Genomic in situ hybridization analysis of Thinopyrum chromatin in a wheat - Th. intermedium partial amphiploid and six derived chromosome addition lines

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1217-1223 ◽  
Author(s):  
Qin Chen ◽  
R L Conner ◽  
A Laroche ◽  
W Q Ji ◽  
K C Armstrong ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Dna Probe ◽  
Addition Lines ◽  
Thinopyrum Intermedium ◽  
Chromosome Addition ◽  
Chromosome Addition Lines ◽  
Partial Amphiploid

The genomic origin of alien chromosomes present in a wheat - Thinopyrum intermedium partial amphiploid TAF46 (2n = 8x = 56) and six derived chromosome addition lines were analyzed by genomic in situ hybridization (GISH) using S genomic DNA from Pseudoroegneria strigosa (2n = 2x = 14, SS) as a probe. The GISH analysis clearly showed that the chromosome complement of the partial amphiploid TAF46 consists of an entire wheat genome plus one synthetic genome consisting of a mixture of six S genome chromosomes and eight J (=E) genome chromosomes derived from Th. intermedium (2n = 6x = 42, JJJsJsSS). There were no Js genome chromosomes present in TAF46. The J genome chromosomes present in TAF46 displayed a unique GISH hybridization pattern with the S genomic DNA probe, in which S genome DNA strongly hybridized at the terminal regions and weakly hybridized over the remaining parts of the chromosomes. This provides a diagnostic marker for distinguishing J genome chromosomes from Js or S genome or wheat ABD genome chromosomes. The genomic origin of the alien chromosomes present in the six derived chromosome addition lines were identified by their characteristic GISH hybridization patterns with S genomic DNA probe. GISH analysis showed that addition lines L1, L2, L3, and L5 carried one pair of J genome chromosomes, while addition lines L4 and L7 each carried one pair of S genome chromosomes. GISH patterns detected by the S genome probe on addition line of L1 were identical to those of the J genome chromosomes present in the partial amphiploid TAF46, suggesting that these chromosomes were not structurally altered when they were transferred from TAF46 to addition lines.Key words: GISH, genomic composition, addition lines, Thinopyrum intermedium, partial amphiploid.

Molecular characterization of a wheat – Thinopyrum ponticum partial amphiploid and its derivatives for resistance to leaf rust

Genome ◽  
2003 ◽  
Vol 46 (5) ◽  
pp. 906-913 ◽  
Author(s):  
Hongjie Li ◽  
Qin Chen ◽  
Robert L Conner ◽  
Beihai Guo ◽  
Yanmin Zhang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Leaf Rust ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Addition Lines ◽  
Thinopyrum Ponticum ◽  
Sequence Tagged Site ◽  
Partial Amphiploid ◽  
Gish Analysis

Leaf rust (caused by Puccinia triticina Eriks.) occurs annually in most wheat-growing areas of the world. Thinopyrum ponticum (Podp.) Z.-W. Liu & R.-C. Wang has provided several leaf rust resistance genes to protect wheat from this fungal disease. Three chromosome substitution lines, Ji806, Ji807, and Ji859, and two chromosome addition lines, Ji791 and Ji924, with a winter growing habit were developed from crosses between wheat (Triticum aestivum L. em Thell.) and the wheat – Th. ponticum partial amphiploid line 693. These lines were resistant to leaf rust isolates from China. Sequence-tagged site (STS) analysis with the J09-STS marker, which is linked to the gene Lr24, revealed that the partial amphiploid line 693 and all of the substitution and addition lines carried gene Lr24. Genomic in situ hybridization (GISH) analysis was carried out on chromosome preparations using total genomic DNA from Pseudoroegneria strigosa (M. Bieb) A. Löve (St genome, 2n = 14) as a probe in the presence of total genomic DNA from T. aestivum 'Chinese Spring' wheat (ABD genomes, 2n = 42). The GISH analysis demonstrated that these lines had a pair of chromosomes displaying the typical pattern of a Js genome chromosome. This indicates that the chromosome that carries gene Lr24 belonged to the Js genome of Th. ponticum. In addition to 40 wheat chromosomes, eight Js and eight J genome chromosomes were also differentiated by GISH in the partial amphiploid line 693. Since most sources of Lr24 have a red grain color, the white-colored seeds in all of these substitution and addition lines, together with high protein content in some of the lines, make them very useful as a donor source for winter wheat breeding programs.Key words: Lr24, genomic in situ hybridization, sequence-tagged site, random amplified polymorphic DNA.

Detection of barley chromatin added to wheat by genomic in situ hybridization

Genome ◽  
1991 ◽  
Vol 34 (3) ◽  
pp. 448-452 ◽  
Author(s):  
Y. Mukai ◽  
B. S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Repeat Sequence ◽  
Genomic Clone ◽  
Genomic In Situ Hybridization ◽  
Addition Lines ◽  
Interphase Nuclei ◽  
Nucleolar Organizing Region ◽  
Chromosome Domains

A technique for in situ hybridization is reported that can be used to detect barley chromatin in wheat background using total genomic DNA as a probe. A 1:2 ratio of biotin-labeled genomic DNA of barley to blocking (unlabeled, sheared) DNA of wheat was sufficient to reveal brownish labeled barley chromosome domains against bluish background of unlabeled wheat chromatin in metaphase, prophase, and interphase nuclei of wheat-barley addition lines. Using this procedure, the behavior of specific barley chromosomes was analyzed in interphase and prophase cells. In prophase cells, the 6H chromosome was always associated with a nucleolus. A genomic clone of α-amylase gene (gRAmy56) that contains a barley-specific dispersed repeat sequence was also used to detect barley chromosomes in a wheat background.Key words: Hordeum vulgare, Triticum aestivum, genomic in situ hybridization, biotin, nucleolar organizing region.

Standard karyotype of Triticum longissimum and its cytogenetic relationship with T. aestivum

Genome ◽  
1993 ◽  
Vol 36 (4) ◽  
pp. 731-742 ◽  
Author(s):  
Bernd Friebe ◽  
Neal Tuleen ◽  
Jiming Jiang ◽  
Bikram S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Addition Line ◽  
Addition Lines ◽  
Substitution Lines ◽  
Chromosome Addition ◽  
C Banding ◽  
Chromosome Addition Lines ◽  
Standard Karyotype ◽  
Fertility Genes

C-banding polymorphism was analyzed in 17 accessions of Triticum longissimum from Israel and Jordan, and a generalized idiogram of this species was established. C-banding analysis was further used to identify two sets of disomic T. aestivum – T. longissimum chromosome addition lines and 13 ditelosomic addition lines and one monotelosomic (6S1L) addition line. C-banding was also used to identify T. aestivum – T. longissimum chromosome substitution and translocation lines. Two major nucleolus organizing regions (NORs) on 5S1 and 6S1 and one minor NOR on 1S1 were detected by in situ hybridization using a 18S–26S rDNA probe. Sporophytic and gametophytic compensation tests were used to determine the homoeologous relationships of T. longissimum chromosomes. The T. longissimum chromosomes compensate rather well and fertility was restored even in substitution lines involving wheat chromosomes 2A, 4B, and 6B that contain major fertility genes. Except for the deleterious gametocidal genes, T. longissimum can be considered as a suitable donor of useful genes for wheat improvement.Key words: Triticum aestivum, Triticum longissimum, homoeology, C-banding, in situ hybridization.

Genomic origins of Thinopyrum chromosomes specifying resistance to wheat streak mosaic virus and its vector, Aceria tosichella

Genome ◽  
1999 ◽  
Vol 42 (2) ◽  
pp. 289-295 ◽  
Author(s):  
Qin Chen ◽  
R L Conner ◽  
A Laroche ◽  
G Fedak ◽  
J B Thomas
Keyword(s):  
In Situ Hybridization ◽  
Mosaic Virus ◽  
Genomic Dna ◽  
Wheat Streak Mosaic Virus ◽  
Genomic In Situ Hybridization ◽  
Alien Chromosome ◽  
Aceria Tosichella ◽  
Resistant Lines ◽  
Partial Amphiploid

The genomic origin of alien chromosomes carrying resistance to wheat streak mosaic virus (WSMV) and the wheat curl mite (WCM), Aceria tosichella Keifer, was evaluated in nine wheat-alien addition, substitution, or translocation lines, derived from hybrids of wheat with Thinopyrum ponticum (Podp.) Barkworth & D.R. Dewey (2n = 10x = 70), or with Th. intermedium (Host) Barkworth & D.R. Dewey (2n = 6x = 42). One of the four wheat × Th. ponticum lines was resistant to WSMV and WCM. The other three lines were resistant to WCM, but susceptible to WSMV. Five wheat × Th. intermedium lines were susceptible to WCM, but were highly resistant to WSMV. Genomic in situ hybridization (GISH) using different genomic DNA probes demonstrated that all WSMV- and WCM-resistant lines carried an alien chromosome(s) related to the Js genome in Th. ponticum and Th. intermedium. The Js alien chromosome(s) displayed a special GISH hybridization pattern with the S genomic DNA probe, in which S genome DNA hybridized strongly in the centromeric regions and occasionally in the terminal regions with no or a weak hybridization signal in the middle of the two arms of the chromosomes. The WCM-resistant wheat × Th. ponticum lines N/5.10.10 and 63-30-2-2-2-8-1 carried the same short arm of chromosome 6Js derived from the partial amphiploid Agrotana, while the WSMV-resistant wheat × Th. intermedium lines T-Ai, CI15092, CI17766, A29-13-3-1 and KS93WGRC27 all had the same alien chromosome or chromosome arm of 4Js from Th. intermedium. The Js genome present in Th. ponticum and Th. intermedium is an important source of WSMV and WCM resistance that can be transferred into wheat.Key words: wheat-Thinopyrum lines, WSMV- and WCM-resistance, genomic in situ hybridization, Js genome.

Genome discrimination by in situ hybridization in Icelandic species of Elymus and Elytrigia (Poaceae: Triticeae)

Genome ◽  
2001 ◽  
Vol 44 (2) ◽  
pp. 275-283 ◽  
Author(s):  
Marian Ørgaard ◽  
Kesara Anamthawat-Jónsson
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Pseudoroegneria Spicata ◽  
Genome Constitution ◽  
Elytrigia Repens ◽  
Elymus Caninus ◽  
Elymus Species

The genome constitution of Icelandic Elymus caninus, E. alaskanus, and Elytrigia repens was examined by fluorescence in situ hybridization using genomic DNA and selected cloned sequences as probes. Genomic in situ hybridization (GISH) of Hordeum brachyantherum ssp. californicum (diploid, H genome) probe confirmed the presence of an H genome in the two tetraploid Elymus species and identified its presence in the hexaploid Elytrigia repens. The H chromosomes were painted uniformly except for some chromosomes of Elytrigia repens which showed extended unlabelled pericentromeric and subterminal regions. A mixture of genomic DNA from H. marinum ssp. marinum (diploid,Xa genome) and H. murinum ssp. leporinum (tetraploid,Xu genome) did not hybridize to chromosomes of the Elymus species or Elytrigia repens, confirming that these genomes were different from the H genome. The St genomic probe from Pseudoroegneria spicata (diploid) did not discriminate between the genomes of the Elymus species, whereas it produced dispersed and spotty hybridization signals most likely on the two St genomes of Elytrigia repens. Chromosomes of the two genera Elymus and Elytrigia showed different patterns of hybridization with clones pTa71 and pAes41, while clones pTa1 and pSc119.2 hybridized only to Elytrigia chromosomes. Based on FISH with these genomic and cloned probes, the two Elymus species are genomically similar, but they are evidently different from Elytrigia repens. Therefore the genomes of Icelandic Elymus caninus and E. alaskanus remain as StH, whereas the genomes of Elytrigia repens are proposed as XXH.Key words: Elymus, Elytrigia, H genome, St genome, in situ hybridization.

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