重瓣百合LiAGL6基因的克隆与表达分析  

Cloning and Expression Analysis of LiAGL6 in Double Lily

作　　者：隋娟娟[1] 李晓昕[2] 吴健[2] 曹兴[3] 吴泽[2] 何俊娜[2] 义鸣放[2] SUI Juanjuan;LI Xiaoxin;WU Jian;CAO Xing;WU Ze;HE Junna;YI Mingfang(Biology and Food Engineering School,Fuyang Normal University,Fuyang 236037,China;Beijing Key Laboratory of Development and Quality Control of Ornamental Crops,Department of Ornamental Horticulture and Landscape Architecture,China Agricultural University, Beijing 100193,China;College of Agronomy,Liaocheng University,Liaocheng 252059,China)

机构地区：[1]阜阳师范学院生物与食品工程学院,安徽阜阳236037 [2]中国农业大学观赏园艺与园林系,花卉发育与品质调控北京市重点实验室,北京100193 [3]聊城大学农学院,山东聊城252059

出　　处：《华北农学报》2018年第2期87-94,共8页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31471904);安徽省教育厅自然科学研究重点项目(KJ2016A871);安徽省高校质量工程项目(2015jyxm221).

摘　　要：为了深入研究重瓣百合花形成的分子机制,以重瓣百合比罗尼卡为试验材料,从中分离得到1个AGL6基因,其开放阅读框为744 bp,共编码247个氨基酸,蛋白质分子量为28.3 ku。亲水性平均系数为-0.748,等电点为8.23。蛋白结构分析显示,百合AGL6蛋白具有典型的MADS-box结构域、K-box区及2个AGL6基序。系统进化树分析结果显示,百合AGL6编码的蛋白属于AP1/AGL9亚家族AGL6分枝的单子叶植物类群,与同为单子叶植物的风信子亲缘关系最近,相似度达到78%,说明克隆得到的基因即为AGL6的同源基因,将其命名为LiAGL6。亚细胞定位表明,LiAGL6在洋葱表皮细胞的细胞核中表达,具备转录因子的基本特征。实时荧光定量PCR结果分析表明,LiAGL6基因在花中表达量最高,在茎、叶中几乎不表达;在整朵花中,LiAGL6在第7轮花瓣中的相对表达量最高,其次依次是第6轮和第3轮,第2轮花瓣中几乎检测不到表达。研究表明,LiAGL6基因可能在百合重瓣花形成方面发挥了调控作用。To explore flower development molecular mechanism of the Double lily,the AGL6 homologous gene was cloned from Double lily cv.Belonica.The open reading frame of LiAGL6 gene was 744 bp,encoded a protein of 247 amino acids.The LiAGL6 protein molecular weight was 28.3 ku,the grand average of hydropathicity was-0.748 and the theoretical pI was 8.23.The protein structure analysis showed that LiAGL6 protein had a typical MADS-box domain,a K-box region and two AGL6 motifs.Phylogenetic tree analysis showed that LiAGL6 belonged to monocotyledon group,AGL6 branch of the AP1/AGL9 subfamily,and had the highest similarity with Hyacinthus orientalis which was also belonged to monocotyledons,the similarity reached 78%.All the results indicated LiAGL6 was the homologous gene of AGL6,so named it LiAGL6.Cellular localization assay revealed LiAGL6 expressed in the nucleus of onion epidemical cells,which was the basic characteristics of the transcription factors.The qRT-PCR result showed the highest expression of LiAGL6 was in flowers while no expression occurred in leaves and stems.LiAGL6 was expressed most in the seventh whorl petals followed by the sixth and third whorl petals,however,there was almost no expression in the second whorl petals.The research showed the LiAGL6 gene might play a regulatory role on formation of Double lily flowers.

关 键 词：重瓣百合 MADS-BOX 基因克隆 LiAGL6基因 表达分析 

分 类 号：Q78[生物学—分子生物学] S644.03[农业科学—蔬菜学]