Flax secoisolariciresinol (SECO) diglucoside (SDG) lignan is an emerging natural product purported to prevent chronic diseases in humans. SECO, the aglycone form of SDG, has shown higher intestinal cell absorption but it is not accumulated naturally in planta. Recently, we have identified and characterized a UDP-glucosyltransferase gene, UGT74S1, that glucosylates SECO into its monoglucoside (SMG) and SDG forms when expressed in yeast. However, whether this gene is unique in controlling SECO glucosylation into SDG in planta is unclear. Here, we report on the use of UGT74S1 in reverse and forward genetics to characterize an ethyl methane sulfonate (EMS) mutagenized flax population from cultivar CDC Bethune and consisting of 1996 M2 families. EMS mutagenesis generated 73 SNP variants causing 79 mutational events in the UGT74S1 exonic regions of 93 M2 families. The mutation frequency in the exonic regions was determined to be one per 28 Kb. Of these mutations, 13 homozygous missense mutations and two homozygous nonsense mutations were observed and all were transmitted into the M3 and M4 generations. Forward genetics screening of the population showed homozygous nonsense mutants completely lacking SDG biosynthesis while the production of SMG was observed only in a subset of the M4 lines. Heterozygous or homozygous M4 missense mutants displayed a wide range of SDG levels, some being greater than those of CDC Bethune. No additional deleterious mutations were detected in these mutant lines using a panel of 10 other genes potentially involved in the lignan biosynthesis. This study provides further evidence that UGT74S1 is unique in controlling SDG formation from SECO and this is the first report of non-transgenic flax germplasm with simultaneous knockout of SDG and presence of SMG in planta.
Mutations are the key drivers for evolution and diversification in plants. In varietal selection, sources for variation are always sought as starting breeding materials. Thus, in the absence of desired natural variations in breeding populations, targeted or random mutagenesis is applied to induce variations. Cultivated potato (Solanum tuberosum L.) is autotetraploid crop species with a narrow and highly heterozygous genetic base, and the complexity of its genome makes its genetic studies more difficult. In the current study, induced mutagenesis was performed in diploid potato using ethyl methane sulfonate (EMS) to enlarge the genetic variability for its use as pre-breeding materials in both polyploid and diploid potato breeding. As starting materials, true potato seeds were treated with 1.2% EMS for 4-6 h along with untreated seeds as controls. A large variation in terms of germination rate, plant, flower, and tuber phenotype was observed in EMS-treated plants compared with their untreated counterparts. In particular, abnormal phenotypes including twisted stem, partial and (or) completely chlorotic leaves and stems, variations in stem colour and weak-stemmed plants with lateral growth habit as well as plants with determinate growth habit were observed along with normal plant characteristics. Moreover, variations in flower colour and tuber colour, shape, and size, as well as yield potential, were observed in EMS-treated lines. The reported phenotypic characterization of EMS mutagenized diploid potato collection is to our knowledge the first in its kind and represents a premium genetic resource for potato breeding programs and plant biologists for genes functional characterization in potato.
Main, D. C., Sanderson, K. R., Fillmore, S. A. E. and Ivany, J. A. 2013. Comparison of synthetic and organic herbicides applied banded for weed control in carrots ( Daucus carota L.). Can. J. Plant Sci. 93: 857–861. The necessity to reduce environmental impact and control the increasing cost of production has many carrot growers seeking new methods of crop management. This research evaluated the potential of applying herbicides in 30-cm bands over the carrot row in combination with between-the-row cultivation to control weeds. Linuron applied in a 30-cm-wide band over the row and linuron applied broadcast provided effective weed control and comparable yield. Clove oil and citrus oil gave slightly better weed control and yield than acetic acid and flaming at the late seeding date. This study demonstrates that acceptable weed control with minimal impact on yield can be achieved with the use of synthetic and organic herbicides applied as a 30-cm-wide band over the row combined with between-row cultivation. This method of weed control provides a 66% reduction in herbicide applied per hectare compared with a broadcast application.
Crop biofortification with Se is widely accepted. However, little is known about the efficiency of selenomethionine (Se‐Met) incorporation into the seed and tuber protein fraction of crops. Here, we report on the efficiency of Se incorporation into the seed and tuber protein fraction of soybean, flax, and potato and the putative identity of some of the proteins that are likely targets for this random pretranslation modification. Soybean, flax, and potato plants received Se using different sodium selenate application methods and rates, and Se‐Met was determined in seed, tuber, and their protein fractions. Selenium content in seed and tuber tissues and in protein fractions was increased by 2 to 86 times. The methionine (Met) pool was increased, as was the proportion of Se‐Met in the Met pool, but the ratio of Se‐Met:total Met was well conserved (1:1) between the treated and nontreated plants. After two‐dimensional (2D) protein electrophoresis, five protein spots were identified as glycinin, trypsin inhibitor (soybean), patatin‐3‐Kuras‐1, patain‐B2, and phosphoenol carboxy kinase (potato), none of which showed S to Se substitution in their primary structure. Linking Se and three stress markers in fresh tuber extracts, glutathione (GSH) level, glutathione peroxidase (GPx), and glutathione reductase (GR) activities were increased by 20 to 35%. The data showed low protein modification in these non‐Se‐accumulator crops and indicated that rates of 5 to 10 g ha–1 Se, through foliar application, may ensure not only an adequate daily intake levels for humans but may also activate antioxidant enzyme systems within the tuber tissue.
Common scab disease in potato has become a widespread issue in major potato production areas, leading to increasing economic losses. Varietal resistance is seen as a viable and long-term scab management strategy. However, the genes and mechanisms of varietal resistance are unknown. In the current study, a comparative RNA transcriptome sequencing and differential gene signaling and priming sensitization studies were conducted in two potato cultivars that differ by their response to common scab ( Streptomyces scabies) , for unraveling the genes and pathways potentially involved in resistance within this pathosystem. We report on a consistent and contrasted gene expression pattern from 1,064 annotated genes differentiating a resistant (Hindenburg) and a susceptible (Green Mountain) cultivars, and identified a set of 273 co-regulated differentially expressed genes in 34 pathways that more likely reflect the genetic differences of the cultivars and metabolic mechanisms involved in the scab pathogenesis and resistance. The data suggest that comparative transcriptomic phenotyping can be used to predict scab lesion phenotype in breeding lines using mature potato tuber. The study also showed that the resistant cultivar, Hindenburg, has developed and maintained a capacity to sense and prime itself for persistent response to scab disease over time, and suggests an immune priming reaction as a mechanism for induced-resistance in scab resistant potato cultivars. The set of genes identified, described, and discussed in the study paves the foundation for detailed characterizations towards tailoring and designing procedures for targeted gene knockout through gene editing and phenotypic evaluation.
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