Cloning and Expression of the EsxA Gene and Analysis of the Growth-Promoting&amp;nbsp;Effects of the Encoded Protein on Rice Seedlings

Yu, Wen; Wang, Xin; Tang, Yi Cong; Yan, Feng Chao; Liu, Wen Zhi; Zheng, Gui Ping; Yin, Dong

doi:10.21203/rs.3.rs-97808/v1

Cited by 2 publications

(4 citation statements)

References 12 publications

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“…On the basis of the transcriptome sequencing data and the results of our previous studies (Yu et al 2021b ), we speculate that EsxA helps plants establish the first defense response by inducing the ROS burst and HR and by activating the transcription of cell structure-related genes to strengthen the cell wall (e.g., increased PAL levels) (Table 1 , Fig. 8 ).…”

Section: Discussionmentioning

confidence: 64%

“…8 ) and protein phosphorylation, resulting in the formation of second messengers. Among which, POD activity was confirmed increased in EsxA treated rice plant (Yu et al 2021b ). The amplified signal will induce the ROS burst in other cells via signal transduction pathways.…”

Section: Discussionmentioning

confidence: 97%

“…In an earlier study, we cloned an EsxA gene from the plant growth-promoting rhizobacterium Paenibacillus terrae NK3-4 and expressed it in Pichia pastoris , with the secreted protein inducing a hypersensitive response (HR) and reactive oxygen species (ROS) burst in leaves as well as enhanced rice plant growth (Yu et al 2021 ). To further assess whether EsxA can induce plant disease resistance, we used various methods to treat rice with EsxA to evaluate its effect on rice resistance to seedling blight.…”

Section: Introductionmentioning

confidence: 99%

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Protein Elicitor EsxA Induces Resistance to Seedling Blight and PR Genes Differential Transcription in Rice

Yan

Zheng

et al. 2021

Rice

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Protein elicitors can induce plant systemic resistance to pathogens. In an earlier study, we cloned an EsxA gene from the plant growth-promoting rhizobacterium Paenibacillus terrae NK3-4 and expressed it in Pichia pastoris. In addition to being important for the pathogenicity of animal pathogens, EsxA can also induce an immune response in animals. While, we found the exogenously expressed EsxA has the activity of elicitor, which can trigger hypersensitive response and reactive oxygen species burst in leaves as well as enhanced rice plant growth. The effects of EsxA on seedling blight (Fusarium oxysporum) resistance and gene transcription, including pathogenesis-related (PR) genes in rice were evaluated. The germination rate was 95.0% for seeds treated with EsxA and then inoculated with F. oxysporum, which was 2.8-times higher than that of F. oxysporum-infected control seeds that were not treated with EsxA (Con). The buds and roots of EsxA-treated seedlings were 2.4- and 15.9-times longer than those of Con seedlings. The plants and roots of seedlings dipped in an EsxA solution and then inoculated with F. oxysporum were longer than those of the Con seedlings. Theplant length, number of total roots, and number of white roots were respectively 23.2%, 1.74-times, and 7.42-times greater for the seedlings sprayed with EsxA and then inoculated with F. oxysporum than for the Con seedlings. The EsxA induction efficiency (spray treatment) on seedling blight resistance was 60.9%. The transcriptome analysis revealed 1137 and 239 rice genes with EsxA-induced up-regulated and down-regulated transcription levels, respectively. At 48 h after the EsxA treatment, the transcription of 611 and 160 genes was up-regulated and down-regulated, respectively, compared with the transcription levels for the untreated control at the same time-point. Many disease resistance-related PR genes had up-regulated transcription levels. The qPCR data were consistent with the transcriptome sequencing results. EsxA triggered rice ISR to seedling blight and gene differential transcription, including the up-regulated transcription of rice PR genes. These findings may be relevant for the use of EsxA as a protein elicitor to control plant diseases.

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

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Protein Elicitor EsxA Induces Resistance to Seedling Blight and PR Genes Differential Transcription in Rice

Yan

Zheng

et al. 2021

Rice

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“…Genomic DNA was isolated from P. terrae NK3-4 using a Bacterial Genomic DNA Extraction kit (Beijing Solarbio Technology Co., Ltd., Beijing, China), and the DNA quality was examined using a ultra-micro ultraviolet spectrophotometer (NanoDrop one, Thermo Fisher Scientific, Waltham, Massachusetts, USA), according to the method described in our previously published article [15]. The full-length EndoAI coding gene sequence was amplified by PCR using the following gene-specific primers (Fig.…”

Section: Cloning and Expression Of Endoaimentioning

confidence: 99%

Antitoxin EndoAI can induce disease resistance in tobacco as a protein elicitor

Yan

Liu

et al. 2021

Chem. Biol. Technol. Agric.

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Background The antitoxin EndoAI is a TA system component that directly inhibits EndoA activity in vitro. The targeted activation of a TA system represents a potentially novel antimicrobial or antiviral strategy. However, whether the antitoxin functions alone and can induce plant disease resistance remain unknown. Results An endoAI was previously identified in the genome of Paenibacillus terrae NK3-4. It underwent a bioinformatics analysis, cloned and expressed in Escherichia coli. Then the functions of EndoAI inducing plant resistance to diseases as an elicitor were evaluated. The results showed that, EndoAI is a stable, alkaline, and hydrophilic protein, with a J-shaped three-dimensional structure in the absence of a ligand. It was clustered on the same branch with an antitoxin from Paenibacillus polymyxa SC2. Ectopically expressed EndoAI triggered a reactive oxygen species burst and a positive hypersensitive response (HR) in tobacco leaves. Moreover, 2 μmol EndoAI induced HR activity in tomato leaf, and it remained active after a 15-min exposure at 4–50 °C, and pH 6–8. Additionally, EndoAI induced plant systemic resistance against Alternaria alternata and tobacco mosaic virus, and the up-regulated transcription of PR genes, including PR1a, PR1b, PR5, PDF1.2, COL1, NPR1, and PAL. Conclusions These results imply that EndoAI may enhance the disease resistance of tobacco by promoting a series of early defense responses and up-regulating PR gene expression. These findings are relevant for future investigations on the mechanism underlying the EndoAI–plant interaction that leads to enhanced disease resistance. Furthermore, the endoAI may be useful for developing effective biocontrol agents to protect plants from diseases. Graphical Abstract

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Cloning and Expression of the EsxA Gene and Analysis of the Growth-Promoting Effects of the Encoded Protein on Rice Seedlings

Cited by 2 publications

References 12 publications

Protein Elicitor EsxA Induces Resistance to Seedling Blight and PR Genes Differential Transcription in Rice

Protein Elicitor EsxA Induces Resistance to Seedling Blight and PR Genes Differential Transcription in Rice

Antitoxin EndoAI can induce disease resistance in tobacco as a protein elicitor

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