Abstract
Resveratrol synthase (RS) is a key enzyme that plays a critical role in the resveratrol synthesis pathway. In this study, six RS genes were isolated and characterized from peanut variety “Zhenzhu Hong” by silico cloning and RT-PCR. Bioinformatics analysis showed that deduced amino acid sequences of the six cloned RS genes were highly conserved with a similarity from 95% to 99% when compared to the RS genes which had been deposited at the GenBank. The results of amino acid sequences analysis showed six RS proteins contained the Chal_Sti_Synt_N and ACP_Syn_III_C domains and can be classified to same family but with different evolutionary distance. Expression pattern analysis by QRT-PCR provided evidence indicating that the mRNA of six RS genes were primarily expressed in the peanut shell at different developmental stages with different expression levels, but only lower levels of them were evident in the peanut kernel. The subcellular localization of RS protein in onion epidermal cell was performed by Agrobacterium tumefaciens-mediated transformation and the green fluorescent was monitored by confocal fluorescence microscopy. The results indicated that, RS1 and RS5 were located in the nucleus and plasma membrane respectively, while the RS2, RS3, RS4 and RS6 were located in both nucleus inner membrane and plasma membrane. The data will provide basic information for elucidating the regulatory mechanisms and enzyme kinetics underlying the RS genes in the resveratrol synthase pathway.
Highlights
Resveratrol (Trans-3,5,4’-trihydroxy stilbene) is a natural plant phytoalexin which produced by plants in response to bacterium, fungal and other biotic or UV irradiation, wounding and other abiotic stress
Primers were designed based on these reference sequences and six cDNA fragments corresponding to the resveratrol synthase (RS) genes from peanut “Zhenzhu Hong” variety were generated by RT-PCR (Figure 2)
Base on the deduced amino acid sequence, six cloned RS genes share with 95% - 99% identity by BLASTP searches, and showing 92% - 98% identity with those peanut RS genes in the public database
Summary
Resveratrol (Trans-3,5,4’-trihydroxy stilbene) is a natural plant phytoalexin which produced by plants in response to bacterium, fungal and other biotic or UV irradiation, wounding and other abiotic stress. It is present in more than 70 dietary plant species and with high concentration reported in grapes, berries and peanuts [1] [2]. The first reported transformation of foreign phytoalexin expression in a novel plant resulted disease resistance, was performed with a peanut RS gene introduced into tobacco, resulted in the rapid accumulation of resveratrol following treatment of cell suspension cultures with fungal elicitor [29]. RS genes have been introduced into a number of plants, including rice, barley and wheat, alfalfa, kiwifruit, grapevine, apple, aspen, papaya, white poplar, oilseed rape, banana, Rehmannia, tomato, Arabidopsis, lettuce, pea, and hop [30]-[33]
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