Welcome to Acta Agronomica Sinica，

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

PDF

2262

Cited

4

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

[1] Okuley J, Lightner J, Feldmann K, Yadav N, Lark E, Browse J. Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis. Plant Cell, 1996, 6: 147-158

[2] Ohlrogge J, Jaworski J G. Regulation of fatty acid synthesis.Plant Mol Biol, 1997, 48: 109-136
[3] Nagahara U. Genome-analysis in Brassica with special reference to the experiments formation of B. napus and peculiar mode of fertilization. Jpn J Bot, 1935, 7: 389-452
[4] Scheffler J A, Sharpe A G, Schmidt H, Sperling P, Parkin I A P, Lühs W, Lydiate D J, Heinz E. Desaturase multigene families of Brassica napus arose through genome duplication.Theor Appl Genet, 1997, 94: 583-591

[5] Lynch M, Conery J S. The evolutionary fate and consequences of duplicate genes. Science, 2000, 290: 1151-1155

[6] Hirotsune S J, Yoshida N, Chen A, Garrett L, Sugiyama F, Takahashi S, Yagami K, Wynshaw-Boris A, Yoshiki A. An expressed pseudogene regulates the messenger-RNA stability of its homologous coding gene. Nature, 2003, 423: 91-96

[7] Frank A C, Amiri H, Andersson S G. Genome deterioration: Loss of repeated sequences and accumulation of junk DNA. Genetica, 2002 115(1): 1-12

[8] Wang G-L(王关林), Fang H-J(方宏筠). Plant Gene Engineering, 2nd edn (植物基因工程·第二版). Beijing: China Science Press, 2002. pp 744-745 (in Chinese)

[9] Wei D S, Li M C, Zhang X X, Ren Y, Xing L J. Identification and characterization of a novel Δ12-fatty acid desaturase gene from Rhizopus arrhizus. FEBS Lett, 2004, 573: 45-50
[10] Romanos M A, Scorer C A, Clare J J. Foreign gene expression in yeast: A review. YEAST, 1992, 8: 423-488

[11] Niu B, Ye H X, Xu Y, Wang S H, Chen P, Peng S M, Ou Y C,Tang L,Chen F. Cloning and characterization of a novel Δ12-fatty acid desaturase gene from the tree Sapium sebiferum.Biotechnol Lett, 2007, 29: 959-964

[12] Guan C-Y(官春云). Advance of high oleic acid in oil seed breeding. Crop Res (作物研究), 2006, 20(1): 1-8 (in Chinese)

[13] Jacq C, Miller J R, Brownlee G G. A pseudogene structure in 5S DNA of Xenopus laevis. Cell, 1977, 13: 109-120

[14] Zhang Z L, Harrison P M, Liu Y, Gerstein M. Millions of years of evolution preserved: A comprehensive catalog of the processed pseudogenes in the human genome.Genome Res,2003,13: 2541-2558

[15] Lynch M, Conery J S. The evolutionary fate and consequences of duplicate genes. Science, 2000, 290: 1151-1155
[16] Balakirev E S, Ayala F J. Pseudogenes: Are they “junk” or “functional” DNA.
Annu Rev Genet, 2003, 37: 123-151
[1] Wang Chu-Rui, Li Kai-Xiang, Zhao Zhi, Xiao Lu, Tang Guo-Yong, Zhao Zhi-Gang, Xu Liang, Du De-Zhi, Liu Hai-Dong. Development and application of KASP markers for functional sites of the early flowering gene BnCRY2 in Brassica napus L. spring rapeseed [J]. Acta Agronomica Sinica, 2026, 52(3): 708-721.
[2] Ma Yi-Na, Wu Xiao-Ming-Yu, Li Ou-Qi, Wang Yuan, Chen Li, Zhang Ying-Chuan, Zhao Lun, Wen Jing, Fu Ting-Dong, Shen Jin-Xiong. Functional analysis of the Bna-miR1040-EIF3A module in regulating flowering time in rapeseed (Brassica napus) [J]. Acta Agronomica Sinica, 2026, 52(2): 349-362.
[3] Zhan Ge-Rui, Yu Wen, Li Feng, Wu Ming-Zhu, Xu Xin, Luo Zhao-Peng, Wu Sheng-Xin, Yang Jun, Zhang Zhi-Qiang, Wang Zhong. Functional study of NtWRKY6 in response to ABA expression and regulation of polyphenol synthesis [J]. Acta Agronomica Sinica, 2026, 52(2): 446-458.
[4] Lou Hong-Xiang, Xing Ren-Peng, Wang Bo, Wang Jing, Xu Zheng-Hua, Zhao Jie, Kuai Jie, Zhou Guang-Sheng. Effects of sowing date on the utilization efficiency of light and temperature resources and yield in Brassica napus in the middle reaches of the Yangtze River [J]. Acta Agronomica Sinica, 2026, 52(2): 539-551.
[5] Jing Xiu-Qing, Cai Yong-Duo, Deng Ning, Zhao Xiao-Dong, Zhai Fei-Hong, Zeng Qun. Identification and expression pattern analysis of RopGEF family genes in Chenopodium quinoa [J]. Acta Agronomica Sinica, 2026, 52(1): 28-43.
[6] WANG Bin, MENG Jiang-Yu, QIU Hao-Liang, HE Ya-Jun, QIAN Wei. Identification and expression pattern analysis of the BnaDUF579 gene family in Brassica napus [J]. Acta Agronomica Sinica, 2025, 51(8): 2100-2110.
[7] XIA Qi, GUO Ying, WANG Kun-Mei, WANG Si-Yi, JU Jian-Ye, PENG Ya-Wen, LIU Zhong-Song, XIA Shi-Tou. Correlation between salicylic acid and anthocyanins accumulation in seeds of different varieties in Brassica napus [J]. Acta Agronomica Sinica, 2025, 51(5): 1189-1197.
[8] WANG Xiao-Lin, LIU Zhong-Song, KANG Lei, YANG Liu. Mapping of silique length and seeds per silique and transcriptome profiling of pod walls in Brassica napus L. [J]. Acta Agronomica Sinica, 2025, 51(4): 888-899.
[9] ZHANG Qin, DAI Cheng, MA Chao-Zhi. Auxin response reporter gene transformation of Brassica napus and dynamic signal analysis of GUS in different tissues [J]. Acta Agronomica Sinica, 2025, 51(3): 667-675.
[10] ZHANG Heng, FENG Ya-Lan, TIAN Wen-Zhong, GUO Bin-Bin, ZHANG Jun, MA Chao. Identification of TaSnRK gene family and expression analysis under localized root zone drought in wheat [J]. Acta Agronomica Sinica, 2025, 51(3): 632-649.
[11] SUN Cheng-Ming, ZHOU Xiao-Ying, CHEN Feng, ZHANG Wei, WANG Xiao-Dong, PENG Qi, GUO Yue, GAO Jian-Qin, HU Mao-Long, FU San-Xiong, ZHANG Jie-Fu. Functional analysis and prediction of long non-coding RNA (lncRNA) in the regulation of branch angle in Brassica napus L. [J]. Acta Agronomica Sinica, 2025, 51(3): 559-567.
[12] XIE Ling-Li, LI Yong-Ling, XU Ben-Bo, ZHANG Xue-Kun. Progress on waterlogging tolerance mechanism and genetic improvement in rapeseed [J]. Acta Agronomica Sinica, 2025, 51(2): 287-300.
[13] ZHUANG Yue-Ming, SONG Yi, CHEN Hang-Hang, LU Zhi-Feng, LIAO Shi-Peng, LI Xiao-Kun, CONG Ri-Huan, REN Tao, LU Jian-Wei. Effects of application of nitrogen on endophytic microbiota and seed quality in rapeseed (Brassica napus L.) seeds [J]. Acta Agronomica Sinica, 2025, 51(12): 3357-3368.
[14] HUANG Rong, ZHOU Qu-Chen, CHEN Chu-Ming, LUO Qian, YI Dong, DU Chang-Huan, HUANG Xiang-Yu, SHENG Feng, DU Xue-Zhu. Effects of BnNRT2.3-like overexpression on nitrogen uptake, utilization efficiency, and yield in Brassica napus L. [J]. Acta Agronomica Sinica, 2025, 51(12): 3184-3197.
[15] JU Jian-Ye, YANG Liu, CHEN Hao, KANG Lei, XIA Shi-Tou, LIU Zhong-Song. Single-nucleus transcriptome analysis reveals the cellular differentiation trajectories and molecular mechanisms underlying yellow seed coat formation in rapeseed [J]. Acta Agronomica Sinica, 2025, 51(11): 2860-2874.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd