An in-frame deletion mutation in the degron tail of auxin coreceptor IAA2 confers resistance to the herbicide 2,4-D in Sisymbrium orientale.

The natural auxin indole-3-acetic acid (IAA) is a key regulator of many aspects of plant growth and development. Synthetic auxin herbicides such as 2,4-D mimic the effects of IAA by inducing strong auxinic-signaling responses in plants. To determine the mechanism of 2,4-D resistance in a Sisymbrium orientale (Indian hedge mustard) weed population, we performed a transcriptome analysis of 2,4-D-resistant (R) and -susceptible (S) genotypes that revealed an in-frame 27-nucleotide deletion removing nine amino acids in the degron tail (DT) of the auxin coreceptor Aux/IAA2 (SoIAA2). The deletion allele cosegregated with 2,4-D resistance in recombinant inbred lines. Further, this deletion was also detected in several 2,4-D-resistant field populations of this species. Arabidopsis transgenic lines expressing the SoIAA2 mutant allele were resistant to 2,4-D and dicamba. The IAA2-DT deletion reduced binding to TIR1 in vitro with both natural and synthetic auxins, causing reduced association and increased dissociation rates. This mechanism of synthetic auxin herbicide resistance assigns an in planta function to the DT region of this Aux/IAA coreceptor for its role in synthetic auxin binding kinetics and reveals a potential biotechnological approach to produce synthetic auxin-resistant crops using gene-editing.

A novel insight into the mode of action of glufosinate: how reactive oxygen species are formed.

Glufosinate targets glutamine synthetase (GS), but its fast herbicidal action is triggered by reactive oxygen species (ROS). The relationship between GS inhibition and ROS accumulation was investigated in Amaranthus palmeri. Glufosinate's fast action is light-dependent with no visual symptoms or ROS formation in the dark. Inhibition of GS leads to accumulation of ammonia and metabolites of the photorespiration pathway, such as glycolate and glyoxylate, as well as depletion of other intermediates such as glycine, serine, hydroxypyruvate, and glycerate. Exogenous supply of glycolate to glufosinate-treated plants enhanced herbicidal activity and dramatically increased hydrogen peroxide accumulation (possibly from peroxisomal glycolate oxidase activity). Glufosinate affected the balance between ROS generation and scavenging. The activity of superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase increased after glufosinate treatment in an attempt to quench the nascent ROS burst. Low doses of atrazine and dinoseb were used to investigate the sources of ROS by manipulating photosynthetic electron transport and oxygen (O2) evolution. ROS formation depended on electron flow and O2 evolution in photosystem II (PSII). Inhibition of GS disrupted photorespiration, carbon assimilation, and linear electron flow in the light reactions. Consequently, the antioxidant machinery and the water-water cycle are overwhelmed in the presence of light and glufosinate. The O2 generated by the splitting of water in PSII becomes the acceptor of electrons, generating ROS. The cascade of events leads to lipid peroxidation and forms the basis for the fast action of glufosinate.

Reactive oxygen species trigger the fast action of glufosinate.

MAIN CONCLUSION: Glufosinate is primarily toxic to plants due to a massive light-dependent generation of reactive oxygen species rather than ammonia accumulation or carbon assimilation inhibition. Glutamine synthetase (GS) plays a key role in plant nitrogen metabolism and photorespiration. Glufosinate (C5H12NO4P) targets GS and causes catastrophic consequences leading to rapid plant cell death, and the causes for phytoxicity have been attributed to ammonia accumulation and carbon assimilation restriction. This study aimed to examine the biochemical and physiological consequences of GS inhibition to identify the actual cause for rapid phytotoxicity. Monocotyledonous and dicotyledonous species with different forms of carbon assimilation (C3 versus C4) were selected as model plants. Glufosinate sensitivity was proportional to the uptake of herbicide between species. Herbicide uptake also correlated with the level of GS inhibition and ammonia accumulation in planta even with all species having the same levels of enzyme sensitivity in vitro. Depletion of both glutamine and glutamate occurred in glufosinate-treated leaves; however, amino acid starvation would be expected to cause a slow plant response. Ammonia accumulation in response to GS inhibition, often reported as the driver of glufosinate phytotoxicity, occurred in all species, but did not correlate with either reductions in carbon assimilation or cell death. This is supported by the fact that plants can accumulate high levels of ammonia but show low inhibition of carbon assimilation and absence of phytotoxicity. Glufosinate-treated plants showed a massive light-dependent generation of reactive oxygen species, followed by malondialdehyde accumulation. Consequently, we propose that glufosinate is toxic to plants not because of ammonia accumulation nor carbon assimilation inhibition, but the production of reactive oxygen species driving the catastrophic lipid peroxidation of the cell membranes and rapid cell death.

Diversity and extent of mutations endowing resistance to the acetolactate synthase (AHAS)-inhibiting herbicides in Indian hedge mustard (Sisymbrium orientale) populations in Australia.

Indian hedge mustard (Sisymbrium orientale) (IHM) is an important broadleaf weed across southern Australia. Resistance to sulfonylurea (SU) herbicides that inhibit acetohydroxyacid synthase (AHAS) is extensive in Australia, but resistance to imidazolinone (IMI) herbicides has only been reported recently. The AHAS-mutation profile of 65 IHM populations collected randomly from cropped fields was investigated to better understand the extent and types of resistance present. Resistance to SU herbicides was present in 40% of the populations and resistance to IMI herbicides in 11%. Mutations were identified in SoAHAS by sequence analysis, and included previously reported amino-acid substitutions at Pro197 and Trp574, but also new substitutions at Pro197 and Asp376 for this species. One population with possible non-target-site resistance was identified. Germination studies with fresh seed found no significant effect by mutations in SoAHAS on germination; however, population factors had a large effect on germination in S. orientale. Resistance to AHAS-inhibiting herbicides in populations of S. orientale is endowed by mutations in SoAHAS in all but one population examined. Mutations at Pro197 conferring resistance to SU herbicides were most common, while mutations at Trp574 that provide resistance to IMI herbicides are also present.

No apparent fitness costs associated with phytoene desaturase mutations conferred resistance to diflufenican and picolinafen in oriental mustard (Sisymbrium orientale L.).

Two mutations Leu498 and Glu425 in the PDS gene were identified as the main cause conferring resistance to diflufenican and picolinafen in two oriental mustard populations P3 and P40. As mutations are suspected to affect fitness, this study was designed to test this hypothesis using the F2 of two crosses P3.2 (P3♂ × S♀) and P40.5 (P40♂ × S♀) of oriental mustard. The F2 plants, which segregated for target-site point mutations of PDS gene (Leu498 and Glu425) grown in monoculture and under competition with wheat in pot-trials and evaluated for growth and fecundity. All F2 individuals were genotyped by using Cleaved Amplified Polymorphic Sequence (CAPS) technique. Regression analysis showed no fitness cost in the resistant plants because no significant difference was identified in seed and biomass production within RR, RS and SS individuals. The absence of measurable negative effects on fitness associated mutations suggests that the frequency of the PDS resistance alleles will not decline in the absence of selection pressure of PDS-inhibitors.

Gene amplification of 5-enol-pyruvylshikimate-3-phosphate synthase in glyphosate-resistant Kochia scoparia.

MAIN CONCLUSION: Field-evolved resistance to the herbicide glyphosate is due to amplification of one of two EPSPS alleles, increasing transcription and protein with no splice variants or effects on other pathway genes. The widely used herbicide glyphosate inhibits the shikimate pathway enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Globally, the intensive use of glyphosate for weed control has selected for glyphosate resistance in 31 weed species. Populations of suspected glyphosate-resistant Kochia scoparia were collected from fields located in the US central Great Plains. Glyphosate dose response verified glyphosate resistance in nine populations. The mechanism of resistance to glyphosate was investigated using targeted sequencing, quantitative PCR, immunoblotting, and whole transcriptome de novo sequencing to characterize the sequence and expression of EPSPS. Sequence analysis showed no mutation of the EPSPS Pro106 codon in glyphosate-resistant K. scoparia, whereas EPSPS genomic copy number and transcript abundance were elevated three- to ten-fold in resistant individuals relative to susceptible individuals. Glyphosate-resistant individuals with increased relative EPSPS copy numbers had consistently lower shikimate accumulation in leaf disks treated with 100 µM glyphosate and EPSPS protein levels were higher in glyphosate-resistant individuals with increased gene copy number compared to glyphosate-susceptible individuals. RNA sequence analysis revealed seven nucleotide positions with two different expressed alleles in glyphosate-susceptible reads. However, one nucleotide at the seven positions was predominant in glyphosate-resistant sequences, suggesting that only one of two EPSPS alleles was amplified in glyphosate-resistant individuals. No alternatively spliced EPSPS transcripts were detected. Expression of five other genes in the chorismate pathway was unaffected in glyphosate-resistant individuals with increased EPSPS expression. These results indicate increased EPSPS expression is a mechanism for glyphosate resistance in these K. scoparia populations.

Arthroscopic assessment of osteochondrosis of the medial humeral condyle treated with debridement and sliding humeral osteotomy.

OBJECTIVE: To document the progression of disease after an unloading procedure when combined with arthroscopic debridement to treat osteochondrosis (OC) lesions affecting the medial humeral condyle in juvenile dogs without medial compartment disease. STUDY DESIGN: Retrospective case series. ANIMALS: Dogs with medial humeral condyle OC (n = 6). METHODS: Dogs with medial humeral condyle OC (9 elbows) were treated by arthroscopic debridement of the OC lesion and sliding humeral osteotomy (SHO). Outcomes were assessed by radiography (n = 9), 2nd-look arthroscopy (9) and clinical examination including lameness assessment (9) at 18-28 months after surgery. RESULTS: Seven elbows had marked progression of pathology affecting the medial compartment, 5 elbows developed subsequent fragmentation of the medial coronoid process of the ulna [FMCP], 2 elbows had fibrocartilaginous infill of the humeral defect without development of coronoid pathology. CONCLUSION: This limited case series suggests that medial compartment disease develops despite unloading of the medial compartment using a distal diaphyseal SHO in juvenile dogs with OC.

Insulinoma: Metastatic Recurrence 38 Years Following Initial Diagnosis in Pregnancy.

A case of recurrent insulinoma spanning 4 decades is described. Following a delayed diagnosis, hyperinsulinemic hypoglycemia was confirmed in a 24-year-old woman during early pregnancy. Initial surgery, culminating in subtotal pancreatectomy, was noncurative. A 1-cm insulinoma was subsequently resected from the head of the pancreas postpartum, with postoperative resolution of hypoglycemia. However, 32 years later, the patient experienced a recurrence of hypoglycemic symptoms. Eventually, a subcentimeter extrapancreatic lesion was identified anterior to the pancreatic head on gallium-68 DOTA-Exendin-4 positron emission tomography/computed tomography. In 2022, a third operation was performed, with excision of a 4 × 3 mm tumor adjacent to the pancreatic head, and histology confirming insulinoma. She was again cured of symptoms.

Gene amplification confers glyphosate resistance in Amaranthus palmeri.

The herbicide glyphosate became widely used in the United States and other parts of the world after the commercialization of glyphosate-resistant crops. These crops have constitutive overexpression of a glyphosate-insensitive form of the herbicide target site gene, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Increased use of glyphosate over multiple years imposes selective genetic pressure on weed populations. We investigated recently discovered glyphosate-resistant Amaranthus palmeri populations from Georgia, in comparison with normally sensitive populations. EPSPS enzyme activity from resistant and susceptible plants was equally inhibited by glyphosate, which led us to use quantitative PCR to measure relative copy numbers of the EPSPS gene. Genomes of resistant plants contained from 5-fold to more than 160-fold more copies of the EPSPS gene than did genomes of susceptible plants. Quantitative RT-PCR on cDNA revealed that EPSPS expression was positively correlated with genomic EPSPS relative copy number. Immunoblot analyses showed that increased EPSPS protein level also correlated with EPSPS genomic copy number. EPSPS gene amplification was heritable, correlated with resistance in pseudo-F(2) populations, and is proposed to be the molecular basis of glyphosate resistance. FISH revealed that EPSPS genes were present on every chromosome and, therefore, gene amplification was likely not caused by unequal chromosome crossing over. This occurrence of gene amplification as an herbicide resistance mechanism in a naturally occurring weed population is particularly significant because it could threaten the sustainable use of glyphosate-resistant crop technology.

Population structure of ALS-inhibiting herbicide-resistant Sonchus oleraceus in South Australia.

BACKGROUND: Annual sowthistle is a weed that is difficult to control in lentil crops in southern Australia due to a lack of herbicide options, widespread herbicide resistance and prolific production of highly mobile seed. This study investigates herbicide resistance in annual sowthistle in the Mid-North (MN) and Yorke Peninsula (YP) regions of South Australia, identifies and characterizes the mechanisms of acetolactate-synthase (ALS)-inhibitor resistance in this amphidiploid species, and combines this with analyses of population structure and gene flow. RESULTS: ALS-inhibitor-resistant annual sowthistle is widespread across the YP and MN of South Australia and is associated with a variety of Proline-197 mutations of the ALS gene, including leucine, alanine, arginine, serine, threonine and histidine. These mutations were found in different combinations on either of the two copies of the ALS gene. An additional 200 tissue samples were collected from across a single field on the YP and the ALS gene was sequenced for all these individuals. Different ALS-inhibitor resistance profiles were evident between mutation combinations and within mutation combinations, possibly mediated by differing subgenome assortment of the mutations, or altered gene experession of the two ALS homeologs. Population genetics analysis showed evidence of long-distance dispersal, resulting in highly mobile resistance genes, and multiple instances of resistance mutation evolution. CONCLUSIONS: Continuing selection of Sonchus oleraceus populations with ALS-inhibiting herbicides has resulted in the accumulation of additional mutations within the ALS gene. New practices to control herbicide-resistant S. oleraceus should be examined, and control should focus on reducing seed set and dispersal to prevent the spread of emerging cases of resistance. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Point mutations including a novel Pro-197-Phe mutation confer cross-resistance to acetolactate synthase (ALS) inhibiting herbicides in Lactuca serriola in Australia.

BACKGROUND: Control of prickly lettuce has become increasingly difficult for lentil growers in southern Australia because of widespread resistance to common herbicides, a lack of alternative herbicide options and the prolific production of highly mobile seed. This study aimed to quantify acetolactate synthase (ALS)-inhibiting herbicide resistance in the Mid North (MN) and Yorke Peninsula (YP) of South Australia, characterize the resistance mutations present and investigate population structure and gene flow in this species. RESULTS: Resistance was identified in all populations tested, with average survival of 92% to chlorsulfuron and 95% to imazamox + imazapyr. Five different amino acid substitutions were identified at proline 197 of the ALS gene. There was no significant difference in the median lethal dose (LD50 ) between plants with these five different substitutions when treated with metsulfuron-methyl; however, the imidazolinone resistance level was higher in plants with a phenylalanine substitution and lower in plants with a serine. Population structure based on 701 single nucleotide polymorphisms and 271 individuals provided evidence for both independent evolution of the same mutation in different populations, as well as frequent short- to medium-distance dispersal accompanied by occasional long-distance dispersal events. The overall inbreeding coefficient (FIS ) was calculated at 0.5174, indicating an intermediate level of outcrossing despite the cross-pollination experiment showing only low outcrossing. In the structure analyses, most individuals from YP were assigned to a single cluster, whereas most individuals from MN were assigned 50% to each of two clusters, indicating some genetic differences between these two regions, but also evidence for dispersal between them. CONCLUSIONS: Use of imidazolinone herbicides has selected for mutations conferring higher levels of resistance, such as the Pro-197-Phe mutation, and resulted in further spread of resistance in this species. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Inheritance and mechanism of glyphosate resistance in annual bluegrass (Poa annua L.).

BACKGROUND: In initial screening, glyphosate was ineffective in controlling five Poa annua populations. These populations were tested for resistance, and studies undertaken to determine resistance mechanisms and inheritance pattern. RESULTS: Dose-response studies conducted at 16/12°C and 27/20°C on the five putative resistant populations showed low-level resistance (1.4- to 2.5-fold) to glyphosate. Shikimic acid accumulation in response to glyphosate confirmed differences among the populations, with greater shikimic acid accumulation in the susceptible population. The EPSPS gene copy number was 0.5- to 5.2-fold greater in one resistant population (HT) than in the susceptible (S) population, but not in the others. EPSPS gene expression was five- to tenfold higher in HT compared with the susceptible population. Target site mutations, differences in glyphosate absorption or translocation or altered expression of aldo-keto reductase (AKR) were not identified in any of the resistant populations. Crosses were successful between one resistant population and the susceptible population (P262-16♂ ✕ S♀) and inheritance of glyphosate resistance appears to be controlled by a single, nuclear dominant gene in this population. CONCLUSION: Our study identified EPSPS gene amplification in a South Australian glyphosate-resistant P. annua population (HT). This mechanism of resistance was not identified in the other four glyphosate-resistant populations, and other common mechanisms were excluded. Although the resistance mechanism in some P. annua populations remains unknown, inheritance studies with one population suggest the involvement of a single dominant gene. © 2021 Society of Chemical Industry.

An investigation of herpes simplex virus promoter activity compatible with latency establishment reveals VP16-independent activation of immediate-early promoters in sensory neurones.

Herpes simplex virus (HSV) type-1 establishes lifelong latency in sensory neurones and it is widely assumed that latency is the consequence of a failure to initiate virus immediate-early (IE) gene expression. However, using a Cre reporter mouse system in conjunction with Cre-expressing HSV-1 recombinants we have previously shown that activation of the IE ICP0 promoter can precede latency establishment in at least 30% of latently infected cells. During productive infection of non-neuronal cells, IE promoter activation is largely dependent on the transactivator VP16 a late structural component of the virion. Of significance, VP16 has recently been shown to exhibit altered regulation in neurones; where its de novo synthesis is necessary for IE gene expression during both lytic infection and reactivation from latency. In the current study, we utilized the Cre reporter mouse model system to characterize the full extent of viral promoter activity compatible with cell survival and latency establishment. In contrast to the high frequency activation of representative IE promoters prior to latency establishment, cell marking using a virus recombinant expressing Cre under VP16 promoter control was very inefficient. Furthermore, infection of neuronal cultures with VP16 mutants reveals a strong VP16 requirement for IE promoter activity in non-neuronal cells, but not sensory neurones. We conclude that only IE promoter activation can efficiently precede latency establishment and that this activation is likely to occur through a VP16-independent mechanism.

Incidence of medial meniscal tears after arthroscopic assisted tibial plateau leveling osteotomy.

OBJECTIVE: To determine the incidence of medial meniscal tears in dogs with naturally occurring cranial cruciate ligament (CCL) disease treated with arthroscopy and tibial plateau leveling osteotomy (TPLO). STUDY DESIGN: Retrospective case series. SAMPLE POPULATION: Canine Stifles (n = 357) with naturally occurring CCL disease. METHODS: Medical records (November 2006-November 2009) were reviewed for all dogs with CCL disease treated with arthroscopic CCL debridement, meniscal probing, and TPLO. We investigated the significance of the preoperative variables; age, weight, tibial plateau angle (TPA), sex, and the preoperative condition of the CCL on the prevalence of concurrent meniscal tears (CMT) and incidence of late onset meniscal tears (LMT). RESULTS: Prevalence of CMT and incidence of LMT was 32.2% and 5.6%, respectively. A significant difference between age of dogs with and without CMT was identified. There was a significantly lower prevalence of CMT and incidence of LMT in dogs that had a partial CCL tear compared with those that had a complete CCL tear. All dogs with LMT treated by partial meniscectomy (PMM) returned to peak postoperative limb function after (PMM) based on client-assessed outcomes. CONCLUSIONS: This study demonstrates the importance of LMT as a complication, the importance of comprehensive meniscal assessment, and may add to the evidence against routinely performing meniscal release in TPLO. The preoperative condition of the CCL should be considered before operating on grossly normal menisci.

Review: evolutionary drivers of agricultural adaptation in Lolium spp.

The genus Lolium comprises many species, of which L. perenne ssp. multiflorum, L. perenne ssp. perenne, and L. rigidum are of worldwide agricultural importance as both pasture crops and as weeds. These three species are inter-fertile, obligate out-crossers with a self-incompatible reproduction system. This combination contributes to high genetic diversity that supplies new variants during expansion to new natural areas and agricultural environments. Human dispersal, de-domestication and crop-weed hybridization events between Lolium spp., or with others such as Festuca spp., are likely associated with their distinct weediness abilities. Furthermore, new introductions followed by introgression may hasten adaptation to new environments. Most Lolium-related weed science studies have focused on adaptation leading to herbicide resistance, but other forms of adaptation may also occur. In this review, we explore how the wide genetic variation among Lolium species and hybridization with other species may contribute to range expansion, and adaptation to both new agricultural practices and future predicted climate change scenarios. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Non-Mendelian inheritance of gene amplification-based resistance to glyphosate in Hordeum glaucum (barley grass) from South Australia.

BACKGROUND: Hordeum glaucum Steud. is an important grass weed species in South Australia that has evolved resistance to glyphosate. This study investigated the mode of inheritance of glyphosate resistance in this species. RESULTS: Hand-pollination of glyphosate susceptible and resistant populations generated two F1 individuals, selfed to yield F2 progenies. In dose-response experiments, the F2 progenies showed intermediate response between the two parent populations. High variation in EPSPS gene copies was observed among F2 individuals, with some individuals possessing more gene copies than the resistant parent population. No evidence of a Mendelian single-gene pattern of inheritance was observed. CONCLUSION: Inheritance of gene amplification in H. glaucum is non-Mendelian. © 2021 Society of Chemical Industry.

Stability of EPSPS gene copy number in Hordeum glaucum Steud (barley grass) in the presence and absence of glyphosate selection.

BACKGROUND: Gene amplification has been shown to provide resistance to glyphosate in several weed species, including Hordeum glaucum populations in South Australia. The stability of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene copies in resistant populations in the presence or absence of glyphosate selection has not been determined. RESULTS: Applying glyphosate to a cloned plant resulted in an increase in resistance and EPSPS copy number in the progeny of that plant compared to the untreated clone. The LD50 (herbicide concentration required for 50% mortality) increased by 75% to 79% in the progeny of the treated clones compared to the untreated in both populations (YP-17 and YP-16). EPSPS copy number estimates were higher in treated individuals compared to untreated individuals with an average of seven copies compared to six in YP-16 and 11 compared to six in YP-17. There was a positive correlation (R2  = 0.78) between EPSPS copy number and LD50 of all populations. CONCLUSION: EPSPS gene copy number and resistance to glyphosate increased in H. glaucum populations under glyphosate selection, suggesting the number of EPSPS gene copies present is dependent on glyphosate selection. © 2021 Society of Chemical Industry.

Increasing the value and efficiency of herbicide resistance surveys.

The scale of herbicide resistance within a cropping region can be estimated and monitored using surveys of weed populations. The current approach to herbicide resistance surveys is time-consuming, logistically challenging and costly. Here we review past and current approaches used in herbicide resistance surveys with the aims of (i) defining effective survey methodologies, (ii) highlighting opportunities for improving efficiencies through the use of new technologies and (iii) identifying the value of repeated region-wide herbicide resistance surveys. One of the most extensively surveyed areas of the world's cropping regions is the Australian grain production region, with >2900 fields randomly surveyed in each of three surveys conducted over the past 15 years. Consequently, recommended methodologies are based on what has been learned from the Australian experience. Traditional seedling-based herbicide screening assays remain the most reliable and widely applicable method for characterizing resistance in weed populations. The use of satellite or aerial imagery to plan collections and image analysis to rapidly quantify screening results could complement traditional resistance assays by increasing survey efficiency and sampling accuracy. Global management of herbicide-resistant weeds would benefit from repeated and standardized surveys that track herbicide resistance evolution within and across cropping regions. © 2021 Society of Chemical Industry.

Evidence for the Application of Emerging Technologies to Accelerate Crop Improvement - A Collaborative Pipeline to Introgress Herbicide Tolerance Into Chickpea.

Accelerating genetic gain in crop improvement is required to ensure improved yield and yield stability under increasingly challenging climatic conditions. This case study demonstrates the effective confluence of innovative breeding technologies within a collaborative breeding framework to develop and rapidly introgress imidazolinone Group 2 herbicide tolerance into an adapted Australian chickpea genetic background. A well-adapted, high-yielding desi cultivar PBA HatTrick was treated with ethyl methanesulfonate to generate mutations in the ACETOHYDROXYACID SYNTHASE 1 (CaAHAS1) gene. After 2 years of field screening with imidazolinone herbicide across >20 ha and controlled environment progeny screening, two selections were identified which exhibited putative herbicide tolerance. Both selections contained the same single amino acid substitution, from alanine to valine at position 205 (A205V) in the AHAS1 protein, and KASP™ markers were developed to discriminate between tolerant and intolerant genotypes. A pipeline combining conventional crossing and F2 production with accelerated single seed descent from F2:4 and marker-assisted selection at F2 rapidly introgressed the herbicide tolerance trait from one of the mutant selections, D15PAHI002, into PBA Seamer, a desi cultivar adapted to Australian cropping areas. Field evaluation of the derivatives of the D15PAHI002 × PBA Seamer cross was analyzed using a factor analytic mixed model statistical approach designed to accommodate low seed numbers resulting from accelerated single seed descent. To further accelerate trait introgression, field evaluation trials were undertaken concurrent with crop safety testing trials. In 2020, 4 years after the initial cross, an advanced line selection CBA2061, bearing acetohydroxyacid synthase (AHAS) inhibitor tolerance and agronomic and disease resistance traits comparable to parent PBA Seamer, was entered into Australian National Variety Trials as a precursor to cultivar registration. The combination of cross-institutional collaboration and the application of novel pre-breeding platforms and statistical technologies facilitated a 3-year saving compared to a traditional breeding approach. This breeding pipeline can be used as a model to accelerate genetic gain in other self-pollinating species, particularly food legumes.

Treatment Outcomes for Periprosthetic Femoral Fractures in Cementless Press-Fit Total Hip Replacement.

OBJECTIVE: The aim of this study was to report outcomes in dogs with periprosthetic femoral fractures associated with a press-fit cementless femoral total hip replacement implant. MATERIALS AND METHODS: Electronic medical records and digital radiographs were used to identify dogs with periprosthetic femoral fractures associated with press-fit cementless total hip replacement. Data collected included signalment, weight, time of fracture, cause of fracture, presence of intra-operative fissure, fracture type, repair technique, and clinical and radiographic outcomes. Long-term patient outcome was assessed by communication with owners or referring veterinarians. RESULTS: Twenty-eight dogs with femoral fracture repair associated with cementless press-fit total hip replacement were identified. Eight of the fractures occurred intraoperatively and 20 occurred at a median of 2 days postoperatively. An oblique or spiral configuration was noted in 19 cases and 15 occurred at the distal end of the femoral stem (type B), with thirteen type B1, one type B2 and one type B3 fractures. Fractures were repaired with non-locking (18/28) or locking-plate fixation (10/28). Cerclage wire was applied around the plate and proximal bone segment in 17/28 dogs. Major complications occurred in 7/28 cases (five deep infection, two mechanical failures). Bone healing was noted in 21/23 cases, for which follow-up radiographic interpretation was available. Return to function was complete in 17 cases, acceptable in 8 cases and unacceptable in 3 cases. CONCLUSIONS: While cementless total hip replacement periprosthetic femoral fractures can be successfully repaired with lateral plate fixation, the risk of infection appears to be high.