Evening primrose (Oenothera biennis) Δ6 fatty acid desaturase gene family: cloning, characterization, and engineered GLA and SDA production in a staple oil crop

Fu, Chun; Chai, Yourong; Ma, Lijuan; Wang, Rui; Hu, Kui; Wu, Jianyong; Li, Jiana; Li, Xue; Lu, Junxing

doi:10.1007/s11032-017-0682-0

Cited by 12 publications

(10 citation statements)

References 79 publications

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“…This suggests that the seed-specific promoter has absolute advantages over the constitutive promoter for seed trait modification, which conforms to the discovery in GLA + SDA engineering in canola using evening primrose Δ6 desaturase genes. 30 In previous reports on engineered PUFA production in plants, multiple cassettes in a plasmid 13,32 and serial retransformation 33 were the main methods for the transformation of multiple genes. The first strategy is a major solution for multigene assembly, but it is easy to produce potential gene silencing resulting from the multiple use of one promoter.…”

Section: ■ Discussionmentioning

confidence: 99%

“…Plasmids containing Pf FAD2 and Pf FAD3 were double-digested with XbaI + HindIII and HindIII + XmaI (Figure S1), respectively, and both plasmids pC2301M1NPB and pC2301M1DPB 30 (containing the seed-specific promoter P NAP and constitutive promoter P D35S , respectively) were double-digested with XbaI + XmaI. Two gene fragments were ligated together with two respective platform vectors to produce two recombinant vectors (Figure S2), pC2301M1NPB-Pf FAD2−LP4-2A−Pf FAD3 (N23) and pC2301M1DPB-Pf FAD2− LP4-2A−Pf FAD3 (C23).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

“…C23 and N23 were individually introduced into B. napus cv. ZS10 by A. tumefaciens (LBA4404)-mediated transformation according to Fu et al 30 Transformants were planted in a greenhouse as described previously. 31 The positive transformation was verified by GUS straining, Basta herbicide resistance and PCR checking, and selfpollinated for five generations.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

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

Xue

Chai

et al. 2023

J. Agric. Food Chem.

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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 :Pf FAD2−Pf FAD3 (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 Pf FAD2−Pf FAD3 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.

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Section: ■ Discussionmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

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

Xue

Chai

et al. 2023

J. Agric. Food Chem.

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“…For HIGS vector construction, the complete RNAi gene cassette sense-intron-antisense from the RNAi pCIT-SsTrx1 vector was digested and inserted into plant expression vectors pBinGlyRed3 (GenScript) and pC2301M1B (Fu et al, 2017). The pBinGlyRed3 vector carries a gene encoding red fluorescent protein.…”

Section: Rnai and Higs Vector Construction And Transformationmentioning

confidence: 99%

Sclerotinia sclerotiorum Thioredoxin1 (SsTrx1) is required for pathogenicity and oxidative stress tolerance

Rana

Ding

Banga

et al. 2021

Molecular Plant Pathology

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Sclerotinia stem rot, caused by the necrotrophic fungus Sclerotinia sclerotiorum, is a destructive plant disease that causes yield losses in vegetable and oilseed brassicas worldwide (Bolton et al., 2006). It is difficult to control S. sclerotiorum as it produces asexual, hard, resting sclerotia that can survive for many years in the soil. These germinate under favourable conditions to initiate a new cycle of the disease (Erental et al., 2007;Xu et al., 2018). Current methods of pathogen control do not provide reliable protection from the disease. It is thus important to develop new management strategies that offer sustainable disease control. Minimizing damage by deploying cultivars with a genetically inherited reduction in the susceptibility to pathogens is the preferred form of disease management. Pathogens in turn

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“…Although various research groups have previously cloned and sequenced the FADS2 genes of many organisms (10)(11)(12)(13)(14)(15)(16)(17)(18), and our own group previously analyzed the molecular mechanism underlying FADS2 substrate specificity (9), it is still not clear which key domain and/or amino acid mediates FADS2 catalytic activity. This is largely a reflection of the fact that, to date, no information is available regarding the FADS2 crystal structure; however, Wang et al (19) and Bai et al (20) recently analyzed the crystal structure of human and mammalian FADS9 (delta 9 desaturase), respectively.…”

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confidence: 99%

Molecular mechanisms underlying catalytic activity of delta 6 desaturase from Glossomastix chrysoplasta and Thalassiosira pseudonana

Shi

粟

et al. 2018

Journal of Lipid Research

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Delta 6 desaturase (FADS2) is a critical bifunctional enzyme required for PUFA biosynthesis. In some organisms, FADS2s have high substrate specificity, whereas in others, they have high catalytic activity. Previously, we analyzed the molecular mechanisms underlying high FADS2 substrate specificity; in this study, we assessed those underlying the high catalytic activity of FADS2s from and To understand the structural basis of this catalytic activity, GcFADS2 and TpFADS2 sequences were divided into nine sections, and a domain-swapping approach was applied to examine the role of each section in facilitating the catalytic activity of the overall protein. The results revealed two regions essential to this process: one that extends from the end of the fourth to the beginning of the fifth cytoplasmic transmembrane domain, and another that includes the C-terminal region that occurs after the sixth cytoplasmic transmembrane domain. Based on the domain-swapping analyses, the amino acid residues at ten sites were identified to differ between the GcFADS2 and TpFADS2 sequences, and therefore further analyzed by site-directed mutagenesis. T302V, S322A, Y375F, and M384S/M385 substitutions in TpFADS2 significantly affected FADS2 catalytic efficiency. This study offers a solid basis for in-depth understanding of catalytic efficiency of FADS2.

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Evening primrose (Oenothera biennis) Δ6 fatty acid desaturase gene family: cloning, characterization, and engineered GLA and SDA production in a staple oil crop

Cited by 12 publications

References 79 publications

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

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

Sclerotinia sclerotiorum Thioredoxin1 (SsTrx1) is required for pathogenicity and oxidative stress tolerance

Molecular mechanisms underlying catalytic activity of delta 6 desaturase from Glossomastix chrysoplasta and Thalassiosira pseudonana

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