Mouse Karyotype Obtained by Combining DAPI Staining with Image Analysis

In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4＇, 6-diamidino-2-phenylindole （DAPI） multiple bands like （J-hands could be produced in mouse. The Meta- Morph software was then used to generate linescans of pixel intensity for the banded chromosomes from short arm to long arm. These linescans were sufficient not only to identify each individual chromosome but also analyze the physical sites of bands in chromosome. Based on the results, the clear and accurate karyotype of mouse metaphase chromosomes was established. The technique is therefore considered to he a new method for cytological studies of mouse.

Chromosome G-banding in SituHybridization of RFLP Marker umc58 Linked with the Gene hm1 Dictating Helminthosporium carbonum Susceptibility1 in Maize

The technique of simultaneous G banding and in situ hybridization has been developed in plants for the first time.Using this technique.RFLP marker umc58 closely linked with the hm1 gene dictating Helminthosporium carbonum susceptibility1 was localized onto 1L3(chromosome 1,long arm,the third band from the centromere to the end of the arm),5L5 and 9L5.Theresults demonstrated that umc58 was a tripli cated sequence.It was deduced that umc58 probably was in a duplicated region that includes a part of Helminthosporium carbonum susceptibility genes(hm1 and hm2),as the hybridization sites of umc58 in chro mosomes 1 and 9 were those at which the genes localize.The techniques of simultaneous G banding and ISH in plants are discussed.

Physical Location of Terminal Markers Belonging to Five Linkage Groups in Maize RFLP Map Using in Situ Hybridization

Ten terminal or subterminal RFLP markers belonging to linkage groups 1, 3, 5, 6, and 10 in maize RFLP map were physically located onto maize mitotic chromosomes with in situ hybridization. All biotinylated probes from 600 to 2 250 bp were detected by DAB staining. The markers belonging to linkage groups 1, 3, 5, 6, and 10 correspondingly located at the chromosomes 1, 3, 5, 6, and 10. All of the tested markers except bnl6.25 and umc44 were duplicated sequences. Each of them was also labeled on another chromosome besides on the chromosome corresponding to its linkage group. The marker bnl3.04 was triplicated sequences and the signals were detected on three nonhomologous chromosomes. In the tested ten markers, there were only four located at the ends of corresponding chromosomes. Others were located at sites midway along the chromosome arms or near the centromeres. The region covered by two terminal or subterminal markers in each of linkage groups 1, 3, 5, and 6 occupied 80.02%, 38.25%, 82.30% and 51.16% of the region of both short and long arms in chromosomes 1, 3, 5,and 6 respectively. Only two terminal markers of linkage group 10 covered the whole chromosome 10. In some linkage groups, two terminal or subterminal markers covered a short genetic distance but were physically distant, while two covering a longer genetic distance were physically closer.

Physical location of rice Gm-6,Pi-5(t) genes in O.officinalis with BAC-FISH

A fluorescence in situ hybridization (FISH) procedure was adopted to physically map two rice BAC clones 24E21 and 4F22 linked to Gm-6 and Pi-5(t) in O. offi-cinalis. FISH results showed that the two BAC clones were located at 4L. The percentage distance from the centromere to the hybridization sites was 72 ± 2.62 for 24E21 and 54± 5.43 for 4F22, the detection rates were 52.70% and 61.2%. The results obtained from the BAC and plasmid clones, RG214 and RZ565 of cultivated rice and O. officinalis were the same. This suggested that the markers, RG214 and RZ565 of cultivated rice and O. officinalis were in the same BAC clones. The homologous sequences of Gm-6 and Pi-5(t) in O. officinalis were positions that signals existed on the 4L. Many signals were observed when no Cot-1 DNA blocked. This also showed that repetitive sequences were some ho-molgous between cultivated rice and O. officinalis. The identification of chromosome 4 of O. officinalis is based on Jena et al. (1994). In our study, we discussed

FISH analysis of the integration patterns in transgenic rice co-transformed by micro-projectile bombardment

Using multi-color fluorescence in situ hybridization (FISH), we localized transferred barnase-psl and pHctinG DNA sequences onto chromosomes of two trans-genie rice plants, named Q12 and Q13, both of which were produced by micro-projectile bombardment. In both Q12 and Q13, each detected cell showed 2-3 signal spots on their chromosomes respectively. The signals of both barnase-psl and pHctinG were mostly detected in the adjacent chromosomal sites in which their signals were overlapped and could be recognized by the signal color on the metaphase chromosomes. Fiber FISH further demonstrated that the multiple copies in each of the two DNA sequences distributed adjacently on the DNA fiber in Q13. Combined with the results of Southern hybridization, the possible integration patterns in transgenic rice co-transformed by micro-projectile bombardment have been discussed.

A correlation method of detecting and estimating interactions of QTLs

More and more studies demonstrate that a great deal of interactions among the quantitative trait loci (QTLs) are far more than those detected by single markers. A correlation method was proposed for estimating the interactions of multiple QTLs detected by multi-markers in several mapping populations. Genetic implication of this method and usage were discussed.