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. 2024 Dec 19;24(1):1211.
doi: 10.1186/s12870-024-05917-5.

Revealing the key role of the folate synthesis regulatory gene DHNA in tobacco leaf yellowing based on BSA-seq, RNA-seq, and proteomic sequencing

Qiuhe Pan  1   2 , Rongli Jia  3 , Kai Pi  4 , Qin Tang  1   2 , Jingyao Zhang  1   2 , Ying Huang  1   2 , Renxiang Liu  5   6
Affiliations

Revealing the key role of the folate synthesis regulatory gene DHNA in tobacco leaf yellowing based on BSA-seq, RNA-seq, and proteomic sequencing

Qiuhe Pan et al. BMC Plant Biol. .

Abstract

Background: The degree of yellowing in tobacco leaves is an important indicator for determining the maturity and harvesting time of tobacco leaves. Decreasing chlorophyll levels helps speed up the ripening process of tobacco leaves for easier mechanical harvesting. Identifying and utilizing genes that regulate yellowing in tobacco leaves are crucial for developing tobacco varieties suitable for mechanized harvesting and understanding the molecular processes that control leaf color changes.

Results: In this study, the phenotypes of the yellow-leaf K326 and K326 varieties were analysed, and it was observed that the yellow-leaf K326 variety exhibited a distinct yellow leaf phenotype with a significant reduction in chlorophyll content. Subsequently, using a combination of BSA-seq, transcriptomic sequencing (RNA-seq), and proteomic sequencing approaches, we identified the candidate gene Nitab4.5_0008674g0010 that encodes dihydroneopterin aldolase as a factor associated with tobacco leaf yellowing. Finally, by measuring the folate content in K326 and Huangye K326, the folate content in Huangye K326 was observed to be significantly lower than that in K326, thus indicating that folate synthesis plays a crucial role in phenotypic changes in tobacco yellow leaves.

Conclusions: This study is the first to use BSA-seq combined with RNA-seq and proteomic sequencing to identify candidate genes in tobacco yellow leaves. The results provide a theoretical basis for the analysis of the mechanism of tobacco yellow leaf mutations.

Keywords: BSA-seq; Chlorophyll content; Leaf yellowing; Proteome; RNA-seq; Tobacco.

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
A Leaf colour phenotypes of K and KY; B The total chlorophyll, chlorophyll a, chlorophyll b, and carotenoid content of K and KY. C Comparison chart of K and KY budding periods
Fig. 2
Fig. 2
A Field growth performance of K and KY; B and C K and KY agronomic traits; C Normal distribution analysis of total chlorophyll content in 1,859 individual plants
Fig. 3
Fig. 3
A Δ(SNP-index) Interval localization map (CI_95 and CI_99 possess 95% and 99% confidence intervals, respectively); B Euclidean distance (ED) interval localization map
Fig. 4
Fig. 4
A Differential expression proteins (DEPs) between K and KY; B GO enrichment analysis of differential proteins; C Differentially expressed genes (DEGs) between K and KY; D VENN analysis of BSA-seq, RNA-seq, and proteomic sequencing; E Sequencing peak plot
Fig. 5
Fig. 5
A Gene expression levels of K and KY; B Correlation between gene expression and chlorophyll content in K and KY; C Protein expression levels of K and KY; D Correlation between protein expression and chlorophyll content in K and KY; E Folate content in K, KY, and Y; F Leaf colour phenotypes of K, KY, and Y
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