title: Isolation of expressed sequences from a specific chromosome of Thinopyrum intermedium infected by BYDV.
abstract: 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.
authors.affiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, Beijing 100101, PR China.
authors.forename: Shu-Mei
authors.initials: SM
authors.lastname: Jiang
authors.forename: Wei-Bo
authors.initials: WB
authors.lastname: Yin
authors.forename: Jun
authors.initials: J
authors.lastname: Hu
authors.forename: Rui
authors.initials: R
authors.lastname: Shi
authors.forename: Ruo-Nan
authors.initials: RN
authors.lastname: Zhou
authors.forename: Yu-Hong
authors.initials: YH
authors.lastname: Chen
authors.forename: Guang-He
authors.initials: GH
authors.lastname: Zhou
authors.forename: Richard R-C
authors.initials: RR
authors.lastname: Wang
authors.forename: Li-Ying
authors.initials: LY
authors.lastname: Song
authors.forename: Zan-Min
authors.initials: ZM
authors.lastname: Hu
mesh_terms: D002874:Chromosome Mapping; D002875:Chromosomes / Q000648:ultrastructure; D032461:Chromosomes, Plant; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D018744:DNA, Plant / Q000378:metabolism; D020224:Expressed Sequence Tags; D005786:Gene Expression Regulation; D017343:Genes, Plant; D018745:Genome, Plant; D017793:Luteovirus / Q000235:genetics; D008957:Models, Genetic; D009841:Oligonucleotides / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D014908:Triticum / Q000235:genetics* / Q000821:virology*
chemical_list: D018076:DNA, Complementary; D018744:DNA, Plant; D009841:Oligonucleotides; D012333:RNA, Messenger