茄子,生长素响应因子,基因克隆," /> 茄子,生长素响应因子,基因克隆,"/> eggplant(Solanum melongena L.),auxin response factor,gene cloning,"/> <font face="Verdana">茄子生长素响应因子SmARF8的克隆与分析</font>

中国农业科学 ›› 2009, Vol. 42 ›› Issue (7): 2434-2441 .doi: 10.3864/j.issn.0578-1752.2009.07.023

• 园艺 • 上一篇    下一篇

茄子生长素响应因子SmARF8的克隆与分析

杜黎明,毛伟海,包崇来,胡天华,朱琴妹,胡海娇   

  1. (浙江省农业科学院蔬菜研究所)
  • 收稿日期:2009-01-07 修回日期:2009-03-26 出版日期:2009-07-10 发布日期:2009-07-10
  • 通讯作者: 毛伟海

Cloning and Characterization of ARF Family Gene SmARF8 in Solanum melongena L.#br#

DU Li-ming, MAO Wei-hai, BAO Chong-lai, HU Tian-hua, ZHU Qin-mei, HU Hai-jiao#br#   

  1. (浙江省农业科学院蔬菜研究所)
  • Received:2009-01-07 Revised:2009-03-26 Online:2009-07-10 Published:2009-07-10
  • Contact: MAO Wei-hai

摘要:

【目的】克隆茄子生长素响应因子新基因,为研究茄子果实形成发育的分子机制提供依据。【方法】采用RT-PCR 和RACE 技术克隆茄子ARF家族相应基因的cDNA全长序列。经序列联配进行与其它物种同源基因的保守域和进化关系分析。实时定量PCR 检测目的基因在茄子不同组织中的表达情况。【结果】将该基因命名为SmARF8,它的cDNA序列全长为3 671 bp,编码阅读框全长2 676 bp。氨基酸序列分析表明,SmARF8蛋白拥有核定位序列,蛋白质结构具有ARF家族的N-端DNA结合域,中间脯氨酸、丝氨酸和苏氨酸残基富集的非保守调控域,C-末端蛋白互作域。氨基酸与拟南芥的调控单性结实基因ARF8相似性较高,进化树分析表明它们聚集在一个分支上,进化关系非常密切。SmARF8基因在幼果中表达最强,在根、茎、叶、花蕾、花朵及成熟果实中表达较强烈。【结论】从茄子中克隆得到一个生长素响应因子家族基因SmARF8。

关键词: 茄子')">茄子, 生长素响应因子, 基因克隆

Abstract:

【Objective】 Cloning of auxin response factor, SmARF8, and its expression analysis can provide a basis for molecular mechanism research of fruit development in eggplant. 【Method】 RT-PCR and RACE were used to amplify full-length cDNA sequence of SmARF8 in eggplant. The analysis of conserved domains and phylogenetic relations among the known ARFs was performed by amino acid sequence alignment method. Real-time PCR was used to analyze the expression patterns of SmARF8 in different tissues. 【Result】 Full-length cDNA sequence of this gene is 3 671 bp with an ORF of 2 676 bp in length. The results of phylogenetic analysis among ARF family genes indicated that this gene was highly homologous with the parthenocarpy-related gene ARF8 in Arabidopsis, so it was named SmARF8. The amino acid sequence analysis showed that it contained a nuclear localization signal. The DNA-binding domain was localized on N-terminal, the middle was un-conserved regulation domain with enriched in proline, serine and threonine residues, and C-terminal was the protein interaction domain. Results of the real-time PCR analysis showed that SmARF8 was detected in the roots, stems, leaves, bud flowers, flowers, anthesis, opening flowers, fruits and old fruits, and it was strongly expressed in immature fruits. 【Conclusion】 Results of this research indicate that SmARF8 is an ARF family gene in eggplant.

Key words: eggplant(Solanum melongena L.)')">eggplant(Solanum melongena L.), auxin response factor, gene cloning