Molecular Biology Reports

, Volume 36, Issue 2, pp 307–314 | Cite as

Characterization of a male sterile related gene BcMF15 from Brassica campestris ssp. chinensis

  • Aimei Tian
  • Jiashu Cao
  • Li Huang
  • Xiaolin Yu
  • Wanzhi Ye


Data from cDNA-AFLP analysis based on the genome-wide transcriptional profiling on the flower buds of the male meiotic cytokinesis (mmc) mutant and its wild-type of Brassica campestris L. ssp. chinensis Makino, syn. B. rapa L. ssp. chinensis, indicated that mutation of the MMC gene resulted in changes in expression of a variety of genes. A transcript-derived fragment specifically accumulated in the wild-type flower buds was isolated, and the corresponding full-length cDNA and DNA was subsequently amplified. Bioinformatical analyses of this gene named BcMF15 (GenBank accession number EF600901) showed that it encoded a protein with 103 amino acids. The BcMF15 had a 88% nucleotide similarity to a lipid transfer protein-like gene. Moreover, sequence prediction indicated that BcMF15 might encode a membrane protein with a signal peptide at the N-terminus. Meanwhile, six domains were predicted in the deduced BcMF15 protein, such as the AAI domain existing in some crucial proteins of pollen development-preferential, signal peptide, transmembrane domain, vWF domain, ZnF_C4 domain, and Tryp_alpha_amyl domain. Spatial and temporal expression patterns analysis by RT-PCR indicated that BcMF15 was exclusively expressed in the fertile line, which indicated this gene is male sterile related. Phylogenetic analysis in Cruciferae revealed that the BcMF15 was relative conservative in evolution. We suppose BcMF15 may be a critical molecule in the transmembrane transportation and signal transduction during microspore development.


BcMF15 Brassica campestris ssp. chinensis Male sterile Microspore development 



cDNA-amplified fragment length polymorphism


Lipid transfer protein


Male meiotic cytokinesis


Polymerase chain reaction


Rapid amplification of cDNA ends


Reverse transcriptase polymerase chain reaction


Transcript-derived fragment


  1. 1.
    Lord EM, Russell SD (2002) The mechanisms of pollination and fertilization in plants. Annu Rev Cell Dev Biol 18:81–105PubMedCrossRefGoogle Scholar
  2. 2.
    Edlund AF, Swanson R, Preuss D (2004) Pollen and stigma structure and function: the role of diversity in pollination. Plant Cell 16:S84–S97PubMedCrossRefGoogle Scholar
  3. 3.
    Twell D (2002) The development biology of pollen. In: O’Neill SD, Roberts JA (eds) Plant reproduction. Annual plant reviews. Sheffield Academic Press, Sheffield, pp 86–153Google Scholar
  4. 4.
    McCormick S (2004) Control of male gametophyte development. Plant Cell 16(Suppl):S142–S153PubMedCrossRefGoogle Scholar
  5. 5.
    Park BS, Park YD, Kim HU, Jin YM, Kim HI (2002) BAN103, a pollen-preferential gene, from Chinese cabbage and its promoter activity. Mol Cell 14:150–157Google Scholar
  6. 6.
    Ye WZ, Cao JS, Xiang X, Zeng GW (2003) Molecular cloning and characterization of the genic male sterility related gene CYP86MF in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino var. communis Tsenet Lee). J Hortic Sci Biotechnol 78:319–323Google Scholar
  7. 7.
    Yu XL, Cao JS, Ye WZ, Wang YQ (2004) Construction of an antisense CYP86MF gene plasmid vector and production of a male-sterile Chinese cabbage transformant by the pollen-tube method. J Hortic Sci Biotechnol 79:833–839Google Scholar
  8. 8.
    Wang YQ, Yu XL, Cao JS (2004) Isolation and characterization of BcMF3, a gene expressed only in maintainer line in Chinese cabbage pak-choi (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee). Acta Genet Sin 31(11):1302–1308PubMedGoogle Scholar
  9. 9.
    Wang YQ, Ye WZ, Cao JS, Yu XL, Xiang X, Lu G (2005) Cloning and characterization of the microspore development related gene BcMF2 in Chinese cabbage-pak-choi (Brassica campestris L. ssp. chinensis Makino). J Integr Plant Biol 47:863–872CrossRefGoogle Scholar
  10. 10.
    Cao JS, Yu XL, Ye WZ, Lu G, Xiang X (2006) Functional analysis of a novel male fertility CYP86MF gene in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). Plant Cell Rep 24:15–23CrossRefGoogle Scholar
  11. 11.
    Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  12. 12.
    Patel O, Dumesny C, Giraud AS, Baldwin GS, Shulkes A (2004) Stimulation of proliferation and migration of a colorectal cancer cell line by amidated and glycine-extended gastrin-releasing peptide via the same receptor. Biochem Pharmacol 68:2129–2142PubMedCrossRefGoogle Scholar
  13. 13.
    Russo GL, Tosto M, Mupo A, Castellano I, Cuomo A, Tosti E (2004) Biochemical and functional characterization of protein kinase CK2 in Ascidian ciona intestinalis oocytes at fertilization. J Biol Chem 279(31):33012–33023PubMedCrossRefGoogle Scholar
  14. 14.
    Motose H, Sugiyam M, Fukuda H (2001) An arabinogalactan protein(s) is a key component of a fraction that mediates local intercellular communication involved intracheary element differentiation of Zinnia Mesophyll cells. Plant Cell Physiol 42(2):129–137PubMedCrossRefGoogle Scholar
  15. 15.
    Clarke A, Gleeson P, Harrison S, Knox RB (1979) Pollen-stigma interactions: identification and characterization of surface components with recognition potential. Proc Natl Acad Sci USA 76(7):3358–3362PubMedCrossRefGoogle Scholar
  16. 16.
    Zakharov A, Muntz K (2004) Seed legumains are expressed in stamens and vegetative legumains in seeds of Nicotiana tabacum L. J Exp Bot 55(402):1593–1595PubMedCrossRefGoogle Scholar
  17. 17.
    Park SY, Jauh G-Y, Mollet J-C, Eckard KJ, Nothnagel EA, Lord EM , Walling LL (2000) A lipid transfer–like protein is necessary for lily pollen tube adhesion to an in vitro stylar matrix. Plant Cell 12:151–163PubMedCrossRefGoogle Scholar
  18. 18.
    Lee Y-H, Chung K-H, Kim H-U, Jin Y-M, Kim H-I, Park B-S (2003) Induction of male sterile cabbage using a tapetum-specific promoter from Brassica campestris L. ssp. pekinensis. Plant Cell Rep 22(44):268–273PubMedCrossRefGoogle Scholar
  19. 19.
    Guerbette F, Grosbois M, Jolliot-Croquin A, Kader J-C, Zachowski A (1999) Lipid-transfer proteins from plants: structure and binding properties. Mol Cell Biochem 192(1–2):157–161PubMedCrossRefGoogle Scholar
  20. 20.
    Arondeo V, Vergnolle C, Cantrel C, Kader J-C (2000) Lipid transfer proteins are encoded by a small mutigene family in Arabidopsis thaliana. Plant Sci 150:1–12CrossRefGoogle Scholar
  21. 21.
    Lu ZX, Gaudet DA, Frick M, Puchalski B, Genswein B, Laroche A (2005) Identification and characterization of genes differentially expressed in the resistance reaction in wheat infected with Tilletia tritici, the common bunt pathogen. J Biochem Mol Biol 38:420–431PubMedGoogle Scholar
  22. 22.
    Broekaert WF, Cammue B, De Bolle M, Thevissen K, De Samblanx GW, Osborn RW (1997) Antimicrobial peptides from plants. Crit Rev Plant Sci 16:297–323CrossRefGoogle Scholar
  23. 23.
    Yang D, Chertov O, Bykovskaia SN, Chen Q, Buffo MJ, Shogan J, Anderson M, Schroder JM, Wang JM, Howard OZ, Oppenheim JJ (1999) Defensins: linkinginnate and adaptive immunity through dendritic and T cell CCR6. Science 286:525–530PubMedCrossRefGoogle Scholar
  24. 24.
    Foster GD, Robinson SW, Blundell RP, Roberts MR, Hodge R, Draper J, Scott RJ (1992) A Brassica napus mRNA encoding a protein homologous to phospholipid transfer proteins, is specifically in the tapetum and developing microspores. Plant Sci 84:187–192CrossRefGoogle Scholar
  25. 25.
    Lauga B, Charbonnel-Campa L, Combes D (2000) Characterization of MZm3-3, a Zea mays tapetum-specific transcript. Plant Sci 157:65–75PubMedCrossRefGoogle Scholar
  26. 26.
    Ariizumi T, Amagai M, Shibata D, Hatakeyama K, Watanabe M, Toriyama K (2002) Comparative study of promoter activity of three anther-specific genes encoding lipid transfer protein, xyloglucan endotransglucosylase/hydrolase and polygalacturonase in transgenic Arabidopsis thaliana. Plant Cell Rep 21:90–96CrossRefGoogle Scholar
  27. 27.
    Toriyama K, Hanaoka K, Okada T, Watanabe M (1998) Molecular cloning of a cDNA encoding a pollen extracellular protein as a potential source of a pollen allergen in Brassica rapa. FEBS Lett 424:234–238PubMedCrossRefGoogle Scholar
  28. 28.
    Annna MK, Hans JB (1999) Cell-specific expression of genes of the lipid transfer protein family from Arabidopsis thaliana. Plant Physiol 40:69–76Google Scholar
  29. 29.
    Wang LP, Cao JS, Ye WZ, Xiang X, Zhou SM (2005) Cloning and evolutionary analysis of homologous sequences of CYP86MF gene in Cruciferae. Hereditas 27(5):395–402PubMedGoogle Scholar
  30. 30.
    Inaba R, Nishio T (2002) Phylogenetic analysis of Brassiceae based on the nucleotide sequences of the S-locus related gene, SLR1. Theor Appl Genet 105:1159–1165PubMedCrossRefGoogle Scholar
  31. 31.
    Kader J-C (1997) Lipid-transfer proteins: a puzzling family of plant proteins. Trends Plant Sci 2:66–70CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Aimei Tian
    • 1
    • 2
  • Jiashu Cao
    • 1
  • Li Huang
    • 1
  • Xiaolin Yu
    • 1
  • Wanzhi Ye
    • 1
  1. 1.Lab of Cell & Molecular Biology, Institute of Vegetable ScienceZhejiang UniversityHangzhouP.R. China
  2. 2.College of Chemistry and Life ScienceThree Gorges UniversityYichangP.R. China

Personalised recommendations