Advertisement

Plant Molecular Biology Reporter

, Volume 29, Issue 4, pp 943–951 | Cite as

Cloning and Expression of an Anther-Abundant Polygalacturonase Gene BcMF17 from Brassica Campestris ssp. Chinensis

  • Aihong Zhang
  • Qizhen Chen
  • Li Huang
  • Lin Qiu
  • Jiashu Cao
Article

Abstract

A polygalacturonase gene, designated BcMF17 (Brassica campestris Male Fertility 17), has been cloned from Brassica campastris. This gene, encoding polygalacturonase (PG), consists of a 1,194 bp open reading frame along with four exons and three introns, and exhibits 79% similarities to BcMF9 (Huang et al. 2009). Detailed multiple amino acid sequence alignment and phylogenetic tree analysis indicate that BcMF17 belongs to a pollen clade and may be involved in pollen wall formation. Multiple alignments against homologous sequences from the genus Brassica indicate that BcMF17 is highly conserved in this genus. Quantitative real-time PCR results indicate that transcripts of BcMF17 are preferentially expressed in reproductive tissues, especially abundant in male floral tissues, but are expressed at low level in vegetative tissues. BcMF17 transcripts in male floral tissues persist from the anther primordia stage till to the mature pollen stage. In situ hybridized signals of BcMF17 are highly detected during anther development. During anther development, expression patterns of BcMF17 are very similar to those of BcMF9; however, unlike BcMF9, BcMF17 is continuously expressed during this period, and its transcripts are detected in the whole plant during its life cycle.

Keywords

BcMF17 Brassica campestris Anther development Polygalacturonase Pollen 

Abbreviations

BcMF17

Brassica campestris Male Fertility 17

BcMF9

Brassica campestris Male Fertility 9

qRT-PCR

quantitative Real-time polymerase chain reaction

PG

Polygalacturonase

cDNA-AFLP

cDNA–amplified fragment length polymorphism

Notes

Acknowledgments

This work was supported by the Chinese Natural Science Foundation (No. 31071805).

Supplementary material

11105_2011_298_MOESM1_ESM.doc (60 kb)
ESM 1 (DOC 60 kb)

References

  1. Alam MM, Sharmin S, Nabi Z, Mondal SI, Islam MS, Bin Nayeem S, Shoyaib M, Khan H (2010) A putative leucine-rich repeat receptor-like kinase of jute involved in stress response. Plant Mol Biol Rep 28:394–402CrossRefGoogle Scholar
  2. Bendtsen JD, Nielsen H, von Heijne G, Brunak S (2004) Improved prediction of signal peptides: signalp 3.0. J Mol Biol 340:783–795PubMedCrossRefGoogle Scholar
  3. Brummell DA, Harpster MH (2001) Cell wall metabolism in fruit softening and quality and its manipulation in transgenic plants. Plant Mol Biol 47:311–340PubMedCrossRefGoogle Scholar
  4. Chen W, Baldwin TC (2007) An improved method for the fixation, embedding and immunofluorescence labeling of resin-embedded plant tissue. Plant Mol Biol Rep 25:27–35CrossRefGoogle Scholar
  5. Dubald M, Barakate A, Mandaron P, Mache R (1993) The ubiquitous presence of exopolygalacturonase in maize suggests a fundamental cellular function for this enzyme. Plant J 4:781–791PubMedCrossRefGoogle Scholar
  6. Emanuelsson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016PubMedCrossRefGoogle Scholar
  7. Hadfield KA, Bennett AB (1998) Polygalacturonases: many genes in search of a function. Plant Physiology 117:337–343PubMedCrossRefGoogle Scholar
  8. Hadfield KA, Rose JKC, Yaver DS, Berka RM, Bennett AB (1998) Polygalacturonase gene expression in ripe melon fruit supports a role for polygalacturonase in ripening-associated pectin disassembly. Plant Physiology 117:363–373PubMedCrossRefGoogle Scholar
  9. Hiller K, Grote A, Scheer M, Munch R, Jahn D (2004) PrediSi: prediction of signal peptides and their cleavage positions. Nucleic Acids Res 32:W375–W379PubMedCrossRefGoogle Scholar
  10. Holmes-Davis R, Tanaka CK, Vensel WH, Hurkman WJ, McCormick S (2005) Proteome mapping of mature pollen of Arabidopsis thaliana. Proteomics 5:4864–4884PubMedCrossRefGoogle Scholar
  11. Honys D, Twell D (2003) Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol 132:640–652PubMedCrossRefGoogle Scholar
  12. Huang L, Cao JS, Ye WZ, Liu TT, Jiang LX, Ye YQ (2008a) Transcriptional differences between the male-sterile mutant bcms and wild-type Brassica campestris ssp. chinensis reveal genes related to pollen development. Plant Biol 10:342–355PubMedCrossRefGoogle Scholar
  13. Huang L, Cao JS, Zhang AH, Ye YQ (2008b) Characterization of a putative pollen-specific arabinogalactan protein gene, BcMF8, from Brassica campestris ssp. Chinensis. Mol Biol Rep 35:631–639PubMedCrossRefGoogle Scholar
  14. Huang L, Ye YQ, Zhang YC, Zhang AH, Liu TT, Cao JS (2009) BcMF9, a novel polygalacturonase gene, is required for both Brassica campestris intine and exine formation. Ann Bot 104:1339–1351PubMedCrossRefGoogle Scholar
  15. Huang L, Zhao XF, Liu TT, Dong H, Cao JS (2010) Developmental characteristics of floral organs and pollen of Chinese cabbage (Brassica campestris L. ssp chinensis). Plant Syst Evol 286:103–115CrossRefGoogle Scholar
  16. Hulzink RJM, Weerdesteyn H, Croes AF, Gerats T, van Herpen MMA, van Helden J (2003) In silico identification of putative regulatory sequence elements in the 5 '-untranslated region of genes that are expressed during male gametogenesis. Plant Physiol 132:75–83PubMedCrossRefGoogle Scholar
  17. Kasahara RD, Portereiko MF, Sandaklie-Nikolova L, Rabiger DS, Drews GN (2005) MYB98 is required for pollen tube guidance and synergid cell differentiation in Arabidopsis. Plant Cell 17:2981–2992PubMedCrossRefGoogle Scholar
  18. Kato H, Xie GS, Sato Y, Imai R (2010) Isolation of Anther-specific gene promoters suitable for transgene expression in rice. Plant Mol Biol Rep 28:381–387CrossRefGoogle Scholar
  19. Kim J, Shiu SH, Thoma S, Li WH, Patterson SE (2006) Patterns of expansion and expression divergence in the plant polygalacturonase gene family. Genome Biol 7:R87PubMedCrossRefGoogle Scholar
  20. Kohli DK, Bachhawat AK (2003) CLOURE: Clustal Output Reformatter, a program for reformatting ClustalX/ClustalW outputs for SNP analysis and molecular systematics. Nucleic Acids Res 31:3501–3502PubMedCrossRefGoogle Scholar
  21. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5:150–163PubMedCrossRefGoogle Scholar
  22. Lau JM, Cooper NG, Robinson DL, Korban SS (2009) Sequence and in silico characterization of the tomato polygalacturonase (PG) promoter and terminator regions. Plant Mol Biol Rep 27:250–256CrossRefGoogle Scholar
  23. Lee BH, Henderson DA, Zhu JK (2005) The Arabidopsis cold-responsive transcriptome and its regulation by ICE1. Plant Cell 17:3155–3175PubMedCrossRefGoogle Scholar
  24. Liu AH, Wang JB (2006) Genomic evolution of Brassica allopolyploids revealed by ISSR marker. Genet Resour Crop Ev 53:603–611CrossRefGoogle Scholar
  25. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  26. Louw C, Young PR, van Rensburg P, Divol B (2010) Regulation of endo-polygalacturonase activity in Saccharomyces cerevisiae. FEMS Yeast Res 10:44–57PubMedCrossRefGoogle Scholar
  27. Malik MR, Wang F, Dirpaul JM, Zhou N, Hammerlindl J, Keller W, Abrams SR, Ferrie AMR, Krochko JE (2008) Isolation of an embryogenic line from non-embryogenic Brassica napus cv. Westar through microspore embryogenesis. J Exp Bot 59:2857–2873PubMedCrossRefGoogle Scholar
  28. Mascarenhas JP (1990) Gene activity during pollen development. Annu Rev Plant Phys 41:317–338CrossRefGoogle Scholar
  29. Meng CM, Zhang TZ, Guo WZ (2009) Molecular cloning and characterization of a novel gossypium hirsutum L. bHLH gene in response to ABA and drought stresses. Plant Mol Biol Rep 27:381–387CrossRefGoogle Scholar
  30. Nielsen H, Engelbrecht J, Brunak S, von Heijne G (1997) A neural network method for identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Int J Neural Syst 8:581–599PubMedCrossRefGoogle Scholar
  31. Park KC, Kwon SJ, Kim PH, Bureau T, Kim NS (2008) Gene structure dynamics and divergence of the polygalacturonase gene family of plants and fungus. Genome 51:30–40PubMedCrossRefGoogle Scholar
  32. Parkin IAP, Gulden SM, Sharpe AG, Lukens L, Trick M, Osborn TC, Lydiate DJ (2005) Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana. Genetics 171:765–781PubMedCrossRefGoogle Scholar
  33. Punwani JA, Rabiger DS, Drews GN (2007) MYB98 positively regulates a battery of synergid-expressed genes encoding filiform apparatus localized proteins. Plant Cell 19:2557–2568PubMedCrossRefGoogle Scholar
  34. Qi JN, Yu SC, Zhang FL, Shen XQ, Zhao XY, Yu YJ, Zhang DS (2010) Reference gene selection for real-time quantitative polymerase chain reaction of mRNA transcript levels in chinese cabbage (Brassica rapa L. ssp pekinensis). Plant Mol Biol Rep 28:597–604CrossRefGoogle Scholar
  35. Qin Y, Leydon AR, Manziello A, Pandey R, Mount D, Denic S, Vasic B, Johnson MA, Palanivelu R (2009) Penetration of the stigma and style elicits a novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil. PLoS Genet 5:e1000621PubMedCrossRefGoogle Scholar
  36. Qiu X, Erickson L (1996) A pollen-specific polygalacturonase-like cDNA from alfalfa. Sex Plant Reprod 9:123–124CrossRefGoogle Scholar
  37. Rhee SY, Osborne E, Poindexter PD, Somerville CR (2003) Microspore separation in the quartet 3 mutants of Arabidopsis is impaired by a defect in a developmentally regulated polygalacturonase required for pollen mother cell wall degradation. Plant Physiol 133:1170–1180PubMedCrossRefGoogle Scholar
  38. Robert LS, Allard S, Gerster JL, Cass L, Simmonds J (1993) Isolation and characterization of a polygalacturonase gene highly expressed in Brassica napus pollen. Plant Mol Biol 23:1273–1278PubMedCrossRefGoogle Scholar
  39. Sitrit Y, Hadfield KA, Bennett AB, Bradford KJ, Downie AB (1999) Expression of a polygalacturonase associated with tomato seed germination. Plant Physiol 121:419–428PubMedCrossRefGoogle Scholar
  40. Taylor JE, Webb STJ, Coupe SA, Tucker GA, Roberts JA (1993) Changes in polygalacturonase activity and solubility of poluronides during ethylene-stimulated leaf abscission in Sambucus-Nigra. J Exp Bot 44:93–98CrossRefGoogle Scholar
  41. Verlent I, Van Loey A, Smout C, Duvetter T, Hendrickx ME (2004) Purified tomato polygalacturonase activity during thermal and high-pressure treatment. Biotechnol Bioeng 86:63–71PubMedCrossRefGoogle Scholar
  42. Villarreal NM, Bustamante CA, Civello PM, Martinez GA (2010) Effect of ethylene and 1-MCP treatments on strawberry fruit ripening. J Sci Food Agr 90:683–689Google Scholar
  43. 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 47:9Google Scholar
  44. Wang J, Tian N, Huang X, Chen LY, Schlappi M, Xu ZQ (2009) The tall fescue turf grass class I Chitinase gene FaChit1 is activated by fungal elicitors, dehydration, ethylene, and mechanical wounding. Plant Mol Biol Rep 27:305–314CrossRefGoogle Scholar
  45. Yoshida O, Nakagawa H, Ogura N, Sato T (1984) Effect of heat-treatment on the development of polygalacturonase activity in tomato fruit during ripening. Plant Cell Physiol 25:505–509Google Scholar
  46. Zhang LL, Chen KS, Xu CJ (2008a) Identification and characterization of transcripts differentially expressed in peel and juice vesicles of immature and ripe orange (Citrus sinensis) fruit. Plant Mol Biol Rep 26:121–132CrossRefGoogle Scholar
  47. Zhang Q, Huang L, Liu TT, Yu XL, Cao JS (2008b) Functional analysis of a pollen-expressed polygalacturonase gene BcMF6 in Chinese cabbage (Brassica campestris L. ssp chinensis Makino). Plant Cell Rep 27:1207–1215PubMedCrossRefGoogle Scholar
  48. Zhu Y, Dun XL, Zhou ZF, Xia SQ, Yi B, Wen J, Shen JX, Ma CZ, Tu JX, Fu TD (2010) A separation defect of tapetum cells and microspore mother cells results in male sterility in Brassica napus: the role of abscisic acid in early anther development. Plant Mol Biol 72:111–123PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Aihong Zhang
    • 1
  • Qizhen Chen
    • 1
  • Li Huang
    • 1
  • Lin Qiu
    • 1
  • Jiashu Cao
    • 1
    • 2
  1. 1.Laboratory of Cell & Molecular Biology, Institute of Vegetable ScienceZhejiang UniversityHangzhouChina
  2. 2.Key Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Ministry of AgricultureHangzhouPeople’s Republic of China

Personalised recommendations