Identification of Haynaldia villosa chromosomes added to wheat using a sequential C-banding and genomic in situ hybridization technique

Abstract

Genomic in situ hybridization (GISH) offers a convenient and effective method for cytological detection, but can not determine the identity of the chromosomes involved. We integrated C-banding with GISH to identify Haynaldia villosa chromosomes in a wheat background. All chromosomes of H. villosa showed C-bands, either in telomeric regions or in both telomeric and centromeric regions, which allowed unequivocal identification of each H. villosa chromosome. The seven pairs of H. villosa chromosomes were differentiated as 1-7 according to their characteristic C-bands. Using a sequential C-banding and GISH technique, we have analyzed somatic cells of F3 plants from the amphiploid Triticum aestivum-H. villosa x 'Yangmai 158' hybrids. Three plants (94009/5-4,94009/5-8 and 94009/5-9) were shown to contain H. villosa chromosome(s). 94009/5-4 (2n = 45) had three H. villosa chromosomes (2, 3 and 4); 94009/5-8 (2n = 45) possessed one chromosome 4 and a pair of chromosome 5, and 94009/5-9 (2n = 43) was found to have one chromosome 6 of H. villosa. The combination of GISH with C-banding described here provides a direct comparison of the cytological and molecular landmarks. Such a technique is particularly useful for identifying and localizing alien chromatin and DNA sequences in plants.

Characterization of a genome-specific Gypsy-like retrotransposon sequence and development of a molecular marker specific for Dasypyrum villosum (L.)

“…With the introduction of alien chromatin into wheat, many useful markers such as morphological markers (Zhong and Qualset 1990), biochemical markers (Li et al 2009), and cytological markers (Zhong et al 1996) have been developed to monitor the behaviour of the alien chromosomes. However, the major limitation of these techniques is that they are highly technical and time-consuming.…”

Introduction of multi-alien chromatins carrying different powdery mildew-resistant genes from rye and Haynaldia villosa into wheat genome.

“…Jiang and Gill (1993) proved that the sequential GISH after N-banding or C-banding gives satisfactory results. Zhong et al (1996) also reported that prior Cbanding has only a slight negative effect on the subsequent GISH. In this study, the sequential C-banding after multicolor fluorescence GISH analysis gave better results compared with the sequential GISH after the C-banding treatment, and enabled us not only to distinguish the H. villosa and rye chromatins from the wheat chromatin simultaneously, but also to identify the alien chromosomes and chromosomal segments.…”

“…In addition, C-banding analysis before or after GISH is effective and useful for identifying alien chromatins (Hutchinson and Seal, 1983;Jiang and Gill, 1993;Zhong et al, 1996). In this study, we report the results of the introduction of multi-alien chromatins with different powdery mildew-resistant genes from S. cereale and H. villosa and the simultaneous identification of these chromatins in a wheat background by multicolor fluorescence GISH analysis.…”

The genetic identity of alien chromosomes in potato breeding lines revealed by sequential GISH and FISH analyses using chromosome-specific cytogenetic DNA markers

“…However, when these two techniques are used separately, it is often difficult to identify the alien chromosomes, especially translocated chromosomes. These two techniques can also be sequentially performed on the same cytological preparations (Jiang and Gill 1993b;Zhong et al 1996;Cai et al 1998;Schubert et al 1998). One problem with sequential analysis is that the quality of subsequent banding or in situ hybridization tends to be less than optimal, owing to the previous treatments.…”