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Single-Molecule Long-Read Sequencing of Purslane (Portulaca oleracea) and Differential Gene Expression Related with Biosynthesis of Unsaturated Fatty Acids.

ABSTRACT: This study aimed to obtain the full-length transcriptome of purslane (Portulaca oleracea); assorted plant samples were used for single-molecule real-time (SMRT) sequencing. Based on SMRT, functional annotation of transcripts, transcript factors (TFs) analysis, simple sequence repeat analysis and long non-coding RNAs (LncRNAs) prediction were accomplished. Total 15.33-GB reads were produced; with 9,350,222 subreads and the average length of subreads, 1640 bp was counted. With 99.99% accuracy, after clustering, 132,536 transcripts and 78,559 genes were detected. All unique SMART transcripts were annotated in seven functional databases. 4180 TFs (including transcript regulators) and 7289 LncRNAs were predicted. The results of RNA-seq were confirmed with qRT-PCR analysis. Illumina sequencing of leaves and roots of two purslane genotypes was carried out. Amounts of differential expression genes and related KEGG pathways were found. The expression profiles of related genes in the biosynthesis of unsaturated fatty acids pathway in leaves and roots of two genotypes of purslane were analyzed. Differential expression of genes in this pathway built the foundation of ω-3 fatty acid accumulation in different organs and genotypes of purslane. The aforementioned results provide sequence information and may be a valuable resource for whole-genome sequencing of purslane in the future.

SUBMITTER: Du H

PROVIDER: S-EPMC8066459 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Purslane is a widespread wild vegetable with both medicinal and edible properties. It is highly appreciated for its high nutritional value and is also considered as a high-quality feed resource for livestock and poultry. In this study, Sanhuang broilers were used to investigate the effect of feeding purslane diets on the growth performance in broilers and their gut microbiota. A total of 48 birds with good growth and uniform weight were selected and randomly allocated to four treatment groups A (control), B, C and D. Dietary treatments were fed with basal diet without purslane and diets containing 1%, 2% and 3% purslane. The 16S rDNA was amplified by PCR and sequenced using the Illumina HiSeq platform to analyze the composition and diversity of gut microbiota in the four sets of samples. The results showed that dietary inclusion of 2% and 3% purslane could significantly improve the growth performance and reduce the feed conversion ratio. Microbial diversity analysis indicated that the composition of gut microbiota of Sanhuang broilers mainly included Gallibacterium, Bacteroides and Escherichia-Shigella, etc. As the content of purslane was increased, the abundance of Lactobacillus increased significantly, and Escherichia-Shigella decreased. LEfSe analysis revealed that Bacteroides_caecigallinarum, Lachnospiraceae, Lactobacillales and Firmicutes had significant differences compared with the control group. PICRUSt analysis revealed bacteria mainly enriched in carbohydrate metabolism pathway due to the additon of purslane in the diet. These results suggest that the addition of purslane to feed could increase the abundance of Lactobacillus in intestine, modulate the environment of gut microbiota and promote the metabolism of carbohydrates to improve its growth performance. This study indicates that the effect of purslane on the growth-promoting performance of broilers might depend on its modulation on gut microbiota, so as to provide a certain scientific basis for the application of purslane in the feed industry.

Project description:Purslane (Portulaca oleraceae L.) is a widespread weed, which is highly appreciated for its high nutritional value with particular reference to the content in omega-3 fatty acids. In the present study, the nutritional value and chemical composition of purslane plants in relation to plant part and harvesting stage were evaluated. Plants were harvested at three growth stages (29, 43 and 52 days after sowing (DAS)), while the edible aerial parts were separated into stems and leaves. Leaves contained higher amounts of macronutrients than stems, especially at 52 DAS. ?-tocopherol was the main isoform, which increased at 52 DAS, as well total tocopherols (values were in the ranges of 197-327 ?g/100 g fresh weight (fw) and 302-481 ?g/100 g fw, for ?-tocopherol and total tocopherols, respectively). Glucose and fructose were the main free sugars in stems and leaves, respectively, whereas stems contained higher amounts of total sugars (values were ranged between 0.83 g and 1.28 g/100 g fw). Oxalic and total organic acid content was higher in leaves, especially at the last harvesting stage (52 DAS; 8.6 g and 30.3 g/100 g fw for oxalic acid and total organic acids, respectively). Regarding the fatty acid content, stems contained mainly palmitic (20.2-21.8%) and linoleic acid (23.02-27.11%), while leaves were abundant in ?-linolenic acid (35.4-54.92%). Oleracein A and C were the major oleracein derivatives in leaves, regardless of the harvesting stage (values were in the ranges of 8.2-103.0 mg and 21.2-143 mg/100 g dried weight (dw) for oleraceins A and C, respectively). Cytotoxicity assays showed no hepatotoxicity, with GI50 values being higher than 400 ?g/mL for all the harvesting stages and plant parts. In conclusion, early harvesting and the separation of plant parts could increase the nutritional value of the final product through increasing the content of valuable compounds, such as omega-3 fatty acids, phenolic compounds and oleracein derivatives, while at the same time, the contents of anti-nutritional compounds such as oxalic acid are reduced.

Dataset Information

Single-Molecule Long-Read Sequencing of Purslane (Portulaca oleracea) and Differential Gene Expression Related with Biosynthesis of Unsaturated Fatty Acids.

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