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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (3): 435-441.doi: 10.3724/SP.J.1006.2010.00435

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Characterization of Six Oleic Acid Desaturase Pseudogenes of Brassica napus

XIAO Gang1,2,ZHANG Zhen-Qian1,WU Xian-Meng1,TAN Tai-Long1,2,GUAN Chun-Yun1*   

  1. 1 Oil Crops Institute/National Oil Crops Improvement Center, Hunau Agricultural University; 2 Pre-State Key Laboratory for Germplasm Innovation and Resource Utilization of Crops, Changsha 410128, China
  • Received:2009-06-30 Revised:2009-12-08 Online:2010-03-12 Published:2010-01-22
  • Contact: GUAN Chun-Yun,E-mail:guancy2000@yahoo.com.cn
  • About author:XIAO Gang,E-mail: sanjian123@yeah.net; Tel: 0731-4617941

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Abstract:

The phenomenon of multi-copy genes is common in plants. Pseudogene is defined as an inactive gene, which can not synthesize functional proteins but share the similar DNA sequences with normal functional genes. In this study, 56 FAD2 DNA clones and 47 FAD2 seed cDNA clones of Brassica napus cv. Xiangyou 15 were investigated, and 6 new copies of FAD2 were detected, designated as FAD2P1-6 respectively. This sequence length of 6 copies ranged from 1 141–1 157 bp and there were no introns in their open reading frames (ORF). These 6 copies share 96.1% identity in nucleotides from one another, and share more than 87% nucleotides identity with AY577313. Deduced amino acid sequences revealed that 1–12 stop codons occurred in the coding region of six copies which will prevent them from coding for a functional protein. These six copies were investigated in vivo in Saccharomyces cerevisiae through being cloned into yeast expression vector pYES2.0, and the 16:2 and 18:2 fatty acids were determined by gas chromatographic analysis. The results revealed that the products of the six copies were not able to synthesize 16:2 and 18:2 fatty acids, suggesting that they are pseudogenes of FAD2. These multiple pseudogenes of FAD2 within the B. napus genome might result from the duplication of large chromosomal segments simultaneously following mutation. Because of the existence of multiple pseudogenes for FAD2 in B. napus genome, we should be careful in genetic research to identify true and false, to avoid wrong conclusions.

Key words: Brassica napus, FAD2, Oleic acid desaturase, Gene expression, Saccharomyces cerevisiae

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