Please wait a minute...
Journal of Integrative Agriculture  2015, Vol. 14 Issue (7): 1251-1260    DOI: 10.1016/S2095-3119(14)60853-4
Crop Genetics · Breeding · Germplasm Resources Advanced Online Publication | Current Issue | Archive | Adv Search |
Molecular evidence for blocking erucic acid synthesis in rapeseed (Brassica napus L.) by a two-base-pair deletion in FAE1 (fatty acid elongase 1)
 WU Lei, JIA Yan-li, WU Gang, LU Chang-ming
Oil Crops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  DNA sequences of fatty acid elongase 1 genes FAE1.1 (EA) and FAE1.2 (EC) were isolated and characterized for 30 commercialized low erucic acid rapeseed (LEAR) cultivars in China. Four types of independent mutation leading to low erucic acid trait were found, i.e., a single-base transition (eA1), a two-base deletion (eC2) and four-base deletion (eC4) as well as single-base transition with a four-base deletion (eA*). Three genotypes, i.e., eA1eA1eC2eC2, eA1eA1eC4eC4 and eA*eA*eC4eC4 were responsible for LEA content in storage lipids of different rapeseed cultivars. Most of the LEAR cultivars had a genotype of eA1eA1eC2eC2, which were descended from the first LEAR cultivar, Oro. Yeast expression analysis revealed that two-base-pair (AA) deletion (eC2) at the base sites of 1422–1423 in the C genome FAE1 gene resulted in the absence of the condensing enzyme and led to the failure to produce erucic acid. Coexpression of FAE1 and ketoacyl-CoA reductase (KCR) or enoyl-CoA reductase (ECR) was found in high erucic acid rapeseed (HEAR) but not in LEAR (eA1eA1eC2eC2 or eA1eA1eC4eC4). Moreover, KCR and ECR were still coordinately regulated in eA1eA1eC2eC2 or eA1eA1eC4eC4 genotypes, suggesting that the expression of two genes was tightly linked. In addition, specific detection methods were developed by high-resolution melting curve analysis in order to detect eA1 and eC4 .

Abstract  DNA sequences of fatty acid elongase 1 genes FAE1.1 (EA) and FAE1.2 (EC) were isolated and characterized for 30 commercialized low erucic acid rapeseed (LEAR) cultivars in China. Four types of independent mutation leading to low erucic acid trait were found, i.e., a single-base transition (eA1), a two-base deletion (eC2) and four-base deletion (eC4) as well as single-base transition with a four-base deletion (eA*). Three genotypes, i.e., eA1eA1eC2eC2, eA1eA1eC4eC4 and eA*eA*eC4eC4 were responsible for LEA content in storage lipids of different rapeseed cultivars. Most of the LEAR cultivars had a genotype of eA1eA1eC2eC2, which were descended from the first LEAR cultivar, Oro. Yeast expression analysis revealed that two-base-pair (AA) deletion (eC2) at the base sites of 1422–1423 in the C genome FAE1 gene resulted in the absence of the condensing enzyme and led to the failure to produce erucic acid. Coexpression of FAE1 and ketoacyl-CoA reductase (KCR) or enoyl-CoA reductase (ECR) was found in high erucic acid rapeseed (HEAR) but not in LEAR (eA1eA1eC2eC2 or eA1eA1eC4eC4). Moreover, KCR and ECR were still coordinately regulated in eA1eA1eC2eC2 or eA1eA1eC4eC4 genotypes, suggesting that the expression of two genes was tightly linked. In addition, specific detection methods were developed by high-resolution melting curve analysis in order to detect eA1 and eC4 .
Keywords:  erucic acid       fatty acid elongase 1       natural mutation       Brassica napus L.  
Received: 21 April 2014   Accepted:
Fund: 

This work was financially supported by the National Natural Science Foundation of China (30471099) and the National High Technology and Development Program of China (2006AA10A113).

Corresponding Authors:  LU Chang-ming, Tel: +86-27-86711501, E-mail: cmlu@oilcrops.cn     E-mail:  cmlu@oilcrops.cn
About author:  WU Lei, E-mail: wulei686@sina.com

Cite this article: 

WU Lei, JIA Yan-li, WU Gang, LU Chang-ming. 2015. Molecular evidence for blocking erucic acid synthesis in rapeseed (Brassica napus L.) by a two-base-pair deletion in FAE1 (fatty acid elongase 1). Journal of Integrative Agriculture, 14(7): 1251-1260.

Bao X, Pollard M, Ohlrogge J. 1998. The biosynthesis of erucicacid in development embryos of Brassica rapa. PlantPhysiology, 118, 183-190

Barret P, Delourme R, Renard M, Domergue F, Lessire R,Delseny M, Roscoe T 1998. A rapeseed FAE1 gene islinked to the E1 locus associated with variation in thecontent of erucic acid. Theoretical and Applied Genetics,96, 177-186

Baud S, Lepiniec L. 2010. Physiological and developmentalregulation of seed oil production. Progress in LipidResearch, 49, 235-249

Blacklock B J, Jaworski J G. 2002. Studies into factorscontributing to substrate specificity of membranebound3-ketoacyl-CoA synthases. European Journal ofBiochemistry, 269, 4789-4798

Dellaporta S L, Wood J, Hicks J B. 1983. A plant DNAminipreparation: Version II . Plant Molecular Biology Report,1, 19-21.

Dyer J M, Mullen R T. 2005. Development and potential ofgenetically engineered oilseeds. Seed Science Research,15, 225-267

Fourmann M, Barret P, Renard M, Pelletie G, Delourme R,Brunel D. 1998. The two genes homologous to ArabidopsisFAE1 co-segregate with the two loci governing erucicacid content in Brassica napus. Theoretical and AppliedGenetics, 96, 852-858

Ghanevati M, Jaworski J G. 2002. Engineering and mechanisticstudies of the Arabidopsis FAE1 β-Ketoacyl-CoA synthaseKCS. European Journal of Biochemistry, 269, 3531-3539

Han J X, Luhs W, Sonntag K, Zahringer U, Borchatrdt D , WolterF P, Heinz E, Frentzen M. 2001. Functional characterizationof β-Ketoacyl-CoA synthase genes from Brassica napus L.Plant Molecular Biology, 46, 229-239

Harvey B L, Downey R K. 1964. The inheritance of erucic acidcontent on rapeseed (Brassica napus L.). Canadian Journalof Plant Science, 44, 104-111

Hu Y P, Wu G, Cao Y L , Wu Y H, Xiao L, Li X D, Lu C M.2009. Breeding response of transcript profiling in developingseeds of Brassica napus. BMC Molecular Biology, 10, 49.

James Jr D W, Lim E, Keller J, Plooy I, Ralston E, Dooner HK. 1995. Directed tagging of the Arabidopsis FATTY ACIDELONGATION1 (FAE1) gene with the maize transposonActivator. The Plant Cell, 7, 309-319

Katavic V, Barton D L, Giblin E M, Reed D W, Kumar A, TaylorD C. 2004. Gaining insight into the role of serine 282 inB. napus FAE1 condensing enzyme. FEBS Letter, 562,118-124

Katavic V, Mietkiewska E, Barton D L, Giblin E M, Reed D W,Taylor D C. 2002. Restoring enzyme activity in nonfunctionallow erucic acid Brassica napus fatty acid elongase 1 bya single amino acid substitution. European Journal ofBiochemistry, 269, 5625-5631

Lassner M W, Lardizabal K, Metz J G. 1996. A jojobaβ-Ketoacyl-CoA synthase cDNA complements the canolafatty acid elongatin mutation in transgenic plants. The PlantCell, 8, 281-292

Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR and the2 -ΔΔCT method. Methods, 25, 402-408

Puyaubert J, Dieryck W, Costaglioli P, Chevalier S, Breton A,Lessire R. 2005. Temporal gene expression of 3-ketoacyl-CoA reductase is different in high and in low erucic acidBrassica napus cultivars during seed development.Biochimica et Biophysica Acta, 1687, 152-163

Puyaubert J, Garbay B, Costaglioli P, Dieryck W, Roscoe TJ, Renard M, Cassagne C, Lessire R. 2001. Acyl-CoAelongase expression during seed development in Brassicanapus. Biochimica et Biophysica Acta, 1533, 141-152

Qiu D, Morgan C, Shi J, Long Y, Liu J, Li R, Zhuang X, Wang Y,Tan X, Dietrich E, Weihmann T, Everett C, Vanstraelen S,Beckett P, Fraser F, Trick M, Barnes S, Wilmer J, SchmidtR, Li J, et al. 2006. A comparative linkage map of oilseedrape and its use for QTL analysis of seed oil and erucic acidcontent. Theoretical and Applied Genetics, 114, 67-80

Rahman M, Sun Z D, McVetty P B, Li G Y. 2008. Highthroughput genome-specific and gene-specific molecularmarkers for erucic acid genes in Brassica napus L. formarker-assisted selection in plant breeding. Theoreticaland Applied Genetics, 117, 895-904

Reed G H, Wittwer C T. 2004. Sensitivity and specificity ofsingle-nucleotide polymorphism scanning by high-resolutionmelting analysis. Clinical Chemistry, 50, 1748-1754

Roscoe T J, Lessire R, Puyaubert J, Renerd M, Delseny M.2001. Mutations in the fatty acid elongation 1 gene areassociated with a loss of β-ketoacyl-CoA synthase activityin low erucic acid rapeseed. FEBS Letter, 492, 107-111

Rossak M, Smith M, Kunst L. 2001. Expression of the FAE1gene and FAE1 promoter activity in developing seeds ofArabidopsis thaliana. Plant Molecular Biology, 46, 717-725

Scarth R, Tang J H. 2006. Modification of Brassica oil usingconventional and transgenic approaches. Crop Science,46, 1225-1236

Stefansson B R, Hougen F W, Downey R K. 1961. Note on theisolation of rape plants with seed oil free from erucic acid.Canadian Journal of Plant Science, 41, 218-219

Wang N, Shi L, Tian F, Ning H C, Wu X M, Long Y, Meng JL. 2010. Assessment of FAE1 polymorphisms in threeBrassica species using EcoTILLING and their associationwith differences in seed erucic acid contents. BMC PlantBiology, 10, 137.Wu G, Wu Y W, Xiao L, Li X, Lu C M. 2008. Zero erucic acid traitof rapeseed (Brassica napus L) results from a deletion offour base pairs in the fatty acid elongase 1 gene. Theoreticaland Applied Genetics, 116, 491-499

Wu Y, Xiao L, Wu G, Lu C M. 2007. Cloning of fatty acidelongase1 gene and molecular identification of A and Cgenome in Brassica species. Science in China (Series C:Life Sciences), 3, 343–349.
[1] ZHU Mei-chen, HU Ran, ZHAO Hui-yan, TANG Yun-shan, SHI Xiang-tian, JIANG Hai-yan, ZHANG Zhi-yuan, FU Fu-you, XU Xin-fu, TANG Zhang-lin, LIU Lie-zhao, LU Kun, LI Jia-na, QU Cun-min. Identification of quantitative trait loci and candidate genes controlling seed pigments of rapeseed[J]. >Journal of Integrative Agriculture, 2021, 20(11): 2862-2879.
[2] Dong Yun, Wang Yi, Jin Feng-wei, Xing Li-juan, Fang Yan, Zhang Zheng-ying, ZOU Jun-jie, Wang Lei, Xu Miao-yun. Differentially expressed miRNAs in anthers may contribute to the fertility of a novel Brassica napus genic male sterile line CN12A[J]. >Journal of Integrative Agriculture, 2020, 19(7): 1731- 1742.
[3] MA Ni, WAN Lin, ZHAO Wei, LIU Hong-fang, LI Jun, ZHANG Chun-lei.
Exogenous strigolactones promote lateral root growth by reducing the endogenous auxin level in rapeseed
[J]. >Journal of Integrative Agriculture, 2020, 19(2): 465-482.
[4] YAN Lei, Tariq Shah, CHENG Yong, Lü Yan, ZHANG Xue-kun, ZOU Xi-ling. Physiological and molecular responses to cold stress in rapeseed (Brassica napus L.)[J]. >Journal of Integrative Agriculture, 2019, 18(12): 2742-2752.
[5] ZENG Liu, CAI Jun-song, LI Jing-jing, LU Guang-yuan, LI Chun-sheng, FU Gui-ping, ZHANG Xue-kun, MA Hai-qing, LIU Qing-yun, ZOU Xi-ling, CHENG Yong . Exogenous application of a low concentration of melatonin enhances salt tolerance in rapeseed (Brassica napus L.) seedlings[J]. >Journal of Integrative Agriculture, 2018, 17(2): 328-335.
[6] XU Ling, Faisal Islam, ZHANG Wen-fang, Muhammad A Ghani, Basharat Ali. 5-Aminolevulinic acid alleviates herbicide-induced physiological and ultrastructural changes in Brassica napus[J]. >Journal of Integrative Agriculture, 2018, 17(03): 579-592.
[7] HAN Pei-pei, QIN Lu, LI Yin-shui, LIAO Xiang-sheng, XU Zi-xian, HU Xiao-jia, XIE Li-hua, YU Chang-bing, WU Yan-feng, LIAO Xing. Identification of suitable reference genes in leaves and roots of rapeseed (Brassica napus L.) under different nutrient deficiencies[J]. >Journal of Integrative Agriculture, 2017, 16(04): 809-819.
[8] Haksong Pak, LI Yu-ling, Hyenchol Kim, JIANG Li-xi . cDNA-Amplified fragment length polymorphism analysis reveals differential gene expression induced by exogenous MeJA and GA3 in oilseed rape (Brassica apus L.) flowers[J]. >Journal of Integrative Agriculture, 2017, 16(01): 47-56.
[9] ZOU Xi-ling, ZENG Liu, LU Guang-yuan, CHENG Yong, XU Jin-song, ZHANG Xue-kun. Comparison of transcriptomes undergoing waterlogging at the seedling stage between tolerant and sensitive varieties of Brassica napus L.[J]. >Journal of Integrative Agriculture, 2015, 14(9): 1723-1734.
[10] WANG Yin, LIU Tao, LI Xiao-kun, REN Tao, CONG Ri-huan, LU Jian-wei. Nutrient deficiency limits population development, yield formation, and nutrient uptake of direct sown winter oilseed rape[J]. >Journal of Integrative Agriculture, 2015, 14(4): 670-680.
[11] Nazim Hussain, Zahra Jabeen, LI Yuan-long, CHEN Ming-xun, LI Zhi-lan, GUO Wan-li, Imran Haider Shamsi, CHEN Xiao-yang , JIANG Li-xi. Detection of Tocopherol in Oilseed Rape (Brassica napus L.) Using Gas Chromatography with Flame Ionization Detector[J]. >Journal of Integrative Agriculture, 2013, 12(5): 803-814.
[12] MA Ni, ZHANG Chun-lei, LI Jun, ZHANG Ming-hai, CHENG Yu-gui, LI Guang-ming, ZHANG Shujie. Mechanical Harvesting Effects on Seed Yield Loss, Quality Traits and Profitability of Winter Oilseed Rape (Brassica napus L.)[J]. >Journal of Integrative Agriculture, 2012, 12(8): 1297-1304.
[13] LU Guang-yuan, ZHANG Fang, ZHENG Pu-ying, CHENG Yong, LIU Feng-lan, FU Gui-ping, ZHANG , Xue-kun . Relationship Among Yield Components and Selection Criteria for Yield Improvement in Early Rapeseed (Brassica napus L.)[J]. >Journal of Integrative Agriculture, 2011, 10(7): 997-1003.
[14] ZOU Juan, LU Jian-wei, LI Yin-shui and LI Xiao-kun. Regional Evaluation of Winter Rapeseed Response to K Fertilization, K Use Efficiency, and Critical Level of Soil K in the Yangtze River Valley[J]. >Journal of Integrative Agriculture, 2011, 10(6): 911-920.
No Suggested Reading articles found!