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园艺学报 ›› 2019, Vol. 46 ›› Issue (1): 74-86.doi: 10.16420/j.issn.0513-353x.2018-0142

• 研究论文 • 上一篇    下一篇

东方百合‘索邦’GA20ox的克隆及表达分析

张旭红1,2,孙美玉1,李靖锐1,王 頔1,张金政1,石 雷1,2,*   

  1. (1中国科学院植物研究所北方资源植物重点实验室/北京植物园,北京 100093;2西北农林科技大学风景园林艺术学院,陕西杨凌 712100)
  • 出版日期:2019-01-25 发布日期:2019-01-25

Cloning and Expression Analysis of GA20ox Gene in Lilium Oriental Hybrids‘Sorbonne’

ZHANG Xuhong1,2,SUN Meiyu1,LI Jingrui1,WANG Di1,ZHANG Jinzheng1,and SHI Lei1,2,*   

  1. (1Key Laboratory of Plant Resources and Beijing Botanical Garden,Institute of Botany,Chinese Academy of Sciences,Beijing 100093,China;2College of Landscape Architecture and Arts,Northwest Agriculture and Forestry University,Yangling,Shaanxi 712100,China)
  • Online:2019-01-25 Published:2019-01-25

摘要: 以东方百合‘索邦’为材料,利用cDNA末端快速扩增技术(RACE)获得编码活性GAs合成过程中双加氧酶的基因GA20ox,对其进行生物信息学分析和表达分析。结果表明,GA20ox全长1 502 bp,包括1个1 167 bp的开放阅读框(ORF),编码388个氨基酸。GA20ox编码的蛋白存在多个糖基化位点、磷酸化位点和酰化位点。‘索邦’GA20ox和其他物种的GA20ox蛋白有很高的相似性,均含有保守的PLN02276结构域。‘索邦’GA20ox与姬蝴蝶兰、铁皮石斛、海枣、油棕和椰子等单子叶植物的GA20ox蛋白具有较高的同源性。实时荧光定量PCR分析表明,GA20ox具有时空表达特异性,在萌芽期之后的茎生根、现蕾期的茎、萌芽期和展叶期的叶片、萌芽期的顶芽等快速发育的组织中有较高的表达。在萌芽期对植株进行高浓度(200和400 mg ? L-1)GA3处理对其生长的影响最大,其中200 mg ? L-1 GA3处理最有利于株高的增加,400 mg ? L-1 GA3处理最有利于地下部分干质量的增加。对萌芽期‘索邦’百合进行200 mg ? L-1 GA3处理,显著抑制其GA20ox在除鳞片外其他部位的表达,表达量均在处理后12 h降至最低,其中在茎和基生根中降幅最大,分别下降了96.79%和94.65%。推测GA20ox的表达受活性GAs的反馈调节,在调控体内活性GAs稳态中发挥重要的作用。

关键词: 百合, GA20ox, 基因克隆, 基因表达, 株高

Abstract: We cloned a cDNA encoding GA20ox from Lilium Oriental Hybrids‘Sorbonne’using rapid amplification of cDNA ends(RACE). The bioinformatics and expression patterns of GA20ox were analyzed in our study. Sequences analysis showed that the cloned GA20ox was 1 502 bp in length with an open reading frame(1 167 bp),which encoded 388 amino acids. The putative protein had many glycosylation,phosphorylation and N-myristoylation sites. Alignment of amino acid sequence showed that GA20ox of‘Sorbonne’shared a PLN02276 binding domain like the GA20ox in other species. Phylogenetic analysis revealed that GA20ox of‘Sorbonne’was highly homologous to GA20ox of other monocots,including Phalaenopsis equestris,Dendrobium catenatum,Phoenix dactylifera,Elaeis guineensis and Cocos nucifera. Quantitative Real-time PCR(qRT-PCR)analysis showed that GA20ox was highly expressed in stem root after germination,stem of squaring stage,leaf of germination stage and leaf expansion stage,and terminal bud of germination stage,implying the temporal and special regulation of GA20ox. The application of high GA3 concentration in germination stage had great influence on plant growth as plant height and biomass of underground were promoted by 200 mg ? L-1 and 400 mg ? L-1 of GA3 treatment,respectively. After treatment of GA3 at 200 mg ? L-1,the expression of GA20ox was significantly suppressed in all tissues(except for bulb scale)and fell to their lowest level at 12 h,especially in the stem and root with decreasing by 96.79% and 94.65% respectively. These results suggest that the expression of GA20ox may be regulated by active gibberellins with a potential feedback mechanism,thus the GA20ox may play a key role for the maintenance of endogenetic active gibberellins homeostasis in lily.

Key words: Lilium, GA20ox, gene cloning, gene expression, plant height

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