Functional identification of oleate 12-desaturase and ω-3 fatty acid desaturase genes from Perilla frutescens var. frutescens

Lee, Kyeong‐Ryeol; Lee, Yong-Jik; Kim, Eunha; Lee, Seul-Bee; Roh, Kyung Hee; Kim, Jong Bum; Kang, Hwan-Ku; Kim, Hyun Uk

doi:10.1007/s00299-016-2053-4

Cited by 41 publications

(34 citation statements)

References 57 publications

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“…Seven AsFAD2 proteins were located in the ER (Fig. 5), which was in agreement with previous findings for cotton FAD2-4:GFP [47], three BnFAD2s:YFP [13] and FrFAD2-1:GFP [48]. The above results were also consistent with the prediction of subcellular localization by Plant-mPLoc 2.0 (Additional file 1: Table S1).…”

Section: Discussionsupporting

confidence: 92%

Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

Miao

zhang

et al. 2019

Preprint

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Background: Linoleic acid is an important polyunsaturated fatty acid, required for all eukaryotes.Microsomal delta-12 (Δ12) oleate desaturase (FAD2) is a key enzyme for linoleic acid biosynthesis.Desert shrub Artemisia sphaerocephala is rich in linoleic acid, it has a large FAD2 gene family with twenty-six members. The aim of this work is to unveil the difference and potentially functionality of AsFAD2 family members. Results: Full-length cDNAs of twenty-one AsFAD2 genes were obtained from A. sphaerocephala. The putative polypeptides encoded by AsFAD2 family genes showed a high level of sequence similarity and were relatively conserved during evolution. The motif composition was also relatively conservative. Quantitative real-time PCR analysis revealed that the AsFAD2-1 gene was strongly expressed in developing seeds, which may be closely associated with the high accumulating ability of linoleic acid in A. sphaerocephala seeds. Although different AsFAD2 family members showed diverse response to salt stress, the overall mRNA levels of the AsFAD2 family genes was stable.Transient expression of AsFAD2 genes in the Nicotiana benthamiana leaves revealed that the encoded proteins were all located in the endoplasmic reticulum. Heterologous expression in Saccharomyces cerevisiae suggested that only three AsFAD2 enzymes, AsFAD2-1, -10, and -23, were Δ12 oleate desaturases, which could convert oleic acid to linoleic acid, whereas AsFAD2-1 and AsFAD2-10 could also produce palmitolinoleic acid. Conclusions: This research reported the cloning, expression studies, subcellular localization and functional identification of the large AsFAD2 gene family. These results should be helpful in understanding fatty acid biosynthesis in A. sphaerocephala, and has the potential to be applied in the study of plant fatty acids traits.

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Section: Discussionsupporting

confidence: 92%

Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

Miao

zhang

et al. 2019

Preprint

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“…In Perilla frutescens , PfrFAD2 and PfrFAD3 genes were expressed in leaves and during all stages of seed development, and their expression levels in 2- to 3-week-old developing seeds were 6.7- and 25-fold higher than their expression in leaves, respectively. In contrast, although the expression of PfrFAD7-1 and PfrFAD7-2 was similar to that of PfrFAD2 and PfrFAD3 genes in leaves, their expressions were much lower in developing seeds [ 55 ]. Thus, the same type of FAD genes from different plants may have different spatial expression patterns, which requires further investigation.…”

Section: Resultsmentioning

confidence: 99%

Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

Chi¹,

Zhang²,

Chen³

et al. 2017

PLoS ONE

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BackgroundFatty acid desaturases are enzymes that introduce double bonds into fatty acyl chains. Extensive studies of fatty acid desaturases have been done in many plants. However, less is known about the diversity of this gene family in peanut (Arachis hypogaea L.), an important oilseed crop that is cultivated worldwide.ResultsIn this study, twelve novel AhFADs genes were identified and isolated from peanut. Quantitative real-time PCR analysis indicated that the transcript abundances of AhFAB2-2 and AhFAD3-1 were higher in seeds than in other tissues examined, whereas the AhADS and AhFAD7-1 transcripts were more abundant in leaves. AhFAB2-3, AhFAD3-2, AhFAD4, AhSLD-4, and AhDES genes were highly expressed in flowers, whereas AhFAD7-2, AhSLD-2, and AhSLD-3 were expressed most strongly in stems. During seed development, the expressions of AhFAB2-2, AhFAD3-1, AhFAD7-1, and AhSLD-3 gradually increased in abundance, reached a maximum expression level, and then decreased. The AhFAB2-3, AhFAD3-2, AhFAD4, AhADS, and AhDES transcript levels remained relatively high at the initial stage of seed development, but decreased thereafter. The AhSLD-4 transcript level remained relatively low at the initial stage of seed development, but showed a dramatic increase in abundance at the final stage. The AhFAD7-2 and AhSLD-2 transcript levels remained relatively high at the initial stage of seed development, but then decreased, and finally increased again. The AhFAD transcripts were differentially expressed following exposure to abiotic stresses or abscisic acid. Moreover, the functions of one AhFAD6 and four AhSLD genes were confirmed by heterologous expression in Synechococcus elongates or Saccharomyces cerevisiae.ConclusionsThe present study provides valuable information that improves understanding of the biological roles of FAD genes in fatty acid synthesis, and will help peanut breeders improve the quality of peanut oil via molecular design breeding.

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“…A single copy of FAD3 is expressed in Arabidopsis, which synthesizes 20% of the ALA in the seeds. Perilla (Perilla frutescens L.) has two copies of FAD3, which synthesize 60% of the ALA in the seeds (Arondel et al, 1992;Lee et al, 2016;Lee et al, 2019). Despite the presence of a higher copy number of FAD3 in soybean than in other plants, the ALA content in soybean seeds is as low as 8%.…”

Section: Discussionmentioning

confidence: 99%

Increased Production of α-Linolenic Acid in Soybean Seeds by Overexpression of Lesquerella FAD3-1

et al. 2020

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Soybean is a major crop that is used as a source of vegetable oil for human use. To develop transgenic soybean with high a-linolenic acid (ALA; 18:3) content, the FAD3-1 gene isolated from lesquerella (Physaria fendleri) was used to construct vectors with two different seed-specific promoters, soybean b-conglycinin (Pb-con) and kidney bean phaseolin (Pphas), and one constitutive cauliflower mosaic virus 35S promoter (P35S). The corresponding vectors were used for Agrobacterium-mediated transformation of imbibed mature half seeds. The transformation efficiency was approximately 2%, 1%, and 3% and 21, 7, and 17 transgenic plants were produced, respectively. T-DNA insertion and expression of the transgene were confirmed from most of the transgenic plants by polymerase chain reaction (PCR), quantitative real-time PCR (qPCR), reverse transcription PCR (RT-PCR), and Southern blot analysis. The fatty acid composition of soybean seeds was analyzed by gas chromatography. The 18:3 content in the transgenic generation T 1 seeds was increased 7-fold in Pb-con:PfFAD3-1, 4-fold in Pphas : PfFAD3-1, and 1.6-fold in P35S:PfFAD3-1 compared to the 18:3 content in soybean "Kwangankong". The increased content of 18:3 in the Pb-con:PfFAD3-1 soybean (T 1 ) resulted in a 52.6% increase in total fatty acids, with a larger decrease in 18:1 content than 18:2 content. The increase in 18:3 content was also maintained and reached 42% in the Pphas : PfFAD3-1 transgenic generation T 2 . Investigations of the agronomic traits of 12 Pb-con:PfFAD3-1 transgenic lines (T 1 ) revealed that plant height, number of branches, nodes, pods, total seeds, and total seed weight were significantly higher in several transgenic lines than those in non-transgenic soybean. Especially, an increase in seed size was observed upon expression of the PfFAD3-1 gene with the b-conglycinin promoter, and 6%-14% higher seed lengths were measured from the transgenic lines.

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Functional identification of oleate 12-desaturase and ω-3 fatty acid desaturase genes from Perilla frutescens var. frutescens

Cited by 41 publications

References 57 publications

Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

Cloning and Functional Analysis of the FAD2 Gene Family from Desert Shrub Artemisia sphaerocephala

Isolation and functional analysis of fatty acid desaturase genes from peanut (Arachis hypogaea L.)

Increased Production of α-Linolenic Acid in Soybean Seeds by Overexpression of Lesquerella FAD3-1

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