scholarly journals Bioinformatics and Expression Pattern Analysis of Soybean Fatty Acid Desaturase Family Gene

2020 ◽  
Author(s):  
Nan Wu ◽  
Long Jiang ◽  
Xiaoming Yu ◽  
Xiangbo Yang ◽  
Dianyuan Chen ◽  
...  

Abstract Background To clarify the homology and structural differences of seven genes in the soybean fatty acid desaturase 2 (FAD2) family and the relationship between the expression level of each gene and oleic acid content at different stages of grain development, the seven genes in this family were studied through informatics analysis and evaluation of expression patterns.Results Database analysis, phylogenetic tree analysis, transmembrane structure prediction, amino acid sequence alignment, intron exon structure analysis, and protein motif analysis were performed for seven genes in the FAD2 family. The results showed that GmFAD2-1 and GmFAD2-2b and Other genes are structurally different, and it is speculated that their functions also have weak changes. By analyzing gene expression patterns at different stages of soybean grain development, the expression levels of the family genes at different stages of grain development first increase, then decrease, and finally decrease. For stability, the correlation analysis showed that the gene expression was negatively correlated with oleic acid content, and the correlation coefficients of GmFAD2-1 and GmFAD2-2b were the largest, showing a significant negative correlation.Conclusions Bioinformatics and expression analysis of genes in the FAD2 family. Identify the structural differences of each gene, and the differences in expression levels at different stages of soybean variety and grain development, and initially identify the differences in function size and catalytic efficiency between genes, in order to create new high-oleic soybean varieties using genetic engineering Quality, providing a theoretical basis.

2020 ◽  
Author(s):  
Nan Wu ◽  
Long Jiang ◽  
Xiaoming Yu ◽  
Xiangbo Yang ◽  
Dianyuan Chen ◽  
...  

Abstract Background To clarify the homology and structural differences of seven genes in the soybean fatty acid desaturase 2 (FAD2) family and the relationship between the expression level of each gene and oleic acid content at different stages of grain development, the seven genes in this family were studied through informatics analysis and evaluation of expression patterns.Results Database analysis, phylogenetic tree analysis, transmembrane structure prediction, amino acid sequence alignment, intron exon structure analysis, and protein motif analysis were performed for seven genes in the FAD2 family. The results showed that GmFAD2-1 and GmFAD2-2b and Other genes are structurally different, and it is speculated that their functions also have weak changes. By analyzing gene expression patterns at different stages of soybean grain development, the expression levels of the family genes at different stages of grain development first increase, then decrease, and finally decrease. For stability, the correlation analysis showed that the gene expression was negatively correlated with oleic acid content, and the correlation coefficients of GmFAD2-1 and GmFAD2-2b were the largest, showing a significant negative correlation.Conclusions Bioinformatics and expression analysis of genes in the FAD2 family. Identify the structural differences of each gene, and the differences in expression levels at different stages of soybean variety and grain development, and initially identify the differences in function size and catalytic efficiency between genes, in order to create new high-oleic soybean varieties using genetic engineering Quality, providing a theoretical basis.


Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1245
Author(s):  
Naoufal Lakhssassi ◽  
Valéria Stefania Lopes-Caitar ◽  
Dounya Knizia ◽  
Mallory A. Cullen ◽  
Oussama Badad ◽  
...  

Soybean is the second largest source of oil worldwide. Developing soybean varieties with high levels of oleic acid is a primary goal of the soybean breeders and industry. Edible oils containing high level of oleic acid and low level of linoleic acid are considered with higher oxidative stability and can be used as a natural antioxidant in food stability. All developed high oleic acid soybeans carry two alleles; GmFAD2-1A and GmFAD2-1B. However, when planted in cold soil, a possible reduction in seed germination was reported when high seed oleic acid derived from GmFAD2-1 alleles were used. Besides the soybean fatty acid desaturase (GmFAD2-1) subfamily, the GmFAD2-2 subfamily is composed of five members, including GmFAD2-2A, GmFAD2-2B, GmFAD2-2C, GmFAD2-2D, and GmFAD2-2E. Segmental duplication of GmFAD2-1A/GmFAD2-1B, GmFAD2-2A/GmFAD2-2C, GmFAD2-2A/GmFAD2-2D, and GmFAD2-2D/GmFAD2-2C have occurred about 10.65, 27.04, 100.81, and 106.55 Mya, respectively. Using TILLING-by-Sequencing+ technology, we successfully identified 12, 8, 10, 9, and 19 EMS mutants at the GmFAD2-2A, GmFAD2-2B, GmFAD2-2C, GmFAD2-2D, and GmFAD2-2E genes, respectively. Functional analyses of newly identified mutants revealed unprecedented role of the five GmFAD2-2A, GmFAD2-2B, GmFAD2-2C, GmFAD2-2D, and GmFAD2-2E members in controlling the seed oleic acid content. Most importantly, unlike GmFAD2-1 members, subcellular localization revealed that members of the GmFAD2-2 subfamily showed a cytoplasmic localization, which may suggest the presence of an alternative fatty acid desaturase pathway in soybean for converting oleic acid content without substantially altering the traditional plastidial/ER fatty acid production.


2019 ◽  
Author(s):  
Nan Wu ◽  
Long Jiang ◽  
Xiaoming Yu ◽  
Xiangbo Yang ◽  
Dianyuan Chen ◽  
...  

Abstract To clarify the homology and structural differences of seven genes in the soybean fatty acid desaturase 2 (FAD2) family and the relationship between the expression level of each gene and oleic acid content at different stages of grain development, the seven genes in this family were studied through informatics analysis and evaluation of expression patterns. Amino acid sequence analysis of the seven FAD2 genes in soybean showed that the genes were located on 6 chromosomes, and the number of amino acids encoded by all the genes ranged between 216 and 387. Isoelectric point analysis showed that the proteins were alkaline. Phylogenetic tree comparison results indicated that the soybean FAD2 genes have higher homology with the Arabidopsis thaliana gene and belong to the same dicotyledon, while GmFAD2-2 and GmFAD2-2a have the highest homology, and GmFAD2-1 and GmFAD2-1b have the same homology, the highest source and less homology with other soybean FAD2 genes, and their function is expected to be slightly different. Transmembrane prediction of each gene in the family showed that GmFAD2-2b contains two transmembrane regions. The remaining genes contain 3-6 different transmembrane structures, all of which are hydrophilic. The amino acid sequences were aligned, and the results showed that the genes in this family contain histidine clusters at the same position, but in GmFAD2-2b, the histidine cluster composition is different from that in other genes. According to the gene 5’ UTR intron cis-regulatory element analysis, the gene contains hormone response elements, light response elements, meristem response elements, and participates in low temperature as a cis-acting element. By analysing the expression patterns of genes in the FAD2 family under low temperature stress, it was found that GmFAD2-2 and GmFAD2-1b exhibited seed-specific expression, while the other genes were constitutively expressed. The genes were up-regulated under low temperature stress. To adapt to low temperature, plants produced more unsaturated fatty acids, and the expression of genes was different at different treatment times. In four soybean genotypes, "Jike Soybean 20", "Ji Midou 3", "Jike Midou No. 1", and "Jike Fresh Bean 1", the soybean FAD content at different stages of grain development was determined, and content was found to increase, then decrease, and finally stabilize. Correlation analysis showed that there was a negative correlation between gene expression and oleic acid content, and the correlation coefficients for GmFAD2-1 and GmFAD2-2b were the largest, showing a significant negative correlation.


2007 ◽  
Vol 84 (3) ◽  
pp. 229-235 ◽  
Author(s):  
Devinder Sandhu ◽  
Jessie L. Alt ◽  
Curtis W. Scherder ◽  
Walter R. Fehr ◽  
Madan K. Bhattacharyya

2019 ◽  
Vol 20 (12) ◽  
pp. 3091 ◽  
Author(s):  
Hao Liu ◽  
Jianzhong Gu ◽  
Qing Lu ◽  
Haifen Li ◽  
Yanbin Hong ◽  
...  

Peanuts with high oleic acid content are usually considered to be beneficial for human health and edible oil storage. In breeding practice, peanut lines with high monounsaturated fatty acids are selected using fatty acid desaturase 2 (FAD2), which is responsible for the conversion of oleic acid (C18:1) to linoleic acid (C18:2). Here, comparative transcriptomics were used to analyze the global gene expression profile of high- and normal-oleic peanut cultivars at six time points during seed development. First, the mutant type of FAD2 was determined in the high-oleic peanut (H176). The result suggested that early translation termination occurred simultaneously in the coding sequence of FAD2-A and FAD2-B, and the cultivar H176 is capable of utilizing a potential germplasm resource for future high-oleic peanut breeding. Furthermore, transcriptomic analysis identified 74 differentially expressed genes (DEGs) involved in lipid metabolism in high-oleic peanut seed, of which five DEGs encoded the fatty acid desaturase. Aradu.XM2MR belonged to the homologous gene of stearoyl-ACP (acyl carrier protein) desaturase 2 (SAD2) that converted the C18:0 into C18:1. Further subcellular localization studies indicated that FAD2 was located at the endoplasmic reticulum (ER), and Aradu.XM2MR was targeted to the plastid in Arabidopsis protoplast cells. To examine the dynamic mechanism of this finding, we focused on the peroxidase (POD)-mediated fatty acid (FA) degradation pathway. The fad2 mutant significantly increased the POD activity and H2O2 concentration at the early stage of seed development, implying that redox signaling likely acted as a messenger to connect the signaling transduction between the high-oleic content and Aradu.XM2MR transcription level. Taken together, transcriptome analysis revealed the feedback mechanism of SAD2 (Aradu.XM2MR) associated with FAD2 mutation during the seed developmental stage, which could provide a potential peanut breeding strategy based on identified candidate genes to improve the content of oleic acid.


2020 ◽  
Vol 21 (3) ◽  
pp. 1104 ◽  
Author(s):  
Nan Wu ◽  
Qiang Lu ◽  
Piwu Wang ◽  
Qi Zhang ◽  
Jun Zhang ◽  
...  

The soybean fatty acid desaturase family is composed of seven genes, but the function of each gene has not been reported. Bioinformatics was used to analyse the structure of genes in this family, as well as the correlation between Δ12-fatty acid desaturase II (FAD2) expression and oleic acid content on different days after flowering of soybean. In the present study, CRISPR/Cas9 technology was used to construct single and double mutant knockout vectors of functional genes in the FAD2 family. Analysis of the molecular biology and expression patterns of genes in the FAD2 family, namely, GmFAD2-1A (Glyma.10G278000) and GmFAD2-2A (Glyma.19G147300), showed that they had little homology with other soybean FAD2 genes, and that their function was slightly changed. Sequencing of the target showed that the editing efficiency of the GmFAD2-1A and GmFAD2-2A genes was 95% and 55.56%, respectively, and that the double mutant editing efficiency was 66.67%. The mutations were divided into two main types, as follows: base deletion and insertion. A near-infrared grain analyser determined the following results: In the T2 generation, the oleic acid content increased from 17.10% to 73.50%; the linoleic acid content decreased from 62.91% to 12.23%; the protein content increased from 37.69% to 41.16%; in the T3 generation, the oleic acid content increased from 19.15% to 72.02%; the linoleic acid content decreased from 56.58% to 17.27%. In addition, the protein content increased from 37.52% to 40.58% compared to that of the JN38 control variety.


Helia ◽  
2015 ◽  
Vol 38 (62) ◽  
Author(s):  
Claudio Ferfuia ◽  
Maurizio Turi ◽  
Gian Paolo Vannozzi

AbstractHigh temperature enhances the oleic acid content in the oil of normal cultivars but conflicting results are reported on temperature effects on oleic acid content in HO cultivars: either no effect or an increase in oleic acid content with temperature. To investigate the effects of temperature on HO genotypes under natural field conditions, a three-year field trial was conducted using two sowing dates and three HO genotypes (two inbred lines and one hybrid). To compare our results with previous works, growing degree-days (GDD) were computed (base temperature=6°C). GDD accumulated during the “flowering – 25 days after flowering” period influenced fatty acid composition of seed. Oleic and linoleic acid contents were affected by accumulated GDD in two HO genotypes (one inbred line and the hybrid). There was an increase of about 3% in oleic acid content as response to more high GDD accumulated. Their content was not modified by GDD in the other inbred line. There was a genotype×environment interaction that we suppose depending on modifier genes. These genetic factors affected oleic acid content. This indicated the importance of breeding targeted to select hybrids with a stable oleic acid content and higher than 90%. Saturated fatty acids (palmitic and stearic) were also influenced by temperature, and there was genetic variability among genotypes.


Agronomy ◽  
2018 ◽  
Vol 8 (10) ◽  
pp. 222 ◽  
Author(s):  
Qinfu Sun ◽  
Jueyi Xue ◽  
Li Lin ◽  
Dongxiao Liu ◽  
Jian Wu ◽  
...  

Rapeseed (Brassica napus L.) with substantial lipid and oleic acid content is of great interest to rapeseed breeders. Overexpression of Glycine max transcription factors Dof4 and Dof11 increased lipid accumulation in Arabidopsis and microalgae, in addition to modifying the quantity of certain fatty acid components. Here, we report the involvement of GmDof4 and GmDof11 in regulating fatty acid composition in rapeseeds. Overexpression of GmDof4 and GmDof11 in rapeseed increased oleic acid content and reduced linoleic acid and linolenic acid. Both qPCR and the yeast one-hybrid assay indicated that GmDof4 activated the expression of FAB2 by directly binding to the cis-DNA element on its promoters, while GmDof11 directly inhibited the expression of FAD2. Thus, GmDof4 and GmDof11 might modify the oleic acid content in rapeseed by directly regulating the genes that are associated with fatty acid biosynthesis.


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