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Powdery Mildew Resistance Evaluation and Resistance Gene Analysis of Sichuan Wheat Varieties (Lines)
YANGJingyuan, GAOJing, SHENGWenting, WANGFei, DUYuting, HUANGShiyu, MUHAMMADSaqlain, MUHAMMADHammad Latif, ZHUWei, ZHOUYonghong, WUDandan, HAOMing, KANGHouyang, LIYinghui
Chin Agric Sci Bull ›› 2025, Vol. 41 ›› Issue (21) : 58-63.
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Abbreviation (ISO4): Chin Agric Sci Bull
Editor in chief: Yulong YIN
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Powdery Mildew Resistance Evaluation and Resistance Gene Analysis of Sichuan Wheat Varieties (Lines)
Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive fungal disease that significantly reduces wheat yield and grain quality worldwide. This study aimed to evaluate powdery mildew resistance in Sichuan wheat varieties (lines) and characterize their resistance genes, which will provide an important theoretical basis and practical guidance for disease-resistant wheat breeding. A total of 168 Sichuan wheat varieties (lines) were screened for seedling powdery mildew resistance under artificial climate chamber, which were inoculated with Bgt isolate E09, and the resistance gene was analyzed by molecular marker detection and genomic in situ hybridization (GISH). The result showed that 35 wheat materials (20.8%) were resistant. Molecular marker analysis showed that 34 materials carries the Pm21 gene, and ‘Shumai 2352’ carries Pm56. GISH analysis further confirmed that wheat cultivar ‘Mianmai 367’ carries a whole-arm chromosomal translocation of V chromosome (6VS·6AL) harboring Pm21, and ‘Shumai 2352’ possesses a whole-arm translocation of R chromosome (6RS·6AL) containing Pm56. The study highlights the limited genetic diversity of powdery mildew resistance in Sichuan wheat germplasm which are mainly relying on the Pm21 resistance gene, and emphasizes the urgent need to diversify resistance sources by introducing more Pm genes to broaden the genetic basis of disease resistance sources.
wheat / powdery mildew / disease resistance gene / wheat breeding / molecular marker / resistance identification / genomic in situ hybridization (GISH)
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Bulk segregant analysis was used to obtain a random amplified polymorphic DNA (RAPD) marker specific for the rye chromosome arm of the 1BL.1RS translocation, which is common in many high-yielding bread wheat varieties. The RAPD-generated band was cloned and end-sequenced to allow the construction of a pair of oligonucleotide primers that PCR-amplify a DNA sequence only in the presence of rye chromatin. The amplified sequence shares a low level of homology to wheat and barley, as judged by the low strength of hybridization of the sequence to restriction digests of genomic DNA. Genetic analysis showed that the amplified sequence was present on every rye chromosome and not restricted to either the proximal or distal part of the 1RS arm. In situ hybridization studies using the amplified product as probe also showed that the sequence was dispersed throughout the rye genome, but that the copy number was greatly reduced, or the sequence was absent at both the centromere and the major sites of heterochromatin (telomere and nucleolar organizing region). The probe, using both Southern blot and in situ hybridization analyses, hybridized at a low level to wheat chromosomes, and no hybridizing restriction fragments could be located to individual wheat chromosomes from the restriction fragment length polymorphism (RFLP) profiles of wheat aneuploids. The disomic addition lines of rye chromosomes to wheat shared a similar RFLP profile to one another. The amplified sequence does not contain the RIS 1 sequence and therefore represents an as yet undescribed dispersed repetitive sequence. The specificity of the amplification primers is such that they will provide a useful tool for the rapid detection of rye chromatin in a wheat background. Additionally, the relatively low level of cross-hybridization to wheat chromatin should allow the sequence to be used to analyse the organization of rye euchromatin in interphase nuclei of wheat lines carrying chromosomes, chromosome segments or whole genomes derived from rye.
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Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive disease worldwide. Host resistance is the preferred method for limiting the disease epidemic, protecting environment, and minimizing the economic losses. In the present study, the reactions to powdery mildew for a collection of 600 wheat cultivars and breeding lines from different wheat growing regions were tested using the Bgt isolate E09. One hundred and sixteen resistant genotypes were identified and then crossed with susceptible wheat cultivars/lines to produce segregating populations for genetic analysis. Among them, 87, 19, and 10 genotypes displayed single, dual, and multiple genic inheritance, respectively. To identify the Pm gene(s) in those resistant genotypes, 16 molecular markers for 13 documented Pm genes were used to test the resistant and susceptible parents and their segregating populations. Of the 87 wheat genotypes that fitted the monogenic inheritance, 75 ones carried the Pm2a allele. Three, two, one, and two genotypes carried Pm21, Pm6, Pm4, and the recessive genes pm6 and pm42, respectively. Four genotypes did not carry any of the tested genes, suggesting that they might have other uncharacterized or new genes. The other 29 wheat cultivars/lines carried two or more of the tested Pm genes and/or other untested genes, including Pm2, Pm5, Pm6, and/or pm42. It was obvious that Pm2 was widely used in wheat production, whereas Pm1, Pm24, Pm33, Pm34, Pm35, Pm45, and Pm47 were not detected in any of these resistant wheat genotypes. This study clarified the genetic basis of the powdery mildew resistance of these wheat cultivars/lines to provide information for their rational utilization in different wheat growing regions. Moreover, some wheat genotypes which may have novel Pm gene(s) were mined to enrich the diversity of resistance source.
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Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most destructive diseases that pose a great threat to wheat production. Wheat landraces represent a rich source of powdery mildew resistance. Here, we report the map-based cloning of powdery mildew resistance gene Pm24 from Chinese wheat landrace Hulutou. It encodes a tandem kinase protein (TKP) with putative kinase-pseudokinase domains, designated WHEAT TANDEM KINASE 3 (WTK3). The resistance function of Pm24 was validated by transgenic assay, independent mutants, and allelic association analyses. Haplotype analysis revealed that a rare 6-bp natural deletion of lysine-glycine codons, endemic to wheat landraces of Shaanxi Province, China, in the kinase I domain (Kin I) of WTK3 is critical for the resistance function. Transgenic assay of WTK3 chimeric variants revealed that only the specific two amino acid deletion, rather than any of the single or more amino acid deletions, in the Kin I of WTK3 is responsible for gaining the resistance function of WTK3 against the Bgt fungus.
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Host resistance conferred by Pm genes provides an effective strategy to control powdery mildew. The study of Pm genes helps modern breeding develop toward more intelligent and customized. Powdery mildew of wheat is one of the most destructive diseases seriously threatening the crop yield and quality worldwide. The genetic research on powdery mildew (Pm) resistance has entered a new era. Many Pm genes from wheat and its wild and domesticated relatives have been mined and cloned. Meanwhile, modern breeding strategies based on high-throughput sequencing and genome editing are emerging and developing toward more intelligent and customized. This review highlights mining and cloning of Pm genes, molecular mechanism studies on the resistance and avirulence genes, and prospects for genomic-assisted breeding for powdery mildew resistance in wheat.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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