使用紫苏 FAD2–FAD3 融合基因对富含 α-亚麻酸的主食油料作物甘蓝型油菜进行工程改造,Journal of Agricultural and Food Chemistry

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使用紫苏 FAD2–FAD3 融合基因对富含 α-亚麻酸的主食油料作物甘蓝型油菜进行工程改造
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2023-05-02 , DOI: 10.1021/acs.jafc.2c09026
Yu-Fei Xue _{1,

2} , Chun Fu ₁ , Cheng-Yan Chai ₁ , Fei-Fei Liao ₁ , Bao-Jun Chen _{1,

2} , Song-Zhen Wei _{1,

2} , Rui Wang _{1,

2} , Huan Gao _{1,

2} , Teng-Teng Fan _{1,

2} , You-Rong Chai _{1,

2}

Affiliation

现代人普遍患有α-亚麻酸（ALA）缺乏症，因为大多数主食油中的ALA含量都很低。因此，增强主要油料作物中的 ALA 非常重要。在这项研究中，使用新设计的双接头 LP4-2A 融合来自 ALA 王种紫苏的FAD2和FAD3编码区，由种子特异性启动子 P _NAP驱动，并将其设计成油菜籽优良品种 ZS10质量背景。_{P NAP}种子油中的平均 ALA 含量：PfFAD2 – PfFAD3(N23) T5 线是对照的 3.34 倍（32.08 对 9.59%），最好的线高达 37.47%。工程构建体对包括含油量在内的背景特征没有明显的副作用。在脂肪酸生物合成途径中，结构基因和调控基因的表达水平在 N23 品系中显着上调。另一方面，编码类黄酮-原花青素生物合成正调节因子但油脂积累负调节因子的基因表达水平显着下调。_{令人惊讶的是，由组成型启动子 P D35S驱动的}PfFAD2 – PfFAD3转基因油菜品系中的 ALA 水平由于外源基因表达水平较低和内源直系同源基因BnFAD2和BnFAD3的下调，没有增加甚至略有下降。

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Engineering the Staple Oil Crop Brassica napus Enriched with α-Linolenic Acid Using the Perilla FAD2–FAD3 Fusion Gene

Modern people generally suffer from α-linolenic acid (ALA) deficiency, since most staple food oils are low in ALA content. Thus, the enhancement of ALA in staple oil crops is of importance. In this study, the FAD2 and FAD3 coding regions from the ALA-king species Perilla frutescens were fused using a newly designed double linker LP4-2A, driven by a seed-specific promoter P_NAP, and engineered into a rapeseed elite cultivar ZS10 with canola quality background. The mean ALA content in the seed oil of P_NAP:PfFAD2–PfFAD3 (N23) T5 lines was 3.34-fold that of the control (32.08 vs 9.59%), with the best line being up to 37.47%. There are no significant side effects of the engineered constructs on the background traits including oil content. In fatty acid biosynthesis pathways, the expression levels of structural genes as well as regulatory genes were significantly upregulated in N23 lines. On the other hand, the expression levels of genes encoding the positive regulators of flavonoid-proanthocyanidin biosynthesis but negative regulators of oil accumulation were significantly downregulated. Surprisingly, the ALA level in PfFAD2–PfFAD3 transgenic rapeseed lines driven by the constitutive promoter P_D35S was not increased or even showed a slight decrease due to the lower level of foreign gene expression and downregulation of the endogenous orthologous genes BnFAD2 and BnFAD3.

更新日期：2023-05-02

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