Cloning and Drought Tolerance Expression Analysis of PtrWRKY51 Gene in Populus trichocarpa

doi:10.7525/j.issn.1673-5102.2022.06.010

Abstract

Abstract:

In order to clarify the function of PtrWRKY51 gene of Populus trichocarpa， the CDS of PtrWRKY51 gene was cloned using P. trichocarpa from Nisqually-1 strain as material. The function of PtrWRKY51 gene was studied by bioinformatics analysis， and assayed by yeast self-activation verification， and subcellular localization and simulated drought stress by real-time qPCR. The results showed that the CDS of PtrWRKY51 was 579 bp and encoded 192 aa. Bioinformatics analysis and subcellular localization experiments showed that PtrWRKY51 was a non-transmembrane alkaline unstable hydrophilic protein， located in the nucleus and contained a conserved domain unique to WRKY family， it was a WRKY transcription factor classified to class IIc. The yeast self-activation verification experiment showed that PtrWRKY51 gene could self-activation activity. qRT-PCR showed that the expression of this gene was significantly induced by 8% PEG6000 and reached the highest expression in stem and leaf after 12 h stress， while the roots appeared after 24 h stress. This study could provide reference for further study on stress tolerance and biological function of PtrWRKY51.

Key words: Populus trichocarpa, PtrWRKY51, bioinformatics, yeast self-activation, subcellular localization, drought tolerance expression analysis

CLC Number:

S792.11

Anying HUANG, Dean XIA, Yang ZHANG, Dongchen NA, Qing YAN, Zhigang WEI. Cloning and Drought Tolerance Expression Analysis of PtrWRKY51 Gene in Populus trichocarpa[J]. Bulletin of Botanical Research, 2022, 42(6): 1005-1013.

Figures/Tables 11

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引物名称 Primer	引物序列 Primer sequence（5′→3′）
PtrWRKY51-F	ATGGAGTACTATCCTCAGAACCCTA
PtrWRKY51-R	CTAGGAAAAGGAAGAAGAGTGGAG
PtrActin-F	AGGCAGGTTTCGCAGGAGATGA
PtrActin-R	TCCATCACCAGAATCCAGCACA
pGBKT7-PtrWRKY51-F	CATATGGCCATGGAGGCCGAATTCATGGAGTACTATCCTCAGAACCCTA
pGBKT7-PtrWRKY51-R	GCCGCTGCAGGTCGACGGATCCCTAGGAAAAGGAAGAAGAGTGGAG
pGBKT7-F	TCATCGGAAGAGAGTAGT
pGBKT7-R	TTCGTTTTAAAACCTAAGAGTC
PBI121-GFP-PtrWRKY51-F	ACGAGCTGTACAAGGGATCCATGGAGTACTATCCTCAGAACCCTA
PBI121-GFP-PtrWRKY51-R	CTCTAGACTCACCTAGGATCCCTAGGAAAAGGAAGAAGAGTGGAG
DL-PtrWRKY51-F	AGTTTCAGCCGTCGGATTATC
DL-PtrWRKY51-R	CTGACTTCTTTCCATACTTCCTCC

元件名称 Element names	基序序列 Motif sequences	数量 /个 Numbers
ABRE	ACGTG	4
ARE	AAACCA	1
CGTCA-motif	CGTCA	3
TCA-element	CCATCTTTTT	2
TC-rich repeats	GTTTTCTTAC	3
TGACG-motif	TGACG	3

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