The Cloning And Expression Of The Related Gene AGL15 Was Extended In The Honeysuckle Period

Lonicera macranthoides from Caprifoliaceae, as one of the sources of Chinese herbal medicine Flos lonicerae. It's dried buds or flowers as medicinal with detoxification, dispelling wind and heat effect. The treatment of carbuncle sore treatment, pharyngitis, erysipelas, heat Duxue dysentery, anemopyretic cold, warm disease. For a long time, how to solve the problem of Lonicera macranthoides bud period short, picking a timely mann er is a diffic?Lt issue in Lonicera macranthoides production and application, named "Xi ang Lei Lonicera japonica " varieties.By the early of Lonicera macranthoides hand were transcriptome de novo sequencing a nalysis, obtained the massive function clear, with independent intellectual property rights of Lonicera macranthoides gene sequence. Two varieties of Unigene gene expression q uantity obvious difference 20709. Among them, an important gene MADS-box gene fa mily plays an important reg?Latory function of 4, we selected the AGL15 homologous gene cloning and verification of the expression.This study to varieties of Lonicera macranthoides "Xianglei" as test material, by homol ogy cloning method, using RT-PCR and race techniques and isolated the f?LI length c DNA of Lonicera macranthoides LmAGL15, and by the method of relative fluorescence quantitative PCR, of the gene in different organs and the process of flower bud differ entiation of the temporal and spatial expression patterns were analyzed. At the same ti me, constructed the gene sense and antisense plant expression vector into Arabidopsis a nd tobacco after RT-PCR, get some positive plants, and the preliminary phenotype obse rved. The main res?Lts are as follows:1. According to the known gene conserved sequence and primer and using with RT-PC R and race methods, first from Lonicera macranthoides c?Ltivars "Xianglei" stems, leav es, of the various stages of bud successfully obtained the ag115 associated gene cDNA sequence, named LmAGL15 (GenBank ???:KR028478). Sequence analysis showed that the K gene contains a unique domain of the MADS domain and the plant,which belongs to MADS-box gene family.2. The relative fluorescence quantitative method using 18S rRNA gene as a reference g ene, of the gene expression differences of Lonicera macranthoides hand in different org ans in the analysis found that different genes in different organs have different expressi on patterns, with some tissue specificity. The main LmAGL15 in bud expression, almos t no expression in leaf and stem.Carries on the analysis to the LmAGL15 gene in Loni cera macranthoides during flower bud differentiation of differentially expressed by relati ve quantitative fluorescence method. The res?Lts show that different genes in theflower bud differentiation process have different trends. The LmAGL15 in the flower bud diff erentiation stage was the highest, the lowest expression in late stage.3. The successf?L construction of LmAGL15 sense and antisense plant expression vector, the inflorescence infection method was transformed Arabidopsis thaliana by genomic PCR detection, some positive plants obtained.4. The LmAGL15 gene in Arabidopsis overexpression by observing the phenotype of transgenic plants, transgenic Arabidopsis can delay flowering. The preliminary res?Lts sho wed that the LmAGL15 gene may play an important role in the reg?Lation of flowering.