Comprehensive analysis of WRKY gene family in Sedum plumbizincicola responding to cadmium stress

doi:10.12302/j.issn.1000-2006.202201015

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2023, Vol. 47 ›› Issue (2): 49-60.doi: 10.12302/j.issn.1000-2006.202201015

Comprehensive analysis of WRKY gene family in Sedum plumbizincicola responding to cadmium stress

WANG Jianchao¹^,²(), QIU Wenmin²(), JIN Kangming², LU Zhuchou², HAN Xiaojiao², ZHUO Renying², LIU Xiaoguang³^,^*(), HE Zhengquan¹^,^*()

1. Key Laboratory of Three Gorges Regional Plant Genetics & Germplasm Enhancement (CTGU),Biotechnology Research Center,China Three Gorges University, Yichang 443002, China
2. Key Laboratory of Tree Breeding of Zhejiang Province, The Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou 311400, China
3. Tangshan Academy of Agriculture Science, Tangshan 063001, China

Received:2022-01-12 Revised:2022-06-25 Online:2023-03-30 Published:2023-03-28

Abstract

Abstract:

【Objective】 WRKY transcription factors play important regulatory roles for a plant abiotic stress response. However, little information is available about WRKY transcription factors in the hyperaccumulating plant Sedum plumbizincicola. The identification of SpWRKY gene family members and analysis of their expression patterns under cadmium stress can provide a reference for molecular cloning of SpWRKYs gene and functional characterization of cadmium tolerance.【Method】 In this study, the genome-wide identification and bioinformatics analysis of WRKY gene family members were carried out. Expression patterns of WRKY genes under cadmium stress were derived from transcriptome data and qPCR, and the roles of SpWRKY69 in cadmium tolerance were assessed by the heterologous expression in Arabidopsis thaliana.【Result】 There were 77 SpWRKYs identified in S. plumbizincicola, unevenly distributed on the chromosomes. Based on the phylogenetic analysis, the SpWRKY proteins were classified into three groups (Ⅰ-Ⅲ), among which the second group was further divided into five subgroups (Ⅱa-e). The synteny analysis showed that there were seven collinear gene pairs in the S. plumbizincicola and A. thaliana WRKY gene family and 53 collinear gene pairs for Kalanchoe fedtschenkoi; 19 pairs of SpWRKYs were identified as the segmental duplication. Cis-regulatory elements related to stress and hormone responses were found in the promoters of SpWRKYs. The expression profiles showed that the expression levels of the seven SpWRKYs were significantly up-regulated under cadmium stress, exhibiting a pattern of increasing initially and then decreasing. Over-expression of SpWRKY69 in A. thaliana showed that SpWRKY69 can enhance the transport rate of Cd ions to the shoot and play a negative role in regulating plant cadmium tolerance.【Conclusion】 Structural features of WRKY gene family members in S. plumbizincicola are similar to those of other species, and the fragment duplication is one of the main evolutionary forces. The relative expression levels of some members change significantly under cadmium stress, and SpWRKY69 can increase the transport rate of Cd ions to the shoot, and so other WRKY members may also be involved in the regulation of plant cadmium tolerance. These results provide a foundation for further functional characterization of SpWRKYs related to Cd tolerance.

Key words: Sedum plumbizincicola, WRKY transcription factor, cadmium stress, gene expression, Arabidopsis transformation

CLC Number:

S718

WANG Jianchao, QIU Wenmin, JIN Kangming, LU Zhuchou, HAN Xiaojiao, ZHUO Renying, LIU Xiaoguang, HE Zhengquan. Comprehensive analysis of WRKY gene family in Sedum plumbizincicola responding to cadmium stress[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2023, 47(2): 49-60.

Figures/Tables 10

Table 1

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基因名 gene ID	序列 sequence	用途 application
SpWRKY42-RT-F	GTTGCCGCAAATGTTTAGCC	伴矿景天筛选基因 qRT-PCR引物与内参引物
SpWRKY42-RT-R	ATAGCTGCTGTGAAATGCGG
SpWRKY72-RT-F	AGGGTTCTTGTTCGGACTGA
SpWRKY72-RT-R	AGCGCTTCTCTGAACCTTCT
SpWRKY40-RT-F	AGTGTAGCTGCTGAATCCGA
SpWRKY40-RT-R	AAAAGTCCAACTCCGGCCTA
SpWRKY46-RT-F	GACAACCGGCACTTTTGACT
SpWRKY46-RT-R	TAGGCTGTGTTGGAGTTGGT
SpWRKY39-RT-F	AGTTTACCGGCGAAGATGGA
SpWRKY39-RT-R	TCCAAATCCGACGACTCTGT
SpWRKY69-RT-F	CCAAACCATAAAGCCACCGA
SpWRKY69-RT-R	TCGCCGGAACTCTTACAACT
SpWRKY29-RT-F	TCACGGCGGAGATATCGAAA
SpWRKY29-RT-R	AAGAGCATCGATACGTCCGT
UBC9-F	TGGCGTCGAAAAGGATTCTGA
UBC9-R	CCTTCGGTGGCTTGAATGGATA
SpWRKY69-F	ATGGCCGTCGACCTCGT	SpWRKY69全长基因克隆引物与拟南芥内参引物
SpWRKY69-R	TCACGACGACTCTAGAATGAGT
actin-F	GCACCCTGTTCTTCTTACCG
actin-R	AACCCTCGTAGATTGGCACA

Comprehensive analysis of WRKY gene family in Sedum plumbizincicola responding to cadmium stress

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