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  • About author: Yang Yuning(1996—),female,postgraduate,Mainly engaged in larch genetic breeding research. Supported by:
    Natural Science Foundation of Heilongjiang Province(C2017008);National Science and Technology Major Project(2018ZX 08020003-001-001)

    摘要:

    为了解转录因子 bHLH 在长白落叶松( Larix olgensis )中的功能,探究该基因在长白落叶松不同组织中及不同逆境胁迫下的表达特性,从长白落叶松根、茎和叶3个不同部位的转录组数据中获得 bHLH34 基因全长序列,并设计引物,克隆得到长白落叶松 bHLH34 基因,其完整的开放阅读框(ORF)长度为696 bp,共编码231个氨基酸。构建亚细胞定位表达载体,瞬时转化毛果杨( Populus trichocarpa )原生质体,在激光共聚焦显微镜下观察显示, LobHLH34 基因定位在细胞核内。系统进化树分析结果显示,长白落叶松与云杉( Picea asperata )、卷柏( Selaginella tamariscina )树种该基因的亲缘关系较近。利用qRT-PCR技术分析了 bHLH34 基因在长白落叶松中的组织表达特异性和应对非生物胁迫的表达。结果表明 LobHLH34 基因在长白落叶松的根、茎、叶中均有表达,其中在茎部表达量最低,在叶中相对表达量最高。 LobHLH34 基因在NaCl、PEG和ABA处理时,不同器官中的表达量也有所不同。推测长白落叶松 bHLH34 基因参与了植物的生长、发育、响应逆境胁迫的过程,且在不同器官中具有特异性。

    Abstract:

    In order to understand the function of transcription factor LobHLH34 and explore its expression pattern in different tissues and under different stress conditions, the full-length sequence of LobHLH34 gene was obtained from transcriptome data of root, stem and leaf of Larix olgensis , by using the primers designed. The complete open reading frame(ORF) of LobHLH34 gene was 696 bp and encoded 231 amino acids. Subcellular expression vector was constructed and transformed into protoplasts of Populus trichocarpa . The result showed that the LobHLH34 gene was located in the cell nucleus under the laser confocal microscope. The results of phylogenetic tree analysis showed that larch had the closest genetic relationship with Picea asperata and Selaginella tamariscina . By qRT-PCR, the tissue-specific expression of LobHLH34 gene and its response to abiotic stress were analyzed respectively. The results showed that LobHLH34 gene was expressed in the roots, stems and leave, and the lowest expression level was found in the stem and the highest relative expression in the leaf respectively. The expression level of LobHLH34 was different in different organs under NaCl, PEG and ABA stress. It was speculated that LobHLH34 gene could be involved in the process of plant growth, development and respond to stress, and it had special expressional pattern in different organs.

    Key words: Larix olgensis , LobHLH34 , gene cloning, bioinformatics analysis, expression analysis

    杨宇宁, 董昊, 董实伟, 王乃锐, 宋跃, 张含国, 李淑娟. 长白落叶松转录因子 LobHLH34 克隆及表达分析[J]. 植物研究, 2022, 42(1): 112-120.

    Yuning Yang, Hao Dong, Shiwei Dong, Nairui Wang, Yue Song, Hanguo Zhang, Shujuan Li. Cloning and Expression Analysis of Transcription Factor LobHLH34 from Larix olgensis [J]. Bulletin of Botanical Research, 2022, 42(1): 112-120.

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