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Genetic Characterization of a Fusarium Head Blight Resistance QTL from Triticum turgidum ssp. dicoccoides

December 2021
Plant Molecular Biology Reporter 39(5)

DOI:10.1007/s11105-020-01277-0

Authors:

Daniela Soresi

Centro de Recursos Naturales Renovables de la Zona Semiárida (CERZOS), Universidad Nacional del Sur (UNS)-CONICET, Bahía Blanca, Argentina

Paolo Bagnaresi

Council for Agricultural Research and Agricultural Economy Analysis

Juan Manuel Crescente

National Scientific and Technical Research Council

Marina Diaz

Universidad Nacional del Sur

Show all 7 authorsHide

Qfhs.ndsu-3AS in wild emmer wheat (Triticum turgidum L. ssp. dicoccoides) is a major quantitative trait locus associated with type II resistance to Fusarium head blight (FHB). It was mapped in a population obtained from durum wheat Langdon (Triticum turgidum L. ssp. durum) and disomic Langdon-T. dicoccoides 3A substitution lines. Langdon(Dic-3A)-10 showed better performance against Fusarium graminearum (Fg) infection than the Langdon variety. To identify the genes underpinning Qfhs.ndsu-3AS, we used RNA-Seq approach in Langdon(Dic-3A)-10 and Langdon at 72 h after inoculation. The reference genomes of Svevo and Zavitan representing ssp. durum and ssp. dicoccoides, respectively, were used in parallel to align the reads and identify the genes expressed during the infection. The size of the Qfhs.ndsu-3AS region spans about 24 Mb and contains about 1000 genes. Fourteen differentially expressed genes (DEGs) were mapped in the delimited region and considered candidates for resistance. Five of them combined a higher expression level in Langdon(Dic-3A)-10 and known function in defense response: three receptor protein kinases, one bZip transcription factor, and one protease inhibitor. The alignment of the reads to the two genomes also allowed the identification of 25 genes expressed but no-DEGs localized in the Qfhs.ndsu-3AS region. Their coding sequences contained 63 non-synonymous changes and one frameshift mutation that could alter the function of the proteins and eventually contribute to resistance against Fg. This list is valuable information towards understanding the mechanism involved in resistance conferred by Qfhs.ndsu-3AS.

Severity progress curves after F. graminearum inoculation in the susceptible durum cultivar Langdon and the derived resistant line Langdon (Dic-3A)-10

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Qfhs.ndsu-3AS region. a Genetic map of Langdon(Dic-3A)-10 (Zhu et al. 2015). b Physical maps of durum wheat Svevo (Chr3A Svevo) and wild emmer Zavitan (Chr3A Zavitan). Marker names correspond to SSRs and EST-derived markers on chromosomes 3A (see Table 1). Markers in bold letters delimit the chromosomal segment defined by Zhu et al. 2015. In Zavitan, the Xgwm2 marker was mapped to ChrUn scaffold

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Chromosome distribution of DEGs using Svevo or Zavitan as the reference genomes (FDR ≤ 0.05, FC ≥ |2|)

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Schematic representation of fourteen differentially expressed genes (DEGs) and nine markers in the Qfhs.ndsu-3AS region of Svevo and Zavitan

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Validation of RNA-Seq expression patterns of five differentially expressed genes (cDNA): DEGs #2, #9, #4, and #6 only expressed in Langdon(Dic-3A)-10 (group I) and DEG#3 only expressed in Langdon (group II) by semi-quantitative RT-PCR. Amplifications of cDNA of genotypes Langdon (L) and Langdon(Dic-3A)-10 (LD) are shown; amplifications of genomic DNA (gDNA) were included as positive controls. PCR-fragments were separated on agarose gels. The differences in molecular weight between bands of the genomic and cDNA of DEG#9 and DEG#6 is due to the presence of an intron of 82 bp and 81 bp, respectively. The expression of the constitutive gene Actin was used as reference level. N: negative control containing the same PCR mixture, but without template cDNA. M: molecular weight marker

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Plant Molecular Biology Reporter (2021) 39:710–726

https://doi.org/10.1007/s11105-020-01277-0

ORIGINAL ARTICLE

Genetic Characterization ofaFusarium Head Blight Resistance QTL

fromTriticum turgidum ssp. dicoccoides

DanielaSoresi1,2 · PaoloBagnaresi3· Juan ManuelCrescente4,5· MarinaDíaz2· LuigiCattivelli3·

LeonardoVanzetti4,5· AliciaCarrera1,6

Received: 5 June 2020 / Accepted: 23 December 2020

Abstract

Qfhs.ndsu-3AS in wild emmer wheat (Triticum turgidum L. ssp. dicoccoides) is a major quantitative trait locus associated

with type II resistance to Fusarium head blight (FHB). It was mapped in a population obtained from durum wheat Langdon

(Triticum turgidum L. ssp. durum) and disomic Langdon-T. dicoccoides 3A substitution lines. Langdon(Dic-3A)-10 showed

better performance against Fusarium graminearum (Fg) infection than the Langdon variety. To identify the genes under-

pinning Qfhs.ndsu-3AS, we used RNA-Seq approach in Langdon(Dic-3A)-10 and Langdon at 72h after inoculation. The

reference genomes of Svevo and Zavitan representing ssp. durum and ssp. dicoccoides, respectively, were used in parallel

to align the reads and identify the genes expressed during the infection. The size of the Qfhs.ndsu-3AS region spans about

24Mb and contains about 1000 genes. Fourteen diﬀerentially expressed genes (DEGs) were mapped in the delimited region

and considered candidates for resistance. Five of them combined a higher expression level in Langdon(Dic-3A)-10 and known

function in defense response: three receptor protein kinases, one bZip transcription factor, and one protease inhibitor. The

alignment of the reads to the two genomes also allowed the identiﬁcation of 25 genes expressed but no-DEGs localized in

the Qfhs.ndsu-3AS region. Their coding sequences contained 63 non-synonymous changes and one frameshift mutation that

could alter the function of the proteins and eventually contribute to resistance against Fg. This list is valuable information

towards understanding the mechanism involved in resistance conferred by Qfhs.ndsu-3AS.

Keywords Durum wheat· Qfhs.ndsu-3AS—Fusarium head blight· Resistance genes· RNA-Seq analysis· Variant

comparison

Introduction

Fusarium graminearum (Fg) is the most common causal

agent of Fusarium Head Blight (FHB), a re-emerging and

destructive wheat disease in warm and humid regions

worldwide (McMullen etal. 1997). FHB has been reported to

cause extensive damage through direct losses in grain yield

and quality mainly because of the presence of mycotoxins

Key message

•Qfhs.ndsu-3AS from T. dicoccoides confers FHB resistance.

RNA-Seq revealed 14 DEGs expressed during infection and 25

no-DEGs harboring non-synonyms SNPs between resistant and

susceptible genomes, representing candidate genes for Qfhs.ndsu-

3AS

* Daniela Soresi

dsoresi@criba.edu.ar

1 CERZOS-CONICET, CCT-Bahía Blanca,

BahíaBlanca8000, Argentina

2 Departamento de Biología, Bioquímica y Farmacia,

Universidad Nacional del Sur, BahíaBlanca8000, Argentina

3 Council forAgricultural Research andEconomics—Research

Centre forGenomics andBioinformatics, via S. Protaso 302,

Fiorenzuolad’Arda(PC)29017, Italy

4 Grupo Biotecnología y Recursos Genéticos, EEA INTA

Marcos Juárez, Ruta 12 km 3, 2580MarcosJuárez, Argentina

5 Consejo Nacional de Investigaciones Cientíﬁcas y Técnicas

(CONICET), BuenosAires, Argentina

6 Departamento de Agronomía, Universidad Nacional del Sur,

8000BahíaBlanca, Argentina

/ Published online: 24 March 2021

1 3

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