Cloning and Expression Analysis of Transcription Factor LobHLH34 from Larix olgensis

doi:10.7525/j.issn.1673-5102.2022.01.012

Abstract

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

CLC Number:

Q781

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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引物名称 Primer	引物序列 Sequence（5′→3′）	用途 Function
LobHLH34-F	CGGGATCCATGATGGGATCACCTCAGAGC	LobHLH34克隆 Cloning of LobHLH34
LobHLH34-R	GCGTCGACGGCAACAGGAGGCCTTAGC	LobHLH34克隆 Cloning of LobHLH34
LobHLH34-FQ	GGCTGTAGCTCATGCATTCTC	qRT-PCR引物 qRT-PCR primer
LobHLH34-RQ	ATTGCCATCCCAGGAAAAC	qRT-PCR引物 qRT-PCR primer
tubulin-F	AGCGACAATGAGGGAGTGC	qRT-PCR内参 Tubulin of qRT-PCR
tubulin-R	TACCGGCACCTGTCTCACTG	qRT-PCR内参 Tubulin of qRT-PCR

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