Chibbar, R.N.

Gangola, M.P.

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eCommons@USASK

Development and utilization of single nucleotide polymorphic markers on udp-glucosyl transferases to develop Fusarium head blight resistant wheat varieties

University of Saskatchewan Research Archive

Development and Utilization of Single Nucleotide Polymorphic Markers on UDP-Glucosyl Transferases to Develop Fusarium Head Blight Resistant Wheat VarietiesManu Pratap GangolaDepartment of Plant SciencesFusarium head blight (FHB) – New sources of resistance are needed Healthy grains FHB infected grainsFHB infected wheat spikeshttp://www.uky.edu/Ag/Wheat/wheat_breeding/FHB.htm• Major disease of wheat• Fusarium graminearum Schwabe– most common species• FHB reduces wheat yield and grain quality• Deoxynivalenol (DON) – virulence factor for FHB• Mutagenesis can be followed to induce variation in the traitToxic to humans2 ppm – Food1 ppm – Baby foodHealth Canada 2016FHB resistance in wheat • FHB resistance – Quantitative trait• B-genome of wheat (AABBDD) participates predominantly in FHB resistance• FHB1 - major QTL (Quantitative Trait Loci)- Present on 3BS • UGT gene - Encodes UDP-glucosyl transferase- Present in close vicinity of Fhb1Hypothesis• Ethylmethane sulfonate can induce variation for FHB resistance using in vitro spike culture technique.• Candidate genes for FHB resistance on B-genome of wheat can be targeted to find single nucleotide polymorphic (SNP) markers.Objectives• To study FHB resistance in the mutant population.• To identify SNP markers associated with FHB resistance.• To utilize FHB resistant genotypes and SNP markers in a breeding program.Spike Culture Derived Variants (SCDV) of AC Nanda - EMSA population of 134 SCDV lines was developedIdentification of FHB resistant and susceptible lines – in vitro spike cultureWheat spikes of resistant and susceptible SCDV linesAC Nanda – FHB susceptible controlSumai3 – FHB resistant controlWorkflow of the projectIdentification of SNPs by comparing gene sequences from AC Nanda and Sumai3SNPs validation in SCDV populationExpression and DON accumulation studyTranscriptome analysisBreeding programField experimentsGenome specificity of the primers2A = Triticum monococcum2B = Aegilops speltoides2D = Aegilops Tauschii2A2B2D = Hexaploid WheatSchematic representation of B-genome specific UGT genesSingle Nucleotide Polymorphic  (SNP) Markers on the genesSNP markers on UGTs can significantly differentiate between FHB -resistant and -susceptible genotypesHigh resolution melt curve (HRM) analysis TaUGT-2BTaUGT-3BTaUGT-ESTBox plot analysis of UGTs and FHB severityExpression of UGTs correlates with accumulation of DeoxynivalenolLocation and TraitMean Sum of SquaresGenotype ReplicationCarmenIncidence 260.5*** 13.3nsSeverity 285.5* 40.8nsVisual rating index (VRI) 265.1** 29.5nsFHB Nursery (University of Saskatchewan)Incidence 651.0** 175.2nsSeverity 76.2**  200.2ns  VRI 72.9***  93.2ns  FHB rating in field experiments confirms the reliability of in vitro spike culture technique***, ** and * are significant at P ≤ 0.001, 0.01 and 0.05, respectively whereas, ns represents non-significant values.Transcriptome analysis during FHB progressionRNA-seq was performed in FHB resistance (5) and susceptible (3) wheat genotypesNew candidate genes and markers will be identifiedBreeding Program• FHB resistant SCDV lines are being crossed with Saskatchewan elite cultivars.• SNP markers are being utilized to screen the population.Conclusions and Future Directions• An in vitro spike culture method was optimized to evaluate FHB resistance in wheat.• New genetic resources of FHB resistance and associated SNP markers were identified that are being used in the breeding program to develop FHB resistant wheat variety.• To understand the FHB resistance mechanism, transcriptomics has been done on FHB resistant and susceptible lines and the data is being analyzed. Acknowledgements• Ravindra N. Chibbar• Seedhabadee Ganeshan• H. Randy Kutcher• Pierre J. Hucl• Pallavi Sharma• Chen Huang

https://harvest.usask.ca/bitstream/10388/8695/1/M.%20Gangola%20and%20R.%20Chibbar%2c%202018.pdf

Development and utilization of single nucleotide polymorphic markers on udp-glucosyl transferases to develop Fusarium head blight resistant wheat varieties

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