Identifying Cytoplasmic Male Sterile Types of Chinese Cabbage (Brassica campestris L.) by Molecular Markers

doi:10.11924/j.issn.1000-6850.casb17020079

Abstract

Abstract: Different sterile cytoplasm types of nine cabbage cytoplasmic male sterile materials were identified by molecular marker in the study in order to better use molecular marker to conduct the assisted breeding in the future. Genomic DNA was isolated from Chinese cabbage by CTAB method. The design of two pairs of specific primers was performed on conserved flanking region of orf138 gene in the GenBank. PCR were performed with genomic DNA of nine Chinese cabbage materials. The bands were sequenced. The homologous comparison was conducted in NCBI, and finally, the type of sterile cytoplasm was determined. The results showed that the bands were amplified only in four Chinese cabbage male sterile materials with two pairs of specific primers PI/PII and PIII/PIV, while the other five materials did not obtain the relative bands. The result was consistent with the field sterility identification. And then four molecular markers of Chinese cabbage Ogura cytoplasmic male sterility (CMS) were obtained. After conducting a homologous comparative analysis with BLAST in GenBank, it was found that the homologous degree was 100% in specific segments of the three sterility materials (L1-CI, L3-CI and L3- F1) and Oguorf138 gene (GenBank accession No.: HQ149728) of the reported broccoli Ogu CMS. The homologous degree of L1-F1 was 99% with a variation point. The type of cytoplasmic male sterility of the other five materials needed further research. Four materials of the nine were identified as the radish cytoplasmic male sterility materials and four molecular markers were obtained.

CLC Number:

S634.1

References

[1] 卢玉华,蔬菜栽培学各论[M],北京：中国农业出版社,2000：175.
[2] 张德双,徐家炳,曹鸣庆,等．大白菜转育新型甘蓝型油菜细胞质雄性不育系的研究[J]．华北农学报,2002,17(1)：60-63．
[3] 刘玉梅,方智远,孙培田,等．十字花科作物雄性不育性获得的主要途径及其利用[J]．中国蔬菜,2002(2)：52-55．
[4] 施展,万正杰,徐跃进,等．大白菜hau胞质雄性不育系的鉴定及不育相关基因结构分析[J]．园艺学报, 2012,39(3)：469-476．
[5] Zhang L, Yasumoto K, Yamagishi H. Identification of cytoplasmic male sterility in Chinese radish following PCR analysis of mitochondrial DNA [J]. Plant Mol Biol Rep, 2012, 30: 817-825.
[6] 励启腾,孟平红,吴跃勇．大白菜细胞质雄性不育系的选育[J]．贵州农业科学,1995(3)：1-4．
[7] 柯桂兰,赵稚雅,宋胭脂,张鲁刚,赵利民．大白菜异源胞质雄性不育系CMS3411-7的选育及应用[J]．园艺学报,1992,19 (4)：333-340．
[8] 王学芳,张彦锋,李殿荣,田建华．不结球白菜异源胞质不育系及其优良组合的选育[J]．安徽农业科学,2004,32 (1)：87-88．
[9] 刘惠吉,王华．同源四倍体白菜异源胞质雄性不育系的选育[J]．南京农业大学学报,2004,27 (23)：30-33．
[10] Ogura H, Studies on the new male-sterility in Japanese radish with special Reference to the utilization of this sterility towards the practical raising of hybrid seeds [J]. Memoirs of the Faculty of Agriculture, 1968, 6(2):39-78.
[11] 侯喜林,曹寿椿,吴志行．十字花科作物几种主要细胞质雄性不育类型及其利用(上)[J]．长江蔬菜,1999(5)：1-3．
[12] 李桂花,张衍荣,曹健,等．雄性不育在十字花科蔬菜中的研究进展[J]．江西农业大学学报,2003,25(2)：181-186．
[13] 张明龙,章徐鸯,林宝刚,等．国内外油菜细胞质雄性不育(CMS)系统的研究进展[J]．黑龙江农业科学,2004(1)：36-38．
[14] Williams P H, Heyn F W. The origin and development of cytoplasmic male sterile Chinese cabbage [C]// Chinese Cabbage, Proceeding of the first international symposium. Taiwan: Asian Vegetable Research and Development Center, 1981: 293-300.
[15] Robertson D, Earle J D P, Mutschler M A, Analysis of organelle genomes in a somatic hybrid derived from cytoplasmic male-sterile Brassica oleracea and atrazine-resistant B. campestris [J]. Theoretical and Applied Genetics, 1987, 74(3):303-309.
[16] 朱玉英,姚文岳,张素琴,等．Ogura细胞质甘蓝雄性不育系选育及其利用[J]．上海农业学报,1988,14(2)：19-24．
[17] 傅廷栋,杨光圣,杨小牛,等．甘蓝型油菜波里马细胞质雄性不育的发现、研究与利用[J]．自然科学进展：国家重点实验室通讯,1995,5(3)：287-293．
[18] 张德双,张凤兰,徐家炳．大白菜CMS96细胞质雄性不育系的特点分析[J]．华北农学报,2005,20(1)：59-62．
[19] 吉立柱,孙德岭,孙立全．十字花科细胞质雄性不育的类型及机理[J]．江苏农业科学,2007(3)：106-110．
[20] Bonhomme S, Budar F, Lancelin D, Small I, Defrance M C, Pelletier G. 1992. Sequence and transcript analysis of the Nco 2.5 Ogura-specific fragment correlation with cyto-plasmic male sterility in Brassica hybrids, Mol Gen Genet, 235: 340-348.
[21] Liu Z Y, Peter S O, Long M H, Weingartner U, Peter S, Kacser O. A PCR assay for rapid discrimination of sterile cytoplasm types in maize. Crop Science,2002, 42: 566-569.
[22] 王春国,宋文芹．花椰菜细胞质雄性不育基因特异PCR标记的筛选[J]．遗传,2005,27(2)：236-240．
[23] Krishnasamy S, Makaroff CA. 1993. Characterization of the radish mitochondrial orfB locus possible relationship with male sterility in Ogura radish. Curr Genet, 24(1-2):156-163.
[24] Yamagishi H, Terachi T. 2001, Intra-and inter-specific variations in the mitochondrial gene orf138 of Ogura-type male-sterile cyto-plasm from Raphanus sativus and R. raphanistrum. Theor Appl Genet, 103(5):725 -732.
[25] Grelon M, Budar F, Bonhomme S, et al. 1994. Ogura cytoplasmic male-sterility (CMS)-associated orf138 is translated into a mito-chondrial membrane polypeptide in male-sterile Brassica cybrids. Mol Gen Genet, 243(5):540 -547.
[26] Hiesel R, Haeseler AV, Brennicke A. 1994. Plant mitochondrial nucleic acid sequences as a tool for phylogenetic analysis. Proc Natl Acad Sci, 91(2):634 -638.