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作物学报 ›› 2012, Vol. 38 ›› Issue (12): 2306-2311.doi: 10.3724/SP.J.1006.2012.02306

• 研究简报 • 上一篇    下一篇

苎麻ACC合酶基因(BnACS1)的克隆和表达分析

周精华1,余伟林1,邢虎成1,2,揭雨成1,2*,钟英丽1,3,敬礼恒1   

  1. 1 湖南农业大学苎麻研究所, 湖南长沙 410128; 2 湖南省种质资源创新与资源利用重点实验室, 湖南长沙 410128; 3 湖南农业大学生物科学与技术学院, 湖南长沙 410128
  • 收稿日期:2012-04-27 修回日期:2012-08-15 出版日期:2012-12-12 网络出版日期:2012-10-08
  • 通讯作者: 邢虎成, E-mail: xhcsoldier@163.com, Tel: 15274806570
  • 基金资助:

    本研究由国家自然科学基金项目(31101098), 湖南省教育厅科学研究优秀人才项目(10B051)和作物种质创新与资源利用国家重点实验室培育基地开放课题(10KFXW09)资助。

Cloning and Characterization of ACC Synthase Gene (BnACS1) from Ramie (Boehmeria nivea)

ZHOU Jing-Hua1, YU Wei-Lin1, XING Hu-Cheng1,2,*, JIE Yu-Cheng1,2, ZHONG Ying-Li1,3,JING Li-Heng1   

  1. 1 Institute of Ramie, Hunan Agricultural University, Changsha 410128, China; 2 Hunan Provincial Key Laboratory of Corp Germplasm Innovation and Utilization, Changsha 410128, China; 3 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China
  • Received:2012-04-27 Revised:2012-08-15 Published:2012-12-12 Published online:2012-10-08
  • Contact: 邢虎成, E-mail: xhcsoldier@163.com, Tel: 15274806570

摘要:

根据苎麻转录组测序中的ACS基因片段, 利用RT-PCR结合RACE技术从湘苎3号中克隆了该基因的全长cDNA序列, 命名为BnACS1, GenBank中的登录号为JQ970520。该基因的cDNA序列全长为1 674 bp, 其开放阅读框长1 470 bp, 编码489个氨基酸多肽, 预测其分子量和等电点分别为54.55 kD6.37, 与苹果(AB034993)、枇杷(GQ370520)、苦瓜(AF248734)、牡丹(DQ337250)、烟草(AY426755)和胡杨(AB033502) ACS基因核苷酸序列的相似性分别为74%74%72%71%70%70%, 氨基酸序列的相似性分别为75%74%71%71%70%74%。半定量RT-PCR分析表明, BnACS1基因在根、茎、茎尖、叶片、雌花和雄花中均有表达, 其中在根和雄花中表达较高, 在茎中表达最低。荧光定量PCR分析表明, BnACS1ABA和干旱的诱导表达上调, 不受高盐诱导表达。

关键词: 苎麻, 乙烯, ACC合成酶, ACS, 克隆表达

Abstract:

According to the ACS gene sequence from ramie transcriptome, we designed primers and cloned a full-length sequence of ACS gene cDNA from Xiangzhu 3 by RT-PCR and RACE methods, named as BnACS1, with the accession number of JQ970520 in GenBank. The full length sequence and the ORF of the BnACS1 gene were 1 674 bp and 1 470 bp, respectively, which encoded 489 amino acids. The molecule weight was 54.55 kD and the pI was 6.37. The similarity comparison revealed that the gene nucleotide sequence shared 74%, 74%, 72%, 71%, 70%, and 70% of homology with the Malus × domestica (AB034993), Eriobotrya japonica (GQ370520), Momordica charantia (AF248734), Paeonia suffruticosa (DQ337250), Nicotiana attenuate (AY426755), and Populus euphratica (AB033502) ACS gene, and the similarity of the amino acid sequences with that of those species was 75%, 74%, 71%, 71%, 70%, and 74%, respectively. The results of semi-quantitative RT-PCR showed that the BnACS1 expressed in root, stem, shoot tip, blade, female flower andmale flower, with the higher expression level in root and male flower, while the lowest expression level in stem. The results of real-time PCR showed that the BnACS1 was induced by dehydration and ABA, but not by high salt.

Key words: Ramie, Ethylene, ACC Synthase, ACS, Cloning and expression

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