| 狗蔷薇细胞分裂素氧化酶基因RcCKX5的克隆及表达分析 |
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| 引用本文： | 王玲,高彬,温超,郗琳,刘凤栾,马男,赵梁军.狗蔷薇细胞分裂素氧化酶基因RcCKX5的克隆及表达分析[J].园艺学报,2014,41(7):1418-1426. |
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| 作者姓名： | 王玲 高彬 温超 郗琳 刘凤栾 马男 赵梁军 |
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| 作者单位： | 中国农业大学观赏园艺与园林系，北京 100193 |
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| 基金项目： | 国家自然科学基金项目（31171993） |
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| 摘 要： | 利用RACE 技术从狗蔷薇（Rosa canina）类根体中克隆得到1 个细胞分裂素氧化酶基因cDNA全长，命名为RcCKX5。序列分析表明，RcCKX5 cDNA 全长1 815 bp，5′ UTR 104 bp，ORF 1 626 bp，3′ UTR85 bp，编码541 个氨基酸，具有CKX 家族典型的FAD（黄素腺嘌呤二核苷酸）结合域和CK（细胞分裂素）结合域。氨基酸序列多重比对和系统进化树分析显示RcCKX5 与可可TcCKX5 亲缘关系最近，与拟南芥AtCKX5、乌拉尔图小麦TuCKX5 同源性较高。荧光定量PCR 分析表明，狗蔷薇RcCKX5 在其根、茎、叶、花、果实中均有表达，根中相对表达量最高，花和果实中相对表达量较高。在狗蔷薇类根体形成发育过程中RcCKX5 的相对表达量不断升高，21 d 时达到最高，随后稍有下降，表明其可能参与了类根体的形成。6-BA 处理结果表明RcCKX5 能够快速响应6-BA 的诱导，表达量显著上调，2.5 mg · L-1 6-BA处理120 h 时，RcCKX5 的相对表达量达到最高峰。
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| 关 键 词： | 狗蔷薇 类根体 细胞分裂素氧化酶 表达分析 |
Isolation and Expression Analysis of Cytokinin Dehydrogenase Gene RcCKX5 in Rosa canina |
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| WANG Ling,GAO Bin,WEN Chao,XI Lin,LIU Feng-Luan,MA Nan,ZHAO Liang-Jun.Isolation and Expression Analysis of Cytokinin Dehydrogenase Gene RcCKX5 in Rosa canina[J].Acta Horticulturae Sinica,2014,41(7):1418-1426. |
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| Authors: | WANG Ling GAO Bin WEN Chao XI Lin LIU Feng-Luan MA Nan ZHAO Liang-Jun |
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| Institution: | Department of Ornamental Horticulture and Landscape Architecture，China Agricultural University，Beijing 100193，;
China |
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| Abstract: | Protocorm-like bodies（PLBs）is a new protocol of efficient regeneration，and the development of rhizoids is a critical stage of PLB regeneration. Recent researches showed that cytokinin regulated the formation of rhizoids. Here，we isolated the full length cDNA of a gene encoding cytokinin dehydrogenase from rhizoids of Rosa canina，named RcCKX5. Sequence analysis indicated that RcCKX5 consisted of 1 815 bp，with ORF of 1 626 bp，5′ UTR of 104 bp，3′ UTR of 85 bp，encoded 541 amino acids. The deduced amino acids possessed conserved domains of CKX family，FAD-binding domain and cytokinin-bining domain. Multiple sequence alignments and phylogenetic analysis showed that RcCKX5 had the highest similarity with TcCKX5 from Theobroma cacao. Real-time PCR analysis indicated that expression level of RcCKX5 in root was the highest，and expression level in flower and fruit was high.Expression level of RcCKX5 keep increasing all along the formation of rhizoids，reached the highest peak in the twenty-first day，then decreased slightly. This indicated RcCKX5 might be involved in rhizoids formation. RcCKX5 was significantly and rapidly induced by 6-BA，expression level of RcCKX5 reached highest with the concentration of 2.5 mg ? L-1 and time of 120 h. |
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| Keywords: | Rosa canina rhizoid cytokinin dehydrogenase expression analysis |
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