilus parasuis. An RDA clone library of 96 isolates was constructed using H. parasuis strains H425(P) (serovar 12) and HS1967 (serovar 4). To screen such a large clone library to determine which clones are strain-specific would typically involved separately labelling each clone for use in Southern hybridisation against genomic DNA from each of the strains. In this study, a novel application of reverse Southern hybridisation was used to screen the RDA library: genomic DNA from each strain was labelled and used to probe the library to identify strain-specific clones. This novel approach represents a significant improvement in methodology that is rapid and efficient.

Identification and immunogenicity of an immunodominant mimotope of Avibacterium paragallinarum from a phage display peptide library

Avibacterium paragallinarum is the causative agent of infectious coryza. The protective antigens of this important pathogen have not yet been clearly identified. In this paper, we applied phage display technique to screen the immunodominant mimotopes of a serovar A strain of A. paragallinarum by using a random 12-peptide library, and evaluated the immunogenicity in chickens of the selected mimotope. Polyclonal antibody directed against A. paragallinarum strain 0083 (serovar A) was used as the target antibody and phage clones binding to this target were screened from the 12-mer random peptide library. More than 50% of the phage clones selected in the third round carried the consensus peptide motif sequence A-DP(M)L. The phage clones containing the peptide motif reacted with the target antibody and this interaction could be blocked, in a dose-dependent manner, by A. paragallinarum. One of the peptide sequences, YGLLAVDPLFKP, was selected and the corresponding oligonucleotide sequence was synthesized and then inserted into the expression vector pFliTrx. The recombinant plasmid was transferred into an expression host Escherichia coli GI826 by electroporation, resulting in a recombinant E. coli expressing the peptide on the bacterial surface. Intramuscular injection of the epitope-expressing recombinant bacteria into chickens induced a specific serological response to serovar A. A. paragallinarum. The chickens given the recombinant E. coli showed significant protection against challenge with A. paragallinarum 0083. These results indicated a potential for the use of the mimotope in the development of molecular vaccines for infectious coryza.

Marker development for genes controlling mango tree architecture

Identification of candidate genes controlling, mango tree architecture.

Development of an avocado rapid library tray system for Hass

The aim of the project is to develop a rapid ripening protocol/test which can be used as a reliable library tray system for Hass Avocados. Fruit will be taken from the end of the packing line to reflect both commercial handling quality and intrinsic fruit quality.

Resting sites, edge effects and dispersion of a polyphagous Bactrocera fruit fly within crops of different architecture

The spot or strip application of poisoned protein bait is a lure-and-kill technique used for the management of fruit flies. Knowledge of where flies occur in the crop environment is an important part of maximizing the efficacy of this tool. Bactrocera tryoni is a polyphagous pest of horticulture for which very little is known about its distribution within crops. With particular reference to edge effects, we monitored the abundance of B. tryoni in two crops of different architecture; strawberry and apple. In strawberries, we found more flies on the crop edge early in the fruiting season, which lessened gradually and eventually disappeared as the season progressed. In apple orchards, no such edge effect was observed and flies were found equally throughout the orchard. We postulated these differences may be due to differences in crop height (high vs. short) and/or crop canopy architecture (opened and branched in apple, dense and closed in strawberry). In a field cage trial, we tested these predictions using artificial plants of different height and canopy condition. Height and canopy structure type had no significant effects on fly oviposition and protein feeding, but the ‘apple’ type canopy significantly influenced resting. We thus postulate that there was an edge effect in strawberry because the crop was not providing resting sites and flies were doing so in vegetation around the field margins. The finding that B. tryoni shows different resting site preferences based on plant architecture offers the potential for strategic manipulation of the fly through specific border or inter-row plantings.

The Genetic Architecture of Climatic Adaptation of Tropical Cattle

Adaptation of global food systems to climate change is essential to feed the world. Tropical cattle production, a mainstay of profitability for farmers in the developing world, is dominated by heat, lack of water, poor quality feedstuffs, parasites, and tropical diseases. In these systems European cattle suffer significant stock loss, and the cross breeding of taurine x indicine cattle is unpredictable due to the dilution of adaptation to heat and tropical diseases. We explored the genetic architecture of ten traits of tropical cattle production using genome wide association studies of 4,662 animals varying from 0% to 100% indicine. We show that nine of the ten have genetic architectures that include genes of major effect, and in one case, a single location that accounted for more than 71% of the genetic variation. One genetic region in particular had effects on parasite resistance, yearling weight, body condition score, coat colour and penile sheath score. This region, extending 20 Mb on BTA5, appeared to be under genetic selection possibly through maintenance of haplotypes by breeders. We found that the amount of genetic variation and the genetic correlations between traits did not depend upon the degree of indicine content in the animals. Climate change is expected to expand some conditions of the tropics to more temperate environments, which may impact negatively on global livestock health and production. Our results point to several important genes that have large effects on adaptation that could be introduced into more temperate cattle without detrimental effects on productivity.

Resting sites, edge effects and dispersion of a polyphagous Bactrocera fruit fly within crops of different architecture

The spot or strip application of poisoned protein bait is a lure-and-kill technique used for the management of fruit flies. Knowledge of where flies occur in the crop environment is an important part of maximizing the efficacy of this tool. Bactrocera tryoni is a polyphagous pest of horticulture for which very little is known about its distribution within crops. With particular reference to edge effects, we monitored the abundance of B. tryoni in two crops of different architecture; strawberry and apple. In strawberries, we found more flies on the crop edge early in the fruiting season, which lessened gradually and eventually disappeared as the season progressed. In apple orchards, no such edge effect was observed and flies were found equally throughout the orchard. We postulated these differences may be due to differences in crop height (high vs. short) and/or crop canopy architecture (opened and branched in apple, dense and closed in strawberry). In a field cage trial, we tested these predictions using artificial plants of different height and canopy condition. Height and canopy structure type had no significant effects on fly oviposition and protein feeding, but the 'apple' type canopy significantly influenced resting. We thus postulate that there was an edge effect in strawberry because the crop was not providing resting sites and flies were doing so in vegetation around the field margins. The finding that B. tryoni shows different resting site preferences based on plant architecture offers the potential for strategic manipulation of the fly through specific border or inter-row plantings. © 2013 Blackwell Verlag GmbH.

Improvement of mango tree architecture

This paper discusses the role of mango canopy architecture in mango productivity and orchard management and considers potential increases on production of high density orchards through improved canopy architecture. Lower tree height, reduced vigour and smaller more open canopies are recognised as important aspects of high density orchards. However, vigour management, light relations, flowering and crop load of high density orchards needs to be better understood if we are to developed sustainable highly productive canopy training and pruning systems that are easy to maintain at high planting densities.