Estimation Of Ploidy And Genome Size On Several Species In Cerasus

Cerasus, a worldwide famous ornamental flowing plant, belongs to the Rosaceae, Prunoidea, and it’s abundant in China. This genus has a high degree of variability and the existence of a large number of hybrid original of polyploidy. There is a large controversy on its infrageneric classification and definition. In this experiment, 13 kinds of Cerasus plants were chosen as the research objects. Ploidy and Genome Size were analyzed by flow cytometry which can provide cytological evidence for the origin and evolution and also provide reference for the whole genome sequencing. The major conclusions are as follows:1. The method of Cerasus nuclear suspension preparationBased on the existing plant flow cytometry studies, a set of economic, fast and efficient process flow cytometry of Cerasus was worked out after optimizing the way of making Cerasus nuclear suspension. By setting the 6 kinds of nuclei lysis control tests, i.e., WPB, Galbraith’s, LB01, GPB, Trisï¹'MgCl2 and Chopping, the WPB buffer was proved to be the most suitable for Cerasus flow cytometry. Respectively discusses the experimental materials, lysis processing time, the choice of the fluorescent dye, rotational speed, the centrifugal time and dyeing time on the result of flow cytometry. It has been detected that the best result is under this circumstance, the seed germination of new leaves as experimental material, cracking liquid extracting 3 min, 4 degrees Celsius temperature centrifugal rotational speed of 800 r?min-1, centrifuge for 5 min, cold dark PI stain 2 h. This paper adopts the BD FACS sortware 1.0.0.650 software drawing analysis.2. Ploidy analysis of CerasusUsing FCM to detect the ploidy of other species, compared with the well-known diploid C.serrulata. We can identify that the following is diploid, C. yedoensis, C. cerasoides, C. jamasakura, C. xueluoensis, C. discoidea, C. campanulata, C. subhirtella ‘Plendula Rosea’, C. subhirtella ‘Plena Rosea’, C. subhirtella var. ascendens; and these are triploid, C. serrulata var.lannesiana ‘Speciosa’, C. subhirtella; while C. pseudocerasus is Tetraploid. The result is the same with Iwatsubo Cerasus chromosome tablet technology statistics. Compared with traditional squash methods, flow cytometry techniques showed great advantages and broad prospects in ploidy identification and genetic breeding in Cerasus.3. Genome Size Study of Cerasus plantIn this study, the genome size of 13 kinds of Cerasus plant was tested by flow cytometry under using soybean(William 82) as an internal standard. It’s the first time to identify the genome size respectively for C. yedoensis, C. serrulata, C. cerasoides, C. campanulata C. jamasakura, C. xueluoensis, C. discoidea, C. subhirtella ‘Plendula Rosea’, C. subhirtella ‘Plena Rosea’, C. subhirtella var. ascendens, C. serrulata var. lannesiana ‘Speciosa’,C. subhirtella, C. pseudocerasus are:320.71 Mb, 366.67 Mb, 371.38,300.94 Mb, 294.23 Mb, 289.83 Mb, 336.83 Mb,294.05 Mb, 274.59 Mb, 302.26 Mb, 551.83 Mb, 973.59 Mb, 1223.47 MB. There is a little difference between diploid genome sizes. The genome increases with the multiple, so does the difference.