Comparative Analysis Of Three CCDD Wild Rice With FISH Technique In The Cell Level

1. In order to study genome difference of O. grandiglumis, O. alta and O. latifolia, analyze the genome of these three CCDD wild rice with FISH technique. With O. sativa and O. officinalis genomic DNA for probe respectively, O.grandiglumis somatic chromosomes on fluorescence in situ hybridization(FISH) combined difference stringency to detect the hybridization signal on chromosomes.The results show that O. sativa genomic DNA is distributed in the C and D genome,But the hybridization signal is much more on the C genome than those on the D genome,which illustrates A and C genome have closer relations than A and D genome With O. officinalis genomic DNA for probe, O. grandiglumis, O. alta and O. latifolia somatic chromosomes on fluorescence in situ hybridization(FISH) combined difference stringency to detect the hybridization signal on chromosomes.The results show that O. officinalis genomic DNA can distinguish the C and D genome of O. grandiglumis under the lower stringency,while C and D genome of O. alta and O. latifolia can be distinguished under the higher stringency,which shows O. alta and O. latifolia have difference with O. grandiglumis in the genome level.2. With O.sativa and O. officinalis Cot-1 DNA for probe respectively, O.grandiglumis somatic chromosomes on fluorescence in situ hybridization combined difference stringency to detect the hybridization signal on chromosomes. The results show that Cot-1 DNA of O. sativa mainly lie on the centromere, near centromere and telomere area of C genome of O.grandiglumis under the 80% stringency, while the signals of these region on D genome is less, which shows that the medium and high repeat sequence on D genome of O.grandiglumis have a certain homology with the medium and high repeat sequence of A genome, but the homology between medium and high repeat sequence of C and A genome is larger. With Cot-1 DNA of O. officinalis for probe, O.grandiglumis somatic chromosomes on fluorescence in situ hybridization, C and D genome of O.grandiglumis can only be separated under higher stringency (80%). With Cot-1 DNA of O. officinalis for probe, O.alta somatic chromosomes on fluorescence in situ hybridization, also found that the C and D genome of O.alta can be distinguished only under the higher stringency, however, the signals on the C genome of O.alta is weaker than those on the C genome of O.grandiglumis, which illustrates that O.alta have difference with O.grandiglumis in the Cot-1 DNA level. 3. With centromere DNA of O.sativa for probe, O. officinalis, O. latifolia and O. alta somatic chromosomes on fluorescence in situ hybridization, the results display that nearly every chromosomes of O. officinalis have hybridization signals, while hybridization signals only distributed on a few chromosomes of O. latifolia and O. alta, and there are 8 chromosomes with signals for O. latifolia,while there are 16 chromosomes with signals for O. alta, which display that the centomere DNA of O. latifolia and O. alta have a certain distincts. According to the centromere DNA difference of O. latifolia and O. alta,we can consider them to be classified the difference subspecies under the same specie.