The development of an improved PCR-based marker system for Sw-5, an important TSWV resistance gene of tomato

Sw-5 is an important disease resistance gene of tomato, providing broad resistance to Tomato spotted wilt virus (TSWV). A cleaved amplified polymorphic sequence (CAPS) marker, closely linked to the gene, has been reported. Although the Sw-5 locus has been characterised, a gene-specific marker has not been developed. This paper presents a PCR-based marker-system that consists of the co-amplification of a dominant marker representing the Sw-5 gene sequence, and the modified CAPS marker as a positive control and indicator of genotype.

Identification of quantitative trait loci for resistance to two species of root-lesion nematode (Pratylenchus thornei and P. neglectus) in wheat

Pratylenchus thornei and P. neglectus are two species of root-lesion nematode that cause substantial yield losses in wheat. No commercially available wheat variety has resistance to both species. A doubled-haploid population developed from a cross between the synthetic hexaploid wheat line CPI133872 and the bread wheat Janz was used to locate and tag quantitative trait loci (QTLs) associated with resistance to both P. thornei and P. neglectus. Wheat plants were inoculated with both species of nematode in independent replicated glasshouse trials repeated over 2 years. Known locations of wheat microsatellite markers were used to construct a framework map. After an initial single-marker analysis to detect marker-trait linkages, chromosome regions associated with putative QTLs were targetted with microsatellite markers to increase map density in the chromosome regions of interest. In total, 148 wheat microsatellite markers and 21 amplified fragment length polymorphism markers were mapped. The codominant microsatellite marker Xbarc183 on the distal end of chromosome 6DS was allelic for resistance to both P. thornei and P. neglectus. The QTL were designated QRlnt.lrc-6D.1 and QRlnn.lrc-6D.1, for the 2 traits, respectively. The allele inherited from CPI133872 explained 22.0-24.2% of the phenotypic variation for P. thornei resistance, and the allele inherited from Janz accounted for 11.3-14.0% of the phenotypic variation for P. neglectus resistance. Composite interval mapping identified markers that flank a second major QTL on chromosome 6DL (QRlnt.lrc-6D.2) that explained 8.3-13.4% of the phenotypic variation for P. thornei resistance. An additional major QTL associated with P. neglectus resistance was detected on chromosome 4DS (QRlnn.lrc-4D.1) and explained a further 10.3-15.4% of the phenotypic variation. The identification and tagging of nematode resistance genes with molecular markers will allow appropriate allele combinations to be selected, which will aid the successful breeding of wheat with dual nematode resistance.

Simple sequence repeat markers associated with three quantitative trait loci for black point resistance can be used to enrich selection populations in bread wheat

Black point in wheat has the potential to cost the Australian industry $A30.4 million a year. It is difficult and expensive to screen for resistance, so the aim of this study was to validate 3 previously identified quantitative trait loci (QTLs) for black point resistance on chromosomes 2B, 4A, and 3D of the wheat variety Sunco. Black point resistance data and simple sequence repeat (SSR) markers, linked to the resistance QTLs and suited to high-throughput assay, were analysed in the doubled haploid population, Batavia (susceptible) × Pelsart (resistant). Sunco and Pelsart both have Cook in their pedigree and both have the Triticum timopheevii translocation on 2B. SSR markers identified for the 3 genetic regions were gwm319 (2B, T. timopheevii translocation), wmc048 (4AS), and gwm341 (3DS). Gwm319 and wmc048 were associated with black point resistance in the validation population. Gwm341 may have an epistatic influence on the trait because when resistance alleles were present at both gwm319 and wmc048, the Batavia-derived allele at gwm341 was associated with a higher proportion of resistant lines. Data are presented showing the level of enrichment achieved for black point resistance, using 1, 2, or 3 of these molecular markers, and the number of associated discarded resistant lines. The level of population enrichment was found to be 1.83-fold with 6 of 17 resistant lines discarded when gwm319 and wmc048 were both used for selection. Interactions among the 3 QTLs appear complex and other genetic and epigenetic factors influence susceptibility to black point. Polymorphism was assessed for these markers within potential breeding material. This indicated that alternative markers to wmc048 may be required for some parental combinations. Based on these results, marker-assisted selection for the major black point resistance QTLs can increase the rate of genetic gain by improving the selection efficiency and may facilitate stacking of black point resistances from different sources.

A multi-microprocessor architecture for solving partial differential equations

This paper presents the architecture of a fault-tolerant, special-purpose multi-microprocessor system for solving Partial Differential Equations (PDEs). The modular nature of the architecture allows the use of hundreds of Processing Elements (PEs) for high throughput. Its performance is evaluated by both analytical and simulation methods. The results indicate that the system can achieve high operation rates and is not sensitive to inter-processor communication delay.

Identification of a mutation in the para-sodium channel gene of the cattle tick Rhipicephalus (Boophilus) microplus associated with resistance to synthetic pyrethroid acaricides

Resistance against synthetic pyrethroid (SP) products for the control of cattle ticks in Australia was detected in the field in 1984, within a very short time of commercial introduction. We have identified a mutation in the domain II S4-5 linker of the para-sodium channel that is associated with resistance to SPs in the cattle tick Rhipicephalus (Boophilus) microplus from Australia. The cytosine to adenine mutation at position 190 in the R. microplus sequence AF134216, results in an amino acid substitution from leucine in the susceptible strain to isoleucine in the resistant strain. A similar mutation has been shown to confer SP resistance in the whitefly, Bemisia tabaci, but has not been described previously in ticks. A diagnostic quantitative PCR assay has been developed using allele-specific Taqman® minor groove-binding (MGB) probes. Using the assay to screen field and laboratory populations of ticks showed that homozygote allelic frequencies correlated highly with the survival percentage at the discriminating concentration of cypermethrin.

Co-evolution of wild rabbits (Oryctolagus cuniculus) and RHDV: resistance and virulence

Rabbit Haemorrhagic Disease Virus (RHDV) was introduced to Australia in 1995 for the control of wild rabbits. Initial outbreaks greatly reduced rabbit numbers and the virus has continued to control rabbits to varying degrees in different parts of Australia. However, recent field evidence suggests that the virus may be becoming less effective in those areas that have previously experienced repeated epizootics causing high mortality. There are also reports of rabbits returning to pre-1995 density levels, Virus and host can be expected to co-evolve. The host will develop resistance to the virus with the virus subsequently changing to overcome that resistance. It has been 12 years since the release of RHDV and it is an opportune time to examine where the dynamic currently stands between RHDV and rabbits. Laboratory challenge tests have indicated that resistance to RHDV has developed to different degrees in populations throughout Australia. In one population a low dose (1:25 dilution) of Czech strain RHDV failed to infect a single susceptible rabbit, yet infected a low to high (up to 73%) percentage across other populations tested. Different selection pressures are present in these populations and will be driving the level of resistance being seen. The mechanisms and genetics behind the development of resistance are also important as the on-going use of RHDV as a control tool in the management of rabbits relies on our understanding of factors influencing the efficacy of the virus. Understanding how resistance has developed may provide clues on how best to use the virus to circumvent these mechanisms. Similarly, it will help in managing populations that have yet to develop high levels of resistance.

Resistance to the root-lesion nematode Pratylenchus thornei of Iranian landrace wheat.

Pratylenchus thornei is widespread throughout the wheat-growing regions in Australia and overseas and can cause yield losses of up to 70% in some intolerant cultivars. The most effective forms of management of P. thornei populations are crop rotation and plant breeding. There have been no wheat accessions identified as completely resistant to P. thornei, therefore breeding programs have used moderately resistant parents. The objective of the present research was to evaluate 274 Iranian landrace wheats for resistance to P. thornei and identify accessions with resistance superior to the current best resistance source (GS50a). Plants were grown in P. thornei inoculated soil under controlled conditions in a glasshouse pot experiment for 16 weeks. Ninety-two accessions found to be resistant or moderately so were retested in a second experiment. From combined analysis of these experiments, 34 accessions were identified as resistant with reproduction factors (final population per kg soil/initial inoculum rate per kg soil) <= 1. In total, 25 accessions were more resistant than GS50a, with AUS28470 significantly (P < 0.05) more resistant. The resistant Iranian landraces identified in the present study are a valuable untapped genetic pool offering improved levels of P. thornei resistance over current parents in Australian wheat-breeding programs.

Evaluation of transgenic bananas expressing anti-apoptotic genes for resistance against Fusarium wilt

The banana industry worldwide is under threat from a fungal disease known as Fusarium wilt, a disease for which there is no chemical control. Conventional breeding approaches to generate resistant banana varieties are lengthy and very difficult. As such, genetic engineering for disease resistance is considered the most viable control option. In this PhD thesis, genetically modified banana plants were generated using several different stress tolerance genes. When challenged with Fusarium wilt in glasshouse trials, some lines showed increased resistance to the disease. The promising elite lines generated in this study will now require testing in field trials.

Resistance to root-lesion nematodes (Pratylenchus thornei and P-neglectus) in synthetic hexaploid wheats and their durum and Aegilops tauschii parents.

Root-lesion nematodes (Pratylenchus thornei Sher and Allen and P. neglectus (Rensch) Filipijev and Schuurmans Stekhoven) cause substantial yield loss to wheat crops in the northern grain region of Australia. Resistance to P. thornei for use in wheat breeding programs was sought among synthetic hexaploid wheats (2n= 6x = 42, AABBDD) produced through hybridisations of Triticum turgidum L. subsp. durum (Desf.) Husn (2n= 4x = 28, AABB) with Aegilops tauschii Coss. (2n= 2x = 14, DD). Resistance was determined for the synthetic hexaploid wheats and their durum and Ae. tauschii parents from the numbers of nematodes in the roots of plants grown for 16 weeks in pots of pasteurised soil inoculated with P. thornei. Fifty-nine (32%) of 186 accessions of synthetic hexaploid wheats had lower numbers of nematodes than Gatcher Selection 50a (GS50a), a partially resistant bread wheat. Greater frequencies of partial resistance were present in the durum parents (72% of 39 lines having lower nematode numbers than GS50a) and in the Ae. tauschii parents (55% of 53 lines). The 59 synthetic hexaploids were re-tested in a second experiment along with their parents. In a third experiment, 11 resistant synthetic hexaploid wheats and their F-1 hybrids with Janz, a susceptible bread wheat, were tested and the F(1)s were found to give nematode counts intermediate between the respective two parents. Synthetic hexaploid wheats with higher levels of resistance resulted from hybridisations where both the durum and Ae. tauschii parents were partially resistant, rather than where only one parent was partially resistant. These results suggest that resistance to P. thornei in synthetic hexaploid wheats is polygenic, with resistances located both in the D genome from Ae. tauschii and in the A and/or B genomes from durum. Five synthetic hexaploid wheats were selected for further study on the basis of (1) a high level of resistance to P. thornei of the synthetic hexaploid wheats and of both their durum and Ae. tauschii parents, (2) being representative of both Australian and CIMMYT (International Maize and Wheat Improvement Centre) durums, and (3) being representative of the morphological subspecies and varieties of Ae. tauschii. These 5 synthetic hexaploid wheats were also shown to be resistant to P. neglectus, whereas GS50a and 2 P. thornei-resistant derivatives were quite susceptible. Results of P. thornei resistance of F(1)s and F(2)s from a half diallel of these 5 synthetic hexaploid wheats, GS50a, and Janz from another study indicate polygenic additive resistance and better general combining ability for the synthetic hexaploid wheats than for GS50a. Published molecular marker studies on a doubled haploid population between the synthetic hexaploid wheat with best general combining ability (CPI133872) and Janz have shown quantitative trait loci for resistance located in all 3 genomes. Synthetic hexaploid wheats offer a convenient way of introgressing new resistances to P. thornei and P. neglectus from both durum and Ae. tauschii into commercial bread wheats.

Uropathogenic Escherichia Coli engages CD14-dependent signaling to enable bladder-macrophage-dependent control of acute urinary tract infection

Background CD14, a coreceptor for several pattern recognition receptors and a widely used monocyte/macrophage marker, plays a key role in host responses to gram-negative bacteria. Despite the central role of CD14 in the inflammatory response to lipopolysaccharide and other microbial products and in the dissemination of bacteria in some infections, the signaling networks controlled by CD14 during urinary tract infection (UTI) are unknown. Methods We used uropathogenic Escherichia coli (UPEC) infection of wild-type (WT) C57BL/6 and Cd14−/− mice and RNA sequencing to define the CD14-dependent transcriptional signature and the role of CD14 in host defense against UTI in the bladder. Results UPEC induced the upregulation of Cd14 and the monocyte/macrophage-related genes Emr1/F4/80 and Csf1r/c-fms, which was associated with lower UPEC burdens in WT mice, compared with Cd14−/− mice. Exacerbation of infection in Cd14−/− mice was associated with the absence of a 491-gene transcriptional signature in the bladder that encompassed multiple host networks not previously associated with this receptor. CD14-dependent pathways included immune cell trafficking, differential cytokine production in macrophages, and interleukin 17 signaling. Depletion of monocytes/macrophages in the bladder by administration of liposomal clodronate led to higher UPEC burdens. Conclusions This study identifies new host protective and signaling roles for CD14 in the bladder during UPEC UTI.

Simulating the evolution of glyphosate resistance in grains farming in northern Australia.

Background and Aims: The evolution of resistance to herbicides is a substantial problem in contemporary agriculture. Solutions to this problem generally consist of the use of practices to control the resistant population once it evolves, and/or to institute preventative measures before populations become resistant. Herbicide resistance evolves in populations over years or decades, so predicting the effectiveness of preventative strategies in particular relies on computational modelling approaches. While models of herbicide resistance already exist, none deals with the complex regional variability in the northern Australian sub-tropical grains farming region. For this reason, a new computer model was developed. Methods: The model consists of an age- and stage-structured population model of weeds, with an existing crop model used to simulate plant growth and competition, and extensions to the crop model added to simulate seed bank ecology and population genetics factors. Using awnless barnyard grass (Echinochloa colona) as a test case, the model was used to investigate the likely rate of evolution under conditions expected to produce high selection pressure. Key Results: Simulating continuous summer fallows with glyphosate used as the only means of weed control resulted in predicted resistant weed populations after approx. 15 years. Validation of the model against the paddock history for the first real-world glyphosate-resistant awnless barnyard grass population shows that the model predicted resistance evolution to within a few years of the real situation. Conclusions: This validation work shows that empirical validation of herbicide resistance models is problematic. However, the model simulates the complexities of sub-tropical grains farming in Australia well, and can be used to investigate, generate and improve glyphosate resistance prevention strategies.

Molecular convergence of hexosamine biosynthetic pathway and ER stress leading to insulin resistance in L6 skeletal muscle cells

Augmentation of hexosamine biosynthetic pathway (HBP) and endoplasmic reticulum (ER) stress were independently related to be the underlying causes of insulin resistance. We hypothesized that there might be a molecular convergence of activated HBP and ER stress pathways leading to insulin resistance. Augmentation of HBP in L6 skeletal muscle cells either by pharmacological (glucosamine) or physiological (high-glucose) means, resulted in increased protein expression of ER chaperones (viz., Grp78, Calreticulin, and Calnexin), UDP-GlcNAc levels and impaired insulin-stimulated glucose uptake. Cells silenced for O-glycosyl transferase (OGT) showed improved insulin-stimulated glucose uptake (P < 0.05) but without any effect on ER chaperone upregulation. While cells treated with either glucosamine or high-glucose exhibited increased JNK activity, silencing of OGT resulted in inhibition of JNK and normalization of glucose uptake. Our study for the first time, demonstrates a molecular convergence of O-glycosylation processes and ER stress signals at the cross-road of insulin resistance in skeletal muscle.

Application of Haar functions for transmission line and transformer differential protection

The development of algorithms, based on Haar functions, for extracting the desired frequency components from transient power-system relaying signals is presented. The applications of these algorithms to impedance detection in transmission line protection and to harmonic restraint in transformer differential protection are discussed. For transmission line protection, three modes of application of the Haar algorithms are described: a full-cycle window algorithm, an approximate full-cycle window algorithm, and a half-cycle window algorithm. For power transformer differential protection, the combined second and fifth harmonic magnitude of the differential current is compared with that of fundamental to arrive at a trip decision. The proposed line protection algorithms are evaluated, under different fault conditions, using realistic relaying signals obtained from transient analysis conducted on a model 400 kV, 3-phase system. The transformer differential protection algorithms are also evaluated using a variety of simulated inrush and internal fault signals.

Resistance to the root-lesion nematode Pratylenchus thornei in wheat landraces and cultivars from the West Asia and North Africa (WANA) region.

Resistance to the root-lesion nematode Pratylenchus thornei was sought in wheat from the West Asia and North Africa (WANA) region in the Watkins Collection (148 bread and 139 durum wheat accessions) and the McIntosh Collection (59 bread and 43 durum wheat accessions). It was considered that landraces from this region, encompassing the centres of origin of wheat and where P. thornei also occurs, could be valuable sources of resistance for use in wheat breeding. Resistance was determined by number of P. thornei/kg soil after the growth of the plants in replicated glasshouse experiments. On average, durum accessions produced significantly lower numbers of P. thornei than bread wheat accessions in both the Watkins and McIntosh Collections. Selected accessions with low P. thornei numbers were re-tested and 13 bread wheat and 10 durum accessions were identified with nematode numbers not significantly different from GS50a, a partially resistant bread wheat line used as a reference standard. These resistant accessions, which originated in Iran, Iraq, Syria, Egypt, Sudan, Morocco, and Tunisia, represent a resource of resistance genes in the primary wheat gene pool, which could be used in Australian wheat breeding programs to reduce the economic loss from P. thornei.

Hip power analysis in individuals with transfemoral amputation: A different strategy different from stabilisation during gait stance

Introduction and Objectives Joint moments and joint powers during gait are widely used to determine the effects of rehabilitation programs as well as prosthetic fitting. Following the definition of power (dot product of joint moment and joint angular velocity) it has been previously proposed to analyse the 3D angle between both vectors, αMw. Basically, joint power is maximised when both vectors are parallel and cancelled when both vectors are orthogonal. In other words, αMw < 60° reveals a propulsion configuration (more than 50% of the moment contribute to positive power) while αMw > 120° reveals a resistance configuration (more than 50% of the moment contribute to negative power). A stabilisation configuration (less than 50% of the moment contribute to power) corresponds to 60° < αMw < 120°. Previous studies demonstrated that hip joints of able-bodied adults (AB) are mainly in a stabilisation configuration (αMw about 90°) during the stance phase of gait. [1, 2] Individuals with transfemoral amputation (TFA) need to maximise joint power at the hip while controlling the prosthetic knee during stance. Therefore, we tested the hypothesis that TFAs should adopt a strategy that is different from a continuous stabilisation. The objective of this study was to compute joint power and αMw for TFA and to compare them with AB. Methods Three trials of walking at self-selected speed were analysed for 8 TFAs (7 males and 1 female, 46±10 years old, 1.78±0.08 m 82±13 kg) and 8 ABs (males, 25±3 years old, 1.75±0.04, m 67±6 kg). The joint moments are computed from a motion analysis system (Qualisys, Goteborg, Sweden) and a multi-axial transducer (JR3, Woodland, USA) mounted above the prosthetic knee for TFAs and from a motion analysis system (Motion Analysis, Santa Rosa, USA) and force plates (Bertec, Columbus, USA) for ABs. The TFAs were fitted with an OPRA (Integrum, AB, Gothengurg, Sweden) osseointegrated implant system and their prosthetic designs include pneumatic, hydraulic and microprocessor knees. Previous studies showed that the inverse dynamics computed from the multi-axial transducer is the proper method considering the absorption at the foot and resistance at the knee. Results The peak of positive power at loading response (H1) was earlier and lower for TFA compared to AB. Although the joint power is lower, the 3D angle between joint moment and joint angular velocity, αMw, reveals an obvious propulsion configuration (mean αMw about 20°) for TFA compared to a stabilisation configuration (mean αMw about 70°) for AB. The peaks of negative power at midstance (H2) and of positive power at preswing / initial swing (H3) occurred later, lower and longer for TFA compared to AB. Again, the joint powers are lower for TFA but, in this case, αMw is almost comparable (with a time lag), demonstrating a stabilisation (almost a resistance for TFA, mean αMw about 120°) and a propulsion configuration, respectively. The swing phase is not analysed in the present study. Conclusion The analysis of hip joint power may indicate that TFAs demonstrated less propulsion and resistance than ABs during the stance phase of gait. This is true from a quantitative point of view. On the contrary, the 3D angle between joint moment and joint angular velocity, αMw, reveals that TFAs have a remarkable propulsion strategy at loading response and almost a resistance strategy at midstance while ABs adopted a stabilisation strategy. The propulsion configuration, with αMw close to 0°, seems to aim at maximising the positive joint power. The configuration close to resistance, with αMw far from 180°, might aim at unlocking the prosthetic knee before swing while minimising the negative power. This analysis of both joint power and 3D angle between the joint moment and the joint angular velocity provides complementary insights into the gait strategies of TFA that can be used to support evidence-based rehabilitation and fitting of prosthetic components.