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Field Performance, Alkaloid Quantities, and Transcriptome Profiles of Nearly Isogenic Tobacco Genotypes Possessing Recessive Allele Combinations at the Nic1, Nic2, and Myc2a Loci

ABSTRACT: Tobacco with modified genetics controlling alkaloid accumulation has been of interest because of the possibility of mandated lowering of nicotine levels in combustible cigarettes by regulatory authorities. Transcription factors coded by the Nic1, Nic2, and Myc2a loci act as positive regulators of genes directly involved in alkaloid accumulation. Different combinations of alternative alleles at these loci were assembled in homozygous nearly isogenic lines (NILs). Recessive nic1/nic2 alleles were found to have greater influence on alkaloid reduction as compared to a mutant myc2a allele. When combined into single genotypes, these alleles operated in an additive manner to further reduce alkaloid levels in field and greenhouse experiments. This was at the expense of reduced cured leaf quality, however. An RNA-seq experiment was carried out to investigate global changes in root expression due to different genotypes at these three loci. Up to 681 differentially expressed genes (DEGs) were identified between the studied NILs, with expression of most DEGs being downregulated by recessive alleles. In general, the mutant myc2a allele appeared to suppress a subset of previously characterized alkaloid biosynthetic genes with relatively weaker effect as compared to the nic1/nic2 alleles. There was little evidence that root expression of Nic1 or Nic2 genes was affected by allelic variability at the Myc2a locus, or vice versa. The list of DEGs influenced by genotypes at these three loci may contain candidate genes coding for currently uncharacterized enzymes involved with tobacco alkaloid accumulation.

ORGANISM(S): Nicotiana tabacum

PROVIDER: GSE284841 | GEO | 2025/05/28

REPOSITORIES: GEO

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Project description:Comparative analysis of tobacco leaves transcriptomes unveils carotenoid pathway potentially determined the characteristics of aroma compounds in different environmental regions. Tobacco (Nicotiana tabacum) is a sensitive crop to environmental changes, and a tobacco with unique volatile aroma fractions always formed in specific ecological conditions. In order to investigate the differential expressed genes caused by environmental changes and reveal the formation mechanism of characteristics of tobacco in three different aroma tobacco regions of Guizhou Province, Agilent tobacco microarray was adapted for transcriptome comparison of tobacco leaves in medium aroma tobacco region Kaiyang and light aroma tobacco regions Weining and Tianzhu. Results showed that there was big difference among the gene expression profiles of tobacco leaves in different environmental conditions. A total of 517 differential expressed genes (DEGs) between Weining and Tianzhu were identified, while 733 and 1,005 genes differentially expressed between Longgang and another two tobacco regions Weining and Tianzhu, respectively. Compared with Longgang, up-regulated genes in Weining and Tianzhu were likely involved in secondary metabolism pathways, especially carotenoid pathway, including PHYTOENE SYNTHASE, PHYTOENE DEHYDROGENASE, LYCOPENE ε-CYCLASE, CAROTENOID β-HYDROXYLASE and CAROTENOID CLEAVAGE DIOXYGENASE 1 genes, while most down-regulated genes played important roles in response to temperature and light radiation, such as heat shock proteins. Gene Ontology and MapMan analyses demonstrated that the DEGs among different environmental regions were significantly enriched in light reaction of photosystem II, response of stimulus and secondary metabolism, suggesting they played crucial roles in environmental adaptation and accumulation of aroma compounds in tobacco plants. Through comprehensive transcriptome comparison, we not only identified several stress response genes in tobacco leaves from different environmental regions but also highlighted the importance of carotenoid pathway genes for characteristics of aroma compounds in specific growing regions. Our study primarily laid the foundation for further understanding the molecular mechanism of environmental adaptation of tobacco plants and molecular regulation of aroma substances in tobacco leaves. In order to investigate the differential expressed genes caused by environmental changes and reveal the formation mechanism of characteristics of tobacco in three different aroma tobacco regions of Guizhou Province, Agilent tobacco microarray was adapted for transcriptome comparison of tobacco leaves in medium aroma tobacco region Kaiyang and light aroma tobacco regions Weining and Tianzhu.

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Field Performance, Alkaloid Quantities, and Transcriptome Profiles of Nearly Isogenic Tobacco Genotypes Possessing Recessive Allele Combinations at the Nic1, Nic2, and Myc2a Loci

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