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

• Published in: Genome Vol. 52; no. 1; pp. 68 - 76
• Main Authors: Jiang, Shu-Mei, Yin, Wei-Bo, Hu, Jun, Shi, Rui, Zhou, Ruo-Nan, Chen, Yu-Hong, Zhou, Guang-He, Wang, Richard R.C, Song, Li-Ying, Hu, Zan-Min
• Format: Journal Article
• Language: English
• Published: Canada NRC Research Press 2009; Canadian Science Publishing NRC Research Press
• Subjects: addition lines; amplification spécifique de séquences hybridées (HSA); Barley; Barley yellow dwarf virus; barley yellow dwarf virus (BYDV); Chromosome Mapping; chromosome microdissection; Chromosomes; Chromosomes - ultrastructure; Chromosomes, Plant; Cloning; Cloning, Molecular; complementary DNA; Deoxyribonucleic acid; disease resistance; DNA; DNA, Complementary - metabolism; DNA, Plant - metabolism; Expressed Sequence Tags; expressed sequences; Gene expression; Gene Expression Regulation; genes; Genes, Plant; genetic techniques and protocols; Genome, Plant; Genomics; Grasses; Hybridization; hybridization-specific amplification; hybridization-specific amplification (HSA); infection; Luteovirus - genetics; microdissection chromosomique; Models, Genetic; nucleic acid hybridization; Oligonucleotides - genetics; RNA, Messenger - metabolism; séquences exprimées; TAi-27; Thinopyrum intermedium; Thinopyrum intermedium subsp. intermedium; Triticum - genetics; Triticum - virology; Triticum aestivum; virus de la jaunisse nanisante de l'orge (BYDV); Viruses; Wheat; Canada
• ISSN: 0831-2796; 1480-3321
• DOI: 10.1139/G08-108

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.