ABSTRACT
Background Transposable elements (TEs) are major components of plant genomes. Despite being regarded as “junk DNA” at first, TEs play important roles for the organisms they are found in. The most obvious and easily recognizable effects caused by TEs result from their mobility, which can disrupt coding sequences or promoter regions. However, with the recent advances in transcriptomics, it is becoming increasingly evident that TEs can act as an additional layer of gene expression regulation through a number of processes, which can involve production of non-coding RNAs. Here, we describe how Tnt1, a stress-responsive LTR-retrotransposon, interferes with gene expression and modulate a number of developmental aspects in tobacco.
Results Through an RNAi approach, we generated tobacco (HP) lines knocked-down for Tnt1 expression. Quantitative RT-PCR experiments confirm that Tnt1 is downregulated in HP lines after ethylene exposure. A RNA-seq experiment was performed and through two independent bioinformatic approaches (with different stringencies) we found 932 and 97 differentially expressed genes in HP lines. A number of phenotypes were observed in such lines, namely lesion mimicry in leaves, underdevelopment of the root system, overproduction of root hairs and early loss of seed viability. Folding prediction of part of the Tnt1 mRNA reveals putative stem-loop secondary structures containing transcriptional regulation sequences, suggesting it could be a source of small RNAs. We also propose a model to explain the Tnt1 expression in both homeostatic and stress conditions, and how it could interact with stress-responsive genes.
Conclusions Our results are consistent that interferences with Tnt1 transcript levels correlate with transcriptomic and phenotypic changes, suggesting a functional role for this element during plant development and stress response.
Competing Interest Statement
The authors have declared no competing interest.
LIST OF ABBREVIATIONS
- cDNA
- complementary DNA
- CPM
- Counts per million
- DEGs
- Differential expression genes
- EFEs
- Ethylene forming enzymes
- FDR
- False discovery rates
- Gag
- Group antigens
- GO
- Gene ontology
- GSI
- germination seed index
- HP
- Hair-pin lines
- INT
- Integrase
- LTR
- Long terminal repeats
- MCE
- Mean conditional entropy
- MGT
- Mean germination time
- miRNA
- micro RNA
- mRNA
- messenger RNA
- ORFs
- Open reading frames
- PR
- pathogenesis-related
- qPCR
- Quantitative polymerase chain reaction
- RNAi
- RNA interference
- RPKM
- Reads per kilo base per million mapped reads
- RT
- Reverse transcriptase
- SGN
- Sol genomics network
- sRNA
- small RNA
- TE
- Transposable Elements
- TMM
- Trimmed means of M values
- WT
- Wild type