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Alternative TitleCloning, Bioinformatics Analysis and Expression Characteristic of Shikimate/Quinate Hydroxycinnamoyl Transferase Gene(GhHCT) in Cotton (Gossypium hirsutum L.)
肖向文; 朱奇朗; 刘海峰; 李雪源; 罗城; 王俊铎; 梁亚军; 陈贤丰; 李晓波
Source Publication农业生物技术学报

莽草酸/奎宁酸羟基肉桂酰转移酶(shikimate/quinate hydroxycinnamoyl transferase,HCT)是木质素生物合成途径中影响木质素G/S-单体生物合成的关键酶之一,棉纤维中木质素含量与纤维品质密切相关.本研究以开花后16 d(16 DPA)的棕色棉(Gossypium hirsutum L.)品种新彩棉6号纤维为材料,克隆了1个木质素代谢相关HCT类似基因,命名为GhHCT (GenBank登录号:KF749428).并通过实时荧光定量PCR技术检测了该基因在棉花不同器官及组织中的相对表达量.序列分析结果表明,该基因的开放读框(open reading frame,ORF)为l500 bp,编码499个氨基酸.生物信息学分析显示,推定的GhHCT蛋白质相对分子质量为55.2 kD,等电点为5.69.该蛋白氨基酸序列同其他物种HCT蛋白显示出很高同源性(82%~95%).GhHCT氨基酸序列包含一个HHAAD酰基转移酶功能结构域以及一个BAHD酰基转移酶基因家族的DFGWG区.系统进化树分析显示,GhHCT与槿麻(Hibiscus cannabinus)HcHCT基因有最近的亲缘关系(94.9%).实时荧光定量PCR(qRT-PCR)检测结果显示,GhHCT在新彩棉6号不同组织中都有表达,但在16 DPA的胚珠中表达最高,其次在花和茎中也有较高表达,在叶片中的表达量最低.本研究为进一步研究GhHCT基因的生物学功能及棉纤维中木质素生物合成代谢的作用机制研究提供前期基础资料,同时也为利用基因工程技术对天然彩色棉纤维品质进行遗传改良提供一定的线索及理论依据.

Other Abstract

Hydroxycinnamoyl CoA: Shikimate/quinate hydroxycinnamoyl transferase (HCT) is a key enzyme involved in the lignin biosynthesis pathway, which affects the biosynthesis of G/S monolignol. In cotton fibers, the content of lignin is closely related to fiber quality. A full- length gene from brown cotton (Gossypium hirsutum L.) cultivar Xincaimian 6 fiber at the 16 day post anthesis (16 DPA) was cloned encoding HCT ortholog, which was named GhHCT (GenBank Accession No. KF749428). The expression of GhHCT in different tissue and organs were examined through Real-time PCR technology. The full-length of the GhHCT ortholog consisted of a 1 500 bp open reading frame (ORF) encoding a polypeptide containing 499 amino acid residues. The molecular weight of deduced amino acids was 55.2 kD, with an isoelectric point (pl) of 5.69. The deduced amino acid sequence showed 82%?95% identities with HCTs of other plants. The deduced amino acid sequence of the GhHCT ortholog has a histidine containing motif (HHAAD), characteristic for acyl transfer catalysis. A second consensus sequence, a DFGWG block, was another acyl transferase of the BAHD gene family. Phylogenetic analysis showed the closest relationship (94.9%) with HCT of Hibiscus cannabinus (HcHCT). According to quantitative Real- time reverse transcription PCR (qRT- PCR) analysis, GhHCT transcript expressed in all the tissues and organs, but the highest expression was observed in ovules at 16 DPA. A relatively high expression was also detected in flowers and stems. The lowest level of expression of GhHCT was found in leaves. This study provides the crucial information for further study on the biological functions of GhHCT gene and molecular mechanism of lignin biosynthesis in cotton fiber. The study also provides clues and theoretical basis for improving fiber quality of naturally colored cotton through genetic engineering.

Keyword天然彩色棉 莽草酸/奎宁酸羟基肉桂酰转移酶 木质素生物合成 实时定量pcr 表达分析 Naturally Colored Cotton Hydroxycinnamoyl Coa Shikimate/quinate Hydroxycinnamoyl Transferase Enzyme (Hct) Lignin Biosynthesis Real-time Pcr Expression Analysis
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Document Type期刊论文
Affiliation中国科学院新疆理化技术研究所干旱区植物资源化学重点实验室;中国彩棉(集团)股份有限公司;新疆农业科学院经济作物研究所;Institute for Green Energy and Clean Environment
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肖向文,朱奇朗,刘海峰,等. 棉花莽草酸/奎宁酸羟基肉桂酰转移酶基因(GhHCT)的克隆、生物信息学分析及表达特性[J]. 农业生物技术学报,2014,22(5):572-579.
APA 肖向文.,朱奇朗.,刘海峰.,李雪源.,罗城.,...&李晓波.(2014).棉花莽草酸/奎宁酸羟基肉桂酰转移酶基因(GhHCT)的克隆、生物信息学分析及表达特性.农业生物技术学报,22(5),572-579.
MLA 肖向文,et al."棉花莽草酸/奎宁酸羟基肉桂酰转移酶基因(GhHCT)的克隆、生物信息学分析及表达特性".农业生物技术学报 22.5(2014):572-579.
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