Comparative Analysis of Deoxynivalenol Biosynthesis Related Gene Expression among Different Chemotypes of Fusarium graminearum in Spring Wheat

Fusarium mycotoxins, deoxynivalenol (DON) and nivalenol (NIV) act as virulence factors and are essential for symptom development after initial infection in wheat. To date, 16 genes have been identified in the deoxynivalenol biosynthesis pathway. However, a comparative gene expression analysis in different chemotypes of F. graminearum in response to fusarium head blight (FHB) infection remains to be explored. Therefore, in this study, nine genes that involved in trichothecene biosynthesis were analysed among 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON) and nivalenol (NIV) producing F. graminearum strains in a time course study. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) revealed that the expression of all examined TRI gene transcripts initiated at two days post-inoculation (dpi), peaked at three to four dpi and gradually decreased at seven dpi. The early induction of TRI genes indicates that presence of high levels of TRI gene transcripts at early stages, is important to initiate the biosynthetic pathway of DON and NIV. Comparison of gene expression among the three chemotypes showed that relative expression of TRI genes was higher in 3-ADON producing strains compared with 15-ADON and NIV strains. Comparatively higher levels of gene expression may contribute to the higher levels of DON produced by 3-ADON strains in infected grains

Construction and Characterization of a cDNA Library from Wheat Infected with Fusarium graminearum Fg 2

Total RNA from wheat spikes infected with F. graminearum Fg2 was extracted and the mRNA was purified. Switching Mechanism at 5′ end of the RNA Transcript (SMART) technique and CDS Ill/3′ primer were used for first-strand cDNA synthesis using reverse transcriptase by RT-PCR. Primer extension polymerase chain reaction was used to construct the double-strand cDNA that was digested by proteinase K, then by Sfi I and fractionated. cDNAs longer than 0.5 kb were collected and ligated to λTriplEx2 vector followed λ phage packaging reaction and library amplification. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. One hundred and sixty five plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. A high quality cDNA library from wheat spikes that have been infected by F. graminearum was successfully constructed

Validating the Strategic Deployment of Blackleg Resistance Gene Groups in Commercial Canola Fields on the Canadian Prairies

Blackleg, caused by the fungal pathogen Leptosphaeria maculans, is a serious threat to canola (Brassica napus L.) production in western Canada. Crop scouting and extended crop rotation, along with the use of effective genetic resistance, have been key management practices available to mitigate the impact of the disease. In recent years, new pathogen races have reduced the effectiveness of some of the resistant cultivars deployed. Strategic deployment and rotation of major resistance (R) genes in cultivars have been used in France and Australia to help increase the longevity of blackleg resistance. Canada also introduced a grouping system in 2017 to identify blackleg R genes in canola cultivars. The main objective of this study was to examine and validate the concept of R gene deployment through monitoring the avirulence (Avr) profile of L. maculans population and disease levels in commercial canola fields within the Canadian prairies. Blackleg disease incidence and severity was collected from 146 cultivars from 53 sites across Manitoba, Saskatchewan, and Alberta in 2018 and 2019, and the results varied significantly between gene groups, which is likely influenced by the pathogen population. Isolates collected from spring and fall stubble residues were examined for the presence of Avr alleles AvrLm1, AvrLm2, AvrLm3, AvrLm4, AvrLm5, AvrLm6, AvrLm7, AvrLm9, AvrLm10, AvrLm11, AvrLepR1, AvrLepR2, AvrLep3, and AvrLmS using a set of differential host genotypes carrying known resistance genes or PCR-based markers. The Simpson’s evenness index was very low, due to two dominant L. maculans races (AvrLm2-4-5-6-7-10-11 and AvrLm2-5-6-7-10-11) representing 49% of the population, but diversity of the population was high from the 35 L. maculans races isolated in Manitoba. AvrLm6 and AvrLm11 were found in all 254 L. maculans isolates collected in Manitoba. Knowledge of the blackleg disease levels in relation to the R genes deployed, along with the L. maculans Avr profile, helps to measure the effectiveness of genetic resistance

Characterization of Callose Deposition and Analysis of the Callose Synthase Gene Family of <i>Brassica napus</i> in Response to <i>Leptosphaeria maculans</i>

Callose plays a critical role in different biological processes including development as well as in the response to multiple biotic and abiotic stresses. In this study, we characterized the callose deposition in cotyledons of different Brassica napus varieties post-inoculated with different Leptosphaeria maculans isolates. Further, members of the callose synthase gene were identified from the whole genome of B. napus using the 12 Arabidopsis thaniana callose synthase protein sequences, and were then classified into three groups based on their phylogenetic relationships. Chromosomal location and duplication patterns indicated uneven distribution and segmental duplication patterns of BnCalS genes in the B. napus genome. Subsequently, gene structures, conserved domains analysis, and protein properties were analyzed for BnCalS genes. In addition, 12 B. napus orthologs of the AtCalS were selected for investigating the tissue expression pattern, indicating diverse expression patterns for these BnCalS genes. Responses of the selected 12 orthologs and all the BnCalS genes were characterized in the different types (AvrLm1-Rlm1, AvrLm4-Rlm4, AvrLepR1-LepR1) of B. napus&#8315;L. maculans interactions and B. napus-Leptosphaeria biglobosa interactions, implying their potential roles in response to Leptosphaeria infection

Detection of Leptosphaeria maculans and Leptosphaeria biglobosa Causing Blackleg Disease in Canola from Canadian Canola Seed Lots and Dockage

Blackleg, caused by Leptosphaeria maculans, is a major threat to canola production in Canada. With the exception of China, L. maculans is present in areas around the world where cruciferous crops are grown. The pathogen can cause trade barriers in international canola seed export due to its potential risk as a seed contaminant. The most recent example is China restricting canola seeds imported from Canada and Australia in 2009. Therefore, it is important to assess the level of Blackleg infection in Canadian canola seed lots and dockage (seeds and admixture). In this study, canola seed lots and dockage samples collected from Western Canada were tested for the presence of the aggressive L. maculans and the less aggressive L. biglobosa. Results showed that both L. maculans and L. biglobosa were present in seed lots and dockage samples, with L. biglobosa being predominant in infected seeds. Admixture separated from dockage had higher levels of L. maculans and L. biglobosa infection than samples from seed lots. Admixture appears to harbour higher levels of L. maculans infection compared to seeds and is more likely to be a major source of inoculum for the spread of the disease than infected seeds

Evolve Roundup Ready high erucic acid, low glucosinolate hybrid summer rape

Evolve is the world’s first ogu INRA CMS hybrid summer rape (Brassica napus L.) Roundup Ready high erucic acid, low glucosinolate cultivar. On average, Evolve yielded 12.4 % more seed, 6 g kg-1 more seed oil and 5 g kg -1 less meal protein than Red River 1861, an open-pollinated Roundup Ready high erucic acid, low glucosinolate summer rape. Evolve has an erucic acid content of 52.3 % in isolated field trials of HEAR genotypes and is adapted to the southern B. napus growing regions of western Canada.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