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Proteomics analysis of a tobacco variety resistant to brown spot disease and functional characterization of NbMLP423 in Nicotiana benthamiana

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Abstract

Background

Tobacco brown spot disease is an important disease caused by Alternaria alternata that affects tobacco production and quality worldwide. Planting resistant varieties is the most economical and effective way to control this disease. However, the lack of understanding of the mechanism of tobacco resistance to tobacco brown spot has hindered progress in the breeding of resistant varieties.

Methods and results

In this study, differentially expressed proteins (DEPs), including 12 up-regulated and 11 down-regulated proteins, were screened using isobaric tags for relative and absolute quantification (iTRAQ) by comparing resistant and susceptible pools and analyzing the associated functions and metabolic pathways. Significantly up-regulated expression of the major latex-like protein gene 423 (MLP 423) was detected in both the resistant parent and the population pool. Bioinformatics analysis showed that the NbMLP423 cloned in Nicotiana benthamiana had a similar structure to the NtMLP423 in Nicotiana tabacum, and that expression of both genes respond rapidly to Alternaria alternata infection. NbMLP423 was then used to study the subcellular localization and expression in different tissues, followed by both silencing and the construction of an overexpression system for NbMLP423. The silenced plants demonstrated inhibited TBS resistance, while the overexpressed plants exhibited significantly enhanced resistance. Exogenous applications of plant hormones, such as salicylic acid, had a significant inducing effect on NbMLP423 expression.

Conclusions

Taken together, our results provide insights into the role of NbMLP423 in plants against tobacco brown spot infection and provide a foundation for obtaining resistant tobacco varieties through the construction of new candidate genes of the MLP subfamily.

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Data availability

Full length sequence of NtMLP423 and NbMLP423 was submitted in GenBank with the accession number OM672246 and OM672247, respectively. All the raw data is publicly available and included in the manuscript also. Further details can be requested at zhangyu02@caas.cn.

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Acknowledgements

Authors are grateful to teachers and the colleagues who have contributed at any level to this research, with special thanks to Shanshan Liu and Chenyu Su for writing most of this manuscript.

Funding

This research was supported by a grant from Sichuan Science and Technology Project (SCYC201902, SCYC202001), Science and Technology Project (110202103014), Guangxi Science and Technology Project (2020450000340001-B01). The funders had no role in study design, data collection and analysis, decision to publish or in preparation of the manuscript.

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Authors and Affiliations

  1. Tobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao, 266101, People’s Republic of China

    Yu Zhang, Jie Wang, Caihong Jiang, Lirui Cheng, Chenggang Luo & Yuanying Wang

  2. Sichuan Tobacco Company, Chengdu, 610000, People’s Republic of China

    Yong Xiao & Shiping Guo

  3. Raw Material Technology Center of Guangxi Tobacco, Nanning, 530000, China

    Haijiang Jia

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  1. Yu Zhang
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Contributions

YZ and JW performed and analyzed most of lab experiment data, YX, CJ, LC and SG performed most of statistical analyses and sequencing data analyses. CL, YW and HJ drafted the manuscript. All authors discussed the results and commented on the paper.

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Correspondence to Chenggang Luo, Yuanying Wang or Haijiang Jia.

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Zhang, Y., Wang, J., Xiao, Y. et al. Proteomics analysis of a tobacco variety resistant to brown spot disease and functional characterization of NbMLP423 in Nicotiana benthamiana. Mol Biol Rep 50, 4395–4409 (2023). https://doi.org/10.1007/s11033-023-08330-7

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