Molecular Identification and Sequence Analysis of Cytoplasmic Male-Sterile Types in Chinese Cabbage

doi:10.7668/hbnxb.2007.06.011

Abstract

Abstract: To study specffic fragments linked to Pol CMS, Ogu CMS, CMS96 and to identify sterile system of CMS96 in Chinese cabbage(Brassica campestris L. ssp. pekznensis), primers of atp6, orf222 and their primer combinations were designed for PCR analysis in 11 Chinese cabbage mtDNAs. Results showed 200 by fragments could only be amplified in all Ogu CMS by atp6 primer; Fragments of 669 bp in CMS96 and 675 by in Pol CMS could only be amplffied by orf222 primer. Further study showed that ORF(open reading frame) in Pol CMS 675 by fragments was ORF224, while it was orf222 in CMS96 669 by fragments. 669 by fragment in CMS96 was named CMS96-orf22.A lignment with nucleotide acids showed CMS96-af222 in Chinese cabbagewas 99% homological to nad6c and orf222 of Nap CMS in Brassaca napus.The hvaluewas 0.0. CMS96-orf222 owned a putative conserved domain YMF19 which was similar to the mito-chonbrial membrane protein YMF19, while 675 by fragments in Pol CMS did not.Meanwhile, by multi-PCR technique with atp6 and orf222 primer combination, polymorphic results were obtained: 200 bp fragments were specific for OguCMS; 800 by fragments were different banck for maintainer, 1 500 and 2 300 by fragments were specffic for Pol CMS; 690 by fragments were specific for CMS96. A rapid molecular technique for distinguishing 3 CMS types in Chinese cabbage were founded. Therefore, 3 CMS systans in Chinese cabbage were different in the reanangement of mtDNAs.Now the chimeric reanagement of mtDNA in CMS96 is obtained and studied.

Key words: Chinesecabbage, Cytoplasmic male sterile, CNIS96, ap6, orf222, Multi-PCR

CLC Number:

S634.1

ZHANG De-shuang, ZHANG Feng-lan, WANG Yong-jian, YU Yang-jun, ZHAO Xiu-yun, XU Jia-bing, FANG Zhi-yuan. Molecular Identification and Sequence Analysis of Cytoplasmic Male-Sterile Types in Chinese Cabbage[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(6): 53-59. doi: 10.7668/hbnxb.2007.06.011.

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