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作物学报 ›› 2009, Vol. 35 ›› Issue (10): 1822-1830.doi: 10.3724/SP.J.1006.2009.01822

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

棉花赤霉素不敏感矮化GID1同源基因的克隆和表达分析

董静,尹梦回,杨帆,赵娟,覃珊,侯磊,罗明,裴炎,肖月华*   

  1. 西南大学生物技术中心,农业部生物技术和作物品质改良重点实验室,重庆400716
  • 收稿日期:2009-03-03 修回日期:2009-06-25 出版日期:2009-10-12 网络出版日期:2009-08-07
  • 通讯作者: E-mail: xiaoyuehua@swu.edu.cn
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2007AA10Z134)资助。

Cloning and Expression Profiling of Gibberellin Insensitive Dwarf GID1 Homologous Genes from Cotton

DONG Jing,YIN Meng-Hui,YANG Fan,ZHAO Juan,QIN Shan,HOU Lei,LUO Ming,PEI Yan,XIAO Yue-Hua*   

  1. Key Laboratory of Biotechnology and Crop Quality Improvement,Ministry of Agriculture,Biotechnology Research Center,Southwest University,Chongqing 400716,China
  • Received:2009-03-03 Revised:2009-06-25 Published:2009-10-12 Published online:2009-08-07
  • Contact: E-mail: xiaoyuehua@swu.edu.cn

摘要:

作为赤霉素(GA)受体,GID1是其重要的信号传导组分。为研究GA在棉花发育(特别是纤维发育)中的作用,本文在EST序列的基础上克隆了6个棉花GID1同源基因(GhGID1-1~6)。多重序列分析表明,棉花GID1同源蛋白GhGID1-1~6与拟南芥和水稻的GID1蛋白高度同源,具有多个与GADELLA蛋白的结合位点以及激素敏感酯酶家族保守域HGGGXSXG。定量RT-PCR分析表明,GhGID1-1~6基因在棉花不同器官和组织中表达水平有差异,其中GhGID1-1GhGID1-2在花器官中高表达,GhGID1-4基因在纤维和根中优势表达。胚珠体外培养基中外加GA使GID1同源基因表达水平发生变化,GhGID1-1GhGID1-2基因的表达水平明显GA抑制。比较发现,在胚珠和纤维中优势表达的GID1同源基因不同,GID1基因表达水平随发育时期变化的趋势也不相同,表明棉花胚珠和纤维具有相对独立的GA信号识别系统。

关键词: GID1, 棉花, 纤维发育, 表达特异性

Abstract:

Gibberellins (GA) play an important role in regulating cotton fiber development. As the GA receptor, GID1 is an essential component of GA signaling pathway. To advance our understanding of GA’s functions in cotton development (especially fiber development), we cloned six cotton GID1 homologous genes (GhGID1-1–6, accession No. are FJ790125FJ790130) by searching EST sequences, extending the flanking sequences by Y-shaped adaptor dependent extension (YADE) method and amplifying the full-length cDNA by 3'-RACE. Multiple sequence alignment indicated that the deduced GhGID1-16proteins shared high sequence identity with GID1s from other species and contained multiple binding sites with GA and DELLA protein as well as HGG and GXSXG domains conserved in hormone-sensitive lipase (HSL) family. Three amino acid residuals (G169, G196, and R251) with the rice GID1 function were also conserved in cotton GID1s. In the phylogenetic tree of GhGID1-16 and other GID1 proteins with known function, GhGID1-16 were clustered with 3 Arabidopsis GID1 proteins, and distantly related to GID1s from rice and S. moellendorffii. Quantitative RT-PCR was employed to detect the expression levels of GhGID1-1–6 in various organs and tissues. The transcripts of GhGID1-1–6 could be detected in all the organs and tissues investigated (including roots, hypocotyls, leaves, petals, anthers, ovules and fibers), although with various levels. GhGID1-1 and GhGID1-2 expressed mainly in floral organs, while GhGID1-4 expressed preferentially in fiber and root. Exogenous GA added in medium led to alteration of gene expression level in the in vitro cultivated ovules, and the expressions of GhGID1-1 and GhGID1-2 were obviously inhibited by GA. These results suggested that GhGID1-1–6 might encode biologically functional GID1 homologues involved in GA signaling in cotton. To further clarify GID1 genes related to fiber development, we compared GhGID1s’ expression in developing ovule (from 6 to 18 days post anthesis, DPA) and the attached fibers by quantitative RT-PCR. GhGID1-1 was predominantly expressed in ovules, and its expression level reached the climax at 10 DPA, and kept the high level at 14 DPA and 18 DPA, while in fibers, GhGID1-4 was the main GID1 homologue expressed, and its highest-level expression occurred at 6 DPA, and declined to a very low level at 14 DPA and 18 DPA.This result strongly suggested that these are relatively independent GA signaling systems in ovules and attached fibers.

Key words: GID1, Cotton, Fiber development, Expression Specificity

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