Canadian Variety Research Articles

CRISPR/Cas9-mediated lipoxygenase gene-editing in yellow pea leads to major changes in fatty acid and flavor profiles.

Although pulses are nutritious foods containing high amounts of protein, fiber and phytochemicals, their consumption and use in the food industry have been limited due to the formation of unappealing flavors/aromas described as beany, green, and grassy. Lipoxygenase (LOX) enzymes are prevalent among pulse seeds, and their activity can lead to the formation of specific volatile organic compounds (VOCs) from certain polyunsaturated fatty acids (PUFAs). As a widespread issue in legumes, including soybean, these VOCs have been linked to certain unappealing taste perception of foods containing processed pulse seeds. To address this problem in pea and as proof of principle to promote the wider use of pulses, a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) construct was designed to create null alleles (knockouts) of PsLOX2 which had been implicated in the generation of VOCs in peas. Successful CRISPR/Cas9-mediated LOX gene editing of stable transgenic pea lines (TGP) was confirmed by DNA sequencing of the wild type (WT) and TGP pslox2 mutant lines. These lines were also assessed for LOX activity, PUFA levels, and VOCs. Compared to WT peas, the TGP lines showed a significant reduction (p < 0.05) in LOX activity and in the concentration of key VOCs, including hexanal, 2-hexenal, heptanal, (E)-2-heptenal, (E,E)-2,4-heptadienal, 1-octen-3-ol, octanal, (E)-2-octenal (E,E)-2,4-nonadienal and furan-2-pentyl. The content of two essential PUFAs, linoleic and α-linolenic acids, the known substrates of LOX in plants, was higher in TGP flours, indicating the efficacy of the CRISPR-mediated gene editing in minimizing their oxidation and the further modification of PUFAs and their products. The collection of VOCs from the headspace of ground pea seeds, using a portable eNose also distinguished the TGP and WT lines. Multiple regression analysis showed that LOX activity correlated with the two VOCs, heptanal and (E,E)-2,4-heptadienal in pea flours. Partial Least Squares Regression (PLS-R) plot for selected PUFAs, VOCs, and sensor responses in WT and TGP lines showed distinct clusters for WT and TGP lines. Together this data demonstrates the utility of CRISPR mediated mutagenesis of PsLOX2 to quickly improve aroma and fatty acid (FA) profiles of pea seeds of an elite Canadian variety.

Defining Mycotoxins Associated with Wheat Grains in Mosul Silo by ELISA

The study aimed to identify fungi-contaminated with some varieties of wheat grains locals, as well as isolate fungi-contaminated with grains imported wheat in Silo of Mosul city, and estimate the level of mycotoxins ochratoxin, zearalenone, and aflatoxin in them by ELISA method. Results showed that the local wheat variety, Ebaa 99, showed the highest percentage of appearance in Aspergillus flavus, was 63.15%, while the lowest appearance of fungi was Mucor spp. With a percentage of 2.47%, the variety Rasheed showed the highest percentage of A. flavus, which was 49.12%, while the lowest appearance was of Rhizopus spp. With a percentage of 2.29%, As well as in the variety Al-Baraka A. flavus showed the highest percentage, which was 5.453%, and the lowest percentage was Cladosporium spp by 9.43%. The contaminated fungi differed from the imported varieties and the percentage of their appearance. In the Canadian variety, the highest appearance of A. flavus was 63.72%, and A. dematatus was the least appearance with 5.42%. The fungus A. flavus also appeared in the American variety, with the highest appearance of 43.17%. The least appearance was Fusarium oxysporum at 5.34%. In the Australian variety, the highest appearance was found in A. flavus, at 54.03%, and the fungus Fusarium spp. was the least appeared fungi, at 1.96%. Estimation of mycotoxins indicates that the aflatoxin amount in the variety Ebaa 99 samples was 34.55 ppm, while its amount of ochratoxin and zearalenone was 21.75 ppm and 0.23 ppm, respectively. Canadian variety grains had the lowest aflatoxin content, with the highest ochratoxin. Australian variety grains had the lowest content of aflatoxin, ochratoxin, and zearalenone amount 12.29 ppm, 0.82 ppm, and 11.51 ppm, respectively.

Physicochemical Properties and Malting Potential of New Canadian Hulless Barley Genotypes

Interest in malting hulless barley (HB) is driven by its potential to produce very high levels of malt extract, but elevated levels of wort ß-glucan and insufficient levels of enzymes in hulless malt have prevented its widespread use. The objective of this study was to highlight the recent advancements in quality improvement of Canadian HB for malting purposes. Five advanced HB lines were evaluated for various barley and malt quality characteristics. All hulless lines exhibited lower grain hardness and slightly lower kernel weight and diameter compared to AAC Synergy (a popular two-rowed covered Canadian variety). The average total starch content of the hulless barley lines (64.2%) was significantly higher than AAC Synergy (59.9%) but slightly lower than that of CDC Clear (65.1%). Despite the low friability values, HB malts had a crumbly texture with good solubility and enzyme accessibility resulting in ample production of fermentable sugars. The average chain length of starch hydrolysis products in wort of new HB lines was similar to that of AAC Synergy, indicating sufficient enzyme activities and a highly effective hydrolysis. The fine-grind extract levels from hulless malt were on average 8% higher than from AAC Synergy, and several new HB lines exceeded the level of CDC Clear extract by 0.7–1.0%. Compared to CDC Clear, the new HB lines showed a significant reduction in concentration of wort ß-glucans. The concentration and the average degree of polymerization of arabinoxylo-oligosaccharides in wort of HB lines were generally higher than in wort of AAC Synergy, partly explaining the higher viscosity of HB worts. In general, the recently developed HB lines exhibited excellent malting potential with significant improvements in the levels of malt enzymes, concentration of free amino nitrogen compounds, and β-glucans in wort.

The prosody of rhetorical questions in English

This article contributes to our knowledge about the prosodic realisation of rhetorical questions (RQs) as compared to information-seeking questions (ISQs). It reports on a production experiment testing the prosody of Englishwh-and polar RQs and ISQs in a Canadian variety. In previous literature, the contribution of prosody to the distinction between the two illocution types has often been limited to the intonational realisation of the terminus of the utterance, i.e. whether it ends in a rise or a fall. Along with edge tones, we tested other phonological and phonetic parameters. Our results are as follows: (i) The intonational terminus was distinctive only for polar questions (rise vs plateau), not forwh-questions (low throughout). (ii) Moreover, the semantic difference between RQs and ISQs is signalled by pitch accents. It is reflected in nuclear pitch accent type forwh-questions, and accent type and position for polar questions. (iii) Phonetically, RQs are produced with longer constituent durations and – forwh-questions – a softer voice quality in thewh-word. Taken together, several intonational categories and phonetic parameters contribute to the distinction between RQs and ISQs. A simple distinction between rising and falling intonation is in any case insufficient.

Features of the Ukrainian soybean market

Purpose. Analyze the trends of soy bean production in the world and identify their impact on the soybean market in Ukraine. Identify the dynamics of submission and registration of applications for inclusion in the State Register of Plant Varieties, suitable for distribution in Ukraine. Compare soybean yields to the leaders of the global soybean market with the yield of soybeans in Ukraine. To highlight information on the best yields of new soybean varieties that are included in the State Register of Plant Varieties, suitable for distribution in Ukraine.Methods. Analytical, mathematical, statistical, graphic.Results The significant influence of trends on the global soybean market on the dynamics of this crop production in Ukraine and the reduction in the backlog of soybean production in Ukraine in dozens of times in comparison with the leaders in this field (USA, Brazil, Argentina, China and India) was discovered. Trends in production growth coincide with the trends of the global soybean market leaders in the USA, Brazil and Argentina. In the period from 2000 to 2014, the trend of the dynamics of production growth is of an expositional nature. The influence of production growth on the process of registration of soybeans new cultivars in the Re­gister of Plant Varieties of Ukraine was revealed. The growth trend in the number of submitted applications is positive. The largest number of soy bean varieties that passed the qualification examination was of Ukrainian descent. Among the varieties of foreign origin are varieties from Canada, France, Austria, USA, Germany, Argentina, Cyprus, Poland, Republic of Serbia, Romania, Croatia, Czech Republic, Switzerland. The new soybean varieties showed the highest yield during the period of qualifying examination in the period from 2010 to 2018.Conclusions. The growth rates of soybean production inUkraine have a similar tendency to growth rates among the leaders of this crop production. The Ukrainian varieties register is dominated by varieties of Ukrainian origin. The average level of yield of soybean cultivars in 2010–2011 corresponds to the average global yield of this crop in the world. The highest yields among the varieties that passed the qualifying exa­mination of plant varieties in the period from 2010 to 2018 were: ‘Silesia’ of French origin, ‘Relay’, ‘Terek’, ‘Avanturine’ of Ukrainian and a Canadian variety of ‘Kofu’.

A new lean no time test baking method with improved discriminating power

In response to customer concerns related to gluten strength in commercial baking, the Canadian Grain Commission assessed whether the Canadian Short Process (CSP) test bake method was generating useful data related to intrinsic strength of wheat varieties. Assessment of CSP loaf volume data for Canadian variety trials spanning 2003 to 2013 showed very little correlation with dough strength parameters as measured by farinograph and extensigraph. A lean no time (LNT) test baking method was developed that can better discriminate genotypes and provide objective indicators of the effect of intrinsic dough strength on baking quality. From early method development, through method validation and verification using diverse sets of samples targeting different Canadian wheat classes and grown in three different crop years, results showed the LNT method to be more discriminating and easily adopted by other laboratories. In 2015, the LNT method was adopted as the method of choice in future Canadian variety registration trials. The LNT method is fast, simple and well-suited to high throughput test baking conditions encountered in the evaluation of large numbers of breeder lines. A new objective parameter, loaf top ratio, was also introduced and found to correlate well with dough strength and dough handling properties.

A new lean no time test baking method with improved discriminating power

In response to customer concerns related to gluten strength in commercial baking, the Canadian Grain Commission assessed whether the Canadian Short Process (CSP) test bake method was generating useful data related to intrinsic strength of wheat varieties. Assessment of CSP loaf volume data for Canadian variety trials spanning 2003 to 2013 showed very little correlation with dough strength parameters as measured by farinograph and extensigraph. A lean no time (LNT) test baking method was developed that can better discriminate genotypes and provide objective indicators of the effect of intrinsic dough strength on baking quality. From early method development, through method validation and verification using diverse sets of samples targeting different Canadian wheat classes and grown in three different crop years, results showed the LNT method to be more discriminating and easily adopted by other laboratories. In 2015, the LNT method was adopted as the method of choice in future Canadian variety registration trials. The LNT method is fast, simple and well-suited to high throughput test baking conditions encountered in the evaluation of large numbers of breeder lines. A new objective parameter, loaf top ratio, was also introduced and found to correlate well with dough strength and dough handling properties.

Review: Shifting patterns in plant cultivar protection for field crops in Canada

Carew, R., Florkowski, W. J. and DePauw, R. 2015. Review: Shifting patterns in plant cultivar protection for field crops in Canada. Can. J. Plant Sci. 95: 813–829. In an era of increased globalization of agricultural trade and rapid biotechnology advances in developing crops with multiple traits, public and private institutions are facing increasing pressure to protect their technologies and develop partnerships to fund research. Under the auspices of international intellectual property rights agreements, Canada adopted stronger intellectual property rights policies to protect new plant cultivars and reward owners for their innovative efforts. This paper illustrates how Plant Breeders’ Rights (PBR) applications for agriculture field crop (cereals, oilseeds, pulses) cultivars and registered cultivars by the Canadian Variety Registration Office of the Canadian Food Inspection Agency (VROCFIA) have evolved over the past 20 yr in response to changes in domestic and international plant cultivar protection policies. The paper also examines the patenting pattern for plant-related traits and technologies. The paper found the bulk of PBR applications for field crops were accounted for by oilseeds followed by cereal and pulse crops such as field pea. In contrast to wheat and canola, which are protected by PBR and registered by the Canadian VROCFIA, pulse crops such as lentil have been developed by Canadian universities and funded by producer levies and released to growers without PBR protection. Among plant-related patents, most of them were credited to corn followed by canola and soybean. The Canadian public sector filed plant patents mainly for canola and concentrated on traits such as insect and disease resistance. The bulk of canola or soybean PBRs applications were by multinational life science seed companies, while the majority of wheat and barley were credited to public institutions such as the Canadian federal department of agriculture and universities.

Potato protein isolates: Recovery and characterization of their properties

An imitation of industrial potato fruit juice (PFJ) was prepared, using Canadian variety of potatoes, and was characterized of being composed of 22.9% patatin, 53.3% protease inhibitors, and 23.7% high MW proteins. To isolate potato proteins from PFJ, several extraction techniques were explored including thermal/acidic combination, acidic, FeCl3, MnCl2, ethanol and (NH4)2SO4 precipitations, and carboxymethyl cellulose complexation. (NH4)2SO4 precipitation led to the highest yield (98.6%) and to the recovery of protein isolates enriched in patatin with high resolubility. FeCl3 precipitation resulted in the highest purification factor (6.2) and isolates with the lowest relative proportion of high MW proteins (<4.6%); however, its optimal isolate showed a wide minimum solubility pH range of 3.0–6.0. FeCl3 and MnCl2 were identified as the best precipitating agents for the enrichment of isolates with >15kDa protease inhibitors. Trypsin inhibiting activities of protease inhibitors were highly preserved upon protein isolation than the chymotrypsin ones. Acidic-based protein isolate showed the highest specific lipid acyl hydrolase activity of patatin towards o-nitrophenyl butyrate, whereas FeCl3-based one exhibited the highest activity towards 4-nitrophenyl laurate.

Salt effect on phenolics and antioxidant activities of Tunisian and Canadian sweet marjoram (Origanum majoranaL.) shoots

Two varieties of Origanum majorana (Canadian and Tunisian) were evaluated for their phenolic, flavonoid and tannin contents, individual phenolic compounds and antioxidant activities under NaCl constraint. The results showed a significant variability in phenolic composition and antioxidant behavior between the two varieties under salt stress. The phenolic composition of methanolic extracts was determined by reversed-phase high-performance liquid chromatography. Amentoflavone was the predominant flavonoid compound; in addition, trans-2-hydrocinnamic acid became the major phenolic acid with salt treatment of the Tunisian variety. In the control, Canadian variety extract was characterized by high levels of gallic acid and amentoflavone. However, under 75 mmol L(-1) NaCl, gallic acid content doubled, whereas amentoflavone content was maintained in the Canadian variety. Stimulation of phenolic acid biosynthesis was observed in these two varieties under salt treatment despite the fact that shoots of the Tunisian variety showed higher antioxidant activities compared to those from the Canadian variety. Tunisian O. majorana might have developed tolerance to salinity and avoided tissue damage by activating enzymes involved in the galactosylation of quercetin into quercetin-3-galactoside and quercetin-3-rhamnoside. Our results confirmed the tolerance of Tunisian O. majorana plants.

Essential oil yield and trichomes structure in two sweet marjoram (Origanum majorana L.) varieties under salt stress

Origanum majorana L. shoots were investigated for their essential oil (EO) yield, composition and the structures responsible for its biosynthesis. Shoots EO was extracted by hydrodistillation and composition was determined by gas chromatography–mass spectrometry (GC-MS) method. Fresh Leaves were observed by light microscopy (LM). Plants were cultivated for 17 days in a basal medium supplemented with 75 mM NaCl. Salt decreased the essential oil yield in Tunisian variety (TV), but an increase was shown in Canadian variety (CV). Salt constraint induced a change in EO chemotype of two varieties. Their anatomical study showed three types of trichomes: (i) non-glandular, simple hairs; (ii) small, capitate glandular trichomes; (iii) and peltate glandular trichomes. Non-glandular trichomes provided of three types at the TV against two at the Canadian one at control. Indeed, with TV we observed; multicellular (M), bicellular (B) and unicellular (U) trichomes. Bicellular trichomes (BT) were observed in CV, and multicellular (MT) and bicellular (UT) in TV at salt constraint. The increase and the decrease of EO yield in CV and TV was respectively due to the presence of three type of EO secretion which seemed to make defect in Tunisian one. Key words: Trichomes, non-glandular, capitate, peltate, glandular, salt, chemotype.

Salt effect on physiological, biochemical and anatomical structures of two Origanum majorana varieties (Tunisian and Canadian)

In this study, we evaluated the salt concentration effect on plant growth, mineral composition, antioxidant responses and anatomical structure of two varieties of Origanum majorana after exposure to NaCl treatment. Our results show an inclusive behaviour of the two varieties, since the majority of sodium was exported and accumulated in their aerial parts. The Canadian variety (CV) appeared relatively more tolerant to salt than the Tunisian one (TV). Transversal section of leaves showed a thickening of dorsal and ventral cuticle, more importantly in CV than in TV, in the presence and in absence of salt. This was accompanied by an increase in the length of palisade cells, and the width of spongy collenchyma lacuna. The stem had a subquadrangular shape in TV and quadrangular in the Canadian variety. At mature stage, the stem pit was reabsorbed in the TV and replaced by a large cavity, whereas it remained unchanged in CV. The relative salt tolerance of the CV was related to: (1) a good selectivity in favour of K + : (2) a strong peroxidase activity and (3) an increase in the lengthening of palisade cell accompanied with an increase of lacunae in spongy parenchyma in CV. Key words: Origanum majorana , salinity, growth, mineral nutrition, leaves, stems, anatomical, antioxidant.

Salt effects on Origanum majorana fatty acid and essential oil composition

The effects of salt on the essential oil yield and fatty acid composition of aerial parts of two marjoram varieties were investigated. Plants with 6 leaves were treated with NaCl (75mM). Salt treatment led to a reduction in aerial part growth. Salinity increased the fatty acid content more significantly in Tunisian variety (TV) than in Canadian variety (CV). CV showed an increase in double-bond index (DBI) and a decrease in malondialdehyde content under salt stress, while the opposite was observed in TV. The DBI was mainly affected by a strong reduction in oleic and linoleic acids in TV, whereas a strong stimulation of linoleic acid in CV was observed. Salt decreased and increased the essential oil yield in TV and CV respectively. The main constituents of the essential oil of TV were trans-hydrate sabinene and terpinen-4-ol, which showed a significant decrease under salt stress. In contrast, the main constituents of the essential oil of CV were sabinene and trans-hydrate sabinene, which showed a significant decrease and increase respectively under salt stress. Marjoram oil is a rich source of many compounds such as essential oils and fatty acids, but the distribution of these compounds differed significantly between the two varieties studied.

Contextualizing St. John’s Youth English within the Canadian Quotative System

Investigations of the Canadian quotative system have to this point focused on mainland urban varieties where General Canadian English is considered to be the linguistic norm. The current analysis seeks to expand our understanding of this system by examining quotative usage among young girls in St. John’s, Newfoundland, where the local vernacular differs in significant ways from the national variety. Variationist methodology is employed on a small corpus of St. John’s Youth English (SJYE), revealing notable similarities in the distribution of quotatives as well as the operation of internal constraints across the paradigm between this variety and that of the mainland. At the same time, there is evidence that be like, the most recent of the quotative cohort, has grammaticalized further in SJYE than in General Canadian, raising questions about the routes by which this change is progressing. The results thus situate SJYEwithin the Canadian quotative system while at the same time highlighting the status of Newfoundland English as a unique Canadian variety.

Mapping and QTL analysis of the barley population Chebec × Harrington

A doubled haploid population of 120 individuals was produced from the parents Chebec, an Australian 2-row barley of feed quality with resistance to the cereal cyst nematode, and Harrington, a 2-rowed, Canadian variety of premium malting quality. This paper describes 18 field and laboratory experiments conducted with the population and summarises the traits mapped and analysed. The genomic location of 25 traits and genes is described and marker–trait associations for 5 traits (malt extract, diastatic power, resistance to cereal cyst nematode, early flowering, resistance to pre-harvest sprouting) important to Australian efforts to improve malting barley varieties have been used in practical breeding programs. Detailed maps for these populations are shown in this paper, while a consensus map incorporating these maps and further experiments on the populations are described elsewhere in this issue.

Study of the LMW glutenin composition of some old Hungarian wheat cultivars using capillary electrophoresis

The LMW glutenin composition of nine old Hungarian wheats (several lines per variety), and one Canadian wheat was studied using one-step, one-dimensional SDS-PAGE (Gupta and MacRitchie, 1991), after which a method was developed for the determination of wheat LMW glutenin patterns using SDS capillary electrophoresis (CE). In both cases the samples were prepared as described by Gupta and MacRitchie (1991), with some modifications in the case of CE. Both methods were used to analyse the homogeneity of the varieties, and the electropherograms obtained with the two methods were compared. It was found that the new method based on CE was suitable for the demonstration of differences and similarities between the wheat LMW glutenins. The method can be used to reveal differences not only between varieties (e.g. between the Hungarian variety Bankuti 1205 and the Canadian variety Marquis), but also between the genotypes making up a variety population (e.g. the sublines of Bankuti 1205). The results indicate that CE is a satisfactory alternative to the traditional PAGE method for the determination of the LMW glutenin subunits of wheat varieties.

The myth of Canadian English

Despite the recent publication of a Canadian style guide and The Oxford Canadian Dictionary, a claim that there is no such thing as a distinctively Canadian variety of English

Selection on pollen for linolenic acid content in rapeseed, Brassica napus L.

AbstractIn order to enhance the economic value of edible rapeseed oil, an improvement of quality is necessary. Mutagenesis of rapeseed resulted in a low linolenic acid content and a low ‘linolenic acid (CIS: 3) level to linoleic acid (CIS: 2) level’ ratio, that is, the linoleic desaturation ratio (LDR), in the seeds of the Canadian variety ‘Stellar’. As an early breeding marker for low linolenic acid content, the pollen fatty acid composition was determined on 80 doubled haploid plants derived from a single F1 hybrid obtained from a cross between ‘Stellar’ and a high CIS: 3 variety ‘Drakkar’. Fatty acid analysis on seed and pollen showed that the low CIS: 3 and the low LDR traits from the ‘Stellar’ variety were expressed in pollen and in seeds, and that a very close correlation (r = 0.88) existed between seed and pollen for these two traits. The inheritance of these traits is controlled by two major genes with additive effects, both in seed and pollen. However, minor genes also appeared to be expressed in pollen and seed. These genes may allow the production of plants with lower CIS: 3 levels than that of the low linolenic acid content parent. The efficiency of this new tool for early screening in breeding programmes is discussed.

Mapping of Molecular Markers on Brassica B‐Genome Chromosomes Added to Brassica napus

AbstractUsing interspecific hybridization among various Brassica species, B‐genome chromosomes from different sources of Brassica, i.e. B. nigra (BB, 2n = 18), B. carinata (BBCC), 2n = 34) and B. juncea (AABB, 2n = 36) were transferred into the Canadian variety ‘Andor’ of B. napus. Monosomic addition lines were selected (AACC + 1B, 2n = 39) by cytological control. For characterization of the alien chromosomes, series of isozymes, RFLPs and RAPD markers were employed. This permitted the identification of a total of 39 lines representing seven of the eight B‐genome chromosomes.

BIRKA BARLEY

Birka is a new two-rowed, nonmalting, predominantly blue aleurone spring barley. It was developed by Weibullsholm Plant Breeding Institute in Sweden and introduced to Canada in 1978. It was tested as W6482 by W. G. Thompson and Sons Ltd. before entering the 1979 Eastern Co-operative 2-row Barley Trial as TB7902. It outyielded Herta by 7 and 5% in Ontario and the Maritimes. Birka is 6 cm shorter and 1 day later in maturity than Herta. Birka has improved lodging resistance and test-weight, but similar disease resistance to Herta.Key words: Barley, Canadian variety