棉花三螺旋转录因子GhGT29的克隆及功能分析

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2015-09-28 DOI:10.16288/J.YCZZ.15-320
L. Yue, L. Xiaodong, D. Yongmei, X. Zongming, Shou-yi Chen
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引用次数: 2

摘要

三螺旋转录因子是植物响应非生物胁迫的重要蛋白质。了解棉花三螺旋结构的分子机制将为通过基因工程提高棉花的抗逆性奠定基础。本研究通过生物信息学分析,利用反转录PCR技术从陆地棉中分离到一个编码三螺旋转录因子的基因。该基因被命名为GhGT29 (GenBank登录号:GhGT29)。JQ013097)全长1 092 bp,包含1 089 bp的开放阅读框,编码363个氨基酸,预测分子量为40.9 kDa,等电点为5.45。SMART分析显示GhGT29含有一个典型的SANT基序。系统发育分析表明,GhGT29属于Trihelix家族的SH4亚家族,与AtSH4-like1和AtSH4-like2亲缘关系最为密切。实时荧光定量PCR (qRT-PCR)分析显示,GhGT29受高盐、干旱、寒冷和脱落酸诱导。表达谱还显示GhGT29在根、茎、叶、花、胚珠(0 DPA)和纤维(12 DPA)等所有组织中均有组成性表达。GhGT29在花中的表达量最高,茎中的表达量最低。利用拟南芥原生质体分析系统,我们发现GhGT29蛋白位于细胞核中,具有反式活化活性。这些结果表明,GhGT29可能参与了旱棉胁迫信号通路中抗逆性相关基因的调控。
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Cloning and functional analysis of the cotton Trihelix transcription factor GhGT29
Trihelix transcription factors are important proteins involved in response to abiotic stresses in plants. Understanding the molecular mechanisms of Trihelix in cottons will lay the foundation to improve stress tolerance by gene engineering. In this study, a gene encoding Trihelix transcription factor was isolated in upland cottons using reverse transcription PCR according to bioinformatic analysis. The gene was named as GhGT29 (GenBank accession No. JQ013097), which was 1 092 bp, contained a 1 089 bp open reading frame and encoded a protein of 363 amino acids with a predicted molecular weight of 40.9 kDa and a isoelectric point of 5.45. SMART analysis showed GhGT29 contained one typical SANT motif. Phylogenetic analysis showed that GhGT29 belonged to the SH4 subfamily of the Trihelix family and was most closely related to AtSH4-like1 and AtSH4-like2. Quantitative real-time PCR (qRT-PCR) analysis revealed that GhGT29 was induced by high salt, drought, cold and abscisic acid. The expression profile also revealed that GhGT29 was constitutively expressed in all tested tissues, such as roots, stems, leaves, flowers, ovules (0 DPA) and fibers (12 DPA). The expression level of GhGT29 was the highest in flowers and the lowest in stems. Using the Arabidopsis protoplasts assay system, we found that the GhGT29 protein was located in cell nuclei and had trans-activation activity. These results revealed that GhGT29 might be involved in the regulation of stress resistance-related genes in stress signaling pathways in upland cottons.
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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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