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

ABSTRACT: The degree of yellowing in tobacco leaves is an important indicator for determining the maturity and harvesting time of tobacco leaves. Reduction in chlorophyll is of utility for promoting the concentrated maturation of tobacco leaves and achieving mechanised harvesting and mining, and utilising tobacco yellow leaf regulatory genes is of great significance for the selection and breeding of tobacco varieties suitable for mechanised harvesting and the resolution of the molecular mechanisms controlling leaf colouration. 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. 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.

ORGANISM(S): Nicotiana tabacum

PROVIDER: GSE270039 | GEO | 2024/10/23

REPOSITORIES: GEO

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Project description:SA0420 is a stable rice (Oryza sativa L.) mutant exhibits multiple traits including a leaf-tip yellowing (YLT), a taro fragrance (or aroma, A), a reduced panicle number (PN), spikelet number (SN), plant height (PH), days to heading (DH), filled grain weight (FGW), and fertility rate (FR), different from those of a local cultivar TNG67. Correlation analysis on F2 progenies of SA0420 and TNG67 revealed that except for aroma which is inheritated independently, phenotypes including YLT, PN, SN, PH, and FGW are tightly associated with each others, indicating a control simultaneously by pleotropic gene. Among those phenotypes, YLT appears the earliest at vegetative stage, therefore may act as a predisposing factor that give rise to other phenotypes. Genetic analysis suggested that YLT is controlled by a single recessive gene. By analyzing the recombinant inbreed lines (RILs) that produce YLT and normal green (G) progenies in a 1:3 ratio, the phenotypes associated with YLT were further confirmed. To investigate the molecular events of leaf-tip yellowing, microarray analyses were performed to compare the gene expression profiles between YLT and G plants of RILs. Clustering of genes according to their involved metabolic pathways revealed that various photosynthesis and carbon fixation gene expressions are lower (~55~75%) in YLT than in G plants, consistent with the observation that photosynthesis rate of SA0420 was only 60% to that of TNG67. Moreover, higher expressions of oxidative phosphorylation-related genes indicated that not only the reduced synthesis rate but also the elevated energy consumptions may synergistically decrease the growth vigor of SA0420. Dissection of the mature leaf revealed that chlorophyll is drastically reduced in the leaf-tip of SA0420 while carotenoids remain stable. This explains the appearance of leaf-tip yellowing and recall an analogy of early senescence. Indeed, many senescence-associated genes (SAG) were found to be up-regulated in SA0420. Keywords: rice (Oryza sativa), yellow and green leaves, molecular functions, signal-microarray, vegetative stage

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

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