Summary Tetraploids have been reported to exhibit increased stress tolerance, but the underlying molecular and physiological mechanisms remain poorly understood. In this study, autotetraploid plants were identified by screening natural seedlings of trifoliate orange ( Poncirus trifoliata ). The tetraploids exhibited different morphology and displayed significantly enhanced drought and dehydration tolerance in comparison with the diploid progenitor. Transcriptome analysis indicated that a number of stress‐responsive genes and pathways were differentially influenced and enriched in the tetraploids, in particular those coding for enzymes related to antioxidant process and sugar metabolism. Transcript levels and activities of antioxidant enzymes (peroxidase and superoxide dismutase) and sucrose‐hydrolysing enzyme (vacuolar invertase) were increased in the tetraploids upon exposure to the drought, concomitant with greater levels of glucose but lower level of reactive oxygen species ( ROS ). These data indicate that the tetraploids might undergo extensive transcriptome reprogramming of genes involved in ROS scavenging and sugar metabolism, which contributes, synergistically or independently, to the enhanced stress tolerance of the tetraploid. Our results reveal that the tetraploids take priority over the diploid for stress tolerance by maintaining a more robust system of ROS detoxification and osmotic adjustment via elevating antioxidant capacity and sugar accumulation in comparison with the diploid counterpart.

Polyploid plants often exhibit enhanced stress tolerance relative to their diploid counterparts, but the physiological and molecular mechanisms of this enhanced stress tolerance remain largely unknown. In this study, we showed that autotetraploid trifoliate orange (Poncirus trifoliata (L.) Raf.) exhibited enhanced salt tolerance in comparison with diploid progenitors. Global transcriptome profiling of diploid and tetraploid plants with or without salt stress by RNAseq revealed that the autotetraploids displayed specific enrichment of differentially expressed genes. Interestingly, the leaves and roots of tetraploids exhibited different expression patterns of a variety of upregulated genes. Genes related to plant hormone signal transduction were enriched in tetraploid leaves, whereas those associated with starch and sucrose metabolism and proline biosynthesis were enriched in roots. In addition, genes encoding different antioxidant enzymes were upregulated in the leaves (POD) and roots (APX) of tetraploids under salt stress. Consistently, the tetraploids accumulated higher levels of soluble sugars and proline but less ROS under salt stress compared to the diploids. Moreover, several genes encoding transcription factors were induced specifically or to higher levels in the tetraploids under salt stress. Collectively, this study demonstrates that the activation of various multifaceted defense systems in leaves and roots contributes to the enhanced salt tolerance of autotetraploids.

The basic helix-loop-helix (bHLH) family of transcription factors (TFs) plays a crucial role in regulating plant response to abiotic stress by targeting a large spectrum of stress-responsive genes. However, the physiological mechanisms underlying the TF-mediated stress response are still poorly understood for most of the bHLH genes. In this study, transgenic pummelo (Citrus grandis) plants overexpressing PtrbHLH, a TF previously identified from Poncirus trifoliata, were generated via Agrobacterium-mediated transformation. In comparison with the wild-type plants, the transgenic lines exhibited significantly lower electrolyte leakage and malondialdehyde content after cold treatment, thereby resulting in a more tolerant phenotype. Meanwhile, the transgenic lines accumulated dramatically lower reactive oxygen species (ROS) levels, consistent with elevated activity and expression levels of antioxidant enzymes (genes), including catalase (CAT), peroxidase and superoxide dismutase. In addition, PtrbHLH was shown to specifically bind to and activate the promoter of PtrCAT gene. Taken together, these results demonstrated that overexpression of PtrbHLH leads to enhanced cold tolerance in transgenic pummelo, which may be due, at least partly, to modulation of ROS levels by regulating the CAT gene.

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