Molecular evidence for blocking erucic acid synthesis in rapeseed (Brassica napus L.) by a two-base-pair deletion in FAE1 (fatty acid elongase 1)

doi:10.1016/S2095-3119(14)60853-4

Journal of Integrative Agriculture ›› 2015, Vol. 14 ›› Issue (7): 1251-1260.DOI: 10.1016/S2095-3119(14)60853-4

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

收稿日期:2014-04-21 出版日期:2015-07-03 发布日期:2015-07-06
通讯作者: LU Chang-ming, Tel: +86-27-86711501, E-mail: cmlu@oilcrops.cn
作者简介:WU Lei, E-mail: wulei686@sina.com
基金资助:
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).

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

Received:2014-04-21 Online:2015-07-03 Published:2015-07-06
Contact: LU Chang-ming, Tel: +86-27-86711501, E-mail: cmlu@oilcrops.cn
About author:WU Lei, E-mail: wulei686@sina.com
Supported by:
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).

摘要/Abstract

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 .

关键词: erucic acid , fatty acid elongase 1 , natural mutation , Brassica napus L.

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 .

Key words: erucic acid , fatty acid elongase 1 , natural mutation , Brassica napus L.

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

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Molecular evidence for blocking erucic acid synthesis in rapeseed (Brassica napus L.) by a two-base-pair deletion in FAE1 (fatty acid elongase 1)

Molecular evidence for blocking erucic acid synthesis in rapeseed (Brassica napus L.) by a two-base-pair deletion in FAE1 (fatty acid elongase 1)

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