Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco

Abstract Background Photosynthetic pigments in higher plants, including chlorophyll (Chl) and carotenoids, are crucial for photosynthesis and photoprotection. Chl-deficient tobacco seedlings generally have a lower photosynthesis rate and higher nitrate-nitrogen (NO3-N) content, which causes a profou...

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Main Authors: Yuqing Feng, Yujing Li, Yuanyuan Zhao, Hongzhi Shi
Format: Article
Language:English
Published: BMC 2023-03-01
Series:BMC Plant Biology
Subjects:
Online Access:https://doi.org/10.1186/s12870-023-04169-z
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author Yuqing Feng
Yujing Li
Yuanyuan Zhao
Hongzhi Shi
author_facet Yuqing Feng
Yujing Li
Yuanyuan Zhao
Hongzhi Shi
author_sort Yuqing Feng
collection DOAJ
description Abstract Background Photosynthetic pigments in higher plants, including chlorophyll (Chl) and carotenoids, are crucial for photosynthesis and photoprotection. Chl-deficient tobacco seedlings generally have a lower photosynthesis rate and higher nitrate-nitrogen (NO3-N) content, which causes a profound influence on tobacco yield and quality. In this study, a stable albino leaf mutant (Al) and slight-green leaf mutant (SG) obtained from the common flue-cured tobacco (Nicotiana tabacum L.) cultivar ‘Zhongyan 100’ (ZY100) by mutagenesis with ethyl methanesulfonate (EMS) were used as materials. The differences between the Chl-deficient mutants and the wild-type (WT) were analyzed in terms of biomass, photosynthetic fluorescence parameters, and carbon- and nitrogen-related physiological parameters. RNA sequencing (RNA-seq) and weighted gene co-expression network analysis (WGCNA) were used to explore the key pathways and candidate genes regulating differentiated chlorophyll and nitrate content. Results The results showed that, when compared to the WT, the Chl content and biomass of mutant plants were considerably lower while the NO3-N content was substantially elevated. The net photosynthetic rate, photosynthetic fluorescence parameters, carbohydrate, soluble protein, and carbon- and nitrogen-related enzyme activities all decreased in leaves of mutants and the development of chloroplasts was abnormal. Applying more nitrogen improved the growth and development of mutants, whereas NO3-N content distinctively increased compared with that of the WT. Through transcriptome sequencing, the downregulated genes in mutants were enriched in plant hormone signal transduction and nitrogen metabolism, which are involved in pigment biosynthesis and the carbon fixation pathway. In addition, two hub genes and seven transcription factors identified from the blue module through WGCNA were likely to be key candidate factors involved in chlorophyll synthesis and nitrate accumulation. Conclusion Our results demonstrated that differences in chlorophyll and nitrate content were caused by the combined effects of chloroplast development, photosynthesis, as well as related biological activity. In addition, transcriptome results provide a bioinformatics resource for further functional identification of key pathways and genes responsible for differences in chlorophyll and nitrate content in tobacco plants.
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spelling doaj.art-f20ac2a5a70542e69945135f2d28e1fc2023-03-22T10:49:55ZengBMCBMC Plant Biology1471-22292023-03-0123111610.1186/s12870-023-04169-zPhysiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobaccoYuqing Feng0Yujing Li1Yuanyuan Zhao2Hongzhi Shi3College of Tobacco, Henan Agricultural UniversityCollege of Tobacco, Henan Agricultural UniversityCollege of Tobacco, Henan Agricultural UniversityCollege of Tobacco, Henan Agricultural UniversityAbstract Background Photosynthetic pigments in higher plants, including chlorophyll (Chl) and carotenoids, are crucial for photosynthesis and photoprotection. Chl-deficient tobacco seedlings generally have a lower photosynthesis rate and higher nitrate-nitrogen (NO3-N) content, which causes a profound influence on tobacco yield and quality. In this study, a stable albino leaf mutant (Al) and slight-green leaf mutant (SG) obtained from the common flue-cured tobacco (Nicotiana tabacum L.) cultivar ‘Zhongyan 100’ (ZY100) by mutagenesis with ethyl methanesulfonate (EMS) were used as materials. The differences between the Chl-deficient mutants and the wild-type (WT) were analyzed in terms of biomass, photosynthetic fluorescence parameters, and carbon- and nitrogen-related physiological parameters. RNA sequencing (RNA-seq) and weighted gene co-expression network analysis (WGCNA) were used to explore the key pathways and candidate genes regulating differentiated chlorophyll and nitrate content. Results The results showed that, when compared to the WT, the Chl content and biomass of mutant plants were considerably lower while the NO3-N content was substantially elevated. The net photosynthetic rate, photosynthetic fluorescence parameters, carbohydrate, soluble protein, and carbon- and nitrogen-related enzyme activities all decreased in leaves of mutants and the development of chloroplasts was abnormal. Applying more nitrogen improved the growth and development of mutants, whereas NO3-N content distinctively increased compared with that of the WT. Through transcriptome sequencing, the downregulated genes in mutants were enriched in plant hormone signal transduction and nitrogen metabolism, which are involved in pigment biosynthesis and the carbon fixation pathway. In addition, two hub genes and seven transcription factors identified from the blue module through WGCNA were likely to be key candidate factors involved in chlorophyll synthesis and nitrate accumulation. Conclusion Our results demonstrated that differences in chlorophyll and nitrate content were caused by the combined effects of chloroplast development, photosynthesis, as well as related biological activity. In addition, transcriptome results provide a bioinformatics resource for further functional identification of key pathways and genes responsible for differences in chlorophyll and nitrate content in tobacco plants.https://doi.org/10.1186/s12870-023-04169-zFlue-cured tobaccoChlorophyll deficientNitrateRNA-sequencingCo-expression network
spellingShingle Yuqing Feng
Yujing Li
Yuanyuan Zhao
Hongzhi Shi
Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco
BMC Plant Biology
Flue-cured tobacco
Chlorophyll deficient
Nitrate
RNA-sequencing
Co-expression network
title Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco
title_full Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco
title_fullStr Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco
title_full_unstemmed Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco
title_short Physiological, transcriptome and co-expression network analysis of chlorophyll-deficient mutants in flue-cured tobacco
title_sort physiological transcriptome and co expression network analysis of chlorophyll deficient mutants in flue cured tobacco
topic Flue-cured tobacco
Chlorophyll deficient
Nitrate
RNA-sequencing
Co-expression network
url https://doi.org/10.1186/s12870-023-04169-z
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AT yuanyuanzhao physiologicaltranscriptomeandcoexpressionnetworkanalysisofchlorophylldeficientmutantsinfluecuredtobacco
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