Molecular cytogenetics for a wheat–Aegilops geniculata 3Mg alien addition line with resistance to stripe rust and powdery mildew

Abstract Background Aegilops geniculata Roth is closely related to common wheat (Triticum aestivum L.) and is a valuable genetic resource for improvement of wheat. Results In this study, the W19513 line was derived from the BC1F10 progeny of a cross between wheat ‘Chinese Spring’ and Ae. geniculata SY159. Cytological examination showed that W19513 contained 44 chromosomes. Twenty-two bivalents were formed at the first meiotic metaphase I in the pollen mother cellsand the chromosomes were evenly distributed to opposite poles at meiotic anaphase I. Genomic in situ hybridization demonstrated that W19513 carried a pair of alien chromosomes from the M genome. Fluorescence in situ hybridization confirmed detection of variation in chromosomes 4A and 6B. Functional molecular marker analysis using expressed sequence tag–sequence-tagged site and PCR-based landmark unique gene primers revealed that the alien gene belonged to the third homologous group. The marker analysis confirmed that the alien chromosome pair was 3Mg. In addition, to further explore the molecular marker specificity of chromosome 3Mg, based on the specific locus amplified fragment sequencing technique, molecular markers specific for W19513 were developed with efficiencies of up to 47.66%. The W19513 line was inoculated with the physiological race E09 of powdery mildew (Blumeria graminis f. sp. tritici) at the seedling stage and showed moderate resistance. Field inoculation with a mixture of the races CYR31, CYR32, CYR33, and CYR34 of the stripe rust fungus (Puccinia striiformis f. sp. triticii) revealed that the line W19513 showed strong resistance. Conclusions This study provides a foundation for use of the line W19513 in future genetic research and wheat improvement.

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Molecular Cytogenetics and SLAF-seq-based Specific Molecular Markers for a Wheat–Aegilops geniculata 3M g Alien Addition Line with Resistance to Stripe Rust and Powdery Mildew

10.21203/rs.3.rs-737578/v1 ◽

2021 ◽

Author(s):

Yongfu Wang ◽

Xiaofang Cheng ◽

Xiaoying Yang ◽

Changyou Wang ◽

Hong Zhang ◽

...

Keyword(s):

Molecular Markers ◽

Powdery Mildew ◽

Molecular Marker ◽

Stripe Rust ◽

Puccinia Striiformis ◽

Addition Line ◽

Field Inoculation ◽

Aegilops Geniculata ◽

Marker Analysis

Abstract Background: Aegilops geniculata Roth is closely related to common wheat (Triticum aestivum L.) and is a valuable genetic resource for improvement of wheat. Results: In this study, the W19513 line was derived from the BC1F10 progeny of a cross between wheat ‘Chinese Spring’ and Ae. geniculata SY159. Cytological examination showed that W19513 contained 44 chromosomes. Twenty-two bivalents were formed at the first meiotic metaphase in the pollen mother cells, and the chromosomes were evenly distributed to opposite poles at meiotic anaphase. Genomic in situ hybridization demonstrated that W19513 carried a pair of alien chromosomes from the M genome. Fluorescence in situ hybridization confirmed detection of variation in chromosomes 4A and 6B. Functional molecular marker analysis using expressed sequence tag–sequence-tagged site and PCR-based landmark unique gene primers revealed that the alien gene belonged to the third homologous group. The marker analysis confirmed that the alien chromosome pair was 3Mg. In addition, to further explore the molecular marker specificity of chromosome 3Mg, based on the specific locus amplified fragment sequencing technique, molecular markers specific for W19513 were developed with efficiencies of up to 47.66%. The W19513 line was inoculated with the physiological race E09 of powdery mildew (Blumeria graminis f. sp. tritici) at the seedling stage and showed moderate resistance. Field inoculation with a mixture of the races CYR31, CYR32, CYR33, and CYR34 of the stripe rust fungus (Puccinia striiformis f. sp. triticii) revealed that the line W19513 showed strong resistance.Conclusions: This study provides a foundation for use of the line W19513 in future genetic research and wheat improvement.

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Molecular Cytogenetic Characterization of a New T2BL·1RS Wheat-Rye Chromosome Translocation Line Resistant to Stripe Rust and Powdery Mildew

Plant Disease ◽

10.1094/pdis-93-2-0124 ◽

2009 ◽

Vol 93 (2) ◽

pp. 124-129 ◽

Cited By ~ 20

Author(s):

Chunmei Wang ◽

Qi Zheng ◽

Lihui Li ◽

Yongchun Niu ◽

Haibo Wang ◽

...

Keyword(s):

In Situ Hybridization ◽

Powdery Mildew ◽

Stripe Rust ◽

Expressed Sequence Tag ◽

Puccinia Striiformis ◽

Wheat Breeding ◽

Translocation Line ◽

Winter Rye ◽

New Genotype

Wheat (Triticum aestivum) genotypes with rye (Secale cereale) 1RS chromosomal translocations are widely used in wheat breeding programs because 1RS carries genes for resistance to several diseases. However, some of the pathogens have evolved into new races that overcome the resistance due to extensive use of cultivars with the resistance genes from rye. Therefore, identification and deployment of new resistance sources with desirable agronomic characteristics are important and urgent. We have used winter rye cultivar German White as a source of genes for desirable traits in wheat improvement. A new genotype named WR04-32 was produced through hybridization and chromosome manipulation between common winter wheat cultivar Xiaoyan 6 and German White. This genotype was highly resistant to a wide spectrum of the wheat stripe rust (Puccinia striiformis f. sp. tritici) and powdery mildew (Blumeria graminis f. sp. tritici) pathotypes prevalent in China. The polymerase chain reaction (PCR) result using EST-STS (expressed sequence tag-site tagged sequence) marker STSWE126 specific to 1RS confirmed 1RS in WR04-32, and it was further proved to be a wheat-rye T2BL·1RS translocation line using sequential genomic in situ hybridization (GISH) and multicolor fluorescence in situ hybridization (FISH) with probes pAs1 and pSc119.2 (or pHvG38). In addition to its resistance to stripe rust and powdery mildew, WR04-32 was genetically stable and had desirable agronomic traits, making it a desirable germplasm for wheat breeding.

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Assessment of resistance to cereal cyst nematode, stripe rust and powdery mildew in the wheat-Thinopyrum intermedium derivatives and their chromosome composition

Plant Disease ◽

10.1094/pdis-10-20-2141-re ◽

2021 ◽

Author(s):

Lei Cui ◽

Yongkang Ren ◽

Yinguang Bao ◽

Hai Nan ◽

Zhaohui Tang ◽

...

Keyword(s):

In Situ Hybridization ◽

Powdery Mildew ◽

Stripe Rust ◽

Puccinia Striiformis ◽

Cyst Nematode ◽

Cereal Cyst Nematode ◽

Thinopyrum Intermedium ◽

Resistant Lines ◽

Multiple Pathogens

Wide hybridization between wheat and wild relatives such as Thinopyrum intermedium is important for broadening genetic diversity and improving disease resistance in wheat. We developed 30 wheat-Th. intermedium derivatives. Here, we report assessments of their resistance to different pathogens including cereal cyst nematode (CCN, Heterodera spp.), Puccinia striiformis causing stripe rust, and Blumeria graminis f. sp. tritici inciting powdery mildew. Under natural field infection, all the wheat-Th. intermedium lines were resistant to at least one of the pathogens, and four lines were resistant to multiple pathogens. Twenty-nine out of 30 tested lines exhibited resistance to H. avenae, a dominant CCN species in wheat fields. Twenty-four lines were resistant to H. filipjevi, an emerging threat to wheat production. Tests of phenotypic responses in the naturally infected field nurseries identified six stripe rust resistant lines and 13 powdery mildew resistant lines. Mitotic observation demonstrated that these newly developed wheat-Th. intermedium derivatives included not only octoploid but also chromosome addition, substitution, and translocation lines. Chromosome compositions of the four lines resistant to multiple pathogens were analyzed by genomic in situ hybridization and fluorescence in situ hybridization. The octoploid lines Zhong 10-68 and Zhong 10-117 carried both intact Th. intermedium chromosomes and translocated chromosomes. Line Zhong 10-149 had 42 wheat chromosomes and two wheat ditelosomes plus a pair of T3BS·J translocated chromosomes. Line Zhong 10-160 carried 41 wheat chromosomes plus one pair of the J genome chromosomes of Th. intermedium. The multiple disease resistant wheat-Th. intermedium derivatives, especially lines with chromosome counts close to common wheat, provide valuable materials for wheat resistance breeding programs.

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Characterization and Evaluation of Resistance to Powdery Mildew of Wheat–Aegilops geniculata Roth 7Mg (7A) Alien Disomic Substitution Line W16998

International Journal of Molecular Sciences ◽

10.3390/ijms21051861 ◽

2020 ◽

Vol 21 (5) ◽

pp. 1861

Author(s):

Yajuan Wang ◽

Deyu Long ◽

Yanzhen Wang ◽

Changyou Wang ◽

Xinlun Liu ◽

...

Keyword(s):

In Situ Hybridization ◽

Powdery Mildew ◽

Common Wheat ◽

Agronomic Traits ◽

Genomic In Situ Hybridization ◽

Substitution Line ◽

Aegilops Geniculata ◽

The Common ◽

Sequential Fish

Aegilops geniculata Roth has been used as a donor of disease-resistance genes, to enrich the gene pool for wheat (Triticum aestivum) improvement through distant hybridization. In this study, the wheat–Ae. geniculata alien disomic substitution line W16998 was obtained from the BC1F8 progeny of a cross between the common wheat ‘Chinese Spring’ (CS) and Ae. geniculata Roth (serial number: SY159//CS). This line was identified using cytogenetic techniques, analysis of genomic in situ hybridization (GISH), functional molecular markers (Expressed sequence tag-sequence-tagged site (EST–STS) and PCR-based landmark unique gene (PLUG), fluorescence in situ hybridization (FISH), sequential fluorescence in situ hybridization–genomic in situ hybridization (sequential FISH–GISH), and assessment of agronomic traits and powdery mildew resistance. During the anaphase of meiosis, these were evenly distributed on both sides of the equatorial plate, and they exhibited high cytological stability during the meiotic metaphase and anaphase. GISH analysis indicated that W16998 contained a pair of Ae. geniculata alien chromosomes and 40 common wheat chromosomes. One EST–STS marker and seven PLUG marker results showed that the introduced chromosomes of Ae. geniculata belonged to homoeologous group 7. Nullisomic–tetrasomic analyses suggested that the common wheat chromosome, 7A, was absent in W16998. FISH and sequential FISH–GISH analyses confirmed that the introduced Ae. geniculata chromosome was 7Mg. Therefore, W16998 was a wheat–Ae. geniculata 7Mg (7A) alien disomic substitution line. Inoculation of isolate E09 (Blumeria graminis f. sp. tritici) in the seedling stage showed that SY159 and W16998 were resistant to powdery mildew, indeed nearly immune, whereas CS was highly susceptible. Compared to CS, W16998 exhibited increased grain weight and more spikelets, and a greater number of superior agronomic traits. Consequently, W16998 was potentially useful. Germplasms transfer new disease-resistance genes and prominent agronomic traits into common wheat, giving the latter some fine properties for breeding.

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Isolation and molecular cytogenetic characterization of a wheat – Leymus mollis double monosomic addition line and its progenies with resistance to stripe rust

Genome ◽

10.1139/gen-2017-0078 ◽

2017 ◽

Vol 60 (12) ◽

pp. 1029-1036 ◽

Cited By ~ 2

Author(s):

Xiaofei Yang ◽

Xin Li ◽

Changyou Wang ◽

Chunhuan Chen ◽

Zengrong Tian ◽

...

Keyword(s):

In Situ Hybridization ◽

Stripe Rust ◽

Rust Resistance ◽

Stripe Rust Resistance ◽

Addition Line ◽

Cytogenetic Characterization ◽

Monosomic Addition ◽

Monosomic Addition Line ◽

Leymus Mollis

A common wheat – Leymus mollis (2n = 4x = 28, NsNsXmXm) double monosomic addition line, M11003-4-3-8/13/15 (2n = 44 = 42T.a + L.m2 + L.m3), with stripe rust resistance was developed (where T.a represents Triticum aestivum chromosome, L.m represents L. mollis chromosome, and L.m2/3 represents L. mollis chromosome of homoeologous groups 2 and 3). The progenies of line M11003-4-3-8/13/15 were characterized by cytological observation, specific molecular markers, fluorescence in situ hybridization (FISH), and genomic in situ hybridization (GISH). Among the progenies, there existed five different types (I, II, III, IV, and V) of chromosome constitution, the formulas of which were 2n = 44 = 42T.a + 1L.m2 + 1L.m3, 2n = 43 = 42T.a + 1L.m2, 2n = 43 = 42T.a + 1L.m3, 2n = 42 = 42T.a, and 2n = 44 = 42T.a + 2L.m2, respectively. Field disease screening showed that types I and III showed high resistance to stripe rust, while types II, IV, and V were susceptible. Leymus mollis was almost immune to stripe rust, whereas the wheat parent, cultivar 7182, was susceptible. Therefore, we concluded that the stripe rust resistance originated from L. mollis. These various lines could be further fully exploited as important disease resistance materials to enrich wheat genetic resources.

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Molecular cytogenetics identification of a wheat – Leymus mollis double disomic addition line with stripe rust resistance

Genome ◽

10.1139/gen-2016-0151 ◽

2017 ◽

Vol 60 (5) ◽

pp. 375-383 ◽

Cited By ~ 5

Author(s):

Aicen Zhang ◽

Wanyue Li ◽

Changyou Wang ◽

Xiaofei Yang ◽

Chunhuan Chen ◽

...

Keyword(s):

In Situ Hybridization ◽

Common Wheat ◽

Stripe Rust ◽

Rust Resistance ◽

Stripe Rust Resistance ◽

Fish Analysis ◽

Addition Line ◽

Disomic Addition Line ◽

Leymus Mollis

Leymus mollis (Trin.) Pilg. (2n = 4x = 28, NsNsXmXm) possesses a number of valuable genes against biotic and abiotic stress, which could be transferred into common wheat background for wheat improvement. In the present study, we determined the karyotypic constitution of a wheat – L. mollis double disomic addition line, M11003-4-4-1-1, selected from the F5 progeny of a stable wheat – L. mollis derivative M39 (2n = 56) × Triticum aestivum cultivar 7182, by morphological and cytogenetic identification, GISH (genomic in situ hybridization), FISH (fluorescent in situ hybridization), molecular markers analysis, and stripe rust resistance evaluation. Cytological studies demonstrated that M11003-4-4-1-1 had a chromosome karyotype of 2n = 46 with 23 bivalents, while GISH and FISH analysis indicated that this line contained 42 common wheat chromosomes and two pairs of L. mollis chromosomes. DNA markers showed that the alien chromosomes from L. mollis belonged to homoeologous groups 5 and 6. Evaluation of the agronomic traits revealed that M11003-4-4-1-1 was resistant to stripe rust at the adult stage, while the plant height was reduced and the 1000-grain weight was increased significantly. Therefore, the new line M11003-4-4-1-1 could be exploited as an important bridge material in chromosome engineering and wheat breeding.

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Identification of an Elite Wheat-Rye T1RS·1BL Translocation Line Conferring High Resistance to Powdery Mildew and Stripe Rust

Plant Disease ◽

10.1094/pdis-02-20-0323-re ◽

2020 ◽

Vol 104 (11) ◽

pp. 2940-2948

Author(s):

Guohao Han ◽

Shiyu Liu ◽

Jing Wang ◽

Yuli Jin ◽

Yilin Zhou ◽

...

Keyword(s):

In Situ Hybridization ◽

Powdery Mildew ◽

Stripe Rust ◽

Translocation Line ◽

Pcr Marker ◽

Pathogen Virulence ◽

Specific Pcr ◽

Secale Cereale L ◽

Molecular Marker Analysis

Wheat-rye T1RS·1BL translocations have been widely used worldwide in wheat production for multiple disease resistance and superior yield traits. However, many T1RS·1BL translocations have successively lost their resistance to pathogens due to the coevolution of pathogen virulence with host resistance. Because of the extensive variation in rye (Secale cereale L.) as a naturally cross-pollinating relative of wheat, it still has promise to widen the variation of 1RS and to fully realize its application value in wheat improvement. In the present study, the wheat-rye breeding line R2207 was characterized by comprehensive analyses using genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization with multiple probes, multicolor GISH, and molecular marker analysis, and then was proven to be a cytogenetically stable wheat-rye T1RS·1BL translocation line. Based on the disease responses to different isolates of powdery mildew and genetic analysis, R2207 appears to possess a novel variation for resistance, which was confirmed to be located on the rye chromosome arm 1RS. Line R2207 also exhibited high levels of resistance to stripe rust at both seedling and adult stages, as well as enhanced agronomic performance, so it has been transferred into a large number of commercial cultivars using an efficient 1RS-specific kompetitive allele specific PCR marker for marker-assisted selection.

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Molecular cytogenetic identification of a wheat–rye 1R addition line with multiple spikelets and resistance to powdery mildew

Genome ◽

10.1139/gen-2015-0129 ◽

2016 ◽

Vol 59 (4) ◽

pp. 277-288 ◽

Cited By ~ 8

Author(s):

Wujuan Yang ◽

Changyou Wang ◽

Chunhuan Chen ◽

Yajuan Wang ◽

Hong Zhang ◽

...

Keyword(s):

Disease Resistance ◽

In Situ Hybridization ◽

Powdery Mildew ◽

Common Wheat ◽

Wheat Chromosome ◽

Triticum Aestivum L ◽

Addition Line ◽

Disease Resistance Screening ◽

High Level

Alien addition lines are important for transferring useful genes from alien species into common wheat. Rye is an important and valuable gene resource for improving wheat disease resistance, yield, and environment adaptation. A new wheat–rye addition line, N9436B, was developed from the progeny of the cross of common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) cultivar Shaanmai 611 and rye (Secale cereal L., 2n = 2x = 14, RR) accession Austrian rye. We characterized this new line by cytology, genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), molecular markers, and disease resistance screening. N9436B was stable in morphology and cytology, with a chromosome composition of 2n = 42 + 2t = 22II. GISH investigations showed that this line contained two rye chromosomes. GISH, FISH, and molecular maker identification suggested that the introduced R chromosome and the missing wheat chromosome arms were 1R chromosome and 2DL chromosome arm, respectively. N9436B exhibited 30–37 spikelets per spike and a high level of resistance to powdery mildew (Blumeria graminis f. sp. tritici, Bgt) isolate E09 at the seedling stage. N9436B was cytologically stable, had the trait of multiple spikelets, and was resistant to powdery mildew; this line should thus be useful in wheat improvement.

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The barley immune receptor Mla recognizes multiple pathogens and contributes to host range dynamics

Nature Communications ◽

10.1038/s41467-021-27288-3 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Jan Bettgenhaeuser ◽

Inmaculada Hernández-Pinzón ◽

Andrew M. Dawson ◽

Matthew Gardiner ◽

Phon Green ◽

...

Keyword(s):

Powdery Mildew ◽

Stripe Rust ◽

Resistance Genes ◽

Host Species ◽

Plant Pathogens ◽

Puccinia Striiformis ◽

Species Specificity ◽

Wheat Stripe Rust ◽

Barley Powdery Mildew ◽

Barley Stripe Rust

AbstractCrop losses caused by plant pathogens are a primary threat to stable food production. Stripe rust (Puccinia striiformis) is a fungal pathogen of cereal crops that causes significant, persistent yield loss. Stripe rust exhibits host species specificity, with lineages that have adapted to infect wheat and barley. While wheat stripe rust and barley stripe rust are commonly restricted to their corresponding hosts, the genes underlying this host specificity remain unknown. Here, we show that three resistance genes, Rps6, Rps7, and Rps8, contribute to immunity in barley to wheat stripe rust. Rps7 cosegregates with barley powdery mildew resistance at the Mla locus. Using transgenic complementation of different Mla alleles, we confirm allele-specific recognition of wheat stripe rust by Mla. Our results show that major resistance genes contribute to the host species specificity of wheat stripe rust on barley and that a shared genetic architecture underlies resistance to the adapted pathogen barley powdery mildew and non-adapted pathogen wheat stripe rust.

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Molecular cytogenetic identification of a wheat – Thinopyrum ponticum substitution line with stripe rust resistance

Genome ◽

10.1139/gen-2017-0099 ◽

2017 ◽

Vol 60 (10) ◽

pp. 860-867 ◽

Cited By ~ 9

Author(s):

Chen Zhu ◽

Yanzhen Wang ◽

Chunhuan Chen ◽

Changyou Wang ◽

Aicen Zhang ◽

...

Keyword(s):

In Situ Hybridization ◽

Stripe Rust ◽

Expressed Sequence Tag ◽

Wheat Breeding ◽

Substitution Line ◽

Stripe Rust Resistance ◽

Fish Analysis ◽

Thinopyrum Ponticum ◽

Expressed Sequence

Thinopyrum ponticum (Th. ponticum) (2n = 10x = 70) is an important breeding material with excellent resistance and stress tolerance. In this study, we characterized the derivative line CH1113-B13-1-1-2-1 (CH1113-B13) through cytological, morphological, genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), expressed sequence tag (EST), and PCR-based landmark unique gene (PLUG) marker analysis. The GISH analysis revealed that CH1113-B13 contained 20 pairs of common wheat chromosomes and one pair of JSt genomic chromosomes. Linkage analysis of Th. ponticum using seven EST and seven PLUG markers indicated that the pair of alien chromosomes belonged to the seventh homeologous group. Nulli-tetrasomic and FISH analysis revealed that wheat 7B chromosomes were absent in CH1113-B13; thus, CH1113-B13 was identified as a 7JSt (7B) substitution line. Finally, adult-stage CH1113-B13 exhibited immunity to wheat stripe rust. This substitution line is therefore a promising germplasm resource for wheat breeding.

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