Stress Tolerance Functional Analysis of the High Expression Heat Shock Protein HbHSP90.4 Gene from the Latex of Hevea brasiliensis

doi:10.7525/j.issn.1673-5102.2022.06.012

Abstract

Abstract:

In order to analyze the role of Heat Shock Protein 90（HSP90） in stress and phytohormone signaling transduction in rubber tree， the full-length cDNA sequence of the HbHSP90.4 gene was cloned from rubber tree Reyan73397 latex by PCR， and HbHSP90.4 gene contained a 2 451 bp open reading frame（ORF） and encoded 816 amino acids. The results of bioinformatics analysis showed that HbHSP90.4 contained the HSP90 superfamily and HATPase superfamily domains， which was a member of the HSP90 family. Phylogenetic analysis revealed that this protein was closely related to cassava MeHSP90. Subcellular localization predictions showed that the HbHSP90.4 gene was localized in endoplasmic reticulum. The qRT-PCR results showed that the HbHSP90.4 gene was mainly expressed in rubber latex. Drought， cold， powdery mildew infection， H₂O₂ and MeJA treatments promoted the up-regulation of the HbHSP90.4 gene in latex respectively， while its expression was significantly down-regulated in ETH， SA and ABA treatments respectively. The plant expression vector HbHSP90.4-mScarlet was also constructed to prepare the material for further transgenic plants. This study laid a solid foundation for the elucidation of the molecular regulation of the latex HbHSP90.4 gene in response to rubber tree stress and the phytohormone signaling pathway.

Key words: Rubber tree, HbHSP90.4, bioinformatics, expression analysis, expression vector construction

CLC Number:

S794.1

Huafeng CHEN, Longjun DAI, Mingyang LIU, Bingbing GUO, Hong YANG, Lifeng WANG. Stress Tolerance Functional Analysis of the High Expression Heat Shock Protein HbHSP90.4 Gene from the Latex of Hevea brasiliensis[J]. Bulletin of Botanical Research, 2022, 42(6): 1023-1032.

Figures/Tables 10

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名称 Name	网址 Website
NCBI ORF Finder	www.ncbi.nlm.nih.gov/orffinder/
NCBI smart BLAST	blast.ncbi.nlm.nih.gov/Blast.cgi
ExPASy ProtParam tool	web.expasy.org/protparam/
TMHMM Server v.2.0	www.cbs.dtu.dk/services/TMHMM-2.0/
DeepLoc1.0	www.cbs.dtu.dk/services/DeepLoc/
SignalP-5.0 Server	services.healthtech.dtu.dk/service.php?SignalP-5.0
SWISS-MODEL	swissmodel.expasy.org/
PSIPRED Workbench	bioinf.cs.ucl.ac.uk/psipred/

引物名称 Primer name	序列 Sequence（5′→3′）	用途 Application
HbHSP90.4-F	ATCTATTGAACGGGAGA	编码区PCR扩增 ORF PCR amplification
HbHSP90.4-R	GCACTAAGGGCCAAG	编码区PCR扩增 ORF PCR amplification
HbHSP90.4-QF	GGACGGGAATGTAAAGCAAAC	荧光定量 Fluorescence quantitative
HbHSP90.4-QR	GCTGTAGATACGGGAAGCAAAA	荧光定量 Fluorescence quantitative
HbActin-F	GATGTGGATATCAGGAAGGA	内参基因 Reference gene
HbActin-R	CATACTGCTTGGAGCAAGA	内参基因 Reference gene
HbHSP90.4-MF	acgggggacgagctcggtaccATGAGGAAGTGGACGATCCCTT	表达载体构建 Construction of expression vector
HbHSP90.4-MR	catgtcgactctagaggatccCAACTCATCCTTGACAGCAGAAGA	表达载体构建 Construction of expression vector

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