Real-time quantitative PCR based method for the quantification of fungal biomass to discriminate quantitative resistance in barley and wheat genotypes to fusarium head blight

Kumar, Arun; Karre, Shailesh; Dhokane, Dhananjay; Kage, Udaykumar; Hukkeri, S. B.; Kushalappa, Ajjamada C.

doi:10.1016/j.jcs.2015.04.005

Cited by 29 publications

(23 citation statements)

References 30 publications

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“…Cultures grown for a week were used to prepare macroconidial suspension for inoculations. The final concentration of spore in the suspension was adjusted to 10 5 ml -1 using sterilized distilled water and 10 μl suspension was inoculated per spikelet [ 20 ]. Plants were inoculated at about 50% anthesis (GS = 60–65) and were covered with plastic bags for 48 hours to maintain high humidity in order to facilitate initial infection.…”

Section: Methodsmentioning

confidence: 99%

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Integrated Metabolo-Transcriptomics Reveals Fusarium Head Blight Candidate Resistance Genes in Wheat QTL-Fhb2

et al. 2016

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BackgroundFusarium head blight (FHB) caused by Fusarium graminearum not only causes severe losses in yield, but also reduces quality of wheat grain by accumulating mycotoxins. Breeding for host plant resistance is considered as the best strategy to manage FHB. Resistance in wheat to FHB is quantitative in nature, involving cumulative effects of many genes governing resistance. The poor understanding of genetics and lack of precise phenotyping has hindered the development of FHB resistant cultivars. Though more than 100 QTLs imparting FHB resistance have been reported, none discovered the specific genes localized within the QTL region, nor the underlying mechanisms of resistance.FindingsIn our study recombinant inbred lines (RILs) carrying resistant (R-RIL) and susceptible (S-RIL) alleles of QTL-Fhb2 were subjected to metabolome and transcriptome profiling to discover the candidate genes. Metabolome profiling detected a higher abundance of metabolites belonging to phenylpropanoid, lignin, glycerophospholipid, flavonoid, fatty acid, and terpenoid biosynthetic pathways in R-RIL than in S-RIL. Transcriptome analysis revealed up-regulation of several receptor kinases, transcription factors, signaling, mycotoxin detoxification and resistance related genes. The dissection of QTL-Fhb2 using flanking marker sequences, integrating metabolomic and transcriptomic datasets, identified 4-Coumarate: CoA ligase (4CL), callose synthase (CS), basic Helix Loop Helix (bHLH041) transcription factor, glutathione S-transferase (GST), ABC transporter-4 (ABC4) and cinnamyl alcohol dehydrogenase (CAD) as putative resistance genes localized within the QTL-Fhb2 region.ConclusionSome of the identified genes within the QTL region are associated with structural resistance through cell wall reinforcement, reducing the spread of pathogen through rachis within a spike and few other genes that detoxify DON, the virulence factor, thus eventually reducing disease severity. In conclusion, we report that the wheat resistance QTL-Fhb2 is associated with high rachis resistance through additive resistance effects of genes, based on cell wall enforcement and detoxification of DON. Following further functional characterization and validation, these resistance genes can be used to replace the genes in susceptible commercial cultivars, if nonfunctional, based on genome editing to improve FHB resistance.

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Section: Methodsmentioning

confidence: 99%

“…The screening is generally done based on spray inoculation which leads to high experimental errors, leading to inconsistent ranking of genotypes over years. Inoculation under controlled conditions can significantly reduce the experimental error and can enable quantification of both spikelet and rachis resistance [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

Integrated Metabolo-Transcriptomics Reveals Fusarium Head Blight Candidate Resistance Genes in Wheat QTL-Fhb2

et al. 2016

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“…RT‐qPCR was performed using fungal‐specific gene ( Tri6 ) primers. The abundance of Tri6 gene was normalized with TaActin, and the results obtained from RT‐qPCR experiment were used to estimate the fungal biomass (Kumar et al ., ).…”

Section: Methodsmentioning

confidence: 99%

Identification and characterization of a fusarium head blight resistance gene TaACT in wheat QTL‐2DL

Kage

Karre

Kushalappa

et al. 2016

Plant Biotechnology Journal

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SummaryFusarium head blight (FHB) resistance in wheat is considered to be polygenic in nature. Cell wall fortification is one of the best resistance mechanisms in wheat against Fusarium graminearum which causes FHB. Metabolomics approach in our study led to the identification of a wide array of resistance‐related (RR) metabolites, among which hydroxycinnamic acid amides (HCAAs), such as coumaroylagmatine and coumaroylputrescine, were the highest fold change RR metabolites in the rachis of a resistant near‐isogenic line (NIL‐R) upon F. graminearum infection. Placement of these metabolites in the secondary metabolic pathway led to the identification of a gene encoding agmatine coumaroyl transferase, herein referred to as TaACT, as a candidate gene. Based on wheat survey sequence, TaACT was located within a FHB quantitative trait loci on chromosome 2DL (FHB QTL‐2DL) between the flanking markers WMC245 and GWM608. Phylogenetic analysis suggested that TaACT shared closest phylogenetic relationship with an ACT ortholog in barley. Sequence analysis of TaACT in resistant and susceptible NILs, with contrasting levels of resistance to FHB, led to the identification of several single nucleotide polymorphisms (SNPs) and two inversions that may be important for gene function. Further, a role for TaACT in FHB resistance was functionally validated by virus‐induced gene silencing (VIGS) in wheat NIL‐R and based on complementation studies in Arabidopsis with act mutant background. The disease severity, fungal biomass and RR metabolite analysis confirmed TaACT as an important gene in wheat FHB QTL‐2DL, conferring resistance to F. graminearum.

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“…Spring barley is grown on ca 250,000 ha, which is 17.5 % of the total area sown to cereals (1,428,171 ha;Ministry of Agriculture, 2015). Grain yield and quality is affected not only by numerous agro-environmental factors (weather, preceding crop, flowering period) but also by pathogens and pests (Mesterházy, 2003;Paul et al, 2005;Šafránková et al 2010;Kumar et al, 2015). Of these latter, the greatest attention over the long term has been dedicated to fungi from the genus Fusarium, and in particular species producing mycotoxins (Semaškienė et al, 2006;Kulík, 2008).…”

Section: Introductionmentioning

confidence: 99%

Occurrence of Fusarium spp. on Malting Barley Grains in the Czech Republic During 2011-2013

Bezděková¹,

Šafránková²,

Víchová³

2016

Acta Univ. Agric. Silvic. Mendelianae Brun.

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frequency of occurrence and species spectrum of Fusarium fungi occurring on grains of five malting varieties of spring barley (Aksamit, Bojos, Malz, Radegast, and Kangoo) were monitored at two locations (Kroměříž and Žabčice, Czech Republic). The effect of three fungicide treatment variants on Fusarium species suppression was also evaluated. During the monitored period, five species were detected: F. poae, F. culmorum, F. graminearum, F. avenaceum, and F. tricinctum. The most frequently isolated species was F. poae. Radegast had the highest frequency of naturally occurring Fusarium fungi, while Kangoo was the least infected variety at both locations. The greatest fungicide effectiveness against Fusarium spp. occurrence on ears was recorded after the variant with application of Hutton at BBCH 39 and Prosaro 250 EC at BBCH 65. During the monitored years, Žabčice had a higher rate of infection by Fusarium fungi. The rates of barley infection by Fusarium pathogens differed among individual years, with the highest rate occurring in 2011 (16 -17 %) and the lowest rate in 2012 (1 -2 %).

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Real-time quantitative PCR based method for the quantification of fungal biomass to discriminate quantitative resistance in barley and wheat genotypes to fusarium head blight

Cited by 29 publications

References 30 publications

Integrated Metabolo-Transcriptomics Reveals Fusarium Head Blight Candidate Resistance Genes in Wheat QTL-Fhb2

Integrated Metabolo-Transcriptomics Reveals Fusarium Head Blight Candidate Resistance Genes in Wheat QTL-Fhb2

Identification and characterization of a fusarium head blight resistance gene TaACT in wheat QTL‐2DL

Occurrence of Fusarium spp. on Malting Barley Grains in the Czech Republic During 2011-2013

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