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Identification of Expressed Sequence Markers for a Major Gene‐Rich Region of Wheat Chromosome Group 1 Using RNA Fingerprinting–Differential Display
Author(s) -
Sandhu Devinder,
Sidhu Deepak,
Gill Kulvinder S.
Publication year - 2002
Publication title -
crop science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.76
H-Index - 147
eISSN - 1435-0653
pISSN - 0011-183X
DOI - 10.2135/cropsci2002.1285
Subject(s) - biology , genetics , gene , genome , chromosome , complementary dna , restriction fragment length polymorphism , microbiology and biotechnology , polymerase chain reaction
This study demonstrates a successful application of RNA fingerprinting–differential display technique in identifying expressed sequence markers for a small targeted region of the wheat ( Triticum aestivum L.) genome. Wheat genes are present in clusters spanning about 10% of the genome. One of the important gene‐rich regions is present on the short arm of wheat homoeologous group 1 chromosomes around fraction length 0.8 (‘1S0.8 region’). The region is about 0.1% of the wheat genome and is flanked by the breakpoints of deletion lines 1BS‐4 and 1BS‐19. The objective of this study was to identify expressed sequence markers for the region. First‐strand cDNA of poly A + mRNA pooled from various developmental stages of the two deletion lines were PCR amplified in the presence of 35 S by means of 90 pair‐wise combinations of 19 primers. Amplification products were size‐separated on a denaturing polyacrylamide urea gel. A total of 6840 fragment bands were amplified, of which 65 were present in the deletion line 1BS‐4, but missing in 1BS‐19. These 65 fragment bands were cut out of the gel, reamplified, and used as probes for gel‐blot DNA analysis of group 1 nullisomic‐tetrasomic lines and the two deletion lines. Nineteen of the 65 fragment bands detected a smear pattern and thus were not mapped. Of the remaining 46 probes, 22 mapped to wheat homoeologous group 1 and seven mapped to the ‘1S0.8 region’. The same approach can be used to target other wheat gene‐rich regions bracketed by deletion breakpoints.