[Analysis of factors influencing GISH used in Triticeae crops' body cells]

Factors influencing genomic in situ hybridization (GISH) used in the body cells of Triticeae crops including Octoploid Tritileymus, Octoploid Trititrigia, Octoploid Tritelytrigia and wheat-Elytrigia intermedium alien disomic addition line were analyzed. The results indicated that ideal GISH effects were based on the high quality chromosome preparations containing more cell mitotic metaphase, better dispersed chromosomes and no foreign matter. The critical factors were suitable ratio of probe DNA to blocker DNA, and the control of elution condition. In addition, some abnormal phenomena, such as chromosome loss, no hybridization signal in alien chromosome, strong or weak hybridization signal, poor or overabundant hybridization signal (high hybridization background), noisy hybridization signal and hybridization spots, were preliminarily analyzed. Proposals and solutions for these problems were further brought forth.