QTL-specific microarray gene expression analysis of wheat resistance to Fusarium head blight in Sumai-3 and two susceptible NILs

Genome ◽  
2009 ◽  
Vol 52 (5) ◽  
pp. 409-418 ◽  
Author(s):  
Saber Golkari ◽  
Jeannie Gilbert ◽  
Tomohiro Ban ◽  
J. Douglas Procunier
Keyword(s):  
Gene Expression ◽  
Fusarium Head Blight ◽  
Subtractive Hybridization ◽  
Triticum Aestivum L ◽  
Defense Responses ◽  
Suppressive Subtractive Hybridization ◽  
Head Blight ◽  
Microarray Gene Expression ◽  
Genes Encoding ◽  
Sumai 3

Fusarium head blight, predominantly caused by Fusarium graminearum (Schwabe) in North America, is a destructive disease that poses a serious threat to wheat ( Triticum aestivum L.) production around the world. cDNA microarrays consisting of wheat ESTs derived from a wheat – F. graminearum interaction suppressive subtractive hybridization library were used to investigate QTL-specific differential gene expression between the resistant Chinese cultivar Sumai-3 and two susceptible near isogenic lines (NILs) following inoculation with F. graminearum. Stringent conditions were employed to reduce the false discovery rate. A total of 25 wheat unigenes were found to express differentially in response to F. graminearum infection. Genes encoding pathogenesis-related (PR) proteins such as β-1,3-glucanase (PR-2), wheatwins (PR-4), and thaumatin-like proteins (PR-5) showed a significant upregulation in genotypes having the Sumai-3 3BS region. For these three genes, the gene activity was significantly less in the genotype (NIL-3) lacking the Sumai-3 3BS segment. Significant upregulation of phenylalanine ammonia-lyase was detected only in the resistant Sumai-3, indicating the importance of both the 2AL and 3BS regions in the activation of effective defense responses to infection by F. graminearum. Differences in gene expression between the resistant Sumai-3 and the susceptible NILs were found to be mainly quantitative in nature.

Induction of wheat defense and stress-related genes in response to Fusarium graminearum

Genome ◽  
2005 ◽  
Vol 48 (1) ◽  
pp. 29-40 ◽  
Author(s):  
Lingrang Kong ◽  
Joseph M Anderson ◽  
Herbert W Ohm
Keyword(s):  
Fusarium Head Blight ◽  
Real Time ◽  
Cdna Library ◽  
Fusarium Graminearum ◽  
Cdna Clone ◽  
Subtractive Hybridization ◽  
Suppressive Subtractive Hybridization ◽  
Nucleotide Sequence Analysis ◽  
Head Blight ◽  
Resistance Response

Fusarium head blight (FHB), caused by species of the fungus Fusarium, is a worldwide disease of wheat (Triticum aestivum L.). The Chinese T. aestivum 'Ning7840' is one of few wheat cultivars with resistance to FHB. To identify differentially expressed genes corresponding to FHB resistance, a cDNA library was constructed using pooled mRNA isolated from glumes of 'Ning7840' harvested at 2, 6, 12, 24, 36, 72, and 96 h after inoculation (hai) with a conidia spore suspension of Fusarium graminearum. Suppressive subtractive hybridization (SSH) cDNA subtraction was carried out using pooled glume mRNAs from the tester and the control. The cDNA library was differentially screened using the forward subtracted cDNAs and the reverse subtracted cDNAs as probes. Twenty-four clones with significant matches to either plant (16 sequences) or fungal (8 sequences) genes were isolated based on their specific hybridization with forward subtracted cDNA and not reverse subtracted cDNA. Six putative defense-related genes were confirmed by real-time quantitative reverse-transcriptase PCR. Many-fold higher induction of three clones (A3F8, B10H1, and B11H3) in the resistant genotypes compared with susceptible genotypes indicates a putative role in the resistance response to Fusarium graminearum. Transcript accumulations of P450, chitinase (Chi1), and one unknown gene (clone B8Q9) in both resistant and susceptible genotypes suggest an involvement in a generalized resistance response to F. graminearum. Nucleotide sequence analysis showed that cDNA clone A4C6 encodes a cytochrome P450 gene (CYP709C3v2), including 14 N-terminal amino acids that have a membrane-associated helical motif. Other domains characteristic of eukaryotic P450 are also present in CYP709C3v2. The deduced polypeptide of cDNA clone B2H2 encodes an acidic isoform of class I chitinase containing a 960-bp coding region. Southern hybridization using aneuploid lines of T. aestivum 'Chinese Spring' indicated that CYP709C3v2 was located on the short arm of chromosomes 2B and 2D.Key words: Fusarium head blight (FHB), suppressive subtractive hybridization, defense response, real-time quantitative RT-PCR.

Multiplexed PCR to screen for a major QTL carrying fusarium head blight resistance in Sumai-3 wheat

2006 ◽  
Vol 86 (3) ◽  
pp. 711-716 ◽  
Author(s):  
M. A. Matus-Cádiz ◽  
C. J. Pozniak ◽  
G. R. Hughes ◽  
P. Hucl
Keyword(s):  
Fusarium Head Blight ◽  
Fusarium Head Blight Resistance ◽  
Triticum Aestivum L ◽  
Wheat Breeding ◽  
Head Blight ◽  
Breeding Lines ◽  
Multiplexed Pcr ◽  
Trait Locus ◽  
Advanced Generation ◽  
Sumai 3

After the initial identification of microsatellites linked to economically important traits of interest, the additional investment to simplify the screening procedure for more routine use is of interest to plant breeders. The objective was to simplify an acrylamide gel based marker-assisted selection (MAS) method to facilitate high-through put screening for Qfhs.ndsu-3BS, a major quantitative trait locus carrying fusarium head blight (FHB) resistance in Sumai-3 wheat (Triticum aestivum L.). Method simplifications included incorporating the use of (1) 96-well DNA extractions, (2) multiplexed PCR reactions using microsatellite primers gwm493 and gwm533, and (3) agarose gels. Our modified FHB-MAS method was verified by screening six resistant (Sumai-3, ND2710, McVey, BacUp, HY644, and Alsen) and 52 susceptible parents and subsequently used to screen 5567 common wheat breeding lines developed from Sumai-3 derivatives. This simple and rapid method allows for the screening of 1000 lines per week, which can be used to skew segregating populations towards more resistant types and thereby advance primarily early and advanced generation lines that carry at least the major FHB QTL. Key words: PCR, Fusarium, microsatellites, gwm493, gwm533, Triticum

scholarly journals Wheat Blast and Fusarium Head Blight Display Contrasting Interaction Patterns on Ears of Wheat Genotypes Differing in Resistance

2016 ◽  
Vol 106 (3) ◽  
pp. 270-281 ◽  
Author(s):  
Xia Ha ◽  
Birger Koopmann ◽  
Andreas von Tiedemann
Keyword(s):  
Gene Expression ◽  
Fusarium Head Blight ◽  
Expression Patterns ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Interaction Patterns ◽  
Head Blight ◽  
Disease Phenotypes ◽  
Wheat Blast ◽  
Sumai 3

The interaction of wheat with two ear pathogens, Magnaporthe wheat blast (MWB) and Fusarium graminearum (Fusarium head blight, FHB), was studied on the phenotypic, histological, and gene expression level. Most of the 27 wheat cultivars inoculated with MWB and F. graminearum displayed inverse disease responses to blast and FHB infection. Two cultivars, Milan and Sumai 3, were selected expressing converse disease phenotypes to blast (Milan, R)/(Sumai 3, S) and FHB (Milan, S)/(Sumai 3, R). Confocal laser scanning microscopy revealed early (12 h postinoculation) colonization of the spikelets by MWB similarly on both cultivars, while F. graminearum infected anthers of the susceptible cultivar earlier. Both pathogens grew much faster in the rachilla of susceptible than resistant cultivars, indicating that resistance is mainly expressed in this part connecting the spikelet with the rachis. In general, O2− and H2O2 levels were unrelated to disease expression in the four studied interactions. The differential disease phenotypes, fungal spread in the rachis, and colonization patterns in the spikelets were confirmed by distinct gene expression patterns. Among the eight genes analyzed, seven were more strongly induced by FHB than by blast. Genes for chitinase (Chi2), β-1,3-glucanase (PR2), a plant defensin homolog (PRPI), and peroxidase (Pox2) were strongly upregulated in Milan in response to both pathogens, while PR2 and PR5 (thaumatin-like protein) were transiently triggered by MWB on both cultivars. Upregulation of cinnamoyl-CoA reductase (CCR), cytochrome P450 (CYP709C1), and UDP-glycosyl transferase (UGT) were more prominent in ears infected with F. graminearum, while upregulation of UGT was higher in Sumai 3 when infected with either pathogen. Cultivar resistance to FHB was reflected by clearly higher expression levels of UGT and CYP709C1 in Sumai 3. The differential responses of wheat to the two ear pathogens demonstrated in this study makes it unlikely that common resistance genes exist for control of FHB and blast, suggesting the need to stack many genes associated with resistance in breeding programs for multiple resistance.

Genetic relationships between resistances to Fusarium head blight and crown rot in bread wheat (Triticum aestivum L.)

2010 ◽  
Vol 121 (5) ◽  
pp. 941-950 ◽  
Author(s):  
Hao Bing Li ◽  
Guo Qiang Xie ◽  
Jun Ma ◽  
Gui Ru Liu ◽  
Shu Min Wen ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Fusarium Head Blight ◽  
Bread Wheat ◽  
Genetic Relationships ◽  
Triticum Aestivum L ◽  
Crown Rot ◽  
Head Blight

scholarly journals Molecular Characterization and Expression of PFT, an FHB Resistance Gene at the Fhb1 QTL in Wheat

2018 ◽  
Vol 108 (6) ◽  
pp. 730-736 ◽  
Author(s):  
Yi He ◽  
Xu Zhang ◽  
Yu Zhang ◽  
Dawood Ahmad ◽  
Lei Wu ◽  
...  
Keyword(s):  
Defense Responses ◽  
Wheat Breeding ◽  
Flanking Sequence ◽  
Head Blight ◽  
Breeding Programs ◽  
Diverse Range ◽  
Fhb Resistance ◽  
Trait Locus ◽  
Sumai 3

Fusarium head blight (FHB) is a destructive fungal disease in wheat worldwide. Efforts have been carried out to combat this disease, and the pore-forming toxin-like (PFT) gene at the quantitative trait locus (QTL) Fhb1 was isolated and found to confer resistance to FHB in Sumai 3. In this study, we characterized PFT in 348 wheat accessions. Four haplotypes of PFT were identified. The wild haplotype of PFT had higher resistance than other haplotypes and explained 13.8% of phenotypic variation in FHB resistance by association analysis. PFT was highly expressed during early flowering and increased after Fusarium graminearum treatment in Sumai 3. Analysis of the 5′ flanking sequence of PFT predicted that the cis elements of the PFT promoter were related to hormones and biological defense responses. However, PFT existed not only in the FHB-resistant accessions but also in some susceptible accessions. These results suggested that FHB resistance in a diverse range of wheat genotypes is partially conditioned by PFT. The profiling of FHB resistance and the PFT locus in this large collection of wheat germplasm may prove helpful for incorporating FHB resistance into wheat breeding programs, although more work is needed to reveal the exact role of the QTL Fhb1 in conferring resistance to fungal spread.

scholarly journals Screening of Differentially Expressed Microsporidia Genes from Nosema ceranae Infected Honey Bees by Suppression Subtractive Hybridization

Insects ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 199
Author(s):  
Zih-Ting Chang ◽  
Chong-Yu Ko ◽  
Ming-Ren Yen ◽  
Yue-Wen Chen ◽  
Yu-Shin Nai
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Differentially Expressed Genes ◽  
Honey Bees ◽  
Physiological Mechanism ◽  
Subtractive Hybridization ◽  
Infection Process ◽  
Nosema Ceranae ◽  
Differentially Expressed ◽  
Suppressive Subtractive Hybridization

The microsporidium Nosema ceranae is a high prevalent parasite of the European honey bee (Apis mellifera). This parasite is spreading across the world into its novel host. The developmental process, and some mechanisms of N. ceranae-infected honey bees, has been studied thoroughly; however, few studies have been carried out in the mechanism of gene expression in N. ceranae during the infection process. We therefore performed the suppressive subtractive hybridization (SSH) approach to investigate the candidate genes of N. ceranae during its infection process. All 96 clones of infected (forward) and non-infected (reverse) library were dipped onto the membrane for hybridization. A total of 112 differentially expressed sequence tags (ESTs) had been sequenced. For the host responses, 20% of ESTs (13 ESTs, 10 genes, and 1 non-coding RNA) from the forward library and 93.6% of ESTs (44 ESTs, 28 genes) from the reverse library were identified as differentially expressed genes (DEGs) of the hosts. A high percentage of DEGs involved in catalytic activity and metabolic processes revealed that the host gene expression change after N. ceranae infection might lead to an unbalance of physiological mechanism. Among the ESTs from the forward library, 75.4% ESTs (49 ESTs belonged to 24 genes) were identified as N. ceranae genes. Out of 24 N. ceranae genes, nine DEGs were subject to real-time quantitative reverse transcription PCR (real-time qRT-PCR) for validation. The results indicated that these genes were highly expressed during N. ceranae infection. Among nine N. ceranae genes, one N. ceranae gene (AAJ76_1600052943) showed the highest expression level after infection. These identified differentially expressed genes from this SSH could provide information about the pathological effects of N. ceranae. Validation of nine up-regulated N. ceranae genes reveal high potential for the detection of early nosemosis in the field and provide insight for further applications.

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