基于奇亚(Salvia hispanica) FAD2和FAD3的油菜(Brassica napus) α-亚麻酸的生物技术

doi:10.1016/j.jia.2023.05.018

Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (12): 3810-3815.DOI: 10.1016/j.jia.2023.05.018

基于奇亚(Salvia hispanica) FAD2和FAD3的油菜(Brassica napus) α-亚麻酸的生物技术

收稿日期:2022-12-29 接受日期:2023-03-30 出版日期:2023-12-20 发布日期:2023-05-16

Biotechnology of α-linolenic acid in oilseed rape (Brassica napus) using FAD2 and FAD3 from chia (Salvia hispanica)

XUE Yu-fei^{1, 2}, INKABANGA TSEKE Alain^{1, 2, 3}, YIN Neng-wen^{1, 2}, JIANG Jia-yi^{1, 2}, ZHAO Yan-ping^{1, 2}, LU Kun^{1, 2}, LI Jia-na^{1, 2}, DING Yan-song^{1, 2}, ZHANG Shi-qing^{1, 2}, CHAI You-rong^{1, 2#}

¹ Integrative Science Center of Germplasm Creation in Western China (Chongqing) Science City and Southwest University/
Chongqing Key Laboratory of Crop Quality Improvement/College of Agronomy and Biotechnology, Southwest University,
Chongqing 400715, P.R.China
² Engineering Research Center of South Upland Agriculture, Ministry of Education/Academy of Agricultural Sciences, Southwest
University, Chongqing 400715, P.R.China
³ Faculty of Agricultural Sciences, National Pedagogic University (UPN), Kinshasa 8815, D.R.Congo

Received:2022-12-29 Accepted:2023-03-30 Online:2023-12-20 Published:2023-05-16
About author:#Correspondence CHAI You-Rong, E-mail: chaiyourong2@163.com, chaiyour@swu.edu.cn
Supported by:
This work was supported by the National Natural Science Foundation of China (31871549, 32001441 and 32272015), the Chongqing Research Program of Basic Research and Frontier Technology, China (cstc2015jcyjBX0143), the Fundamental Research Funds for the Central Universities, China (XDJK2020C038), the National Key R&D Program of China (2016YFD0100506), and the Young Eagles Program of Chongqing Municipal Commission of Education, China (CY220219).

摘要/Abstract

摘要： α-亚麻酸(ALA, 18:3^Δ9,12,15)是人类必需的脂肪酸，因为它是合成ω-3长链多不饱和脂肪酸(LC-PUFA)的前体物质。当今世界，人们普遍缺乏ALA，因为大多数大宗食用油中ALA含量较低或缺乏，基于生物技术提升大宗油料作物的ALA含量是一种很有前途的策略。在已知的油料作物中，奇亚(Salvia hispanica)的种子油中ALA含量最高。在本研究中，基于连接肽LP4-2A，我们构建了奇亚FAD2和FAD3的融合基因，然后构建了其种子特异性启动子P_NapA驱动的植物表达载体，通过根癌农杆菌介导成功转化到大宗油料作物甘蓝型油菜(Brassica napus)中。在T0、T1和T2株系的种子中，ALA的平均含量分别为20.86%、23.54%和24.92%，分别为未转化材料（对照）的2.21、2.68和3.03倍（含量分别为9.42%、8.78%和8.22%）。T0、T1和T2植株的种子中，最高ALA含量分别为38.41%、35.98%和39.19%，是对照的4.10—4.77倍。转基因株系中，脂肪酸（FA）途径结构基因BnACCD、BnFATA、BnSAD、BnSCD、BnDGAT1、BnDGAT2和BnDGAT3以及正调控转录因子的编码基因BnWRI1、BnLEC1、BnL1L、BnLEC2、BnABI3、BnbZIP67和BnMYB96均显著上调，而油脂积累负调控因子、次生代谢正调控因子的编码基因BnTT1、BnTT2、BnTT8、BnTT16、BnTTG1和BnTTG2均显著下调，这表明外源融合基因ShFAD2-ShFAD3直接和间接地重塑了转基因油菜种子中FA相关的整个代谢网络的正、负效应位点。

Abstract: α-Linolenic acid (ALA, 18:3Δ9,12,15) is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids (LC-PUFA). Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content. Biotechnological enrichment of ALA in staple oil crops is a promising strategy. Chia (Salvia hispanica) has the highest ALA content in its seed oil among known oil crops. In this study, the FAD2 and FAD3 genes from chia were engineered into a staple oil crop, oilseed rape (Brassica napus), via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter PNapA. In seeds of T0, T1, and T2 lines, the average ALA contents were 20.86, 23.54, and 24.92%, respectively, which were 2.21, 2.68, and 3.03 folds of the non-transformed controls (9.42, 8.78, and 8.22%), respectively. The highest seed ALA levels of T0, T1, and T2 plants were 38.41, 35.98, and 39.19% respectively, which were 4.10–4.77 folds of the respective controls. FA-pathway enzyme genes (BnACCD, BnFATA, BnSAD, BnSCD, BnDGAT1, BnDGAT2, and BnDGAT3) and positive regulatory genes (BnWRI1, BnLEC1, BnL1L, BnLEC2, BnABI3, BnbZIP67, and BnMYB96) were all significantly up-regulated. In contrast, BnTT1, BnTT2, BnTT8, BnTT16, BnTTG1, and BnTTG2, encoding negative oil accumulation regulators but positive secondary metabolism regulators, were all significantly down-regulated. This means the foreign ShFAD2-ShFAD3 fusion gene, directly and indirectly, remodeled both positive and negative loci of the whole FA-related network in transgenic B. napus seeds.

Key words: biotechnology , α-linolenic acid , oilseed rape (Brassica napus) , , FAD2 , FAD3 , chia (Salvia hispanica)

. 基于奇亚(Salvia hispanica) FAD2和FAD3的油菜(Brassica napus) α-亚麻酸的生物技术[J]. Journal of Integrative Agriculture, 2023, 22(12): 3810-3815.

XUE Yu-fei, INKABANGA TSEKE Alain, YIN Neng-wen, JIANG Jia-yi, ZHAO Yan-ping, LU Kun, LI Jia-na, DING Yan-song, ZHANG Shi-qing, CHAI You-rong. Biotechnology of α-linolenic acid in oilseed rape (Brassica napus) using FAD2 and FAD3 from chia (Salvia hispanica)[J]. Journal of Integrative Agriculture, 2023, 22(12): 3810-3815.

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基于奇亚(Salvia hispanica) FAD2和FAD3的油菜(Brassica napus) α-亚麻酸的生物技术

Biotechnology of α-linolenic acid in oilseed rape (Brassica napus) using FAD2 and FAD3 from chia (Salvia hispanica)

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