Abstract
Resveratrol (Res) is a type of natural plant stilbenes and phytoalexins that only exists in a few plant species. Studies have shown that the Res could be biosynthesized and accumulated within plants, once the complete metabolic pathway and related enzymes, such as the key enzyme resveratrol synthase (RS), existed. In this study, a RS gene named PNRS1 was cloned from the peanut, and the activity was confirmed in E. coli. Using transgenic approach, the PNRS1 transgenic rice was obtained. In T3 generation, the Res production and accumulation were further detected by HPLC. Our data revealed that compared to the wild type rice which trans-resveratrol was undetectable, in transgenic rice, the trans-resveratrol could be synthesized and achieved up to 0.697 μg/g FW in seedlings and 3.053 μg/g DW in seeds. Furthermore, the concentration of trans-resveratrol in transgenic rice seedlings could be induced up to eight or four-fold higher by ultraviolet (UV-C) or dark, respectively. Simultaneously, the endogenous increased of Res also showed the advantages in protecting the host plant from UV-C caused damage or dark-induced senescence. Our data indicated that Res was involved in host-defense responses against environmental stresses in transgenic rice. Here the results describes the processes of a peanut resveratrol synthase gene transformed into rice, and the detection of trans-resveratrol in transgenic rice, and the role of trans-resveratrol as a phytoalexin in transgenic rice when treated by UV-C and dark. These findings present new outcomes of transgenic approaches for functional genes and their corresponding physiological functions, and shed some light on broadening available resources of Res, nutritional improvement of crops, and new variety cultivation by genetic engineering.
Highlights
Resveratrol (3,40,5-trihydroxy-stilbene, Res) is a non-flavonoid polyphenolic compound in plant phenylpropanoide related secondary metabolism
Resports showed that the resveratrol synthase (RS) gene from Arachis hypogaea that transformed into Nicotiana tabacum cold lead to the production of Res [17], and many other RS genes of different species have been transformed into various plant species and microorganisms [14], which suggested that genetic engineering in this field is feasible and effective
The results of SDS-PAGE were shown in Fig 1A, 6× His tag fused RS (46 kD) or Maltose binding protein (MBP) fused 4-coumarate-CoA ligase (4CL) (103 kD) emerged in the cell lysate of E. coli transformed with pET-28a-PNRS1 or pMAL-c2x-At4CL2, compared with the protein brands of related empty vectors
Summary
Resveratrol (3,40,5-trihydroxy-stilbene, Res) is a non-flavonoid polyphenolic compound in plant phenylpropanoide related secondary metabolism. In many plant species such as rice, it is konwn to contain the synthetic substrates of Res, such as coumaryl Co-A and malonyl Co-A, but lack of the key synthetic enzyme of RS, so it is still unable to synthesize Res within the plants [14]. Since there are such an important role of the RS gene in the Res synthesis process, so it has been intensively and globally investigated [15, 16]. In view of the existence of endogenous processing and modification, the metabolites of trans-resveratrol and its isomers or derivates could be newly synthesized and detected in transgenic plants
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