Screening and analysis of differentially expressed genes from an alien addition line of wheat Thinopyrum intermedium induced by barley yellow dwarf virus infection

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1114-1121 ◽  
Author(s):  
Shu-Mei Jiang ◽  
Long Zhang ◽  
Jun Hu ◽  
Rui Shi ◽  
Guang-He Zhou ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Expressed Sequence Tag ◽  
Barley Yellow Dwarf Virus ◽  
Differentially Expressed ◽  
Addition Line ◽  
Alien Chromosome ◽  
Thinopyrum Intermedium ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus ◽  
Expressed Sequence

The alien addition line TAI-27 contains a pair of chromosomes of Thinopyrum intermedium that carry resistance against barley yellow dwarf virus (BYDV). A subtractive library was constructed using the leaves of TAI-27, which were infected by Schizaphis graminum carrying the GAV strain of BYDV, and the control at the three-leaf stage. Nine differentially expressed genes were identified from 100 randomly picked clones and sequenced. Two of the nine clones were highly homologous with known genes. Of the remaining seven cDNA clones, five clones matched with known expressed sequence tag (EST) sequences from wheat and (or) barley whereas the other two clones were unknown. Five of the nine differentially expressed sequences (WTJ9, WTJ11, WTJ15, WTJ19, and WTJ32) were highly homologous (identities >94%) with ESTs from wheat or barley challenged with pathogens. These five sequences and another one (WTJ18) were also highly homologous (identities >86%) with abiotic stress induced ESTs in wheat or barley. Reverse Northern hybridization showed that seven of the nine differentially expressed cDNA sequences hybridized with cDNA of T. intermedium infected by BYDV. Three of these also hybridized with cDNA of line 3B-2 (a parent of TAI-27) infected by BYDV. The alien chromosome in TAI-27 was microdissected. The second round linker adaptor mediated PCR products of the alien chromosomal DNA were labeled with digoxygenin and used as the probe to hybridize with the nine differentially expressed genes. The analysis showed that seven differentially expressed genes were homologous with the alien chromosome of TAI-27. These seven differentially expressed sequences could be used as ESTs of the alien chromosome of TAI-27. This research laid the foundation for screening and cloning of new specific functional genes conferring resistance to BYDV and probably other pathogens.Key words: suppression subtractive hybridization (SSH), expressed sequence tag (EST), linker adaptor mediated polymerase chain reaction (LA-PCR), chromosome microdissection.

Isolation of expressed sequences from a specific chromosome of Thinopyrum intermedium infected by BYDV

Genome ◽  
2009 ◽  
Vol 52 (1) ◽  
pp. 68-76 ◽  
Author(s):  
Shu-Mei Jiang ◽  
Wei-Bo Yin ◽  
Jun Hu ◽  
Rui Shi ◽  
Ruo-Nan Zhou ◽  
...  
Keyword(s):  
Large Scale ◽  
Barley Yellow Dwarf Virus ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Addition Line ◽  
Alien Chromosome ◽  
Thinopyrum Intermedium ◽  
Specific Chromosome ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus

To map important ESTs to specific chromosomes and (or) chromosomal regions is difficult in hexaploid wheat because of its large genome size and serious interference of homoeologous sequences. Large-scale EST sequencing and subsequent chromosome localization are both laborious and time-consuming. The wheat alien addition line TAi-27 contains a pair of chromosomes of Thinopyrum intermedium (Host) Barkworth & D.R. Dewey that carry the resistance gene against barley yellow dwarf virus. In this research, we developed a modified technique based on chromosome microdissection and hybridization-specific amplification to isolate expressed sequences from the alien chromosome of TAi-27 by hybridization between the DNA of the microdissected alien chromosome and cDNA of Th. intermedium infected by barley yellow dwarf virus. Twelve clones were selected, sequenced, and analyzed. Three of them were unknown genes without any hit in the GenBank database and the other nine were highly homologous with ESTs of wheat, barley, and (or) other plants in Gramineae induced by abiotic or biotic stress. The method used in this research to isolate expressed sequences from a specific chromosome has the following advantages: (i) the obtained expressed sequences are larger in size and have 3′ end information and (ii) the operation is less complicated. It would be an efficient improved method for genomics and functional genomics research of polyploid plants, especially for EST development and mapping. The obtained expressed sequence data are also informative in understanding the resistance genes on the alien chromosome of TAi-27.

PCR-based molecular marker for the Bdv2 Thinopyrum intermedium source of barley yellow dwarf virus resistance in wheat

2001 ◽  
Vol 52 (12) ◽  
pp. 1383 ◽  
Author(s):  
P. Stoutjesdijk ◽  
S. J. Kammholz ◽  
S. Kleven ◽  
S. Matsay ◽  
P. M. Banks ◽  
...  
Keyword(s):  
Molecular Marker ◽  
Barley Yellow Dwarf Virus ◽  
Wheat Breeding ◽  
Addition Line ◽  
Thinopyrum Intermedium ◽  
Barley Yellow Dwarf ◽  
Pcr Product ◽  
Partial Amphiploid ◽  
Yellow Dwarf Virus ◽  
Pcr Based Molecular Marker

Because of the importance of BYDV in wheat production worldwide, and given the difficulties of bioassaying for resistance, a molecular marker was developed for the resistance known as Bdv2 that originates on the long arm of chromosome 7Ai1 of Thinopyrum intermedium. This resistance was identified in a partial amphiploid line TAF46, a disomic addition line to wheat (L1), a telosomic addition line (7Ai1 L), and a series of recombinants and translocations. A RAPD (random amplified polymeric DNA) marker for the resistant germplasm was cloned and sequenced, and primers were designed against that sequence to produce a sequence characterised amplified region (SCAR) marker. A single PCR product is produced only with genotypes carrying the resistance from any of the available recombinants. The cloned sequence, recommended primers, and PCR protocols are described. The usefulness of the marker has been demonstrated for following Bdv2 in segregating wheat breeding germplasm, with the imminent release of a BYDV-resistant cultivar.

Structural organization of an alien Thinopyrum intermedium group 7 chromosome in U.S. soft red winter wheat (Triticum aestivum L.)

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 716-722 ◽  
Author(s):  
M. G. Francki ◽  
O. R. Crasta ◽  
H. C. Sharma ◽  
H. W. Ohm ◽  
J. M. Anderson
Keyword(s):  
Winter Wheat ◽  
Structural Organization ◽  
Barley Yellow Dwarf Virus ◽  
Repetitive Sequences ◽  
Alien Chromosome ◽  
Thinopyrum Intermedium ◽  
Soft Red Winter Wheat ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus ◽  
Red Winter Wheat

Barley yellow dwarf virus (BYDV) resistance in soft red winter wheat (SRWW) cultivars has been achieved by substituting a group 7 chromosome from Thinopyrum intermedium for chromosome 7D. To localize BYDV resistance, a detailed molecular genetic analysis was done on the alien group 7 Th. intermedium chromosome to determine its structural organization. Triticeae group 7 RFLP markers and rye specific repetitive sequences used in the analysis showed that the alien chromosome in the P29 substitution line has distinguishing features. The 350–480 bp rye telomeric sequence family was present on the long arm as determined by Southern and fluorescence in situ hybridization. However, further analysis using a rye dispersed repetitive sequence indicated that this alien chromosome does not contain introgressed segments from the rye genome. The alien chromosome is homoeologous to wheat chromosomes 7A and 7D as determined by RFLP analysis. Presence of the waxy gene on chromosomes 7A, 7B, and 7D but its absence on the alien chromosome in P29 suggests some internal structural differences on the short arm between Th. intermedium and wheat group 7 chromosomes. The identification of rye telomeric sequences on the alien Thinopyrum chromosome and the homoeology to wheat chromosomes 7A and 7D provide the necessary information and tools to analyze smaller segments of the Thinopyrum chromosome and to localize BYDV resistance in SRWW cultivars.Key words: barley yellow dwarf virus, Thinopyrum intermedium, rye repetitive sequences, RFLP, homoeologous group 7.

Disomic Thinopyrum intermedium addition lines in wheat with barley yellow dwarf virus resistance and with rust resistances

Genome ◽  
1995 ◽  
Vol 38 (2) ◽  
pp. 385-394 ◽  
Author(s):  
P. J. Larkin ◽  
P. M. Banks ◽  
E. S. Lagudah ◽  
R. Appels ◽  
Chen Xiao ◽  
...  
Keyword(s):  
Stripe Rust ◽  
Stem Rust ◽  
Rust Resistance ◽  
Barley Yellow Dwarf Virus ◽  
Yellow Dwarf ◽  
Addition Lines ◽  
Thinopyrum Intermedium ◽  
Barley Yellow Dwarf ◽  
Group 2 ◽  
Yellow Dwarf Virus

Zhong 5 is a partial amphiploid (2n = 56) between Triticum aestivum (2n = 42) and Thinopyrum intermedium (2n = 42) carrying all the chromosomes of wheat and seven pairs of chromosomes from Th. intermedium. Following further backcrossing to wheat, six independent stable 2n = 44 lines were obtained representing 4 disomic chromosome addition lines. One chromosome confers barley yellow dwarf virus (BYDV) resistance, whereas two other chromosomes carry leaf and stem rust resistance; one of the latter also confers stripe rust resistance. Using RFLP and isozyme markers we have shown that the extra chromosome in the Zhong 5-derived BYDV resistant disomic addition lines (Z1, Z2, or Z6) belongs to the homoeologous group 2. It therefore carries a different locus to the BYDV resistant group 7 addition, L1, described previously. The leaf, stem, and stripe rust resistant line (Z4) carries an added group 7 chromosome. The line Z3 has neither BYDV nor rust resistance, is not a group 2 or group 7 addition, and is probably a group 1 addition. The line Z5 is leaf and stem rust resistant, is not stripe rust resistant, and its homoeology remains unknown.Key words: Agropyron, intermediate wheatgrass, leaf rust, stem rust, stripe rust, luteovirus.

Varying chromosome composition of 56-chromosome wheat × Thinopyrum intermedium partial amphiploids

Genome ◽  
1993 ◽  
Vol 36 (2) ◽  
pp. 207-215 ◽  
Author(s):  
P. M. Banks ◽  
S. J. Xu ◽  
R. R.-C. Wang ◽  
P. J. Larkin
Keyword(s):  
Barley Yellow Dwarf Virus ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Molecular Evidence ◽  
Thinopyrum Intermedium ◽  
Meiotic Analysis ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus ◽  
Head Morphology ◽  
Partial Amphiploids

Thinopyrum intermedium (2n = 42) is a source of many potentially useful genes for wheat improvement. Many partial amphiploids have been produced between Th. intermedium and Triticum aestivum that are fertile and stable. These partial amphiploids all have 56 chromosomes, including seven pairs of chromosomes from Th. intermedium. To explore the genomic composition of these lines, meiotic analysis was conducted on 32 hybrid combinations between eight different partial amphiploids. All but two of the chosen parents were distinguishable on the basis of perenniality, head morphology, and reactions to leaf, stripe, and stem rusts and to barley yellow dwarf virus. Chromosome pairing in the hybrids clearly indicated that all but two of the partial amphiploids differed in their composition of Thinopyrum chromosomes. The differences varied from one to five chromosomes. This confirms molecular evidence that the extra genome of the octoploid partial amphiploids is a variable synthetic genome combining chromosomes of the three Thinopyrum genomes E, J, and X. Though the extra synthetic genomes vary widely between different octoploids, they are nevertheless stable once formed. It is argued that the failure to establish these octoploid amphiploids as a new crop is a consequence of their differing chromosome complements, which makes it impractical to interbreed them.Key words: Thinopyrum, Agropyron, agrotriticum, wheat, amphiploid, octoploid, barley yellow dwarf virus, rust.

Expressed-sequence-tag approach to identify differentially expressed genes following peripheral nerve axotomy

1999 ◽  
Vol 64 (1) ◽  
pp. 34-40 ◽  
Author(s):  
Katsuhisa Tanabe ◽  
Saya Nakagomi ◽  
Sumiko Kiryu-Seo ◽  
Kazuhiko Namikawa ◽  
Yuji Imai ◽  
...  
Keyword(s):  
Peripheral Nerve ◽  
Differentially Expressed Genes ◽  
Expressed Sequence Tag ◽  
Differentially Expressed ◽  
Expressed Sequence

scholarly journals Expression of Thinopyrum intermedium-Derived Barley yellow dwarf virus Resistance in Elite Bread Wheat Backgrounds

2001 ◽  
Vol 91 (1) ◽  
pp. 55-62 ◽  
Author(s):  
L. Ayala ◽  
M. van Ginkel ◽  
M. Khairallah ◽  
B. Keller ◽  
M. Henry
Keyword(s):  
Bread Wheat ◽  
Seed Quality ◽  
Barley Yellow Dwarf Virus ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thinopyrum Intermedium ◽  
Mendelian Segregation ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus

Resistance to Barley yellow dwarf virus (BYDV) is not found in wheat but is available in a Thinopyrum intermedium translocation (Ti) carried on chromosome 7DL of bread wheat recombinant lines. We used one of those lines (TC14/2*Spear) to introgress the Ti into bread wheat cultivars and to determine the influence of wheat backgrounds, with and without known tolerance to BYDV, on the expression of resistance. Two single and three backcross populations, segregating for the presence of the alien fragment, were tested under field conditions and artificial inoculation with BYDV isolates MAV-Mex and PAV-Mex. Lines containing the fragment were identified using the microsatellite marker gwm37. Tillering, biomass, grain yield, thousand-kernel weight, and seed quality were evaluated in inoculated and noninoculated plots. Resistance was assessed by enzyme-linked immunosorbent assay. In early generations, the alien fragment followed expected Mendelian segregation, whereas in the advanced ones a slight bias against its transmission was observed. No positive nor negative effects of Ti on agronomic performance and quality were found. A significant optical density reduction in individuals carrying the fragment was observed after PAV infection in crosses with lines Anza and Baviacora but not with Milan. In addition, the fragment was associated with a lower frequency of infected plants for both PAV and MAV isolates. The reduced yield loss associated with the presence of the translocation was due largely to the lower infection rate.

Molecular characterization of a Thinopyrum intermedium Group 2 chromosome (2Ai-2) conferring resistance to barley yellow dwarf virus

Genome ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 1129-1135 ◽  
Author(s):  
Z Y Zhang ◽  
Z Y Xin ◽  
P J Larkin
Keyword(s):  
Barley Yellow Dwarf Virus ◽  
Rapd Marker ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Rflp Markers ◽  
Thinopyrum Intermedium ◽  
Barley Yellow Dwarf ◽  
Wheat Group ◽  
Group 2 ◽  
Yellow Dwarf Virus

The wheat – Thinopyrum intermedium addition lines Z1 and Z2 carry 21 pairs of wheat chromosomes and one pair of Th. intermedium chromosomes (2Ai-2) conferring resistance to barley yellow dwarf virus (BYDV). GISH results using the genomic DNA of Pseudoroegneria strigosa (S genome) as the probe indicated that the 2Ai-2 chromosome in Z1 and Z2 is an S–J intercalary translocation. Most of the 2Ai-2 chromosome belongs to the S genome, except for about one third in the middle region of the long arm that belongs to the J genome. The results of detailed RFLP analyses confirmed that the 2Ai-2 chromosome is extensively homoeologous to wheat group 2 chromosomes. Some new RFLP markers specific to the 2Ai-2 chromosome were identified. A RAPD marker, OP-R16340, specific to the 2Ai-2 chromosome, was screened. We converted the RAPD marker into a sequence-characterized amplified region (SCAR) marker (designated SC-R16). The study establishes the basis for selecting translocation lines with small segments of the 2Ai-2 chromosome and localizing the BYDV resistance gene when introgressed into a wheat background.Key words: Thinopyrum intermedium, barley yellow dwarf virus (BYDV), genomic in situ hybridization (GISH), RFLP, SCAR, homoeologous group 2.

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