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Molecular cloning and characterization of three isoprenyl diphosphate synthase genes from alfalfa

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Abstract

Isoprenoid is the precursor for the biosynthesis of saponins, abscisic acid, gibberellins, chlorophylls and many other products in plants. Saponins are an important group of bioactive plant natural products. The alfalfa (Medicago sativa L.) saponins are glycosides of different triterpene aglycones and possess many biological activities. We isolated three genes (MsFPPS, MsGPPS and MsGGPPS) encoding isoprenyl diphosphate synthases (IDS) from alfalfa via a homology-based PCR approach. The enzyme activity assay of purified recombined MsFPPS and MsGGPPS expressed in Escherichia coli indicated that they all had IDS activity. Expression analysis of the three genes in different alfalfa tissues using real time PCR displayed that they were expressed in all tissues although they had a different expression patterns. MsFPPS and MsGPS displayed a significant increase in transcript level in response to methyl jasmonate, but the transcript level of MsGGPPS decreased obviously. To elucidate the functions of the three IDSs, their overexpression driven by a constitutive cauliflower mosaic virus-35S promoter in tobacco plants was applied and analyzed. The T0 transgenic plants of MsFPPS showed high levels of squalene content when compared with control. However, no differences were detected in T0 transgenic plants of MsGPPS and MsGGPPS. In addition, the overexpression of MsFPPS induced senescence response in transgenic plant leaves. This result may indicate that MsFPPS performs a role not only in phytosterol and triterpene biosynthesis, but also in growth regulation.

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Acknowledgments

This work was supported by Industry System of Modern Agriculture Construction Funds, China (No. CARS-35).

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Authors and Affiliations

  1. College of Animal Science and Technology, China Agriculture University, Bei**g, 100193, China

    Yan Sun & He Zhou

  2. Institute of Agriculture and Life Science, Chongqing University, Chongqing, 400030, China

    Ruicai Long & Ze Zhang

  3. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Bei**g, 100193, China

    Junmei Kang, Tiejun Zhang & Qingchuan Yang

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  1. Yan Sun
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  2. Ruicai Long
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  6. He Zhou
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  7. Qingchuan Yang
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Correspondence to Qingchuan Yang.

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Sun, Y., Long, R., Kang, J. et al. Molecular cloning and characterization of three isoprenyl diphosphate synthase genes from alfalfa. Mol Biol Rep 40, 2035–2044 (2013). https://doi.org/10.1007/s11033-012-2262-z

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  • DOI: https://doi.org/10.1007/s11033-012-2262-z

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