Abstract

Alternaria sp. MG1, an endophytic fungus previously isolated from Merlot grape, produces resveratrol from glucose, showing similar metabolic flux to the phenylpropanoid biosynthesis pathway, currently found solely in plants. In order to identify the resveratrol biosynthesis pathway in this strain at the gene level, de novo transcriptome sequencing was conducted using Illumina paired-end sequencing. A total of 22,954,434 high-quality reads were assembled into contigs and 18,570 unigenes were identified. Among these unigenes, 14,153 were annotated in the NCBI non-redundant protein database and 5341 were annotated in the Swiss-Prot database. After KEGG mapping, 2701 unigenes were mapped onto 115 pathways. Eighty-four unigenes were annotated in major pathways from glucose to resveratrol, coding 20 enzymes for glycolysis, 10 for phenylalanine biosynthesis, 4 for phenylpropanoid biosynthesis, and 4 for stilbenoid biosynthesis. Chalcone synthase was identified for resveratrol biosynthesis in this strain, due to the absence of stilbene synthase. All the identified enzymes indicated a reasonable biosynthesis pathway from glucose to resveratrol via glycolysis, phenylalanine biosynthesis, phenylpropanoid biosynthesis, and stilbenoid pathways. These results provide essential evidence for the occurrence of resveratrol biosynthesis in Alternaria sp. MG1 at the gene level, facilitating further elucidation of the molecular mechanisms involved in this strain's secondary metabolism.

Highlights

  • Resveratrol is a phenolic compound that was found to have multiple functions, including neuroprotection, with no major adverse effects (Park and Pezzuto, 2015)

  • Each sequenced sample yielded 2 × 100 bp independent reads from either end of a cDNA fragment

  • 18,570 unigenes were identified from the contigs data set with an N50 length of 2153 bp and a mean length of 1122.38 bp, of which 6471 genes (34.85%) were >1 kb

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Summary

Introduction

Resveratrol is a phenolic compound that was found to have multiple functions, including neuroprotection, with no major adverse effects (Park and Pezzuto, 2015). High quantities of resveratrol are urgently needed for application in functional food processing (Bhullar and Hubbard, 2015). Resveratrol is mainly produced by extraction from plant materials, which is highly limited by plant growth times and low yields (Kiselev, 2011). Many efforts have been made to construct resveratrol-producing Escherichia coli or yeast strains through genetic modification (Kiselev, 2011). Most genes used in currently reported methods are plant based. Identifying microbial genes would provide new genes for metabolic engineering of resveratrol production using

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