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CELL BIOLOGY & MOLECULAR GENETICS

Identification of Expressed Sequence Markers for a Major Gene-Rich Region of Wheat Chromosome Group 1 Using RNA Fingerprinting–Differential Display

Dep. of Agronomy and Horticulture, 362H Plant Science, P.O. Box 830915, Univ. of Nebraska-Lincoln, Lincoln, NE 68583-0915

* Corresponding author (KGILL1{at}UNL.EDU)

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 ³⁵S 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.

This article has been cited by other articles:

				M. Erayman, D. Sandhu, D. Sidhu, M. Dilbirligi, P. S. Baenziger, and K. S. Gill Demarcating the gene-rich regions of the wheat genome Nucleic Acids Res., July 7, 2004; 32(12): 3546 - 3565. [Abstract] [Full Text] [PDF]