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| 甜樱桃(Prunus avium L.)LEAFY基因的克隆及表达模式分析 |
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段续伟1,2, 王晶1,3, 张晓明1,3, 闫国华1,2, 周宇1,3, 倪杨1,3, 张开春1,2
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| (1.北京市林业果树科学研究院, 北京 100093;2.农业部都市农业(华北)重点实验室, 北京 100093;3.北京市落叶果树工程技术研究中心, 北京 100093) |
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| 摘要: |
| 为探索LEAFY基因在甜樱桃花芽分化过程中的表达及作用，以甜樱桃（Prunus avium）‘红蜜’（Hongmi）为试材，利用同源克隆和RACE方法克隆甜樱桃中LEAFY同源基因cDNA全序列，对其序列进行分析，利用实时荧光定量PCR（qRT-PCR）对该基因在花芽分化过程中的表达量进行鉴定。结果显示：在甜樱桃中具有3中不同长度类型，分别为1 532、1 498和1 410 bp，主要由3'UTR区长度不同所导致，开放阅读框长1 233 bp，编码410个氨基酸，推测蛋白分子量约为45.6 ku，等电点约为6.88。结构预测NCBI序列比对发现其氨基酸序列与其他植物中同源基因相似性均在70%以上，故命名为PaLEAFY。qRT-PCR分析发现，在果实成熟至成熟后33 d，PaLEAFY在花芽中的表达量由高变低，果实成熟后45 d，PaLEAFY表达量开始急剧升高并维持至69 d，69 d之后表达量再次升高，并于果实成熟后99 d达到最高。此外，PaLEAFY在甜樱桃花的柱头中的表达量最高，其次是花瓣、雄蕊和花萼，在甜樱桃的茎和叶中的表达量非常低。说明PaLEAFY参与甜樱桃花芽分化及花发育过程。 |
| 关键词: 甜樱桃 LEAFY 克隆分析 花芽分化 |
| DOI：10.11841/j.issn.1007-4333.2018.04.03 |
| 投稿时间：2017-06-23 |
| 基金项目:国家自然科学基金青年基金项目（31401827）；北京市农林科学院科技创新能力建设专项（KJCX2016）；中国博士后科学基金项目（2016M590063） |
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| Molecular cloning and expression profile of LEAFY gene in Prunus avium L. |
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DUAN Xuwei1,2, WANG Jing1,3, ZHANG Xiaoming1,3, YAN Guohua1,2, ZHOU Yu1,3, NI Yang1,3, ZHANG Kaichun1,2
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| (1.Beijing Academy of Forestry and Pomology Sciences, Beijing 100093, China;2.Key Laboratory of Urban Agriculture(North China), Ministry of Agriculture, Beijing 100093, China;3.Beijing Engineering Research Center for Deciduous Fruit Trees, Beijing 100093, China) |
| Abstract: |
| To study the expression and function of LEAFY in process of flower bud differentiation into sweet cherry,the full-length cDNA encoding LEAFY was obtained from the flower bud of the sweet cherry (Prunus avium L.) ‘Hongmi’ and amplified by homologous cloning and rapid-amplification of cDNA ends (RACE).LEAFY expression in the process of bud differentiation was identified by real-time fluorescence quantitative PCR (qRT-PCR).The results showed that there were 3 length types LEAFY in ‘Hongmi’,which were 1 532,1 498 and 1 410 bp respectively.The length differences were mainly caused by different length of 3'UTR regions,containing a 1 233 bp open reading frame (ORF),which encoded a protein of 410 amino acids and had more than 70% amino acid sequence similarity with LEAFY homologous protein in other plants.The deduced protein molecular weight was 45.6 ku and its theoretical isoelectric point was 6.88,namely PaLEAFY.Real-time RT-PCR analysis indicated that:After fruit mature till 33 days,PaLEAFY expression changing trend was from high to low in flower bud.And at 45 days after fruit mature.PaLEAFY expression began to rise and maintained still till 69 days after fruit mature,then rose again at 69 days after fruit mature and reached highest at 99 days after fruit mature.In addition,the expression of PaLEAFY gene in stem and leaf were lower than that in the flowers.The expression of PaLEAFY was highest in stigma,and then were petal,stamen and sepal of flower suggesting that PaLEAFY was involved in flower bud differentiation and floral development process in sweet cherry. |
| Key words: Prunus avium L. LEAFY cloning and expression analysis flower bud differentiation |
