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.

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.

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.

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.

Comparison of Thinopyrum intermedium derivatives carrying barley yellow dwarf virus resistance in wheat

Genome ◽  
2009 ◽  
Vol 52 (6) ◽  
pp. 537-546 ◽  
Author(s):  
L. Ayala-Navarrete ◽  
E. Tourton ◽  
A. A. Mechanicos ◽  
P. J. Larkin
Keyword(s):  
Barley Yellow Dwarf Virus ◽  
Yellow Dwarf ◽  
Substitution Line ◽  
Dwarf Virus ◽  
Translocation Line ◽  
Thinopyrum Intermedium ◽  
Barley Yellow Dwarf ◽  
Genetic Stocks ◽  
Chromosome Arm ◽  
Yellow Dwarf Virus

Resistance to both barley yellow dwarf virus (BYDV) and cereal yellow dwarf virus (CYDV) has been demonstrated in wheat genetic stocks with Thinopyrum intermedium chromatin. A number of resistance-bearing translocations have been reported on chromosome arm 7DL from two independent Th. intermedium sources; one source is the addition line L1 and the other is the spontaneous substitution line P29. Another source of resistance in wheat cytogenetic stocks is available as a 2Ai(2D) substitution line. We used a set of 38 molecular markers and the available deletion stocks to compare the size of the 7DL translocations more comprehensively than has been done previously. We also compared the efficacy of BYDV resistance of the various genetic stocks both before and after transfer to a common genetic background. TC14 was confirmed as carrying the smallest translocation, replacing about 20% of the distal end of 7DL. TC5 and TC10 had 90% of the chromosome arm replaced by Th. intermedium chromatin; the proximal 10% corresponded to wheat chromatin. YW642 appeared to have the whole 7DL replaced by Th. intermedium chromatin, as confirmed by the co-dominant marker cfd68 mapping on the bin nearest the centromere. Translocation line P961341 had bins 3, 7, and 8 replaced by Th. intermedium chromatin, making this the second smallest translocation with BYDV and CYDV resistance. The translocation sizes reported here differ from some of the previous estimates. The translocated Th. intermedium segments appeared to be bigger than the replaced wheat 7DL fragments. All the resistances derived from the L1 and P29 group 7 chromosomes and the 2Ai#2 chromosome were effective in reducing the number of infected plants and the mean virus titre, regardless of the background. Some evidence is discussed suggesting the long arm of the Th. intermedium group 7 chromosome 7Ai#1 carries two resistances, the distal Bdv2 and a proximal second gene.

Identification and characterization of wheat-wheatgrass translocation lines and localization of barley yellow dwarf virus resistance

Genome ◽  
2000 ◽  
Vol 43 (4) ◽  
pp. 698-706 ◽  
Author(s):  
O R Crasta ◽  
M G Francki ◽  
D B Bucholtz ◽  
H C Sharma ◽  
J Zhang ◽  
...  
Keyword(s):  
Repetitive Sequence ◽  
Barley Yellow Dwarf Virus ◽  
Rflp Analysis ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Thinopyrum Intermedium ◽  
Barley Yellow Dwarf ◽  
Translocation Lines ◽  
Yellow Dwarf Virus ◽  
Alien Chromatin

Stable introgression of agronomically important traits into crop plants through wide crossing often requires the generation and identification of translocation lines. However, the low efficiency of identifying lines containing translocations is a significant limitation in utilizing valuable alien chromatin-derived traits. Selection of putative wheatgrass-wheat translocation lines based on segregation ratios of progeny from γ-irradiated seed using a standard phenotypic analysis resulted in a low 4% success rate of identifying barley yellow dwarf virus (BYDV) resistant and susceptible translocation lines. However, 58% of the susceptible progeny of this irradiated seed contained a Thinopyrum intermedium chromosome-specific repetitive sequence, which indicated that γ-irradiation-induced translocations occurred at high rate. Restriction fragment length polymorphism (RFLP) analysis of susceptible lines containing alien chromatin, their resistant sister lines and other resistant lines showed that more than one third of the progeny of γ-irradiated double monosomic seeds contained wheatgrass-wheat translocations. Genomic in situ hybridization (GISH) analysis of selected lines confirmed that these were wheatgrass-wheat translocation lines. This approach of initially identifying BYDV susceptible deletion lines using an alien chromosome-specific repetitive sequence followed by RFLP analysis of their resistant sister lines efficiently identified resistant translocation lines and localized the BYDV resistance to the distal end of the introgressed Th. intermedium chromosome.Key words: gene introgression, wide crosses, chromosome, repetitive elements, RFLP, Thinopyrum intermedium.

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