Subjects: Biology >> Botany >> Applied botany submitted time 2019-08-26 Cooperative journals: 《广西植物》
Abstract： To study the effect of cycloartenol synthase gene (CAS) in the sterol biosynthesis of Paeonia delavayi, using the RT-PCR technology, a complete open reading frame of PdCAS gene was cloned from Paeonia delavayi for the first time. The full-length cDNA of PdCAS gene was 2 274 bp, encodes 757 amino acids. The PdCAS protein sequence of P. delavayi had more than 86% similarty with the Prunus persica, Prunus yedoensis var. nudiflora and Vitis vinifera. Sequence alignment analysis showed that PdCAS had a typical conserved DCTAE motif of oxidosaualene cyclase and the marker sequence DGSWYGSWGVCFTYG of triterpenoid synthase．The phylogenetic analysis revealed that PdCAS and Malus domestica (XP 008391430.1), Pyrus bretschneideri (XP 009370034.1), Rosa chinensis(XP 024193310.1), Prunus yedoensis var. Nudiflora (PQQ11009.1), Prunus persica (XP 007225240.1)clustered together. Transcription pattern analysis showed that PdCAS expressed significantly in different tissues and had the highest transcript profile in flowers. PdCAS was closely related to the synthesis of sterols from Paeonia delavayi.
Subjects: Biology >> Botany >> Applied botany submitted time 2019-08-26 Cooperative journals: 《广西植物》
Abstract： In order to reveal the molecular mechanism of terpenoid metabolism in Zanthoxylum armatum and the effect of grafting on its flavor, we designed specific primers based on transcriptome data and cloned a novel full-length cDNA sequence of geranylgeranyl pyrophosphate synthase (GGPPS) gene from Zanthoxylum armatum by RT-PCR. Named ZaGGPPS. The ZaGGPPS gene was analyzed by using NCBI, ProParam, SignalP 4.1 server, DNAMAN and MEGA7.0 software. The expression of ZaGGPPS gene in grafted and seedling trees was compared. The results showed that ZaGGPPS contains a complete open reading frame (OFR), consisting of 1 086 bp, encoding 361 amino acids. The relative molecular weight of the protein is 39 079.14 Da and the theoretical isoelectric point pI is 6.38. Blast comparison results showed that the protein belongs to the GGPPS family and contains two specific aspartic acid enrichment motifs, namely "DDXXXXD" and "DDXXD", and five characteristic functional domains. Phylogenetic tree results showed that Zanthoxylum armatum has close relationships with sweet orange (Citrus sinensis), clementine mandarin (C. clementina) and pomelo (C. maxima). Fluorescence quantitative PCR showed that the expression level of ZaGGPPS gene in Zanthoxylum armatum range from high to low as follows: leaf of seedling tree, leaf of grafted tree, stem of seedling tree and stem of grafted tree. Geranylgeranyl pyrophosphate synthase is a key enzyme in terpenoid biosynthesis pathway of Zanthoxylum armatum. Grafting can affect the expression of ZaGGPPS gene in leaves and stems. The ZaGGPPS gene of Zanthoxylum armatum was cloned and analyzed in order to provide theoretical basis for further study on the molecular mechanism of aroma formation of Zanthoxylum armatum and selection of excellent varieties by molecular biological means.
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