Skip to main content
Log in

Functional Analysis of a Novel Male Fertility Lipid Transfer Protein Gene in Brassica campestris ssp. chinensis

  • Original Paper
  • Published:
Plant Molecular Biology Reporter Aims and scope Submit manuscript

Abstract

In our previous work, a novel gene (BcMF15) encoding a lipid transfer protein (LTP) was isolated from floral buds of Chinese cabbage pak choi (Brassica campestris ssp. chinensis, syn. B. rapa ssp. chinensis) by cDNA-amplified fragment length polymorphism transcript profiling and rapid amplification of cDNA ends PCR. This LTP gene was exclusively expressed in the fertile line, indicating that this gene was related to male sterility. Functional analysis by RNA interference showed aberrant development and precocious degeneration of the tapetum. Pollen germination tests indicated that only 36 % of pollen from the transgenic lines can normally germinate. Morphological investigations showed that the transgenic plants developed smaller floral organs with thin anthers and lower amounts of pollen. Scanning electron microscopy revealed that the transgenic plants had higher numbers of abnormal pollen grains. Histological observations also demonstrated that microsporogenesis was suppressed and resulted in the failure of microspore maturation. These results demonstrated that BcMF15 played a major role in tapetum development, thereby providing valuable information on the biological function of LTP genes during pollen development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • 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 21:90–96

    Article  CAS  Google Scholar 

  • Broekaert WF, Cammue BPA, De Bolle MFC, Thevissen K, De Samblanx GW, Osborn RW (1997) Antimicrobial peptides from plants. Crit Rev Plant Sci 16:297–323

    CAS  Google Scholar 

  • Cao JS, Yu XL, Huang AJ, Xu SY (2000) Enhancement of plant regeneration frequency of in vitro cultured Chinese cabbage. Acta Hortic Sin 27:452–454

    Google Scholar 

  • 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 24:15–23

    Article  Google Scholar 

  • Chae K, Kieslich CA, Morikis D, Kim SC, Lord EM (2009) A gain-of-function mutation of Arabidopsis lipid transfer protein 5 disturbs pollen tube tip growth and fertilization. Plant Cell 21:3902–3914

    Article  PubMed  CAS  Google Scholar 

  • Choi YO, Kim SS, Lee S, Kim S, Yoon GB, Kim H, Lee YP, Yu GH, Hyung NI, Sung SK (2010) Isolation and promoter analysis of anther-specific genes encoding putative arabinogalactan proteins in Malus x domestica. Plant Cell Rep 29(1):15–24

    Article  PubMed  CAS  Google Scholar 

  • Chuang CF, Meyerowttz EM (2000) Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci 97:4985–4990

    Article  PubMed  CAS  Google Scholar 

  • Doughty J, Wong HY, Dickinson HG (2000) Cysteine-rich pollen coat proteins (PCPs) and their interactions with stigmatic S (incompatibility) and S-related proteins in Brassica: putative roles in SI and pollination. Ann Bot 85:161–169

    Article  CAS  Google Scholar 

  • EI-Shehawi AM, Elseehy MM, Hedgcoth C (2011) Isolation and sequence analysis of wheat tissue-specific cDNAs by differential display. Plant Mol Biol Rep 29(1):135–148

    Article  Google Scholar 

  • 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 84:187–192

    Article  CAS  Google Scholar 

  • Huang L, Cao JS, Ye WZ, Liu TT, Jiang LH, Ye YQ (2008) Transcriptional difference between the male-sterile mutant bcms and wild type Brassica campestris ssp. chinensis reveal genes related to pollen development. Plant Biol 10:342–355

    Article  PubMed  CAS  Google Scholar 

  • Huang L, Zhao X, Liu TT, Dong H, Cao JS (2010) Developmental characteristics of floral organs and pollen of Chinese cabbage (Brassica campestris L. ssp. chinensis). Plant Syst and Evol 286:103–115

    Article  Google Scholar 

  • Huang L, Ye W, Liu T, Cao J (2009a) Characterization of the male-sterile line ‘Bcajh97-01A/B’ and identification of candidate genes for genic male sterility in Chinese cabbage-pak-choi. J Am Soc Hortic Sci 134:632–640

    Google Scholar 

  • Huang MD, Wei FJ, Wu CC, Hsing YI, Huang AH (2009b) Analyses of advanced rice anther transcriptomes reveal global tapetum secretory functions and potential proteins for lipid exine formation. Plant Physiol 149:694–707

    Article  PubMed  CAS  Google Scholar 

  • Kato H, Xie G, Sato Y, Imai R (2010) Isolation of anther-specific gene promoters suitable for transgene expression in rice. Plant Mol Biol Rep 28:381–387

    Article  CAS  Google Scholar 

  • Lauga B, Charbonnel-Campa L, Combes D (2000) Characterization of MZm3-3, a Zea mays tapetum-specific transcript. Plant Sci 157:65–75

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−△△CT method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Li YF, Qiu L, Huang L, Cao JS (2012a) BcJMJ30, the gene encoding jmjC Domain-containing histone demethylase is associated with pollen development and fertilization in Brassica campestrisssp. Chinensis Plant Mol Biol Rep 30:529–538

    Article  CAS  Google Scholar 

  • Li YF, Qiu L, Huang L, Cao JS (2012b) Molecular cloning and characterization of BcMF22, a novel gene related to pollen development and fertilization in Brassica campestrisssp. Chinensis Plant Mol Biol Rep 30:860–866

    Article  CAS  Google Scholar 

  • Lord EM, Russell SD (2002) The mechanisms of pollination and fertilization in plants. Annu Rev Cell Dev Biol 18:81–105

    Article  PubMed  CAS  Google Scholar 

  • 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 38:420–431

    Article  PubMed  CAS  Google Scholar 

  • Park SY, Jauh GY, Mollet JC, 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 12:51–163

    Google Scholar 

  • Peng H, Feng YM, Zhang H, Wei XK, Liang SC (2012) Molecular cloning and characterisation of genes coding for glycine- and proline-rich proteins (GPRPs) in soybean. Plant Mol Biol Rep 30:566–577

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor, Cold Spring Harbor Laboratory

    Google Scholar 

  • Smith NA, Singh SP, Wang MB, Stoutjesdijk PA, Green AG, Waterhouse PM (2000) Total silencing by intron-spliced hairpin RNAs. Nature 407:319–320

    Article  PubMed  CAS  Google Scholar 

  • Tian AM, Cao JS, Li Huang YXL, Ye WZ (2009) Characterization of a male sterile related gene BcMF15 from Brassica campestris ssp. Chinensis Mol Biol Rep 36(2):307–314

    Article  CAS  Google Scholar 

  • Toriyama K, Hanaoka K, OkadaT WM (1998) Molecular cloning of a cDNA encoding a pollen extracellular protein as a potential source of a pollen allergen in Brassica rapa. FEBS Lett 424:234–238

    Article  PubMed  CAS  Google Scholar 

  • Wu LJ, Wang XT, Wu LC, Wang PG, Chen YH (2011) Molecular cloning and expression analysis of an HINT1 homologue from maize (Zea mays L.). Plant Mol Biol Rep 29:1006–1012

    Article  CAS  Google Scholar 

  • Xue Y, Collin S, Davis DR, Thomas CM (1994) Differential screening of mitochondrial cDNA libraries from male-fertile and cytoplasmic male-sterile sugar-beet reveals genome rearrangements at atp6 and atpA loci. Plant Mol Biol 25:91–103

    Article  PubMed  CAS  Google Scholar 

  • Yang D, Chertov O, Bykovskaia SN, Chen Q, Buffo MJ, Shogan J, Anderson M, Schroder JM, Wang JM, Howard OZ, Oppenheim JJ (1999) Defensins: linking innate and adaptive immunity through dendritic and T cell CCR6. Science 286:525–530

    Article  PubMed  CAS  Google Scholar 

  • Ying ZT, Yu X, Davis MJ (1999) New method for obtaining transgenic papaya plants by Agrobacterium-mediated transformation of somatic embryos. Proc Fla state Hortic Soc 112:201–205

    Google Scholar 

  • Zhang DS, Liang WQ, Yin CS, Zong J, Gu FW, Zhang DB (2010) OsC6, encoding a lipid transfer protein, is required for postmeiotic anther development in rice. Plant Physiol 154:149–162

    Article  PubMed  CAS  Google Scholar 

  • Zhang Q, Huang L, Liu TT, Yu XL, Cao JS (2008) Functional analysis of a pollen-expressed polygalacturonase gene BcMF6 in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). Plant Cell Rep 27:1207–1215

    Article  PubMed  CAS  Google Scholar 

  • Zhao DQ, Zhou CH, Sheng YL, Liang GH, Tao J (2011) Molecular cloning and expression of phytoene synthase, lycopene beta-cyclase, and beta-carotene hydroxylase genes in Persimmon (Diospyros kaki L.) fruits. Plant Mol Biol Rep 29:345–351

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Natural Science Foundation of China (no. 30671426).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jiashu Cao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tian, A., Jiang, J. & Cao, J. Functional Analysis of a Novel Male Fertility Lipid Transfer Protein Gene in Brassica campestris ssp. chinensis . Plant Mol Biol Rep 31, 775–782 (2013). https://doi.org/10.1007/s11105-012-0552-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11105-012-0552-1

Keywords