The Researches On Blossom Biology And The Analysis On Genetic Relationship Using ISSR Of The Series Prunus Varieties Of'Ziguiqiulu'

P. salicina Lindl is a deciduous arbors, which belongs to the Roseceae Prunus.L plant. It flowering in spring, the new saplings began to bear only two years after planting, and arrived full fruiting period4-5years after planting. It is a good non-wood tree specie. Prunus Salicina "ziqiu", Prunus Salicina "zilu", Prunus Salicina "qiulu" are new varieties breeded out for ten years which are suitable for the half high mountain areas which the elevation is from700m to1500m of the west of Hubei regions. Prunus Salicina "ziqiu" is a middle-late mature specie which fruit maturation is middle-August. But Prunus Salicina "zilu" and Prunus Salicina "qiulu" are late mature species which ripe in middle-September. This paper using the Prunus Salicina "ziqiu", Prunus Salicina "zilu", Prunus Salicina "qiulu" as the materials, using of paraffin section, fluorescence microscopy and other methods to investigate the biology characteristics of flowering phenophase, the quality of pollen, the rate of cross-pollination and self-pollination, acception of stigma, the process of pollination and fertilization. And use ISSR markers to study the genetic relationships of the14Prunus samples from zigui hubei which include Prunus Salicina "ziqiu", Prunus Salicina "zilu" and Prunus Salicina "qiulu". Its findings can be summarized as follows:1. The study on flowering phenophase. The anthesis of Prunus Salicina "ziqiu", Prunus Salicina "zilu" and Prunus Salicina "qiulu" are always in late-March or early-April. The florescence of Prunus Salicina "zilu" and Prunus Salicina "qiulu" are the same, but the florescence of Prunus Salicina "ziqiu" is later4to7days than theirs. The whole florescence can continue7to10days. Flowering process of single flower can be divided into3periods of pink bud stage, abducent bud stage and drop bud stage.2. The study on pollination and fertilization. The fresh pollen germination rate of Prunus Salicina "ziqiu", Prunus Salicina "zilu" and Prunus Salicina "qiulu" are79.3%,56.8%and70.5%. The observation of stigma, three Prunus species's effective pollination period is two days before flowering to four days after flowering, a total of six days. But the best pollination period is two days before flowering to flowering the first two days, a total of four days. The fruit-bearing rate of self-pollination of Prunus Salicina "ziqiu", Prunus Salicina "zilu" and Prunus Salicina "qiulu" are8%,5.5%and6.5%. The fruit-bearing rate of polycross of theirs are15.5%,23.5%and18.5%. The pollens of Prunus Salicina "ziqiu" begun to germinate after pollinating1h, and the pollen tube can grow to the base of stylus after pollinating36h. After pollinating72h, the double fertilization process was completed.3. The establishment of ISSR-PCR reaction systerm. Through the usage of the orthogonal design, an optimized ISSR-PCR reaction systerm was established:20μL reaction systerm containing lOXBuffer (Mg2+free)2.0uL,1.5U Tap DNA polymerase,0.5μmol/Lprimer,0.2mmol/L dNTPs,2.0mmol/L MgC12,20ng template DNA. The PCR reactions program was pre-denaturing at94℃for5minutes,.38cycles of denaturation at94℃for45seconds, annealing at55-56℃for1minute, extension at72℃for1.5minutes. With a10minutes final extension at72℃. Then saved at4℃.4. The analysis of genetic relationships of the14Prunus samples. By the optimized ISSR-PCR reaction systerm, ten ISSR primers were selected which can yield specific bands with moderate band numbers, clear background and well reproducibility. In14expermental varieties,101DNA bands were amplified, in which had83polymorphic bands, the PPB was82.18%. The software of POPGENE1.31and NTSYS2.10e were used to analysis the results of ISSR-PCR. The results showed that the genetic distance ranged from0.1467to0.5027, the genetic identity ranged from0.6022to0.8601, the genetic similarity coefficients ranged from0.65to0.89,and the average was0.78. Using UPGMA clustering method,14Prunus species were placed into two different cluster: the first cluster inclus Prunus Salicina "ziqiu", Prunus Salicina "zilu" and Prunus Salicina "qiulu" and Prunus American "meilinvhuang" which was introduced from American. The other local varieties were classified the second cluster.