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园艺学报 ›› 2012, Vol. 39 ›› Issue (10): 1975-1982.

• 观赏植物 • 上一篇    下一篇

月季RhRTE1 的克隆及其对乙烯响应特性分析

孙翠慧, 刘道凤, 张帅, 李娜, 马男, 薛璟祺, 高俊平   

  1. (1 中国农业大学观赏园艺与园林系,北京 100193;2 中国农业科学院蔬菜花卉研究所,北京 100081)
  • 出版日期:2012-10-25 发布日期:2012-10-25

Isolation of RhTRE1 and Its Expression in Response to Ethylene During Flower Opening of Roses

 SUN  Cui-Hui, LIU  Dao-Feng, ZHANG  Shuai, LI  Na, MA  Nan, XUE  Jing-Qi, GAO  Jun-Ping   

  1. (1Department of Ornamental Horticulture and Landscape Architecture,China Agricultural University,Beijing 100193,China;2Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China)
  • Online:2012-10-25 Published:2012-10-25

摘要: 利用数据库中已发表的拟南芥AtRTE1基因序列,从构建的月季花朵乙烯响应EST数据库中筛选得到一个RTE1同源片段,结合RACE技术,从月季切花品种‘Samantha’花瓣中克隆得到全长为1 233 bp的RTE1同源基因。该基因包含一个732 bp的开放阅读框、339 bp的5非翻译区和162 bp的3′非翻译区,编码一个243 aa的肽链,其理论分子量为27.6 kD,等电点为6.87。该基因推定的氨基酸序列与拟南芥AtRTE1同源性较高,属于RTE家族中的第1组别基因,命名为RhRTE1。该基因在月季花朵自然开放中表达逐步增强,并能够被外源乙烯和切伤处理所显著诱导。将35S::GFP-RhRTE1融合蛋白转入拟南芥,发现RhRTE1蛋白主要定位于细胞膜上,并且转化植株降低了对乙烯的敏感性。以上结果说明RhRTE1可能参与了乙烯介导的月季花朵开放过程,并可能参与了花朵响应伤害的修复过程。

关键词: 月季, 花朵开放, 乙烯, RTE, 基因表达, 功能验证

Abstract: A homologous fragment with RTE1 was obtained through screening the rose EST database,then the full length(1 233 bp)of this gene was cloned from rose petals. This gene contained a 732 bp open reading frame,a 339 bp 5′ untranslated region,and a 162 bp 3′ untranslated region. The predicted protein consisted of 243 amino acids,had a calculated molecular mass of 27.6 kD,and its isoelectric point is 6.87. This gene shared high identity with AtRTE1 inArabidopsis,and it belonged to group one in RTE gene families,so it was named as RhRTE1. The expression of RhRTE1 enhanced gradually during the flower opening process,and it could also be induced by ethylene and wounding treatments. Through transferring 35S::GFP-RhRTE1 fusion protein into Arabidopsis,we found that RhRTE1 was located on the cell membranes,and over expression of RhRTE1 reduced the ethylene sensitivity of Arabidopsis. The results above indicated that RhRTE1 may be involved in ethylene regulated flower opening and damage recovery in rose flowers.

Key words: rose, Rosa hybrida, flower opening, ethylene, RTE, gene expression, functional verification

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