Genetic Transformation and Function Analysis of PsnHB13 and PsnHB15 of Populus simonii × Populus nigra

doi:10.7525/j.issn.1673-5102.2023.03.003

Abstract

Abstract:

Transcriptome analysis in Populus simonii × P. nigra overexpressing PsnCycD1；1showed PsnHB3 and PsnHB15 are differential genes. Analysis of PsnHB13 and PsnHB15 conservative domains through InterPro tool. The STRING software was utilized to explore PsnHB13 and PsnHB15 proteins interaction network analysis. Yeast two hybrid was used to verify the interaction proteins，calculate the ratio of leaf length to width，the ratio of dry weight to fresh weight of leaves and stems of transgenic plants，and analyze the transcriptome of PsnHB13 overexpression plants. Conserved domain analysis showed that PsnHB13 and PsnHB15 genes belonged two subfamilies，showing that PsnHB13 mainly contains the HDZip I domain，and PsnHB15 mainly contains the HDZip Ⅲ domain. ，Each gene is screened to 10 interacting genes. Moreover，PsnHB15 has a higher probability of interaction with family proteins. Yeast two-hybrid assays demonstrated that PsnHB15 interacts with PsnHB13 and PsnHB13 interacts with PsnCycD1；1. The aspect ratio of leaves in transgenic lines overexpressing PsnHB13 and PsnHB15 increased in the early seedling stage.The ratio of dry weight to fresh weight in stems of PsnHB13 overexpression lines increased significantly（P<0.05）. The analysis of the transcriptome revealed that Gene Ontologies（GO） enrichment analysis identified 3 significantly enriched GO terms，including response to chemical，response to organic substance，regulation of RNA metabolic process. KEGG pathway enrichment analysis demonstrated that transcription factors，plant hormone signal transduction and Cytochrome P450 were significantly enriched. These differential genes，including MADS-box transcription factor，MYBP transcription factor，ERF1 transcription factor，GH3 auxin responsive genes，SAUR protein family. PsnHB13 and PsnHB15 play an important role in the growth and development of Populus simonii × P. nigra，and are key genes to explore the growth law of plants and reveal the relationship between cell cycle and growth regulation.

Key words: PsnHB13, PsnHB15, Populus simonii × P. nigra, bioinformatics-based analyses, transcriptome analysis

CLC Number:

S792.11

Zhanmin ZHENG, Yubing SHANG, Guangbo ZHOU, Di XIAO, Yi LIU, Xiangling YOU. Genetic Transformation and Function Analysis of PsnHB13 and PsnHB15 of Populus simonii × Populus nigra[J]. Bulletin of Botanical Research, 2023, 43(3): 340-350.

Figures/Tables 11

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引物名称 Primer name	碱基序列 Sequence（5′→3′）	用途 Application
PsnHB13-XbaⅠ-F	5′-ATCTCTAGACACTAGTCTAGGTGATGACGTGC-3′	克隆PsnHB13基因 Cloning of the gene PsnHB13
PsnHB13-KpnⅠ-R	5′ATCGGTACCGGCATTCCAGGCTCATTTTCTTAGTTG3′	克隆PsnHB13基因 Cloning of the gene PsnHB13
PsnHB15-NdeⅠ-F	5′ATCCATATGATGATGGCAATGTCCTGCAAGG3′	克隆PsnHB15基因 Cloning of the gene PsnHB13
PsnHB15-BamHⅠ-R	5′ATCGGATCCTCAAACAAAAGACCAGTTTATAAAC3′	克隆PsnHB15基因 Cloning of the gene PsnHB13
pROKⅡ-F	5′AAGACCGGCAACAGGATTC3′	pROKⅡ载体引物 Primer of pROKⅡ vector
pROKⅡ-R	5′CGCACAATCCCACTATCCTT3′	pROKⅡ载体引物 Primer of pROKⅡ vector
PsnCycD1；1-RT-F	5′CCTCTGGTTCCTTCTCTATTGG3′	qRT-PCR检测表达量引物 Detection primer of qRT-PCR
PsnCycD1；1-RT-R	5′CAGAAACCCAGTATAGGCTCC3′	qRT-PCR检测表达量引物 Detection primer of qRT-PCR
PsnHB13-RT-F	5′TGAAAAGCAGAGAACCGACTG3′	qRT-PCR检测表达量引物 Detection primer of qRT-PCR
PsnHB13-RT-R	5′GCAATACCTGTAGGCCTGATAG3′	qRT-PCR检测表达量引物 Detection primer of qRT-PCR
PsnHB15-RT-F	5′GAATCCTCAACTGTACTCGCTC3′	qRT-PCR检测表达量引物 Detection primer of qRT-PCR
PsnHB15-RT-R	5′AGAGCCTCATTAAAACCCCTG3′	qRT-PCR检测表达量引物 Detection primer of qRT-PCR
Psnactin-RT-F	5′AATACCCCATTGAGCACGG3′	qRT-PCR内参基因引物 Reference primer of qRT-PCR
Psnactin-RT-R	5′ACTCACACCATCACCAGAATC3′	qRT-PCR内参基因引物 Reference primer of qRT-PCR

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