Effects of the D3 gene on transcriptional regulation and its role in defense responses

doi:10.3969/j.issn.1004-1524.20221667

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (12): 2763-2774.DOI: 10.3969/j.issn.1004-1524.20221667

• Crop Science • Previous Articles Next Articles

Effects of the D3 gene on transcriptional regulation and its role in defense responses

XU Yue¹^,²(), WANG Shaomin³, TAN Xiaojing¹^,², LUO Yingjie¹^,², CHANG Jingyi², DENG Hui², LIU Xiuli², CUI Weijun², ZHOU Jie², WU Yueyan¹, YAN Chengqi¹^,⁴^,^*(), WANG Xuming²^,^*()

1. College of Biology and Environment, Zhejiang Wanli University, Ningbo 315100, Zhejiang, China
2. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Agricultural Technology Extension and Service Station of Yuyao City, Yuyao 315400, Zhejiang, China
4. Ningbo Academy of Agricultural Sciences, Ningbo 315100, Zhejiang, China

Received:2022-11-22 Online:2023-12-25 Published:2023-12-27

Abstract

Abstract:

Bacterial leaf blight is a bacterial disease caused by Xanthomonas oryzae pv. oryzae (Xoo). Bacterial leaf blight can severely reduce rice yields. With the development of molecular biotechnology, editing potential disease resistance genes has become a method to improve rice disease resistance and reduce the loss of the disease. In this study, D3 gene edited lines and wild-type inoculation experiments were used to determine D3 gene resistance and high-throughput sequencing technology was employed to analyze the transcriptomes of three different D3 gene-edited lines, and the results showed that compared with the wild type, D3 gene was significantly induced, while the D3 edited lines showed different degrees of blight resistance and a total of 8 184 DEGs were identified at the transcriptome level, including 4 201 up-regulated genes and 3 983 down-regulated genes, among which 359 DEGs were found simultaneously differentially expressed in the three groups. After that, this part of differentially expressed genes (DEGs) was verified by quantitative PCR. GO functional enrichment analysis showed that the genes related to biological pathways, cellular components and molecular functions were significantly differentially expressed in D3 gene-edited lines compared with the wild-type. KEGG pathway enrichment analysis found that DEGs were significantly enriched in plant-pathogen interaction, plant hormone signal transduction and phenylpropane biosynthesis pathways. Further experiments showed that, compared with the wild type, pathogen-related genes (PRs) in D3 gene-edited lines were significantly induced after inoculation with Xoo, and D3 gene-edited lines showed different degrees of bacterial blight resistance, thus further confirmed the role of D3 in bacterial blight resistance. This study enriches the role of the rice D3 gene in affecting transcriptional regulation, especially its impact on plant disease resistance-related genes. At the same time, this study expands the ideas for the research on the disease resistance mechanism of D3 gene.

Key words: rice, bacterial blight, D3 gene, transcriptome analysis, differentially expressed genes

CLC Number:

S435.111.1

XU Yue, WANG Shaomin, TAN Xiaojing, LUO Yingjie, CHANG Jingyi, DENG Hui, LIU Xiuli, CUI Weijun, ZHOU Jie, WU Yueyan, YAN Chengqi, WANG Xuming. Effects of the D3 gene on transcriptional regulation and its role in defense responses[J]. Acta Agriculturae Zhejiangensis, 2023, 35(12): 2763-2774.

Figures/Tables 10

Table 1 D3 gene-edited transgenic GEM line

株系 Line	突变方式 Mutation	处理组 Treatment group	对照组 Control group(Nip)
GEM1	-AT	A-1、A-2、A-3	CK-1
GEM2	-AGT	B-1、B-2、B-3	CK-2
GEM3	-CAGTTGCG	C-1、C-2、C-3	CK-3

Table 2 Primers used for qRT-PCR

基因Gene	正向引物Forward primer(5'→3')	反向引物Reverse primer(3'→5')
Os02g0462800	AAGGCTCGCCTTATCCACGC	CAGTCGTCCTGTGCTCCGTC
Os01g0884300	GAAGCCGGTGGCGATCAAGA	GAAGCCGGTGGCGATCAAGA
Os04g0118800	TGGTTTTGCAAACGAGCGAG	ACCTTCACACCTGCCAACAA
Os06g0521900	CACAGCTGCGAGCACTACGA	TTGTCGAACTTGGCTGGCGT
Os12g0528801	CGCCACCTTCTCTCCGGTTC	GCTGAGCAGCAGGAGACACT

Fig.1 The expression levels of the D3 gene induced by Xoo at different time points

Fig.2 Edit plots of disease spot phenotypes between lines and Nip

Fig.3 Overall characterization of sample relationship A, Venn diagrams of differentially expressed genes among the gene editing groups and the control group; B, Numbers of differentially expressed genes among the gene editing groups and the control group; C, qRT-PCR analysis of relative expression levels of differentitally genes among the gene editing groups and the control group.

Fig.4 GO enrichment analysis of DEGs with significant expression difference These genes with significant difference are summarized into three main GO categories (biological processes, molecular function, and cellular components). Orange is the up regulated, green is the down regulated.

Fig.5 KEGG enrichment pathway analysis of differentially expressed genes among gene editing treatment groups and control group The size of the circle represents the number of DEG enriched in the corresponding pathway, and the color of the circle represents the corresponding pathway.

Fig.6 Analysis of differentially expressed genes among gene editing treatment groups and control group in the phenylpropane biosynthetic pathway

Fig.7 Differentially expressed gene analysis among gene editing treatment groups and control group in plant pathogen interactions

Fig.8 Analysis of differentially expressed genes among gene editing treatment groups and control group in signal transduction pathways

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Effects of the D3 gene on transcriptional regulation and its role in defense responses

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