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43 result(s) for "Luckett, David J."
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Genome-Wide Delineation of Natural Variation for Pod Shatter Resistance in Brassica napus
Resistance to pod shattering (shatter resistance) is a target trait for global rapeseed (canola, Brassica napus L.), improvement programs to minimise grain loss in the mature standing crop, and during windrowing and mechanical harvest. We describe the genetic basis of natural variation for shatter resistance in B. napus and show that several quantitative trait loci (QTL) control this trait. To identify loci underlying shatter resistance, we used a novel genotyping-by-sequencing approach DArT-Seq. QTL analysis detected a total of 12 significant QTL on chromosomes A03, A07, A09, C03, C04, C06, and C08; which jointly account for approximately 57% of the genotypic variation in shatter resistance. Through Genome-Wide Association Studies, we show that a large number of loci, including those that are involved in shattering in Arabidopsis, account for variation in shatter resistance in diverse B. napus germplasm. Our results indicate that genetic diversity for shatter resistance genes in B. napus is limited; many of the genes that might control this trait were not included during the natural creation of this species, or were not retained during the domestication and selection process. We speculate that valuable diversity for this trait was lost during the natural creation of B. napus. To improve shatter resistance, breeders will need to target the introduction of useful alleles especially from genotypes of other related species of Brassica, such as those that we have identified.
Protease Inhibitors Purified from the Canola Meal Extracts of Two Genetically Diverse Genotypes Exhibit Antidiabetic and Antihypertension Properties
Valorization of vegetable oil waste residues is gaining importance due to their high protein and polyphenol contents. Protease inhibitors (PIs), proteins from these abundantly available waste residues, have recently gained importance in treating chronic diseases. This research aimed to use canola meal of genetically diverse Brassica napus genotypes, BLN-3347 and Rivette, to identify PIs with diverse functionalities in therapeutic and pharmacological applications. The canola meal PI purification steps involved: native PAGE and trypsin inhibition activity, followed by ammonium sulfate fractionation, anion exchange, gel filtration, and reverse-phase chromatography. The purified PI preparations were characterized using SDS-PAGE, isoelectric focusing (IEF), and N terminal sequencing. SDS-PAGE analysis of PI preparations under native reducing and nonreducing conditions revealed three polymorphic PIs in each genotype. The corresponding IEF of the genotype BLN-3347, exhibited three acidic isoforms with isoelectric points (pI) of 4.6, 4.0, and 3.9, while Rivette possessed three isoforms, exhibiting two basic forms of pI 8.65 and 9.9, and one acidic of pI 6.55. Purified PI preparations from both the genotypes displayed dipeptidyl peptidase-IV (DPP-IV) and angiotensin-converting enzyme (ACE) inhibition activities; the BLN-3347 PI preparation exhibited a strong inhibitory effect with lower IC50 values (DPP-IV 37.42 µg/mL; ACE 129 µg/mL) than that from Rivette (DPP-IV 67.97 µg/mL; ACE 376.2 µg/mL). In addition to potential human therapy, these highly polymorphic PIs, which can inhibit damaging serine proteases secreted by canola plant pathogens, have the potential to be used by canola plant breeders to seek qualitative trait locus (QTLs) linked to genes conferring resistance to canola diseases.
Extraction, Chemical Characterization, In Vitro Antioxidant, and Antidiabetic Activity of Canola (Brassica napus L.) Meal
Canola (Brassica napus L.) meal is a by-product after oil extraction from canola seed and is of relatively low value. This meal may have additional value in the biotechnology, food, and pharmaceutical industries if health-promoting useful bioactive compounds can be identified. Hence, seven canola meal extracts (CMEs) were generated using different organic solvents for two genotypes. HPLC and LCMS analyses were employed for the determination of the phenolic and antioxidant activity of meal extracts, including recovery of major biological compounds. When comparing genotype-1 with genotype-2, the latter had higher antioxidant activity in acetone extract (AE). This study also indicated seven major glucosinolates in CMEs in which water (WE) appeared to be the best solvent for the recovery of glucosinolates. Higher quantities of phenolic, glucosinolate, and antioxidant were present in genotype-2 compared with genotype-1. Using HPLC-DAD and LC-MS analysis 47 compounds were detected. We could identify 32 compounds in canola meal extracts: nine glucosinolates and twenty-three phenolic derivatives. Phenolic compounds in canola meal were conjugates and derivatives of hydroxycinnamic acid (sinapic, ferulic, and caffeic acids). Among phenolics, kaempherol as conjugate with sinapic acid was found; sinapine and trans-sinapic acid were the most abundant, as well as major contributors to the antioxidant and free radical scavenging activities of canola meal extracts. Some samples exhibited mild to moderate in-vitro antidiabetic activity in a Dipeptidyl Peptidase-IV inhibition assay.
Impact of Sowing Time and Genotype on Water Use Efficiency of Lentil (Lens culinaris Medick.)
Productive water use can be an effective adaptation strategy for improving crop performance. A 2-year field study was undertaken in 2018 and 2019 to investigate the effect of sowing date and genotype on water-use efficiency of lentils grown in diverse locations in Australia. Above-ground dry matter accumulation, grain yield, soil evaporation, water use, and water-use efficiency (WUE) were measured and/or calculated at crop maturity. Early sowing (SD1/mid-April), late maturity and supplementary irrigation increased water use. The long growth cycle resulting from early sowing influenced WUE for dry matter production and grain yield. WUE ranged from 10.5 to 18.8 kg dry matter ha−1 mm−1 (WUEET (evapotranspiration)) and 17.1 to 28.3 kg dry matter ha−1 mm−1 (WUET (transpiration)) for dry matter production. For grain yield, WUE ranged from 2.11 to 5.65 kg grain ha−1 mm−1 (WUEET) and 4.71 to 9.19 kg grain ha−1 mm−1 (WUET). There was more water loss through soil evaporation in SD1 compared to the other sowing dates. Excessive or limited availability of water did not translate to more dry matter accumulation and grain yield. The study concluded that SD1 gives the maximum water productivity for biomass, and SD2 (end of April) and SD3 (mid-May) for grain yield.
Phenolic Compounds with Antioxidant Properties from Canola Meal Extracts Inhibit Adipogenesis
The extraction of phenolic compounds from canola meal produces functional health products and renders the canola meal a more digestible animal feed. The extracted phenolics may have novel bioactivity worth investigation. In this study, several solvents were evaluated for their ability to extract phenolic compounds from canola meal: water (WE) and various 80% organic solvent/water mixtures of methanol (ME), acetone (AE), ethanol (EE), butanol (BE), chloroform (CE) and hexane (HE). The in vitro antioxidant and anti-obesity properties of various extracts were investigated. Anti-obesity properties were studied using adipogenic differentiation inhibition of a murine mesenchymal stem cell line (C3H10T1/2) and a pancreatic lipase inhibition assay. AE, ME, and BE showed significant (p < 0.05) adipogenesis and pancreatic lipase inhibitory activities and may have more pharmacological properties. AE down-regulated the gene expression of the major adipogenic transcription factor, peroxisome proliferator-activated receptor gamma (PPARγ), correlating to phenolic content in a dose-dependent manner. The chemical characterization of AE revealed the presence of sinapic acid, ferulic acid, and kaempferol derivatives as main bioactive phenols.
Least absolute shrinkage and selection operator regression used to select important features when predicting wheat yield from various genotype groups
Bread wheat and durum wheat genotypes were grown in field experiments at two locations in New South Wales, Australia across several years and using two sowing times (‘early’ v. ‘late’). Genotypes were grouped based on genetic similarity. Grain yield, grain size, soil characteristics and daily weather data were collected. The weather data were used to calculate water and heat stress indices for four key growth periods around flowering. Least absolute shrinkage and selection operator (LASSO) was used to predict grain yield and to identify the most influential features (a combination of index and growth period). A novel approach involving the crop water supply–demand ratio effectively summarized water relations during growth. LASSO predicted grain yield quite well (adjusted R 2 from 0.57 to 0.98), especially in a set of durum genotypes. However, the addition of other important variables such as lodging score, disease incidence, weed incidence and insect damage could have improved modelling results. Growth period 2 (30 days pre-flowering up to flowering) was the most sensitive for yield loss from heat stress and water stress for most features. Although one group of bread wheat genotypes was more sensitive to water stress (drought) in period 3 (20 days pre-flowering to 10 days post-flowering). Evapotranspiration was a significant positive feature but only in the vegetative phase (pre-flowering, period 1). This study confirms the usefulness of LASSO modelling as a technique to make predictions that could be used to identify genotypes that are suitable candidates for further investigation by breeders for their stress-tolerance ability.
Multi-environment QTL studies suggest a role for cysteine-rich protein kinase genes in quantitative resistance to blackleg disease in Brassica napus
Background Resistance to the blackleg disease of Brassica napus (canola/oilseed rape), caused by the hemibiotrophic fungal pathogen Leptosphaeria maculans , is determined by both race-specific resistance ( R ) genes and quantitative resistance loci (QTL), or adult-plant resistance (APR). While the introgression of R genes into breeding material is relatively simple, QTL are often detected sporadically, making them harder to capture in breeding programs. For the effective deployment of APR in crop varieties, resistance QTL need to have a reliable influence on phenotype in multiple environments and be well defined genetically to enable marker-assisted selection (MAS). Results Doubled-haploid populations produced from the susceptible B. napus variety Topas and APR varieties AG-Castle and AV-Sapphire were analysed for resistance to blackleg in two locations over 3 and 4 years, respectively. Three stable QTL were detected in each population, with two loci appearing to be common to both APR varieties. Physical delineation of three QTL regions was sufficient to identify candidate defense-related genes, including a cluster of cysteine-rich receptor-like kinases contained within a 49 gene QTL interval on chromosome A01. Individual L. maculans isolates were used to define the physical intervals for the race-specific R genes Rlm3 and Rlm4 and to identify QTL common to both field studies and the cotyledon resistance response. Conclusion Through multi-environment QTL analysis we have identified and delineated four significant and stable QTL suitable for MAS of quantitative blackleg resistance in B. napus , and identified candidate genes which potentially play a role in quantitative defense responses to L. maculans .
Modelling the effects of cold temperature during the reproductive stage on the yield of chickpea (Cicer arietinum L.)
Abstract During the reproductive stage, chilling temperatures and frost reduce the yield of chickpea and limit its adaptation. The adverse effects of chilling temperature and frost in terms of the threshold temperatures, impact of cold duration, and genotype-by-environment-by-management interactions are not well quantified. Crop growth models that predict flowering time and yield under diverse climates can identify combinations of cultivars and sowing time to reduce frost risk in target environments. The Agricultural Production Systems Simulator (APSIM-chickpea) model uses daily temperatures to model basic crop growth but does not include penalties for either frost damage or cold temperatures during flowering and podding stages. Regression analysis overcame this limitation of the model for chickpea crops grown at 95 locations in Australia using 70 years of historic data incorporating three cultivars and three sowing times (early, mid, and late). We modified model parameters to include the effect of soil water on thermal time calculations, which significantly improved the prediction of flowering time. Simulated data, and data from field experiments grown in Australia (2013 to 2019), showed robust predictions for flowering time (n = 29; R2 = 0.97), and grain yield (n = 22; R2 = 0.63–0.70). In addition, we identified threshold cold temperatures that significantly affected predicted yield, and combinations of locations, variety, and sowing time where the overlap between peak cold temperatures and peak flowering was minimal. Our results showed that frost and/or cold temperature–induced yield losses are a major limitation in some unexpected Australian locations, e.g., inland, subtropical latitudes in Queensland. Intermediate sowing maximise yield, as it avoids cold temperature, late heat, and drought stresses potentially limiting yield in early and late sowing respectively.
Phytosterol, Tocopherol and Carotenoid Retention during Commercial Processing of Brassica napus (Canola) Oil
Brassica napus (canola) seed is a rich source of phytosterols, tocopherols and carotenoids, which all have recognized health benefits, although these are reduced or lost during crude oil refinement. Many studies are now outdated, so new research to monitor bioactive retention through current processing techniques is warranted. In this work, canola seed, in-process seed, and oil samples were collected from the major stages of five commercial canola oil processes. Analysis of phytosterols, tocopherols and carotenoids indicated seed pre-treatment enhanced bioactive concentrations in the crude oil. Although the bleaching step in each process eliminated all carotenoids, high concentrations of phytosterols and tocopherols remained in the refined oil across all processes, with losses notably lower than those found in previous reports. Moreover, crude oil samples from a two-stage cold pressing process showed greatly enriched concentrations of tocopherols (+122%), sterols (+140%) and carotenoids (+217%). The results show that modern Australian canola oil processing retains high phytosterol and tocopherol concentrations and warrants further investigation into bioactive enrichment strategies. Given the growing interest in health-enhanced foods, this study provides opportunities for nutrition and health-enhanced oil products and the potential for adding value in the edible oil industry.
Evaluation of resistance to Phomopsis stem blight (caused by Diaporthe toxica) in Lupinus albus
Historically, in Australia, Phomopsis stem blight in Lupinus albus crops is rare. However, in 2004 an outbreak of this disease occurred in southern New South Wales affecting stems, pods and seeds of the cultivar Kiev-Mutant. This virulent outbreak represents a potential threat to the Australian lupin industry. The current research was therefore initiated to optimise disease screening protocols for evaluation of the disease. Screening for resistance to Phomopsis stem blight is important because stubble is a potential high source of inoculum in no-tillage systems, and grazing of Phomopsis infected stubble can cause lupinosis in stock animals. A single spore isolate of the fungal pathogen Diaporthe toxica collected from the 2004 outbreak was used to assess the levels of disease resistance in stem tissue in L. albus cultivars in some experiments and in other experiments field-infected stubble was used as inoculum. Resistance of current L. albus cultivars, breeding lines, and landraces to D. toxica was assessed in both glasshouse and field experiments in 2007 and 2008. The results showed that resistance existed in some cultivars and several germplasm accessions. Environmental effects and possible differences in pathogen race structure lead to some poor correlations of resistance ratings between different experiments (ranging from r  = −0.489 to 0.800, depending on the experiment and tissue assessed). In field experiments consistent expression of the disease was dependent on rainfall. Screening in an irrigated disease nursery improved the methodology. Nevertheless, results suggest that it will be possible to develop new L. albus varieties that are resistant to Phomopsis stem blight should the disease become widespread in south-eastern Australia.