Genetic Identifications On Cytoplasmic Male Sterile Lines Of Hong Cai-tai Eru-3A (Brassica Campestrisl. Ssp. Chinensisl. Var.Utilistsen Et Lee.)

A new kind of cytoplasmic male sterility of Brassica napus (eru CMS) was transfered to purple cai-tai by the way of distant hybridization occurring between different genuses, So we obtain corresponding male sterile line of purple cai-tai. Analysises on botany,cytology, isoenzyme and PCR amplification were carried out.The results were as follow:1. Analysis on botany of the cytoplasmic male sterility of Hong Cai-tai. The field survey and agronomic characters observed that the cytoplasmic sterile lines in the growing period of morphological index (for example:plant height, open measure,leaflength and width etc) has been close to the male parent Hong Cai-ta Yuyou,but in the flower organ formation, there were very significantly differences in stamen length,silk length and height difference of pistil and stamen,the cytoplasmic male sterility of Hong Cai-tai of flower organ formation was in the middle. Through the determination of the pollen vigor and observation in past years, show that the sterile lines is can genetic, and thoroughly abortion cytoplasmic male sterile lines.2. The Cytomorphology Study of Anther Abortion in new Male-Sterile Lines of Hong Cai-tai(Brassica campestrisL.ssp.chinensisL.var.utilisTsen et Lee.).Cytological observations of anther in new Hong Cai-tai cytoplasmic male sterile line and maintainer line Yuyou was made by means of paraffin section in order to determine the period of anther abortion, methods and characteristics. The results showed that the abortion occurred at the stage of Microspore early stage,observing the tapetum cell abnormal and Microspore early stage being extruded.In the end the tapetum cell and Microspore early stage disaggregated.3. Analysis on isoenzyme of the cytoplasmic male sterility of Hong Cai-tai. The isozyme(POD,COD,EST) in leaf, leaf stalk, young stem,flower bud and flower of Hong Cai-tai was analyzed by PEGM electrophoresis.The results showed that CMS lines of Hong Cai-Tai in middle flower bud POD isozyme more than the corresponding maintainer line Yuyou1enzyme belt (Rf=0.257), there may be relate to the sterility, and in leaves and young stem separately less than maintainer line Yuyou Rf=0.227and Rf=0.272and0.788of enzyme belts. CMS lines of Hong Cai-Tai of COD isozyme in leaves and petioles respectively less Rf=0.476and Rf=0.754enzyme belt than the corresponding maintainer line Yuyou, but on the Big flower bud and middle flower bud more than corresponding maintainer line Yuyou1enzyme belt (Rf=0.411), this shows that it is characteristics of the isozyme bands in the bud. In isozyme EST, maintainer line of Hong Cai-tai Yuyou have more enzyme belts relative to the cytoplasmic male sterility of Hong Cai-tai. The maintainer line of Hong Cai-tai Yuyou have more one enzyme belt than CMS lines of Hong Cai-Tai in leaves, petioles and middle flower bud,just the opposite in little flower bud.4.Molecular identification of cytoplasmic male sterile types in Hong Cai-tai. In this study,primers were designed according to the reported fertility-related gene atp6,orf138,orf224-atp6,atp6-orf263,orf222-nad5-orfl39and the DNA amplification of1CMS lines of Hong Cai-tai and its maintainer line and4CMS lines of Brassica napus carried out by PCR.The result indicated that cytoplasmic male sterile types of Hong Cai-tai belong to the Ogu CMS.