Development of disease resistant fenugreek for Western Canada.

Cercospora leaf spot (CLS) caused by Cercospora traversiana is an important phyto-pathological problem of fenugreek (Trigonella foenum graecum), a multiuse legume crop. Knowledge about the inheritance of genes controlling CLS resistance is essential when selecting suitable breeding approaches while information about epidemiological factors affecting the disease can help develop new control strategies. Our greenhouse and field experiments showed CLS resistance in fenugreek (L3717 and PI138687) to be governed by a single dominant gene which is moderately heritable (46% narrow sense heritability). This indicates a relatively simple pathway for transfer of genes to adapted fenugreek cultivars. Rapid screening techniques (detached leaf assay and whole plant assay) were developed to identify the degree of resistance to C. traversiana in fenugreek genotypes. Several epidemiological factors such as temperature, physical injury (wounding), level of host resistance, plant age and inoculum concentration were found influencing CLS severity in controlled environment conditions

Evaluation of virus-induced gene silencing methods for forage legumes including alfalfa, sainfoin and fenugreek

Virus-induced gene silencing (VIGS) is a rapid reverse genetics tool that has been developed in a wide variety of plant species for assessing gene functions. However, while VIGS has been utilized successfully in the diploid model leguminous species Medicago truncatula (Gaertn.) (barrel medic), such a platform has yet to be established in forage legume crop species. Therefore, we evaluated the effectiveness of this method in forage legumes using a previously developed PEBV (pea early browning virus) system whereby a fragment of the pea (Pisum sativum L.) PHYTOENE DESATURASE (PDS) gene was transferred into a range of alfalfa (Medicago sativa L.), sainfoin (Onobrychis viciifolia Scop.), and fenugreek (Trigonella foenum-graecum L.) cultivars using leaf infiltration and apical meristem injection. Barrel medic was used as a positive control. Gene silencing was observed after 10–15 d through the presence of a leaf bleaching phenotype, and was confirmed using quantitative real-time RT-PCR. Silencing of PDS was achieved in a selection of cultivars in all species assessed, with the highest silencing efficiency apparent in fenugreek. The introduction of a highly homologous gene fragment from a heterologous plant species to target endogenous genes for transient VIGS-based silencing in a range of species of interest represents a potentially useful strategy for the rapid functional characterization of candidate genes in forages.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

TECHNIQUES FOR SCREENING CERCOSPORA LEAF SPOT RESISTANT FENUGREEK GENOTYPES

Cercospora leaf spot (CLS) caused by Cercospora traversiana is an important phyto-pathological problem of fenugreek (Trigonella foenum graecum L), a multiuse legume crop. Field screenings for resistant plants, although accurate and effective, demand significant time and a sizable workforce to accomplish the goal. Also, weather conditions in the field may not always be favourable for uniform disease spread which eventually may lead to failure of the overall experiment. Whole plant assays (WPA) and detached leaf assays (DLA) with artificial inoculation not only help in scaling up the number of plants screened but also reduce the space, time and the amount of inoculum needed for the experiment. The results from our two experiments indicate that both WPA and DLA methods can be used reliably to differentiate the resistant and susceptible genotypes of fenugreek. In addition, the correlation coefficient (r = 0.875, pThe accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Hybridization between resistant and susceptible fenugreek accessions and evaluation of Cercospora leaf spot resistance in segregating generations

Cercospora leaf spot (CLS), caused by Cercospora traversoana, is an important phyto-pathological problem of self-pollinated fenugreek (Trigonella-foenum graecum). Developing resistant genotypes in crop plants has been considered the best option to control diseases for economic, environmental, and social reasons. However, before this can be accomplished, knowledge about the inheritance of disease-resistant genes is necessary for creating high-yielding resistant genotypes. One susceptible fenugreek cultivar, Tristar, and two resistant accessions L3717 and PI138687 were used in two-way crosses using hand emasculation and pollination technique in a greenhouse. F1 plants were grown in a greenhouse and allowed to grow till maturity to produce F2 seeds. Some flowers from F1 plants were crossed back to both resistant and susceptible parents separately to generate backcross (BC1) seeds. Parents, F1, F2, and BC1 populations were grown in the greenhouse using a RCBD with four replications. Plants were inoculated 30 d after sowing with a suspension of C. traversoana at 2 × 105 conidia mL−1. Symptoms were observed and rated on individual plants 25 d after inoculation, and plants were categorized according to susceptible or resistant reactions based on rating scores. Mean disease score was significantly different (p < 0.0001) among generations. In both the cross combinations, results showed CLS resistance in fenugreek (from L3717 and PI138687) was governed by a single dominant gene which is moderately heritable (46% narrow sense heritability). This indicates a relatively simple pathway for transfer of genes to adapted fenugreek cultivars.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Elucidation of Physiological, Transcriptomic and Metabolomic Salinity Response Mechanisms in Medicago sativa

Alfalfa (Medicago sativa L.) is a widely grown perennial leguminous forage crop with a number of positive attributes. However, despite its moderate ability to tolerate saline soils, which are increasing in prevalence worldwide, it suffers considerable yield declines under these growth conditions. While a general framework of the cascade of events involved in plant salinity response has been unraveled in recent years, many gaps remain in our understanding of the precise molecular mechanisms involved in this process, particularly in non-model yet economically important species such as alfalfa. Therefore, as a means of further elucidating salinity response mechanisms in this species, we carried out in-depth physiological assessments of M. sativa cv. Beaver, as well as transcriptomic and untargeted metabolomic evaluations of leaf tissues, following extended exposure to salinity (grown for 3–4 weeks under saline treatment) and control conditions. In addition to the substantial growth and photosynthetic reductions observed under salinity treatment, we identified 1233 significant differentially expressed genes between growth conditions, as well as 60 annotated differentially accumulated metabolites. Taken together, our results suggest that changes to cell membranes and walls, cuticular and/or epicuticular waxes, osmoprotectant levels, antioxidant-related metabolic pathways, and the expression of genes encoding ion transporters, protective proteins, and transcription factors are likely involved in alfalfa’s salinity response process. Although some of these alterations may contribute to alfalfa’s modest salinity resilience, it is feasible that several may be disadvantageous in this context and could therefore provide valuable targets for the further improvement of tolerance to this stress in the future.Science, Faculty ofNon UBCChemistry, Department ofReviewedFacultyResearcherGraduat

Identification of Differential Drought Response Mechanisms in Medicago sativa subsp. sativa and falcata through Comparative Assessments at the Physiological, Biochemical, and Transcriptional Levels

Alfalfa (Medicago sativa L.) is an extensively grown perennial forage legume, and although it is relatively drought tolerant, it consumes high amounts of water and depends upon irrigation in many regions. Given the progressive decline in water available for irrigation, as well as an escalation in climate change-related droughts, there is a critical need to develop alfalfa cultivars with improved drought resilience. M. sativa subsp. falcata is a close relative of the predominantly cultivated M. sativa subsp. sativa, and certain accessions have been demonstrated to exhibit superior performance under drought. As such, we endeavoured to carry out comparative physiological, biochemical, and transcriptomic evaluations of an as of yet unstudied drought-tolerant M. sativa subsp. falcata accession (PI 641381) and a relatively drought-susceptible M. sativa subsp. sativa cultivar (Beaver) to increase our understanding of the molecular mechanisms behind the enhanced ability of falcata to withstand water deficiency. Our findings indicate that unlike the small number of falcata genotypes assessed previously, falcata PI 641381 may exploit smaller, thicker leaves, as well as an increase in the baseline transcriptional levels of genes encoding particular transcription factors, protective proteins, and enzymes involved in the biosynthesis of stress-related compounds. These findings imply that different falcata accessions/genotypes may employ distinct drought response mechanisms, and the study provides a suite of candidate genes to facilitate the breeding of alfalfa with enhanced drought resilience in the future