Plant Cell Reports

, Volume 37, Issue 7, pp 1003–1009 | Cite as

The putative pectin methylesterase gene, BcMF23a, is required for microspore development and pollen tube growth in Brassica campestris

  • Xiaoyan Yue
  • Sue Lin
  • Youjian Yu
  • Li Huang
  • Jiashu Cao
Original Article


Key message

BcMF23a contributes to pollen wall development via influencing intine construction, which, in turn, influences pollen tube growth.


Pollen wall, the morphological out face of pollen, surrounds male gametophyte and plays an important role in plant reproduction. Pectin methylesterases (PMEs) are involved in pollen wall construction by de-esterifying pectin of the intine. In this study, the function of a putative pectin methylesterase gene, Brassica campestris Male Fertility 23a (BcMF23a), was investigated. Knockdown of BcMF23a by artificial microRNA (amiRNA) technology resulted in abnormal pollen intine formation outside of the germinal furrows at the binucleate stage. At the trinucleate stage, 20.69% of pollen possessed the degradation of nuclei, cytoplasm and the intine, resulting in shrunken pollen, whereas the remaining 75.86% were wall-disrupted with degrading cytoplasm and broken exine inside the germinal furrows. In addition, pollen abortion in transgenic plants caused germination percentage reduction by 19% in vitro and pollen tube growth disruption in natural stigma in vivo. Taken together, BcMF23a is involved in pollen development and pollen tube growth, possibly via participating in intine construction. This study may contribute towards understanding the function of pollen-specific PMEs and the molecular regulatory network of pollen wall development.


Brassica campestris Pectin methylesterases PMEs Pollen development Intine Pollen tube 



This work was funded by the National Natural Science Foundation of China (Grant number 31471877) and the Grand Science and Technology Special Project of Zhejiang Province (Grant number 2016C02051-6-1).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Bosch M, Hepler PK (2006) Silencing of the tobacco pollen pectin methylesterase NtPPME1 results in retarded in vivo pollen tube growth. Planta 223:736–745. CrossRefPubMedGoogle Scholar
  2. Bosch M, Cheung AY, Hepler PK (2005) Pectin methylesterase, a regulator of pollen tube growth. Plant Physiol 138:1334–1346. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Coimbra S, Costa M, Jones B et al (2009) Pollen grain development is compromised in Arabidopsis agp6 agp11 null mutants. J Exp Bot 60:3133–3142. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Cutler S, Mccourt P (2005) Dude, where’s my phenotype? Dealing with redundancy in signaling networks. Vis Statement Plant Physiol 138:558–559. CrossRefGoogle Scholar
  5. Dardelle F, Lehner A, Ramdani Y et al (2010) Biochemical and immunocytological characterizations of Arabidopsis pollen tube cell wall. Plant Physiol 153:1563–1576. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Di Matteo A, Giovane A (2005) Structural basis for the interaction between pectin methylesterase and a specific inhibitor protein. Plant Cell 17:849–858. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Francis KE, Lam SY, Copenhaver GP (2006) Separation of Arabidopsis pollen tetrads is regulated by QUARTET1, a pectin methylesterase gene. Plant Physiol 142:1004–1013. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Gu Z, Steinmetz LM, Gu X et al (2003) Evolution of duplicate genes versus genetic robustness against null mutations. Trends Genet 19:354–356CrossRefPubMedGoogle Scholar
  9. Hongo S, Sato K, Yokoyama R, Nishitani K (2012) Demethylesterification of the primary wall by PECTIN METHYLESTERASE35 provides mechanical support to the Arabidopsis stem. Plant Cell 24:2624–2634. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Honys D, Twell D (2004) Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol 5:R85.1–R.85.13. CrossRefGoogle Scholar
  11. Honys D, Reňák D, Twell D (2006) Male gametophyte development and function. Plant Biotechnol 1:209–224. CrossRefGoogle Scholar
  12. Huang L, Cao J, Zhang A et al (2009a) The polygalacturonase gene BcMF2 from Brassica campestris is associated with intine development. J Exp Bot 60:301–313. CrossRefPubMedGoogle Scholar
  13. Huang L, Ye Y, Zhang Y et al (2009b) BcMF9, a novel polygalacturonase gene, is required for both Brassica campestris intine and exine formation. Ann Bot 104:1339–1351. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Jiang L, Yang S-L, Xie L-F et al (2005) VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract. Plant Cell Online 17:584–596. CrossRefGoogle Scholar
  15. Jiang J, Zhang Z, Cao J (2013) Pollen wall development: the associated enzymes and metabolic pathways. Plant Biol 15:249–263. CrossRefPubMedGoogle Scholar
  16. Jimenez-Lopez JC, Kotchoni SO, Rodríguez-García MI, Alché JD (2012) Structure and functional features of olive pollen pectin methylesterase using homology modeling and molecular docking methods. J Mol Model 18:4965–4984. CrossRefPubMedGoogle Scholar
  17. Leroux C, Bouton S, Kiefer-Meyer M-C et al (2015) PECTIN METHYLESTERASE48 is involved in Arabidopsis pollen grain germination. Plant Physiol 167:367–380. CrossRefPubMedGoogle Scholar
  18. Lin S, Dong H, Zhang F et al (2014) BcMF8, a putative arabinogalactan protein-encoding gene, contributes to pollen wall development, aperture formation and pollen tube growth in Brassica campestris. Ann Bot 113:777–788. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Lin S, Huang L, Yu X et al (2017) Characterization of BcMF23a and BcMF23b, two putative pectin methylesterase genes related to pollen development in Brassica campestris ssp. chinensis. Mol Biol Rep 44:139–148. CrossRefPubMedGoogle Scholar
  20. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Markovič O, Janeček Š (2004) Pectin methylesterases: sequence-structural features and phylogenetic relationships. Carbohydr Res 339:2281–2295. CrossRefPubMedGoogle Scholar
  22. Mcdonald BA, Martinez JP (1990) Restriction fragment length polymorphisms in Septoria tritici occur at a high frequency. Curr Biol 17:133–138Google Scholar
  23. Mollet J-C, Leroux C, Dardelle F, Lehner A (2013) Cell wall composition, biosynthesis and remodeling during pollen tube growthGoogle Scholar
  24. Pelloux J, Rusterucci C, Mellerowicz E (2007) New insights into pectin methylesterase structure and function. Trends Plant Sci 12:267–277. CrossRefPubMedGoogle Scholar
  25. Rockel N, Wolf S, Kost B et al (2008) Elaborate spatial patterning of cell-wall PME and PMEI at the pollen tube tip involves PMEI endocytosis, and reflects the distribution of esterified and de-esterified pectins. Plant J 53:133–143. CrossRefPubMedGoogle Scholar
  26. Sablok G, Pérez-quintero ÁL, Hassan M et al (2011) Artificial microRNAs (amiRNAs) engineering—on how microRNA-based silencing methods have affected current plant silencing research. Biochem Biophys Res Commun 406:315–319. CrossRefPubMedGoogle Scholar
  27. Schwab R, Voinnet O (2010) RNA silencing amplification in plants: size matters. Proc Natl Acad Sci 107:14945–14946. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Shi J, Cui M, Yang L et al (2015) Genetic and biochemical mechanisms of pollen wall development. Trends Plant Sci 20:1–13. CrossRefGoogle Scholar
  29. The Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408:796–815. CrossRefGoogle Scholar
  30. Tian GW, Chen MH, Zaltsman A, Citovsky V (2006) Pollen-specific pectin methylesterase involved in pollen tube growth. Dev Biol 294:83–91. CrossRefPubMedGoogle Scholar
  31. Wang H, Zhuang X, Cai Y et al (2013) Apical F-actin-regulated exocytic targeting of NtPPME1 is essential for construction and rigidity of the pollen tube cell wall. Plant J 76:367–379. CrossRefPubMedGoogle Scholar
  32. Wolf S, Greiner S (2012) Growth control by cell wall pectins. Protoplasma 249:169–175. CrossRefGoogle Scholar
  33. Yu X, Cao J, Ye W, Wang Y (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 79:833–839. CrossRefGoogle Scholar
  34. Zhang D, Shi J, Yang X (2016) Role of lipid metabolism in plant pollen exine development. In: Nakamura Y, Li-Beisson Y (eds) Lipids in plant and algae development. Springer, Cham, pp 315–337CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Xiaoyan Yue
    • 1
  • Sue Lin
    • 2
  • Youjian Yu
    • 3
  • Li Huang
    • 1
  • Jiashu Cao
    • 1
  1. 1.Laboratory of Cell and Molecular Biology, Institute of Vegetable ScienceZhejiang UniversityHangzhouChina
  2. 2.Institute of Life SciencesWenzhou UniversityWenzhouChina
  3. 3.College of Agriculture and Food ScienceZhejiang A & F UniversityLin’anChina

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