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

Genome ◽  
2016 ◽  
Vol 59 (4) ◽  
pp. 277-288 ◽  
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.

Random chromosome elimination in synthetic Triticum–Aegilops amphiploids leads to development of a stable partial amphiploid with high grain micro- and macronutrient content and powdery mildew resistance

Genome ◽  
2010 ◽  
Vol 53 (12) ◽  
pp. 1053-1065 ◽  
Author(s):  
Vijay K. Tiwari ◽  
Nidhi Rawat ◽  
Kumari Neelam ◽  
Sundip Kumar ◽  
Gursharn S. Randhawa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Powdery Mildew ◽  
Powdery Mildew Resistance ◽  
Chromosome Elimination ◽  
Triticum Aestivum L ◽  
Crop Evolution ◽  
High Fertility ◽  
Mildew Resistance ◽  
Partial Amphiploids

Synthetic amphiploids are the immortal sources for studies on crop evolution, genome dissection, and introgression of useful variability from related species. Cytological analysis of synthetic decaploid wheat ( Triticum aestivum L.) –   Aegilops kotschyi Boiss. amphiploids (AABBDDUkUkSkSk) showed some univalents from the C1 generation onward followed by chromosome elimination. Most of the univalents came to metaphase I plate after the reductional division of paired chromosomes and underwent equational division leading to their elimination through laggards and micronuclei. Substantial variation in the chromosome number of pollen mother cells from different tillers, spikelets, and anthers of some plants also indicated somatic chromosome elimination. Genomic in situ hybridization, fluorescence in situ hybridization, and simple sequence repeat markers analysis of two amphiploids with reduced chromosomes indicated random chromosome elimination of various genomes with higher sensitivity of D followed by the Sk and Uk genomes to elimination, whereas 1D chromosome was preferentially eliminated in both the amphiploids investigated. One of the partial amphiploids, C4 T. aestivum ‘Chinese Spring’ – Ae. kotschyi 396 (2n = 58), with 34 T. aestivum, 14 Uk, and 10 Sk had stable meiosis and high fertility. The partial amphiploids with white glumes, bold seeds, and tough rachis with high grain macro- and micronutrients and resistance to powdery mildew could be used for T. aestivum biofortification and transfer of powdery mildew resistance.

Characterization of an Agropyron elongatum chromosome conferring resistance to cephalosporium stripe in common wheat

Genome ◽  
1996 ◽  
Vol 39 (1) ◽  
pp. 56-62 ◽  
Author(s):  
Xiwen Cai ◽  
Stephen S. Jones ◽  
Timothy D. Murray
Keyword(s):  
In Situ Hybridization ◽  
Wheat Chromosome ◽  
Triticum Aestivum L ◽  
Chromosome Substitution ◽  
Agropyron Elongatum ◽  
Breeding Lines ◽  
C Banding ◽  
Small Band ◽  
Cephalosporium Gramineum

Related wheat (Triticum aestivum L.) breeding lines, PI 561033, REA 9232, REA 9257, and CI 13113 were analyzed cytogenetically to characterize the association of resistance to cephalosporium stripe (caused by Cephalosporium gramineum Nis. & Ika.) with Agropyron elongatum chromatin. One pair of A. elongatum chromosomes was detected in PI 561033, REA 9232, and CI 13113 by genomic in situ hybridization. The sib line of PI 561033 and REA 9232, REA 9257, which is not resistant to this disease, lacked this pair of A. elongatum chromosomes. PI 561033 was characterized as a disomic T. aestivum – A. elongatum 6Ae#2(6A) chromosome substitution line using test crosses and C-banding. In situ hybridization and test crosses showed that the donor parent, CI 13113, also had chromosome 6A substituted by A. elongatum chromosome 6Ae#2. The C-banding pattern of 6Ae#2 showed two subterminal bands on the long arm and one small band proximal to the centromere on the short arm. Based on chromosome pairing and compensation, chromosome 6Ae#2 shows a close homoeologous relationship with wheat chromosome 6A. Key words : Cephalosporium gramineum, Agropyron elongatum, in situ hybridization, C-banding, chromosome substitution.

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

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.

Molecular cytogenetic analysis of intergeneric chromosomal translocations between wheat (Triticum aestivum L.) and Dasypyrum villosum arising from tissue culture

Genome ◽  
2000 ◽  
Vol 43 (5) ◽  
pp. 756-762 ◽  
Author(s):  
Hong-Jie Li ◽  
Bei-Hai Guo ◽  
Yi-Wen Li ◽  
Li-Qun Du ◽  
Xu Jia ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Tissue Culture ◽  
In Situ Hybridization ◽  
Wheat Chromosome ◽  
Triticum Aestivum L ◽  
Average Frequency ◽  
Dasypyrum Villosum ◽  
Chromosome Translocations ◽  
Chromosome Arm

Fluorescence in situ hybridization (FISH) was applied with total genomic DNA extracted from Dasypyrum villosum (L.) Candargy as a probe to characterize chromosome translocations arising from tissue culture in hybrids of Triticum aestivum × (T. durum - D. villosum, amphiploid). Chromosome translocations between wheat and D. villosum occurred in callus cells at an average frequency of 1.9%. Translocations existed not only in callus cells but also in regenerants. Three plants with translocation chromosomes were characterized among 66 regenerants of T. aestivum 'Chinese Spring' × 'TH1W' and 'NPFP' × 'TH1'. One of them proved to be a reciprocal translocation with an exchange of about one third of a wheat chromosome arm with about one half of a chromosome arm of D. villosum. The breakpoints of the other two translocations were located at, or near centromeres. The results are similar for both callus cells and regenerants and provide further evidence that translocations take place in tissue culture. Other structural chromosomal changes, for example, fragments, telocentrics, dicentromeres, and deletions, as well as numerical alterations including aneuploidy and polyploidy were recorded both in callus cells and regenerants.Key words: wheat, Dasypyrum villosum, translocation, genomic in situ hybridization, tissue culture.

scholarly journals New Insights Into the Introgression Between Agropyron Cristatum P Genome and Wheat Genome

2021 ◽  
Author(s):  
Zhi Zhang ◽  
Shenghui Zhou ◽  
Weihua Liu ◽  
Liqiang Song ◽  
Jinpeng Zhang ◽  
...  
Keyword(s):  
Disease Resistance ◽  
In Situ Hybridization ◽  
Common Wheat ◽  
Agronomic Traits ◽  
Wheat Genome ◽  
Agropyron Cristatum ◽  
Translocation Line ◽  
Gene Markers ◽  
Physiological Races

Abstract Agropyron cristatum (2n = 4x = 28, PPPP) is an important wild relative of common wheat and confers desirable agronomic traits to common wheat. A previous report showed that the wheat-A. cristatum 6P translocation line WAT655 carrying A. cristatum 6PS (0.81–1.00) exhibited high resistance to prevalent physiological races (CYR32 and CYR33). In this study, three disease resistance-related transcriptomes, which were mapped to A. cristatum 6PS (0.81–1.00) through the analysis of specific molecular markers, were searched from among A. cristatum full-length transcriptomes. Then, three disease resistance-related gene markers, A. cristatum P genome-specific markers, and fluorescence in situ hybridization (FISH)/genomic in situ hybridization (GISH) probes made from the DNA of three bacterial artificial chromosome (BAC) clones, three genes, and A. cristatum “Z559” were used to analyze the BC5F2 and BC5F2:3 genetic populations of the translocation line WAT655. The results revealed the introgression can spontaneously occur between A. cristatum P genome and wheat genome, and indicated the three genes could constitute a gene cluster according to the positions of their FISH signals. Additionally, kompetitive allele-specific PCR (KASP) markers of the three genes were developed to detect and acquire 24 wheat-A. cristatum breeding materials, which showed resistance to physiological races (CYR32 and CYR33) and other desirable agronomic traits according to the field investigation. In conclusion, our study not only provides new insights into the introgression between A. cristatum P genome and wheat genome, but also provides the desirable breeding materials for breeding practice.

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.

Genomic and chromosomal distribution patterns of various repeated DNA sequences in wheat revealed by a fluorescence in situ hybridization procedure

Genome ◽  
2013 ◽  
Vol 56 (3) ◽  
pp. 131-137 ◽  
Author(s):  
Shirabe Komuro ◽  
Ryota Endo ◽  
Kaori Shikata ◽  
Akio Kato
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dna Sequences ◽  
Wheat Chromosome ◽  
Triticum Aestivum L ◽  
Fluorescent Labeling ◽  
Homologous Sequence ◽  
Chromosomal Distribution ◽  
Discrete Signals

Wheat (Triticum aestivum L.) is an allohexaploid, in which each of the three genomes has a high 1C content. This indicates the presence of multiple tandemly repeated sequences, which should be detectable using in situ hybridization. Some repeats have already been described, but others remain to be recognized. To discover others, 2000 plasmid wheat clones were examined for signal presence after fluorescence in situ hybridization and microscopic signal observation. Among them, 47 clones produced strong discrete signals on wheat chromosomes. Two of the newly identified clones (pTa-535 and pTa-713) were determined to have especially valuable sequences for chromosome identification. In combination with pTa-86 (the pSc119 homologous sequence), these probes enable unambiguous discrimination of all wheat chromosomes including orientation. Four newly identified sequences (pTa-465, pTa-k566, pTa-s120, and pTa-s126) were useful in that they produced discrete signals on various wheat chromosome arms. Two other clones (pTa-k288 and pTa-k229) produced GISH-like (genomic in situ hybridization) signals because they allowed the A, B, and D genomes to be distinguished simultaneously. In addition, centromere, centromere-related, and ribosomal DNA clones were identified. Also described are improvements on slide preparation and reprobing procedures. To enhance discrete signal detection, a new direct fluorescent-labeling procedure, namely the VentR (exo-) terminal extension method, was employed.

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

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yongfu Wang ◽  
Xiaofang Cheng ◽  
Xiaoying Yang ◽  
Changyou Wang ◽  
Hong Zhang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
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 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.

scholarly journals Human neutrophilic and eosinophilic granulocytes display different levels of c-fos proto-oncogene expression: an in situ hybridization study.

1987 ◽  
Vol 35 (8) ◽  
pp. 837-842 ◽  
Author(s):  
H Kreipe ◽  
H J Radzun ◽  
K Heidorn ◽  
C Mäder ◽  
M R Parwaresch
Keyword(s):  
In Situ Hybridization ◽  
Neutrophilic Granulocytes ◽  
Blood Monocytes ◽  
In Vitro Differentiation ◽  
Monocytic Differentiation ◽  
Eosinophilic Granulocytes ◽  
Fos Expression ◽  
High Level

The cellular homologue of the retroviral oncogene v-fos has been shown to be involved in cell differentiation of hematopoietic cells. By use of the human promyelocyte cell line HL-60, several in vitro differentiation studies suggested a selective activation of c-fos during monocytic differentiation of myeloid precursor cells. In contrast to these observations, we found high levels of c-fos mRNA in purified normal human granulocytes, whereas c-fos was only faintly expressed in blood monocytes. In situ hybridization revealed that the high level of c-fos expression is restricted to neutrophilic granulocytes, whereas c-fos transcription is not detectable in eosinophilic granulocytes. These results indicate that in vitro differentiation systems can be misleading and may not reflect the in vivo situation. The high level of c-fos expression in neutrophilic granulocytes may be caused by superinduction due to the reduced capacity for protein synthesis in these cells.

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