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

Genome ◽  
2017 ◽  
Vol 60 (10) ◽  
pp. 860-867 ◽  
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.

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

Plant Disease ◽  
2009 ◽  
Vol 93 (2) ◽  
pp. 124-129 ◽  
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.

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

Genome ◽  
2017 ◽  
Vol 60 (5) ◽  
pp. 375-383 ◽  
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.

scholarly journals Molecular cytogenetics and St-chromosome-specific molecular markers development of novel stripe rust resistant wheat–Thinopyrum intermedium as well as wheat–Thinopyrum ponticum substitution lines

2021 ◽  
Author(s):  
Siwen Wang ◽  
Changyou Wang ◽  
Xianbo Feng ◽  
Jixin Zhao ◽  
Pingchuan Deng ◽  
...  
Keyword(s):  
Molecular Marker ◽  
Stripe Rust ◽  
Rust Resistance ◽  
Snp Array ◽  
Kernel Weight ◽  
Stripe Rust Resistance ◽  
Substitution Lines ◽  
Fish Genome ◽  
Thinopyrum Ponticum

Abstract Background Owing to the excellent resistance to abiotic and biotic stress, Thionpyrum intermedium (2n = 6x = 42, JJJsJsStSt) and Thinopyrum ponticum (2n = 10x = 70) are both widely utilized in wheat germplasm innovation programs. Disomic substitution lines (DSLs) carrying one pair of alien chromosomes are valuable bridge materials for novel genes transmission, FISH karyotype construction and specific molecular marker development. Results Six wheat–Thinopyrum DSLs derived from crosses between Abbondanza nullisomic lines (2n = 40) and two octoploid Trititrigia lines (2n = 8x = 56), were characterized by a sequential fluorescence in situ hybridization (FISH)–genome in situ hybridization (GISH), a multicolor GISH (mc-GISH), and an analysis of wheat 15K SNP array combined with molecular marker selection. ES-9 (DS2St (2A)) and ES-10 (DS3St (3D)) are wheat–Th. ponticum DSLs, while ES-23 (DS2St (2A)), ES-24 (DS3St (3D)), ES-25(DS2St (2B)), and ES-26 (DS2St (2D)) are wheat–Th. intermedium DSLs. ES-9, ES-23, ES-25 and ES-26 conferred higher thousand-kernel weight and stripe rust resistance at adult stages, while ES-10 and ES-24 performed highly resistant to stripe rust at all stages. Furthermore, cytological analysis showed that the alien chromosomes (2St/3St) belonging to the same homoeologous group derived from different donors carried the same FISH karyotype and could normally form a bivalent. Based on specific-locus amplified fragment sequencing (SLAF-seq), two 2St-chromosome-specific markers (PTH-005 and PTH-013) and two 3St-chromosome-specific markers (PTH-113 and PTH-135) were developed. Conclusions The six wheat–Thinopyrum disomic substitution lines conferring stripe rust resistance will be used as bridging parents for valuable resistant genes transmission. And the utility of PTH-113 and PTH-135 in a BC1F2 population showed the newly developed markers could be useful tools for efficient identification of St chromosomes in a common wheat background.

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

Genome ◽  
2017 ◽  
Vol 60 (12) ◽  
pp. 1029-1036 ◽  
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.

Cytogenetic and molecular markers for detecting Aegilops uniaristata chromosomes in a wheat background

Genome ◽  
2014 ◽  
Vol 57 (9) ◽  
pp. 489-497 ◽  
Author(s):  
Wenping Gong ◽  
Guangrong Li ◽  
Jianping Zhou ◽  
Genying Li ◽  
Cheng Liu ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Expressed Sequence Tag ◽  
Triticum Turgidum ◽  
Wheat Breeding ◽  
Addition Lines ◽  
Chromosome Engineering ◽  
Aegilops Uniaristata ◽  
Expressed Sequence ◽  
Simple Sequence

Aegilops uniaristata has many agronomically useful traits that can be used for wheat breeding. So far, a Triticum turgidum – Ae. uniaristata amphiploid and one set of Chinese Spring (CS) – Ae. uniaristata addition lines have been produced. To guide Ae. uniaristata chromatin transformation from these lines into cultivated wheat through chromosome engineering, reliable cytogenetic and molecular markers specific for Ae. uniaristata chromosomes need to be developed. Standard C-banding shows that C-bands mainly exist in the centromeric regions of Ae. uniaristata but rarely at the distal ends. Fluorescence in situ hybridization (FISH) using (GAA)8 as a probe showed that the hybridization signal of chromosomes 1N–7N are different, thus (GAA)8 can be used to identify all Ae. uniaristata chromosomes in wheat background simultaneously. Moreover, a total of 42 molecular markers specific for Ae. uniaristata chromosomes were developed by screening expressed sequence tag – sequence tagged site (EST–STS), expressed sequence tag – simple sequence repeat (EST–SSR), and PCR-based landmark unique gene (PLUG) primers. The markers were subsequently localized using the CS – Ae. uniaristata addition lines and different wheat cultivars as controls. The cytogenetic and molecular markers developed herein will be helpful for screening and identifying wheat – Ae. uniaristata progeny.

scholarly journals New ND-FISH-Positive Oligo Probes for Identifying Thinopyrum Chromosomes in Wheat Backgrounds

2019 ◽  
Vol 20 (8) ◽  
pp. 2031 ◽  
Author(s):  
Wei Xi ◽  
Zongxiang Tang ◽  
Shuyao Tang ◽  
Zujun Yang ◽  
Jie Luo ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Sequences ◽  
Triticum Aestivum L ◽  
Wheat Breeding ◽  
Fish Analysis ◽  
Breeding Programs ◽  
Gish And Fish

Thinopyrum has been widely used to improve wheat (Triticum aestivum L.) cultivars. Non-denaturing fluorescence in situ hybridization (ND-FISH) technology using oligonucleotides (oligo) as probes provides a convenient and efficient way to identify alien chromosomes in wheat backgrounds. However, suitable ND-FISH-positive oligo probes for distinguishing Thinopyrum chromosomes from wheat are lacking. Two oligo probes, Oligo-B11 and Oligo-pThp3.93, were designed according to the published Thinopyrum ponticum (Th. ponticum)-specific repetitive sequences. Both Oligo-B11 and Oligo-pThp3.93 can be used for ND-FISH analysis and can replace conventional GISH and FISH to discriminate some chromosomes of Th. elongatum, Th. intermedium, and Th. ponticum in wheat backgrounds. The two oligo probes provide a convenient way for the utilization of Thinopyrum germplasms in future wheat breeding programs.

Molecular Cytogenetics of Chromosome 2St as Well as Chromosome 3St Derived from Thinopyrum Intermedium and Thinopyrum Ponticum

2021 ◽  
Author(s):  
Siwen Wang ◽  
Changyou Wang ◽  
Xianbo Feng ◽  
Jixin Zhao ◽  
Pingchuan Deng ◽  
...  
Keyword(s):  
Stripe Rust ◽  
Snp Array ◽  
Molecular Cytogenetics ◽  
Kernel Weight ◽  
Stripe Rust Resistance ◽  
Substitution Lines ◽  
Fish Genome ◽  
Thinopyrum Ponticum ◽  
Marker Selection

Abstract Owing to the excellent resistance to abiotic and biotic stress, Thionpyrum intermedium (2n = 6x = 42, JJJsJsStSt) and Thinopyrum ponticum (2n = 10x = 70) are both widely utilized in wheat germplasm innovation programs. Disomic substitution lines (DSLs) carrying one pair of alien chromosomes are valuable bridge materials for novel genes transmission. In this study, six wheat-Thinopyrum DSLs were derived from crosses between Abbondanza nullisomic lines (2n = 40) and two octoploid Trititrigia lines (2n = 8x = 56), characterized by a sequential fluorescence in situ hybridization (FISH)-genome in situ hybridization (GISH), a multicolor GISH (mc-GISH), and an analysis of wheat 15K SNP array combined with molecular marker selection. ES-9 and ES-10 were two wheat- Th. ponticum disomic substitution lines, DS2St (2A) and DS3St (3D). While ES-23, ES-24, ES-25, and ES-26 were four wheat- Th. intermedium disomic substitution lines, DS2St (2A), DS3St (3D), DS2St (2B), DS2St (2D). The FISH karyotypes of Th. ponticum 2St/3St chromosomes were well coincident with the ones of Th. intermedium. The chromosome configurations of F1 hybrids derived from crosses between ES-23 and ES-9, as well as ES-24 and ES-10 were mostly formed 21Ⅱ. Four St-chromosome-specific markers were developed by specific-locus amplified fragment sequencing (SLAF-seq). Additionally, the substitution lines containing chromosome 2St conferred higher thousand-kernel weight and stripe rust resistance at adult stages, while the substitution lines containing chromosome 3St were highly resistant to stripe rust at all stages. Therefore, these six substitution lines could serve as useful bridging parents for wheat genetic improvement.

Fluorescent in situ hybridization (FISH): A useful diagnostic tool for childhood conjunctival melanoma

2021 ◽  
pp. 112067212110307
Author(s):  
Raquel María Moral ◽  
Carlos Monteagudo ◽  
Javier Muriel ◽  
Lucía Moreno ◽  
Ana María Peiró
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Pediatric Population ◽  
Diagnostic Technique ◽  
Histopathological Diagnosis ◽  
Fish Analysis ◽  
Conjunctival Melanoma ◽  
Adequate Treatment ◽  
First Case

Introduction: Conjunctival melanoma is extremely rare in children and has low rates of resolution. Definitive histopathological diagnosis based exclusively on microscopic findings is sometimes difficult. Thus, early diagnosis and adequate treatment are essential to improve clinical outcomes. Clinical case: We present the first case in which the fluorescent in situ hybridization (FISH) diagnostic technique was applied to a 10-year-old boy initially suspected of having amelanotic nevi in his right eye. Based on the 65% of tumor cells with 11q13 (CCND1) copy number gain and 33% with 6p25 (RREB1) gain as measured by the FISH analysis, and on supporting histopathological findings, the diagnosis of conjunctival melanoma could be made. Following a larger re-excision, adjuvant therapy with Mitomycin C (MMC), cryotherapy and an amniotic membrane graft, the patient has remained disease-free during 9 years of long-term follow-up. Case discussion: Every ophthalmologist should remember to consider and not forget the possibility of using FISH analyses during the differential diagnosis of any suspicious conjunctival lesions. Genetic techniques, such as FISH, have led to great advances in the classification of ambiguous lesions. Evidence-based guidelines for diagnosing conjunctival melanoma in the pediatric population are needed to determine the most appropriate strategy for this age group.

Rapid identification of a stripe rust resistance gene YrXK in Chinese wheat line Xike01015 using specific locus amplified fragment (SLAF) sequencing

Plant Disease ◽  
2021 ◽  
Author(s):  
Cai Sun ◽  
Yike Liu ◽  
Qiang Li ◽  
Baotong Wang ◽  
Shuhui Chen ◽  
...  
Keyword(s):  
Stripe Rust ◽  
Puccinia Striiformis ◽  
Gene Annotation ◽  
Dominant Gene ◽  
Rapid Identification ◽  
Wheat Breeding ◽  
Stripe Rust Resistance ◽  
Phenotypic Variance ◽  
F2 Population ◽  
Chinese Wheat

Wheat stripe rust, an airborne fungal disease and caused by Puccinia striiformis Westend. f. sp. tritici (Pst), is one of the most devastating diseases on wheat. It is the most effective and economical measure for the diseases control to identify high-level resistance genes and apply in wheat breeding. Chinese wheat cultivar Xike01015 presents high levels of all stage resistance (ASR) to the current predominant Pst race CYR33. In this study, a single dominant gene, designated as YrXk, was identified in Xike01015 conferring resistance to CYR33 with genetic analysis of F2 and BC1 population from cross of Mingxian169 (susceptible) and Xike01015. The specific length amplified fragment sequencing (SLAF-seq) strategy was used to construct linkage map in the F2 population. QTL analysis mapped YrXk to a 12.4 Mb segment on chromosome1BS, explaining over 86.96% phenotypic variance. Gene annotation in the QTL region identified three differential expressed candidate genes , TraesCS1B02G168600.1, TraesCS1B02G170200.1, and TraesCS1B02G172400.1. The qRT-PCR results displayed that TraesCS1B02G170200.1 and TraesCS1B02G168600.1 significantly up-regulated and down-regulated, respectively, and TraesCS1B02G170200.1 slightly up-regulated after changed with CYR33 in the seedling stage, which indicating these genes may function in wheat resistance to stripe rust. The results of this study can be used in wheat breeding for improving resistance to stripe rust.

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