Transfer Of Ogura Cytoplasmic Male Sterility To Cauliflower (Brassica Oleracea Var.Botrytis L.) By Asymmetric Protoplast Fusion

Asymmetric protoplast fusion is a practical and important approach for the genetic improvement of crops.Through the technique, we can transfer cytoplasmic male sterility into crop species,save time and manpower,and make full use of heterosis efficiently.Also, some genes from wild species could be transferred into the genome of crop plants via asymmetric protoplast fusion and improve its resistance is further improved.The key to successful plant protoplast culture and fusion techniques is to select those genotypes with strong cell divison and high frequency of shoots as recipient parents.In this paper, desirable genotypes among four Brassica oleracea L. varieties were selected by shoots induced through the explant of hypocotyl and protoplast culture of hypocotyl.Asymmetric protoplast fusion between cauliflower cv. Aoxue cauliflowerl (recipient)and Brassica napus of Ogura Cytoplasmic Male Sterility(donor) was performed.The results were listed as follows:1 We have selected three genotypes, Jingyansiji(cabbage),Tianwangxuebai and Aoxue cauliflowerl (cauliflower),whose frequency of shoot inducing was high by means of hypocotyl and protoplast culture among Aoxue cauliflowerl,Tianwangxuebaijingyansiji and Green (?) (broccoli).Compared to protoplast culture,we can also select desirable genotypes as recipient parents through hypocotyls culture.2 We obtained 32 somatic hybrids between Brassica oleracea var.botrytis L. and Ogura Brassica napus L.by PEG method.3 The gene orfl38 of Ogura mitochondrial DNA was introduced into cauliflower.4 Morphological,cytological and molecular identification,we obtained 32 somatic hybrids among 68 plantlets regenerated .And this shown that it was necessary to use comprehensive methods for obtaining true results among asymmetric somatic hybrids identifying.5 Within different growth periods of donor and receptor,peroxidase isozyme(POX) and phosphoglucomutase(PGM) were selected to identify somatic hybrids.6 We combined 46 RAPD 10-mer-primers which showed polymorphism among Brassica napus L. and Brassica oleracea L..Primer H and Primer3 among these primers showed distinctions between donor and recipient parents.7 STS(Sequence Tagged Site) marker in orf138 of Ogura mitochondrial DNA was closely related to Ogura cytoplasmic male sterility,which meant that it could be used as molecular marker in the identification of the cauliflower somatic hybrids on the Ogura cytoplasmic male sterility.8 In this study, it was shown that the fused cell division did not depend on the irradiation...